proceedings of the south dakota academy of science volume 77 … · 2017. 7. 12. · proceedings of...

Proceedingsof the

South Dakota Academy of Science

Volume 771998

Published by the South Dakota Academy of ScienceAcademy Founded November 22, 1915

EditorKenneth F. Higgins

Terri Symens, Wildlife and Fisheries, SDSUand

Tom Holmlund, Ag Communications, SDSUProvided Secretarial and Computer Application Assistance

TABLE OF CONTENTS

SDAS Executive Committee ................................................................................1SDAS Membership 1998......................................................................................2Presidential Address. Science in Rural America. Royce C. Engstrom ..............3Transport of Nutrients and Phytoplankton Into Two Glacial Prairie

Lakes. Lois Haertel and Nels H. Troelstrup, Jr..........................................13Ornamental Traits in Hyalella Azteca as Indicators of Water Quality;

Implications for Biological Monitoring. Eric A. Miller and Nels H. Troelstrup, Jr. .................................................................................27

Comparative Energy Flow to the Fish Community in a Prairie Systemand a Forested Stream Using Growth Rate and Stable Isotope Analysis. Thayne A. Munce, Kyle N. Seifert and Craig N. Spencer..........................29

A Field-Oriented General Chemistry Community Project: Rapid Creek Water Quality. Martha M. Tremblay and Cathleen J. Webb ...........41

Strawberry Clover and Its Use in the Amelioration of Cadmium Contaminated Sites. Peter Jauert and R. Neil Reese .................................43

Lead Soil Levels at Four Elementary School Playgrounds in Rapid City. Derek Thirstrup and Cathleen J. Webb .......................................................45

A Study of Herbicide Retention in Two Different Humin Soil Types.Walter Whiteside, Justin Kurcirek, Christine J. Guetzloff,James C. Sorenson and Thomas F. Guetzloff .............................................47

Numerical Treatment of a Mathematical Model of Two Competitors in a Partially Periodically Muddy Lake. Alicia Vander Weyst and A.S. Elkhader........................................................................................57

Hydrology of Glacial Lakes, Fort Sisseton Area. Perry H. Rahn .....................59Some Model Selection Procedures in Discriminant Analysis. Lu Zhang.........67Abortion in America. Joel Vander Kooi ............................................................69Utilization of Hands-On Activities in the Science Classroom:

An Assessment of Student Performance. Nels H. Troelstrup, Jr. and Dennis Clark.........................................................................................71

Implementation of Multimedia Instruction in Biology and Chemistry Courses: Student Reactions. Thomas F. Guetzloff and James C. Sorenson.......................................................................................73

Taste and Odor Aversion Conditioning in MusMusculus and AssociatedChanges in Heart Rate and Fluid Consumption. Linda R. Alzieblerand Thomas P. Cox......................................................................................83

Decreased Aggression in Gonadectomized Male Fighting Fish (BettaSplendens) Systematically Related to Increasing Levels of Estrogen. Michael J. Huxford and Thomas P. Cox .....................................................91

Obligatory Winter Dipause in Statoblasts of Plumatella Casmania(Bryozoa: Phylactolaemata). Melissa A. Albers and Jonathan C. Wright ......................................................................................93

Chromosomal Analysis of Deformed Frogs. Angela Reister, Sara Horner and Debra Carlson ..................................................................95

Multivariate Morphometric Variation in the Coffee Snake, Ninia Diaemata. Brian E. Smith ..........................................................................97

Separation of Maternal and Embryo Contributions to Reproductive Failure in Yellow Mice (Ay/a; C57BL/6J). Gerald A. Dickens, Maureen R. Diggins and Nels H. Granholm...............................................99

Bovine Agouti Molecular Analysis. Marcus D. Johansen, Raymond R.R. Rowland, Carl A. Westby, Donald M. Marshalland Nels H. Granholm...............................................................................101

Comparison of Adult Echinococcus Multilocuraris Excretory/Secretory(ES) Antigens to Those of Protoscoleces. Troy L. Lackey,Margaret Perry and Mike B. Hildreth .......................................................103

Gangliosidosis in Sheep Caused By a Defective b Galactosidase Gene.K. Heidebrink, R. Rowland, L. Holler and T. Cheesbrough.....................105

Comparison of the Cellular and Humoral Immune Response as a Predictor for Protection in a Type II Bovine Viral Diarrhea VirusChallenge. A.A. Ahmed, A.A. Salama, D.J. Hurley, L.J. Braun and C.C.L. Chase.......................................................................................107

The Nucleocapsed Protein of Porcine Reproductive and Respiratory Syndrome Virus Localizes to the Nubleolus. Christopher J. Kuckleburg, Bobby L. Kervin, David A. Benfield and Raymond R.R. Rowland.....................................................................109

Analysis of the Structural Genes of a PRRSV Strain That Grows Poorly in Macrophages. K.L. Kauers, M.S. Steffen, D. Shah, D.A. Benfield and R.R.R. Rowland ..........................................................111

Congenital PRRSV: Sites of Virus Replication During Acute and Persistent Infection. S.R. Lawson, D.A. Benfield and R.R.R. Rowland.........................................................................................113

Nitrogen Partitioning in a Manured Field in Southeast South Dakota.H. Smeltekop, D. Malo and D. Clay .........................................................115

Spatial and Temporal Differences of Herbicide Mineralization: Relationship to Weed Control. Z. Liu, S.A. Clay and D.E. Clay ............117

Root Morphology and Endogenous Cytokinin Levels of Flood Tolerant and Flood Susceptible Soybeans. Steven B. Ortmeier and R. Neil Reese ......................................................................................119

Effects of Outcrossing on Kernel Traits and Grain Yield in Hybrid Maize (Zea Mays L.). C.T. Mack, Z.W. Wicks III and P.B. Beauzay .....121

Extraction and Analysis of DNA from Recalcitrant Plant Leaf Tissue.E.M. Galloway and M.R. Duvall ..............................................................123

Mutagenesis and Analysis of Aureobasidium Pullulans. Anna R. Ollerand Carl A. Westby....................................................................................129

Allelopathic Potential of Echinacea Angustifolia D.C.’s Root Extracts.Kimberly Piechowski and R. Neil Reese ..................................................131

A Study of Fossil and Modern Members of the Paniceae (Poaceae) from North and South America. J.J. Smith and M.L. Gabel ...................133

A Study of Lycopodium Dendroideum in the Black Hills of South Dakota.Terri Hildebrandt, Mark Gabel, Cecilie Steib and Shane Sarver..............135

Dynamics of Source-Sink Landscapes Relative to Waterfowl Populationsin the Northern Great Plains. Kenneth F. Higgins, Rex R. Johnson,David E. Naugle, Matthew M. Holland, Thomas R. Cooperand Michael E. Estey.................................................................................137

Goldeye Recruitment and Growth in Two Missouri River Backwaters.Daniel N. Moon, Shannon J. Fisher and David W. Willis ........................139

Spring Migration of Raptors in Moody County, South Dakota.Any E. Gabbert and Natoma A. Schneider ...............................................145

White Crappie Biology in an Upper Missouri River Backwater.Randy A. Sheik, Shannon J. Fisher and David W. Willis .........................151

Resource Overlap and Partitioning Between Larval Yellow Perchand Adult Fathead Minnows in a Prairie Glacial Lake.Shannon J. Fisher, Gene F. Galinat and David W. Willis .........................163

Quaternary Ammonium Research. Dianna Wineinger....................................171Quaternary Ammonium Synthesis. Michael Hanson ......................................175Stabilities of Non-G•U Mismatches in Internal Loops of RNA Hairpins.

Peter deLannoy, LaHoma Easterwood and Stephanie Price .....................177Effect of Ligand Deuteration on the EU3+ (5D0) Lifetime in Tris

(2,2,6,6-Tetramethyl-3-5-Heptanedionato) Europiuni (III).Todd C. Schwendemann, Paul S. May and Mary T. Berry.......................187

Product Analysis for the Photochemical Addition of Ethanol toMaleic Acid and Its Esters. M. Robert Stoner, Thomas Tranand Roy C.Prebble.....................................................................................189

Isomerization of Trimethyl Phosphite. Drew J. Paulson andArlen Viste.................................................................................................191

FTIR Spectra of Polymers in Reflectance and Transmission.John D. Gilbertson and Arlen Viste ..........................................................193

Fourier Image Analysis. John P. Berdahl and Karel Vander Lugt ..................195An Inexpensive Device for Measurement and Presentation of Atomic

Spectra. James L. Lefferts ........................................................................197Electronic States in Semimagnetic Quantum Wells.

James Niggeman and A.G. Petukhov........................................................199Temperature Controller for a Microcalorimeter Control System.

L.W. Watson and T. Ashworth...................................................................201Self-Consistent Perturbation Theory Calculations for Two Dimensional

Superconductors. John J. Diesz................................................................203Climatological Trends in the Black Hills. Stephen D. Trimarchi,

James R. Miller, Jr. and L. Ronald Johnson..............................................205Senegal’s Theory of Atomic Particles. Claffey J. Senegal, Jr. ........................219Patterns in Avian Community Structure and Non-Point Source

Disturbance Potential Along the Land-Water Interface of aPrairie Pothole Lake. Kristel K. Bakker and Nels H. Troelstrup, Jr........221

Minutes of the Eighty-Third Annual Meeting of the South DakotaAcademy of Science..................................................................................231

PROC. S.D. ACAD. SCI., VOL. 77 (1998) 1

SOUTH DAKOTA ACADEMY OF SCIENCE1998-1999 EXECUTIVE COMMITTEE

PRESIDENT Royce Engstrom, USD Chemistry,677-5370 or [email protected]

PRESIDENT-ELECT R. Neil Reese, SDSU Biology, [email protected]

FIRST VICE-PRESIDENT Lenore Koczon, NSU Chemistry, [email protected]

SECOND VICE-PRESIDENT Charles Lamb, BHSU Biology, [email protected]

SECRETARY-TREASURER Bill Soeffing, USF Natural Sciences,331-6759, Fax [email protected]

PROCEEDINGS EDITOR Kenneth F. Higgins, SDSU Wildlife andFisheries, 688-4779, Fax [email protected]

FIRST PAST-PRESIDENT Sharon Clay, SDSU Plant Science,[email protected]

SECOND PAST-PRESIDENT John Thomas, USD Biochemistry,[email protected]

MEMBERS-AT-LARGE Audrey Gabel, BHSU Biology, [email protected]

Gary Larson, SDSU Biology, [email protected]

Donna Hazelwood, DSU Natural Sciences,[email protected]

Tim Sorenson, Augustana Mathematics,[email protected]

2 PROC. S.D. ACAD. SCI., VOL. 77 (1998)

SOUTH DAKOTA ACADEMYOF SCIENCE MEMBERSHIP 1998

Regular and AssociateMembership

Albers, Melissa*Barondeau, MikeBerdahl, John*Berg, Sherwood O.Bunkers, Matthew J.Card, CurtisCarlson, DebraChase, ChrisClay, SharonCox, Thomas P.DeBoer, Darrell W.Deisz, JohndeLannoy, PeterDetwiler, AndrewDoolittle, JamesEarl, GaryElkhader, A.S.Engstom, RoyceGabel, AudreyGaines, RobertaGalloway, Elizabeth*Gilbertson, John*Granholm, NelsHaertel, LoisHampton, Douglas*Hansen, Michael*Hempel, RobertHiggins, KennethHildebrand, TerriHorner, Sara*Hornstra, Fred Jr.Hurley, DavidJacobson, CarrieJohansen, Marcus*Johnson, GaryJohnson, L. RonaldJohnson, LelandKauers, Kevin*Kawatski, JosephKihm, AllenKing, Kevin

Klawiter, W.F.Knapp, EmilKreber, RobertLarson, GaryLawson, Steven*Liu, ZhuojingMenzel, RichardMiller, JamesNaughten, JohnNiggemann, James*Norton, JamesOde, DavidOller, Anna*Palmer, JeffreyParadise, MichaelPaulson, Drew*Rahn, PerryReister, Angela*Rickerl, DianeSandberg, PhilipSawyer, JohnScalet, CharlesSchneider, Natoma*Shore, Sid*Simmons, RickSmeltekop, Hugh*Smith, BrianSmolik, JamesSoeffing, WilliamSpencer, CraigSpinar, LeoStene, JohnStetler, LarryStoner, M. RobertTatina, RobertThomas, JohnVander Kooi, Joel*Vander Lugt, KarelViste, ArlenWatson, Laura*Weisshaar, DuaneWessel, DouglasWineiger, Dianna*Winters, Wade

Wright, JonathanYocum, KennethZhang, Lu

*denotes associatemember

Life Members

Beaty, MarjorieCobb, Henery V.Dillon, Ray D.Estee, Charles R.Foss, HaroldFroiland, S. GordonGaalswyk, ArieGoodell, Harold B.Green, MortonGrieb, EdnaGries, John PaulHalstead, Al L.Houk, William F.Hugghins, Ernest J.Kintner, Robert RoyKlug, Harlean L.Landborg, Richard JohnLoomer, JerryMcGregor, Ralph H.Mickelson, John C.Miller, NormanNelson, Ronald V.Olson, Oscar E.Partlo, Fay L.Pengra, Robert M.Redin, Robert D.Rissky, Roy W.Schmulbach, James C.Smith, Harold L.Van Bruggen, TheodoreWebster, Victor S.Wicks, Zeno W. III


PRESIDENTIAL ADDRESS

Science in Rural America

Address to the South Dakota Academy of SciencesBlack Hills State University

April 17, 1998

Presented by Royce C. EngstromUniversity of South Dakota

I. Introduction

The South Dakota Academy of Sciences (SDAS) represents the broadest spec-trum of scientists in the state, representing virtually every discipline, every levelof education, and every school in South Dakota. Our state is one of the mostrural in the country, with all of the benefits and problems that affords.Consequently, this audience may contain the world’s most expert group on thetopic of this talk, “Science in Rural America.” Therefore, this is a most appro-priate group to discuss the topic with and this is the group that is, perhapsunknowingly, charged with guiding this rural state into the future.

Our country’s attitude toward and willingness to support science has changedfrequently over recent years, and not always in a predictable direction. Forexample, just two years ago, the budgets of science-related federal agencies wereunder considerable attack, and now, due in large measure to an active collectivevoice of the science community, we are looking toward some of the healthiestincreases we’ve seen in many of our careers. It is clear that we are workingunder different conditions than we have known in the past, and those conditionshave especially profound implications to states that, like South Dakota, have his-torically held a restricted view as to the role of university-based science. Bothnationally and at home, there are increasing expectations of our educational sys-tem, and that certainly applies to science. Students have an increasing array ofchoices for their education and the distinction between those choices is increas-ingly blurred. To be sure, the demands upon each of us as scientists and educa-tors continue to increase as we try to fulfill the expectations of our peers, oursupervisors, and ourselves.

This talk will be divided into three parts: 1) background information and data thatshow how the rural states compare with respect to a variety of parameters; 2) adiscussion of expectations of scientific research and education that come fromvarious national dialogues; and 3) some considerations as to how South Dakotamight respond to national expectations and state needs.


II. Background

The National Science Foundation has prepared an extensive graphical databasein which states are profiled. This database is available on the web and makes useof a geographic information system to show trends and contrasts. I’ve picked outsome of the data from that “Almanac” to illustrate how the rural states, and SouthDakota in particular, look with respect to some defining characteristics. Theparameters cover a wide range, from simple population distribution to technolo-gy-related economic development trends.

First, let’s look at a few demographic issues. State population is the primary cri-teria I want to use to define “rural” in the context of this discussion. Certainly,there are many areas of the country that use the term rural to describe parts oftheir states, particularly in the context of advertising a certain quality of life.However, total state population will be used here to define rural, because it seemsto be a state’s population base that, in large measure, determines the magnitudeof its science and technology enterprise. The central plains and the mountainwest constitute the major rural section of the country, with South Dakota rightnear the top of the “most rural” list. Furthermore, population changes over thepast decade show that many of these areas are becoming more, not less, rural.Exceptions are a few of the mountain states, which have shown substantialgrowth in recent years. The distribution of gross state products (GSP) by statelargely mirrors state populations, as does the distribution of appropriations to thestate higher education systems. There are some notable exceptions to these dis-tributions, however. For example, two rural states, Wyoming and Nevada, areamong the highest in per capita GSP. Several of the plains states are among thehighest in per capita state appropriations for higher education. In fact, all of thestates surrounding South Dakota except Montana are in the top third with respectto per capita appropriations. South Dakota and Montana are in the lower third,with $163 and $144, respectively, compared to $251 and higher for the sur-rounding states.

The Southern Technology Council is an alliance that has been formed to improveeconomic conditions in the south through science and technology. They haveworked aggressively to study and strengthen the interactions between southernuniversities and the private sector of the state. One of the parameters they haveinvestigated has to do with student retention and migration into and out of thestates. They looked at four trends: 1) retention of high school graduates; 2)retention of people with degrees in higher education; 3) migration of high schoolgraduates; and 4) migration of people with degrees. The net migration index isa measure of how people with degrees move into and out of a state. An index of1 indicates an overall balance between people leaving the state and entering thestate after receiving their highest degree. A value below 1 indicates a netexporter of human resources; above 1 indicates a next importer. Many of therural states are net exporters of the product of education, with South Dakota one


of the most obvious. With respect to the cumulative of all four parameters, thestates with the overall highest index (greatest importers) tend to be along a coast.This trend is especially relevant to South Dakota. For example, Gateway 2000has made the decision to move management of its operation to New York andCalifornia partly because it can’t fill its employment demands from the locallabor pool.

Let’s look specifically at the distribution of some factors related to science andtechnology. For example, the number of scientists and engineers in each of thestates has been examined by the National Science Foundation, and shows that thestates with the lower populations, not suprisingly, have fewer scientists and engi-neers. However, even on a per capita basis, there are indications that the morerural the state, the lower the proportion of its population engaged in science andengineering. That observation correlates well with the number of high-technol-ogy establishments present in the rural states, which again, is often dispropor-tionately low. Interestingly, the number of graduate students in science and engi-neering on a per capita basis doesn’t clearly show the same trends as population,suggesting that we educate our respective “share” of science and technology peo-ple, then send them out of the state for employment.

Let’s finish this descriptive section with some research trends, because there aresome nice surprises there. As a country, we concentrate our research resourcesinto a relatively small number of states. For example, approximately 50% of thenation’s federal R&D resources go to only 6 states. There are a large number ofstates that make up the “tail” of the federal R&D picture, with 20 states receiv-ing the bottom 5%. The picture with respect to the distribution of NSF dollarshas changed significantly in recent years, however. Let’s look specifically nowat South Dakota. As a state, we have moved from being essentially at the bot-tom of all the states to being in the middle third of the country on a per capitabasis of NSF awards and dollars. In fact, as of 1997, no other state in the coun-try has shown a faster relative increase in the number of NSF awards than hasSouth Dakota and the state is among the leaders in relative increase in NSF fund-ing. That may say more about where we were than where we are, but is nonethe-less a fact that has attracted the attention of NSF officials. Likewise, SouthDakota has been on the move with respect to funding through the Small BusinessResearch Innovation program, which provides funding for private sector enter-prises to development new products and processes. Through the concertedefforts of Mel Ustad at DSU, South Dakota has dramatically increased its awardsthrough the SBIR program.

The overall picture of South Dakota in this discussion of demographics and per-formance shows a state that is one of the least equipped to move into the tech-nological economy, but that is nonetheless making impressive gains. How doesthat position us to respond to what the nation demands from its higher educationcommunity?


III. National Expectations of Higher Education in the Sciences

During the past several years, there have been a number of special task forces toexamine the effectiveness of higher education and the ability of the country tomove into our technological future. You are probably all aware of CongressmanVern Ehler’s Congressional Committee on Science and Technology. Ehlers is aPh.D. physicist and former faculty member at Calvin College in Michigan. Hehas assembled an impressive group of educators and private sector scientists,(with barely any representation from the rural states) to examine how our scien-tific infrastructure needs to change to keep the country competitive. They are inthe early stages of their work.

The National Science Foundation conducted an intensive study recently to exam-ine the status of undergraduate science education and to make recommendationsas to how we, as a nation, can improve it. The study resulted in the report,“Shaping the Future: New Expectations for Undergraduate Education inScience, Mathematics, Engineering, and Technology. The committee waschaired by Melvin George, then of the University of Minnesota. The committeedid not have any members from the low population states. The report has, how-ever, been discussed widely, even in the rural states. For example, in May thereis a conference at the University of Nebraska to conduct follow-up activity relat-ed to the report. The goal to be derived from the review is:

“All students have access to supportive, excellent undergraduateeducation in science, mathematics, engineering, and technology,and all students learn these subjects by direct experience with meth-ods and processes of inquiry.”

While at first this sounds like apple pie, the report emphasizes that certain keywords in the statement call for a change in the way we, as educators, approachour work. “All students” is calling for us to stop the perceived practice of con-centrating on cloning ourselves; “supportive” suggests that we currently practicethe weeding out approach to teaching; “inquiry” emphasizes the process and thewonder of investigation as opposed to mastering just factual material. The com-mittee formulated a series of recommendations targeted at a variety of groups.

• Faculty should believe and affirm that every student can learn, should startthe learning process with the student’s experience, and have high expecta-tions within a supportive climate, build inquiry, a sense of wonder, and theexcitement of discovery.

• Departments should set goals and accept responsibility for undergraduatelearning with measurable expectations for all students, should work collab-oratively with departments of education and the K-12 sector, the businessworld, and provide pedagogical skill development for those graduate stu-dents intending on becoming teachers.


• Governing Boards and Administrators need to reexamine institutional mis-sions in light of needs in undergraduate education, hold accountable anddevelop reward systems for departments and programs, provide for strongfaculty development and reduce organizational rigidities.

• Accrediting agencies need to focus on student learning, not just organiza-tional and process issues.

The report continues with recommendations to business and industry, the gov-ernment, and the National Science Foundation itself, urging that the agencyaccept leadership for implementing all the recommendations.

At the same time, the National Research Council was conducting its own studyof undergraduate education in science, mathematics, engineering, and technolo-gy, also with a select committee, and also without any representation from a ruralstate. The recommendations of this group sound quite similar:

• Students should have access to supportive, excellent programs, and all stu-dents should acquire literacy by direct experience with the methods andprocesses of inquiry.

• Departments and programs should define their missions and establish spe-cific educational goals; they should be evaluated against those goals by fairassessments that are as rigorous as the ones applied for research.

• Institutions must promote a new balance and a new linkage between teach-ing and research so that teaching is enlivened by investigation and researchis defined more broadly, so that faculty can be rewarded for educationalscholarship as well as for other kinds of scholarship.

• Institutions and departments should promote educational innovation boththrough cultural change and through providing resources.

I have been involved in the same kind of self-examination of the subdiscipline ofanalytical chemistry that involved a small group of faculty, private sector scien-tists, and government laboratory scientists. This one was organized by TedKuwana from Kansas University, so it involved at least a handful of people fromrural states.

The main point is that the nation, through the work of these committees andmany others, is examining the effectiveness of the way we teach science andengineering and recommendations are being made and listened to that haveimportant impact on our lives.

Research is under equally intense pressure, as educational institutions are lookedto more and more to be the engines of economic development and to be relevantto the daily lives of taxpayers and policy-makers. For example, I just recentlyattended a workshop sponsored by NASULGC entitled, “The Research Businessof the University,” which focuses on how individual faculty as well as wholeinstitutions could move into the private sector through technology transfer, intel-


lectual property considerations, initiation of research parks, and industry-univer-sity partnerships. A NASULGC publication called, “For Every DollarInvested…. The economic impact of public universities,” describes how invest-ments in research result in both direct and indirect returns to the economy.

Another recent publication entitled, “University Research, Touching the Lives ofall Americans,” describes a number of research activities around the nation undertopics such as health, the information age, the environment, automobiles, and theenergy supply. In fact, the National Science Foundation has recently changed itsreview criteria to increase emphasis on the broader impact of the proposedresearch.

IV. South Dakota’s response

How does South Dakota’s rural nature fit in with the increasing expectationsfrom both the national and local public? I think we are well-suited to respond tosome areas, and it will be a struggle responding to others. These are my thoughtson some of these areas.

The Research-Education Connection Within our system of higher education,we have the potential for integrating research and education to a greater degreethan most states. The research expectations of faculty vary considerably fromcampus to campus in South Dakota, as they should. But nowhere are they sostringent as to preclude the involvement of undergraduates as is often the case atlarge, research-intensive universities. The relatively modest settings that our sci-ence programs are housed in and the instrumentation facilities we have usuallydemand the overlap of teaching and research, making it relatively easy to com-bine the two. In fact, one could argue that in order to carry out research, therehad better be a strong overlap with teaching because of the relatively largeamount of time we spend teaching. For the most part, South Dakota faculty don’tenjoy the luxury of large blocks of time dedicated solely to research. Our classand program size, compared to most other states, is small, so that it is not a dif-ficult task to learn who the capable and interested students are. Those are the stu-dents that we can encourage into our research laboratories. The intimacy of ourenvironments should work to our advantage in integrating teaching and research.

Relevancy of Science to Lives of Citizens The rural nature of our state resultsin our population being quite aware of, if not directly involved in, the land andnatural features around us. One could say that the Black Hills and the Badlandsare a part of the everyday lives of the people of West River, the Missouri Riveris a unique feature that most people in the central and southeastern parts of ourstate have dealt with or enjoyed on some level, and the prairie pothole region ofthe northeast represents one of the most productive habitats in the region, if notthe country. Why not make increasing use of these natural laboratories in ourcourses and seminars, not only in those courses that pertain in an obvious way tothem, such as geology or ecology, but in physics, chemistry, and mathematics as


well? There are programs around the country that focus on marine science, theDesert Research Institute, the Great Lakes Center, and so on. These institutes alltake an interdisciplinary approach to scientific research and education. Could weconsider and promote teaching and research efforts that focus on our own natur-al wonders?

Public Awareness Despite the seemingly wide gulf between those of us in aca-demics and those in public policy or those in the private sector, there exists thepotential to bridge that gap more easily than in many other states. The lines ofcommunication in South Dakota are not that difficult to overcome. It isn’t inmany states where I could say my local legislature was my hardware man, oneof my U.S. Senators did the contract for deed when I bought my house, or I canget on the program of the local service club just about any time I want. An exam-ple of interest on the part of policy-makers is the student poster session in theCapitol in Pierre, which incidentally was cosponsored by the Academy ofScience. Nearly half the legislature showed up!

V. Conclusion The SDAS is in an excellent position to become a strong voicefor science education in South Dakota. It is made up of scientists from all of theschools in the state, public and private, it has ties to industry, and has strong con-nections to the K-12 sector. Its membership runs the spectrum from nationallyprominent research scientists to excellent classroom teachers, and there aren’ttoo many members that don’t serve in both roles. Many of our members arewell-connected to national societies and therefore keep abreast of innovations inthe nation. The Academy, as a whole, has a broad view of the world of researchand education, and has the stature to not only respond to the forces acting uponus, but to help define those forces and shape the local and national responses tothem.

SENIOR RESEARCH PAPERS

PRESENTED AT

THE 83RD ANNUAL MEETING

OF THE

SOUTH DAKOTA ACADEMY OF SCIENCE


TRANSPORT OF NUTRIENTS AND PHYTOPLANKTON INTO

TWO GLACIAL PRAIRIE LAKES

Lois Haertel and Nels H. Troelstrup, Jr. Department of Biology and Microbiology

South Dakota State UniversityBrookings, SD 57007

ABSTRACT

The objectives of this study were to describe and compare inflow rates,nutrient concentrations and phytoplankton taxa in two South Dakota prairie lakesof different trophy: Eutrophic Lake Cochrane (Deuel County) and hypertrophicOak Lake (Brookings County).

Stream flows, total phosphorus (TP), total Kjeldahl nitrogen (TKN), nitrite-nitrate-N, iron, silica, manganese, calcium, sodium, conductivity and algalspecies composition and density were measured from Lake Cochrane on threedates in 1997. Sampling sites included a natural inflow below a sediment con-trol dam, an artificial diversion pipe from Lake Oliver to Lake Cochrane, an arti-ficial outflow from Lake Cochrane and a mid-basin site. Lake Oak was sampledon two dates in 1997 from major inflows from the west and north, spring seep-age flowing into the lake on the west shore, and a mid-basin site.

Mean concentrations of TP in Lake Cochrane were highest in the inflowfrom the sediment control dam, intermediate in the inflow from Lake Oliver andlowest midbasin. TN concentrations were highest midbasin and in the inflowfrom Lake Oliver, and lowest in the inflow from the sediment control dam.Average P loading rates were 116.3 g/da from Lake Oliver and 8.2 g/da from thesediment dam. Outlet removal was estimated at 24.8 g/da. TN loading rateswere 43.7 g/da from the sediment control dam and 2005.6 g/day from LakeOliver. Outlet removal was estimated at 919.2 g/day. Populations of green algaeand cryptophyte flagellates were highest in the in the inflow from the sedimentcontrol dam. Dinoflagellates and Botryococcus were highest midbasin.Chrysophyte flagellates and coccoid bluegreen algae were most abundant in theoutlet.

In Oak Lake, mean concentrations of both TP and TKN were highest mid-basin and lowest in the inflows. Average P loading rates were 25.5 g/da from thewest inflow, 2.6 g/da from the west seepage area and 68.3 g/da from the northinflow. Average N loading rates were 94.0 g/da from the west inflow, 67.7 g/dayfrom the west seepage and 715.3 g/day from the north inflow. Populations ofmost taxonomic groups of algae were greatest at the midbasin site.

Loading of P into Lake Cochrane may threaten the high water quality cur-rently present in that lake. The Lake Oliver inflow is of particular concern. Oaklake may be generating much of its P internally. N-fixing bluegreen algae are


stimulated by high P levels and contribute to high TKN levels measured midlake.Restoration of higher water quality to Oak Lake may require sediment removal.

INTRODUCTION

Prairie lake water quality has been degraded by cultural eutrophication andformation of blooms of bluegreen algae (Cyanobacteria). Inflows of nutrients,particularly phosphorus (P) into lakes has been assumed to stimulate algalblooms (Thomas 1969) and removal of P from inflowing waters has resulted indramatic improvement of water quality (Thomas, 1973, Edmondson 1991).Algae increases in prairie lakes have been associated with increases in both nitro-gen (N) and P (Haertel 1976, Smith 1982, Buskerud and Haertel 1992). Iron (Fe)may also be limiting to many algae and may prevent N-fixation by bluegreens(Goldman and Horne 1983). Silicon (Si) is a major nutrient for diatoms and lackof Si may trigger a change from a more desirable diatom flora to a less desirablebluegreen flora (Schelske and Stoermer 1972). Manganese (Mn) can also some-times limit algae (Lange 1971) and limits the rate of photosynthesis (Vymazal1995). Calcium (Ca) influences algal nutrient uptake (Rigby et al 1980) and alsomay help precipitate P from the water column (Danen-Louwerse et al l995).Sodium (Na) may benefit N metabolism in bluegreen algae (Ward and Wetzel1975).

The purpose of this study was to compare concentrations of nutrients andphytoplankton, in inflows and midbasin waters of Lakes Oak and Cochranelocated in eastern South Dakota. Loading rates were also calculated for P and N.

STUDY AREA AND METHODS

Oak Lake is hypertrophic (mean midbasin total P of 277 ug/l, mean Secchidepth of 0.2 m, and mean chlorophyll a of 86 ug/l), has a mean specific conduc-tance of 472 uS/cm, a surface area of 1.6 km2, a mean depth of 1.1 m, a maxi-mum depth of 2 m, and drains a watershed of 16.8 km2 (Troelstrup, unpublisheddata). Intermittent streams flow into Oak lake from the west, north and south.Above ground springs also contribute water to the west shore of Oak lake and tothe west inflowing stream. Sampling stations in Oak Lake (Figure 1) were in thenorth inlet, west inlet, cattail littoral zone at the entrance of the west inlet, down-stream from one of the major seepages and midbasin. The west inlet drains anarea of 2.4 km2 and the north inlet drains an area of 6.0 km2.

Lake Cochrane is eutrophic (mean midbasin total P of 24 ug/l, mean Secchidepth of 1.3 m, and mean chlorophyll a of 10 ug/l), has a mean specific conduc-tance of 2119 uS/cm, a surface area of 1.5 km2, a mean depth of 3.9 m, a maxi-mum depth of 7.9 m, and drains a natural watershed of 3.6 km2. Intermittentstreams enter Lake Cochrane from the south and west. The lake has no naturaloutflow. The south inlet to Cochrane was not flowing in 1997. The west inlet toCochrane enters a constructed open water sediment retention pond before flow-ing through cattail-bulrush littoral zone. Beginning in 1993, an artificial diver-


sion was constructed to allow water from Lake Oliver to enter Lake Cochrane,adding runoff from the Oliver watershed of 1.3 km2 to Lake Cochrane. An arti-ficial outlet from Cochrane had previously been constructed. Sampling stationsin Lake Cochrane (Figure 2) were in the west inlet just below the sediment reten-tion pond, in the artificial inlet from Lake Oliver just above or below the finalculvert entering Lake Cochrane, in the central lake basin, and just below the arti-ficial outlet culvert.

Water Chemistry

Oak Lake was sampled June 17 and July 14, 1997. The west littoral zonestation was not sampled July 14. Replicate samples were collected from LakeCochrane June 11, July 14 and August 26, 1997. After June 11, only current flowmeasurements were taken in the outlet. Two replicate samples of each variablewere taken at each location. Samples for total P were frozen in polycarbonatebottles, and samples for Total Kjeldahl N (TKN), nitrite-nitrate N, and chloro-phyll a were refrigerated prior to return to the laboratory. Nitrogen sampleswere processed the day after collection.

The following methods of the U. S. Environmental Protection Agency(1983) were used for laboratory analysis: TKN 351.3 (colorimetric), nitrite-nitrate N 300A (ion chromatography), total P 365.1 (persulfate digestion, colori-metric), and cations by atomic absorption, direct aspiration (Ca, 215.1, and Na273.1). Total nitrogen was determined by summing TKN and nitrite-nitratenitrogen. Nitrite N was assumed to be negligable at the pH levels measured(Mortimer 1941-1942). Chlorophyll samples were filtered and frozen the sameday as collected. Chlorophyll a was measured colorimetrically following ace-tone extraction (APHA 1985).

Field measurements of temperature were made with a bucket thermometer.Electrical conductivity was measured with a LaMotte DA DS conductivity meter.pH was determined colorimetrically (Hach Chemical Corp., Loveland, CO,method 17-N). Turbidity was measured with a Hach 2100 P turbidometer.Water transparency was determined with the use of a 20 cm white and blackSecchi disk. Field chemical measurements were performed for Fe (Hach IR-21)Mn (Hach Mn-PAN) and Si (Hach SI-7 or SI-5) to enable same day analysis.Stream Discharge

Inlet and outlet stream flows were estimated from measurements of streamwidth, depth and current velocity (@60% of maximum depth) from severalpoints within each channel (Carter and Dividian 1969). Oak Lake stream flowswere measured on chemistry sampling dates (except for outlet and south inletsites). Additional stream flow data were obtained from the Oak Lake FieldStation Monitoring database (Troelstrup, unpublished data) to improve estimatesof critical and actual loading to Oak Lake. These data were collected biweekly,during the ice-free season from 1996-1998. Lake Cochrane stream flows weremeasured only on chemistry sampling dates (1997). No other discharge datawere collected for this basin.


Phytoplankton

Algal samples were preserved in Lugol’s solution and counted, after settling,in a Sedgwick-Rafter Cell at both 100x and 300x, using a Whipple Disc. Two ormore crosswise swipes were counted at low power for larger forms and one ormore swipes at high power for all forms. All samples were counted until at least100 units of the most abundant species were encountered (Lund et al. 1958).Eukaryotic algae were counted as cells. Large colonial bluegreen algae werecounted by measuring the number of squares of the Whipple Disc grid coveredby the colony. Values were then converted to cells by counting average numbersof cells per square and multiplying by the number of squares covered. Length offilamentous bluegreen algae was measured using the Whipple Disc and similar-ly converted to number of cells.

Algae were identified and counted as species whenever possible, but forsimplicity, some taxa were analyzed as genera. Nitzschia and Synedra could notbe reliably separated in a Sedgwick-Rafter cell and were counted by size cate-gories. Identification of selected individuals on a regular microscope slideenabled determination of the more abundant forms. Infrequently encounteredgenera were added into higher taxonomic groups totals. Eukaryotic algae wereidentified according to keys in Smith (1950), Prescott (1962, 1978), Tiffany andBritton (1971), and Komarkova-Legnerova (1969). Most bluegreen algae wereidentified according to Drouet (1959), where the traditional genera Microcystisand Aphanothece are changed to Anacystis and Coccochloris, respectively.Gleocapsa and Calothrix were identified according to Rippka et al (1979).Statistical Analysis

Means of measured concentrations of variables were compared between sta-tions within lakes using one-way analysis of variance (PROC GLM, SASInstitute Inc. 1989).

RESULTS

Nutrient levels

In Oak Lake, turbidity, pH, total P and total N concentrations were muchhigher in the midbasin station than in any of the tributaries (Table 1). Becauseof the larger flow rate, most of the incoming N and P was entering the lake fromthe north inlet (Table 2). Far more N and P was leaving the lake through the out-let than entering from the inflows.

Si was highest in the west stream and seepage sites and showed depletion inthe midbasin site (Table 3). Fe and Mn were highest in the west stream and lit-toral zone sites and showed depletion in the midbasin site but not to levels thatwere likely to limit the growth of algae. Mn concentrations were below detec-tion (0.05 mg/l) in the north inlet and may have limited the growth of algae. Ca,Na and electrical conductivity were high in the seepage area and nearby west lit-toral zone.


Lake Cochrane P concentrations showed a reverse pattern from that in Oakwith high concentrations measured in the inflows and low concentrations mea-sured midbasin (Table 3). TKN was high in both midbasin and the Oliver inflowwhereas nitrate+nitrite-N was highest in the west inflow. Because of the higherflow rates in the Oliver inlet, both N and P loading into Cochrane was highest inthat inflow (Table 4). Also, the littoral zone downstream from the sediment out-let removes N and P from the water before it enters the lake (Haertel et al 1995).There is no littoral zone at the opening of the culvert that dumps Oliver waterinto Cochrane; nutrient loading shown in Table 4 is the nutrient loading enteringthe lake. In Cochrane, much more P and N were entering the lake than leavingby the outlet. Cochrane appears to be retaining N and P while Oak is exportingboth nutrients downstream.

Concentrations of Si, Fe, Mn and Ca were highest in the west inlet anddepleted downstream (Table 3). Conversely, both Na and electrical conductivi-ty levels were highest in the midbasin and outlet sites, reflecting natural concen-tration processes when the lake had no outlet. Midbasin conductivity levelsdecreased from 2556 uS June 11 to 1805 uS August 26, 1998, possibly as a resultof flushing Lake Cochrane with water from Lake Oliver.

Phytoplankton

In Oak Lake, coccoid bluegreens, N-fixing filamentous bluegreens, centricdiatoms, green algae, and unidentified small flagellates (probably mostly chrys-ophytes), showed increased concentrations at the midbasin station relative to theinflows (Table 5). Intermediate concentrations of several greens and N-fixingbluegreens were found at the west littoral zone station, probably as a result ofmixing with midlake water. Conversely, pennate diatoms, and cryptophyte andeuglenophyte flagellates were most abundant at the littoral zone station and lowor intermediate at the midbasin station.

Algal taxa not shown in Table 5 included the bluegreens Calothrix sp. (mostabundant at the midlake station) and Nodularia harveyensis, Lyngbya contortaand L. versicolor (not different between stations); and the greens Staurastrumgracile (abundant midlake), Chlamydomonas sp. (abundant in the seepageinflow), and Oocystis sp. (not different between stations). Pediastrum spp.included P. duplex + P. boryanum. Nitzschia/Synedra was probably mostly N.acicularis.

In Lake Cochrane, N-fixing bluegreens, green algae, pennate diatoms andfilamentous bluegreens were most abundant in water flowing out of the sedimentdam. Dinoflagellates and Botryococcus were most abundant at the midbasin sta-tion. Coccoid bluegreens and chrysophyte flagellates were most abundant in theoutflow. (Table 6).

Algal taxa not shown in Table 6 and not differing between stations includedthe N-fixing bluegreens A. holsatica and Calothrix sp., the centric diatoms S.niagarae, Chaetoceras elmoreii and M. varians, the green algae, Monoraphidiumsp. and Oocystis sp. and the dinoflagellate Glenodinium sp. Lyngbya spp. includ-


ed L. contorta and L. versicolor. Nitzschia/Synedra in Lake Cochrane was prob-ably mostly N. holsatica. Pediastrum spp. included P. duplex + P. boryanum.Scenedesmus spp. included S. quadridans and S. dimorpha)

DISCUSSION

Oak Lake and Lake Cochrane process nutrients differently. Cochrane is notexperiencing the same midbasin TP concentration as Oak. Cochrane is too deepfor wind resuspension of P-rich sediments whereas Oak is not (Haertel 1976). Inaddition, more abundant algae in Oak raise the pH through photosynthesis.Raised pH increases the solubility of Fe-bound P (Anderson 1975), increasingthe probability that suspended sediments will desorb P and stimulate even greateralgal growth. This, further raises pH and accelerates the process. Respiration ofthe very large algal biomass may also lower oxygen levels at the sediment waterinterface, encouraging P-release (Hosper 1980, 1997). Abundant N-fixing algaein midbasin (table 5) are stimulated by the high P levels.

Once this process has begun, it becomes self-perpetuating; it requires lowernutrient loading to trigger the change back to a less eutrophic stage than isrequired to trigger the change to a hypertrophic stage (Hosper 1997). Oak hasreached this self-perpetuating stage as documented by increased levels of TP,TKN, N-fixing bluegreens and many other algae midbasin. Cochrane has not yetreached this stage, however, if present rates of P loading continue, it may do so.

Vollenwieder (1976) developed an equation for calculating the critical loadof P that is likely to trigger the change to a more eutrophic state.Lc = Ec x qs (1 + (md/qs)0.5)Where:Ec – excessive loading concentration (20 ug/L as per Dillon 1975)qs - hydraulic load (m/yr) = md/TwTw - hydraulic residence time (yr) = Lake Volume (m3)/Annual Inflow (m3/yr)md - mean depth (m)

For Lake Cochrane the critical rate of P loading for 1997 flows measured thusbecomes 38.8 mg/m2/yr. Using our 1997 loading rate averages (Table 4), LakeCochrane is receiving 28.3 mg/m2/yr from Lake Oliver and 2.0 mg/m2/yr fromthe sediment control dam, for a total loading of 30.3 mg/m2/yr, 0.8 times the crit-ical rate. Non-point source loading (shoreline erosion, fertilizer use etc.) mayfurther contribute to actual loadings, pushing the lake over the critical rate.Groundwater inflows and outflows are also not considered in the above calcula-tion. However, groundwater is more likely to transport N than P.

Applying Vollenwieder’s (1976) formula to our 1997 data, the critical ratefor Oak Lake is 15.7 mg/m2/yr and the actual loading rate estimated is 22.0mg/m2/yr, 1.4 times the critical rate. Using the longer term stream dischargemeasurements available for Oak Lake, inflow and outflow rates become abouttenfold greater (Table 2). Recalculating the critical rate for Oak using the high-er average flows measured during the entire 1996-1998 time period, the criticalrate for Oak becomes 72.4 mg/m2/yr. If the P concentrations measured in the


inlets in 1996-1998 were the same as those measured in 1997, then the actualloading of P for Oak lake for the 1996-1998 period would become 236.9mg/m2/yr, or 3.3 times the critical rate. Thus our estimate of the ratio betweenthe loading rates and the critical rates measured for both Oak and Cochrane dur-ing 1997 may be underestimates. During periods of higher inflows, Cochranemay also be exceeding the critical rate. In addition, concentrations of inflowingnutrients may be higher in wetter years. Average total P concentrations of 161ug/l were measured in the west inlet to Lake Cochrane in 1993 (Haertel et al1995), slightly higher than the 154 ug/l measured in this study.

Because of high midlake concentrations of nutrients, Oak lake exported473.2 g/d more P to downstream water bodies than the lake received in 1997(Table 2). Nutrient release from wind-suspended midlake sediments was proba-bly the source of the additional P. Using the higher average flows measured inthe 1996-1998 period, and assuming 1997 nutrient concentrations, the estimateof P export rises to 8079.7 g/d. Allowing the outlet to flow naturally thusremoves nutrients from Oak Lake.

The situation is reversed in Lake Cochrane. Despite P export through theartificial outlet, Cochrane retained 12.7 g/d of the P coming in the inlets (Table4). Much of this excess P is probably incorporated into organic material.Increased midbasin concentrations of Botryococcus and dinoflagellate planktonsupport this hypothesis. Since most of this P is coming in from an artificialinflow (Lake Oliver, Table 4) and since the flow data in this paper are based ononly three dates of measurement, more nutrient loading data should be collectedfrom Lake Cochrane to properly evaluate the eutrophication risk of the P loadingfrom the Oliver diversion.

Hosper (1997) suggests that nutrient levels need to be much lower to reversethe eutrophication process than to initiate it. Because the algal bloom is self-sus-taining in Oak and abundant nutrients are likely to exist in the sediments,improvement in water quality may only be possible through dredging. However,the present high water quality in Lake Cochrane could be more inexpensivelypreserved by limiting nutrient inputs.

REFERENCES

Buskerud, S. T. and L. Haertel. 1992. Explanation of water transparency andplankton species abundance in a multibasin prairie lake. p. 75-90. In:Aquatic Ecosystems in Semiarid Regions: Implications for resource man-agement. R. D. Robarts and M. L. Bothwell (eds.) N. H. R. I. SymposiumSeries 7. Environment Canada.

Carter, R.W. and J. Davidian. 1969. General procedure for gaging streams. Book3, Chapter A6 of Techniques of water-resources investigations of the UnitedStates Geological Survey. United States Government Printing Office,Washington, D.C. 13p.


Danen-Louwerse, J. J., L. Lijklema and M. Coenraats. 1995. Co-precipitationof phosphate with calcium carbonate in Lake Veluwe. Water Res. 29:1781-1785.

Dillon, P.J. 1975. The phosphorus budget of Cameron Lake, Ontario: Theimportance of flushing rate to the degree of eutrophy of lakes. Limnologyand Oceanography 20(1): 28-39.

Drouet, F. 1959. Myxophyceae. In: W. T. Edmondson (ed.) Freshwater biolo-gy, 2nd edition. John Wiley and Sons New York.

Goldman, C. R. and A. J. Horne. 1983. Limnology. McGraw Hill, New York,464p.

Haertel, L. 1976. Nutrient limitation of algal standing crops in shallow prairielakes. Ecology 57:664-678.

Haertel, L., W. G. Duffy and D. E. Kokesh. 1995. Influence of vegetated wet-lands on the water quality of two glacial prairie lakes. J. Minn. Acad. Sci.59(4):1-10.

Komarkova-Legnerova J. 1969. The systematics and ontogenesis of the generaAnkistrodesmus Corda and Monoraphidium Gen. Nov. In Studies inPhycology. B. Fott, Ed. E. Schweizerbart’sche Verlagsbuchhandlung,Stuttgart.

Lange, W. 1971. Limiting elements in filtered Land Erie water. Water Res.5:1031-1048.

Lund, J. W. G., C. Kipling and E. D. LeCren. 1958. The inverted microscopemethod of estimating algal numbers and the statistical basis of estimationsby counting. Hydrobiology 11:143-170.

Mortimer, C. H. 1941-1942. The exchange of dissolved substances betweenmud and water in lakes. J. Ecol. 29:280-329; 30:147-201.

Prescott, G. W. 1962. Algae of the Western Great Lakes Area. Wm. C. BrownCompany Publishers, Dubuque, Iowa.

Prescott, G. W. 1978. How to know the freshwater algae. 3rd edition. Wm. C.Brown Company Publishers, Dubuque, Iowa.

Rigby, C. H., S. R. Craig and K. Budd. 1980. Phosphate uptake bySynechococcus lipoliensis (Cyanophyceae): Enhancement by calcium ion.J. Phycol. 16:389-393.


Rippka, R., J/ Deruelles, J.B. Waterbury, M. Herdman and R. Y. Stanier. 1979.Generic assignments, strain histories and properties of pure cultures ofCyanobacteria. J. General Microbiol. 3:1-61.

Round, F. E., R. M. Crawford and D. G. Mann. 1990. The diatoms. Biology andmorphology of the genera. Cambridge University Press, New York.

SAS Institute Incorporated. 1989. SAS/STAT users guide version 6. 4th edi-tion. Volume 2. SAS Institute Inc., Cary NC, USA.

Schelske, C. L. and E. F. Stoermer. 1972. Phosphorus, silica, and eutrophica-tion of Lakes Michigan. Limnol. Oceanogr. Special Symp. 1:157-170.

Smith, V. H.. 1982. The nitrogen and phosphorus dependence of biomass inlakes: an empirical and theoretical analysis. Limnol. Oceanogr. 34:1162-1173.

Tiffany, L. H. and M. E. Britton. 1971. The algae of Illinois. Hafner PublishingCo. New York.

Vollenwieder, R. A. 1976. Advances in defining critical loading levels for phos-phorus in lake eutrophication. Mem. Ist. Ital. Idrobiol. 33: 53-83

Vymazal, J. 1995. Algae and element cycling in wetlands. Lewis Publishers,Ann Arbor.

Ward, A. K. and R. G. Wetzel. 1975. Sodium: Some effects in Blue-Green algalgrowth. J. Phycol. 4:357-363.


Figure 2. Sampling locations within LakeCochrane, Deuel County, South Dakota during1997.Figure 1. Sampling locations with-

in the Oak Lake basin, BrookingsCounty, South Dakota during 1997.


ORNAMENTAL TRAITS IN HYALELLA AZTECAAS INDICATORS OF WATER QUALITY:

IMPLICATIONS FOR BIOLOGICALMONITORING

Eric A. Miller and Nels H. Troelstrup Jr.Department of Biology and Microbiology


Ornamental traits are sexually selected traits that an organism uses to attracta mate or defend against a rival. Studies have shown that ornamental traits aremore sensitive to environmental changes during development relative to natural-ly selected traits and tend to develop smaller in size or more asymmetrical inresponse to disturbance. Hyalella azteca is a freshwater benthic macroinverte-brate that has been used in numerous studies to establish water quality criteria infreshwater ecosystems. Male H. azteca use their antennae and gnathopods(claws) during precopulatory struggles. Hence, these traits are considered“armaments”. Thus, ornamental traits in H. azteca provide a unique opportuni-ty to examine the response of these traits to common disturbances affectingaquatic ecosystems such as nutrient enrichment. The objectives of the currentstudy were to 1) experimentally assess population level responses of H. azteca tonitrogen enrichment, 2) experimentally assess the ornamental trait response sig-nature of H. azteca to nitrogen enrichment and 3) characterize the ornamentaltrait response signature of natural populations of H. azteca in mesotrophic,eutrophic and hypereutrophic basins.

Amphipods were reared in 9 aquaria over a 7 week experimental period andwere randomly dosed with ammonium nitrate (NH4NO3). Three control tanksreceived blank doses (O mg/L NH4NO3) three aquaria received low level doses(10 mg/L NH4NO3) and three tanks received high level doses (50mg/L NH4NO3):Morphological characters of test organisms were measured with a computerimaging system and morphometry program. Morphological characters were alsomeasured on amphipods collected from 9 randomly selected lakes in theNorthern Glaciated Plain ecoregion.

Survival of amphipods was significantly different in all three treatmentswith highest survival in control amphipods. Male morphological traits (totalbody, mean 2nd antenna and mean gnathopod length) and 2nd antenna asymme-try seemed to respond to a subsidy-stress condition in that largest response sig-natures were observed in low treatment tanks and smallest response signatureswere observed in high treatment tanks. Male 2nd antennae length was signifi-cantly smaller in high treatment tanks relative to controls. Female morphologi-cal traits (total length and mean 2nd antenna length) were progressively smallerin the treatment tanks relative to the control tanks and the only females with eggspresent in the brood pouch were from control tanks. Smallest ornamental trait


responses (mean 2nd antenna and mean gnathopod length) in male amphipodssampled from lakes were observed in hypereutrophic basins while largestresponses were observed in mesotrophic basins. However, total body length wasgreatest in hypereutrophic basins. Mean second antenna length was again theonly trait which varied significantly among lake classes. Total body length,mean 2nd antenna length and brood size in female amphipods sampled fromlakes were progressively smaller in more enriched lakes.

Results from this study indicate that 2nd antenna of H. azteca may be themost sensitive to ammonia stress and nutrient enrichment. In addition, overallgrowth and fecundity of H. azteca may be reliable indicators of stress in theaquatic environment.


COMPARATIVE ENERGY FLOW TO THE FISHCOMMUNITY IN A PRAIRIE STREAM AND AFORESTED STREAM USING GROWTH RATE

AND STABLE ISOTOPE ANALYSIS

Thayne A. Munce, Kyle N. Seifert, and Craig N. SpencerDepartment of Biology

Augustana CollegeSioux Falls, SD 57197

We compared energy flow to the fish community in two contrasting streamsin northwest Iowa. Analysis focused on creek chubs (Semotilus atromaculatus)collected from an open prairie stream (Anderson Creek) and a forested stream(School Creek). Creek chubs from the prairie stream had significantly highergrowth rates than fish from the forested stream. Mean weights for age 0+ fishincreased from 7.4g in the forested stream to 10.3g in the prairie stream. Meanweights for age 1+ fish increased from 12.5g to 22.3g. By contrast, age 2+ fishwere comparable in size in the two streams.

These results are consistent with the well-known river continuum conceptwhich predicts increased energy flow to food webs in open-canopied streamsdue to increased in-stream primary (autochthonous) production resulting fromhigher light availability compared to heavily-shaded forested streams which relymore on detritus-based (allochthonous) energy sources from the watershed. Weattempted to trace shifts in food sources to food webs of the study streams usingstable isotopes, however the results were mixed. The δ15N values for creekchubs from the prairie stream were significantly higher than from the forestedstream, although the magnitude of the increase was small (0.5 %o). There wasno difference in δ13C values in fish from the two streams.

INTRODUCTION

In the upper midwest, prairie streams with open canopies predominate.However, there are scattered forested areas where closed-canopied streams arefound. Small forested streams typically have allochthonous-based food webs(Minshall 1967; Fisher and Likens 1973; Cummins et al. 1982), while open-prairie streams have autochthonous-based food webs. Allochthonous-basedfood webs are supported by substantial inputs of terrestrial plant material withlimited autochthonous production due to shading by terrestrial vegetation.Autochthonous-based food webs rely primarily on internal carbon sources suchas periphyton as their chief energy source (Minshall 1978; Busch and Fisher1981). Greater autochthonous production and decreased inputs of terrestrialmaterials can influence consumer trophic levels within the stream (Murphy and


Meehan 1991). Invertebrates that consume autochthonous energy sources ben-efit from improved food resources when more sunlight reaches the stream chan-nel, nutrient concentrations are elevated, and algal growth is stimulated (Weber1981). Such enrichment may also have an impact on higher trophic levels, suchas fish.

Analysis of fish growth can be an indirect method of comparing energy flowto contrasting stream ecosystems. In the present study, we compared fish popu-lations from a prairie stream and a forested stream. We focused on creek chubs(Semotilus atromaculatus), the most common predatory fish in the studystreams. In addition to standard analysis of fish growth and food habits, we alsoused stable isotope analysis as an indirect means of studying the flow of organ-ic material through stream food webs (Peterson and Fry 1987). In previouswork, Rosenfeld and Roff (1992) found that algal carbon was significantly more13C depleted (-35.5%o) than terrestrial carbon (-27%o), and Junger and Planas(1994) found that stable isotope values of δ13C can be used as tracers in provid-ing signatures consistent with allochthonous carbon sources in small shadedstreams, while autochthonous carbon signatures are more prevalent in openstreams. Despite these findings, other studies have demonstrated an absence ofa distinct isotopic signature or an extensive variability among various plant types(Fry and Sherr 1984; Ehrlinger and Rundel 1989). In effect, this may limit theaccuracy or usefulness of δ13C as a tracer in food webs. In a recent review,France (1994) analyzed 803 published measurements of δ13C for allochthonouslitter and both lotic attached algae and consumers, and concluded that for 50%of the fishes and 70% of the invertebrates it was impossible to discriminatebetween allochthonous and autochthonous carbon dependency based upon δ13Canalysis. In some cases, an enhanced analysis of food sources can be accom-plished through the use of δ15N in addition to δ13C (Fry 1991). Therefore, weconducted dual isotope analyses in the present study in attempting to trace theflow of allochthonous and autochthonous food sources in two contrastingstreams.

STUDY SITES

We selected two second-order study streams in northwest Iowa near thetown of Estherville. School Creek, a relatively clear, rocky-bottomed streamruns through the central valley of Fort Defiance State Park, an area characterizedby steep valleys with heavy forest cover of deciduous type. It is important tonote that at the time when samples were taken from this stream (mid-October,1997), free-flowing conditions did not exist. Rather, the stream was character-ized by intermittent pools separated by stretches of dry stream bed. These con-ditions were quite different than those of a preliminary sampling effort in thespring of 1997, when School Creek flowed continuously through Fort DefiancePark.

Above the valley in the grassland/cropland area of this region, a few milesfrom School Creek, is an state preserve called Anderson Prairie. It is character-


ized by little to no tree cover, and gently rolling hills dominated by smoothbrome (Bromus inermis), and switchgrass (Panicum virgatum). Through thispreserve runs a small stream similar in size to School Creek, which is identifiedas Anderson Creek. This creek was free-flowing during both of our samplingperiods.

METHODS

A preliminary study was conducted on May 3, 1997 focusing on larger fish.Based on results from this preliminary study, a more comprehensive study wasdesigned for the following fall. Field collections for the second phase of the pro-ject were conducted on October 15, 1997. Fish were collected from the studysites using seines (mesh size 4-6mm). In the 150m tract where most of our sam-pling was done on School Creek, only three large pools existed in the Octobersampling. These pools ranged in length from 12.5m to 14.0m, with an averagewidth of approximately 3m.

Back at the laboratory, fish were sorted by species using a taxonomic keyby Eddy and Hodson (1982). The creek chubs were measured for total length andweighed. Scales were removed from both the medial and posterior flank of thelargest 25 creek chubs collected from each of the two creeks. An approximategrowth rate analysis can be made by aging fish through an interpretation ofannual layers laid down in the hard parts of fish, most commonly scales, in con-junction with current size measurements (Graham 1929, Everhart et al. 1953).In order to increase accuracy of our age estimates, each fish was aged indepen-dently by two different researchers until both agreed on the correct age. In somecases, large numbers of scales were examined before a useful sample was found.

The stomach contents of several of the Creek Chubs from each site wereexamined qualitatively under a dissecting scope. We also measured stable iso-tope values of both carbon and nitrogen on the 15 largest chubs from eachstream. Analyses were made on white muscle tissue following lipid removalusing a chloroform/methanol extraction (Bligh and Dyer 1959). The remainingmuscle tissue was dried overnight at room temperature, freeze dried, and thenground into a fine powder using a mortar and pestle. Samples were then treatedwith 1 N HCl for 24 hrs. to remove any potential carbonates (Rounick et al.1982), rinsed with distilled water, and dried at room temperature prior to isotopicanalysis. Samples were weighed into foil containers and combusted in a CarloErba CHN Analyzer. The remaining CO2 and nitrogen gasses were then analyzedfor stable isotopes with a SIRA-10 isotope ratio mass spectrometer. Carbon sam-ples were standardized to Peedee Belmnite and the nitrogen samples to atmos-pheric nitrogen. Precision was better than 0.2%o for C and 0.5%o for N. Isotopevalues were calculated as:


Xsample - Xstandard

δ13C or δ15N = _______________ x 1000Xstandard

where X= 13C/12C or 15N/14N.

RESULTS AND DISCUSSION

Growth Data

Creek chubs collected from Anderson Creek and School Creek fell into 3 to4 age groups (Figure 1). These age groups ranged from the young-of-the -year(YOY), or 0+ age group, to two 3+ age group fish from School Creek.Comparative statistical analyses using 2-way ANOVA were performed to com-pare size of creek chubs of various ages from the two creeks based upon lengthas well as weight (Table 1). Overall growth rates were significantly higher inAnderson Creek than School Creek (Figures 2&3, Table 1). There was a signif-icant interaction between age and weight (p<0.05), indicating that the differencein size between the two creeks was not uniform across all ages (Table 1).Comparison of individual age class indicated significantly higher growth ratesfor 0+ and 1+ fish in Anderson Creek but no difference for 2+ fish (Figures2&3). Mean lengths for age 0+ fish increased from 9.7cm in School Creek to10.8cm in Anderson Creek, while the mean weights for these fish increased from7.4g to 10.3g. For age 1+ fish, mean lengths increased from 11.6cm to 13.8cm,while mean weights increased from 12.5g to 22.3g.

The increased growth of creek chubs in the open prairie stream (AndersonCreek) may have resulted from increased invertebrate production, the primaryfood source of creek chubs, due to the generally higher protein content anddigestibility of algae and algal-based detritus compared to most allochthonousplant material in the forested stream (Triska et al. 1975). Moreover, increasedlight availability in the open-prairie stream may have stimulated autochthonousproduction, as reported in other studies (Bilby and Bisson 1992). In addition,immediately upstream of the study site, Anderson Creek flows through a pastureheavily laden with grazing cattle, which may also have stimulated autochtho-nous production through increased nutrient loadings from animal waste materi-al.

Our growth rate data suggests there may be a period of critical growth forcreek chubs climaxing near the end of the second growing season, when nutri-tional variability and enrichment may have its greatest impact on creek chubgrowth. After the first two years of growth, the nutritional benefits of the prairiestream may diminish, allowing those fish from a forested stream such as SchoolCreek to catch up in size to their autochthonous counterparts (Figures 2&3). Theolder surviving chubs are likely more mobile and may move to favorable feed-ing areas whereas younger fish may be restricted to smaller feeding areas.


Our field data lends further evidence of a more favorable environment forcreek chubs in Anderson Creek. We noted greater survival among the young ageclasses in Anderson Creek than School Creek (Figure 1). We actually collectedmore age 1+ chubs from Anderson Creek than age 0+ chubs, while our collec-tions in School Creek showed a marked reduction of age 1+ chubs compared toage 0+ chubs (Figures 1a & 1b). Additionally, the reduced water volume inSchool Creek observed in the Fall of 1997 likely limited the feeding areas avail-able to the creek chubs creating an intensified competitive feeding situation inwhich the increased mobility of the older fish would have been a great advan-tage. Finally, it is also possible that the small sample size (n=10) of the 2+ agegroups limited our ability to discern growth rate differences in these older fish.We had a much larger sample size for the two younger age classes (0+, n= 137).

In a preliminary study in the Spring of 1997, stream flows were higher, andwe collected a larger number of older creek chubs from both creeks. Analysisof these fish also provides evidence for enhanced fish growth in Anderson Creek(Figure 4). For age 3+ creek chubs, the weight of creek chubs from AndersonCreek was significantly greater than School Creek (p<0.05, t-test). Although wecaught significant numbers of smaller creek chubs in the spring of 1997, wereleased most of them and thus did not have sufficient numbers for statisticalcomparison.

There was no appreciable difference in length vs. mass ratios for the creekchubs from the two streams (Figure 5), indicating that creek chubs reaching agiven length have similar mass in both streams. Although nutritional statusappeared to differ in the two streams, the basic developmental growth patternsof the fish appeared unaffected. Although the 0+ and 1+ fish in School Creekwere smaller, their condition factor was nearly identical to fish from AndersonCreek.

Stable Isotope Analysis

Mean δ13C values for creek chubs were not statistically different for the twostreams (Figure 6). The mean δ13C value for Anderson Creek chubs was -23.82%o compared to -23.74%o for School Creek (Figure 6). By contrast, themean δ15N value was significantly higher in Anderson Creek (mean = 13.65%o)than School Creek (mean = 13.17%o) (Figure 7).

There are several possible explanations for the nearly identical δ13C valuesin fish from the two streams. First, the diet of the chubs from both streams couldbe based on similar carbon sources which would result in analogous δ13C valuesin the fish. Such an explanation would be in conflict with previous conjecturesconcerning the relative importance of allochthonous and autochthonous foodsources in prairie and forested streams. However, in a recent review, France(1995) concluded that the overlap in δ13C values for allochthonous andautochthonous carbon sources makes it impossible to discriminate between car-bon dependencies in many stream ecosystems. Therefore, an alternative expla-nation could be that the carbon sources of the fishes’ diets from our two streams


are different, but that the different food sources possess nearly identical δ13Cvalues. This possibility remains open as we did not measure isotope ratios inpotential food sources in this study.

Previous studies have shown δ15N values to be an effective indicator oftrophic level with an increase of 1.5-5% per trophic position due to a mass-dependent isotopic fractionation during nitrogen excretion (Cabana andRasmussen 1994). The slightly higher δ15N for chubs in Anderson Creek couldindicate higher trophic placement for these fish (Figure 7). This interpretationlends further support to our previous conclusions about the Anderson Creek foodwebs in that a broader trophic spectrum with additional intermediate levels mayindicate a more productive system capable of supporting a more elaborate troph-ic system. Nevertheless, one must be careful not to place too much emphasis onthis point as δ15N values only differed by 0.5%o between the two streams.

Stomach Contents

In addition to stable isotope analysis, we also analyzed stomach contents ofchubs as an indicator of food sources. Many components encountered in fishstomachs are either unidentifiable (e.g. detritus) or non-plant items (e.g., ani-mals) and thus contain no apparent information about the original plant carbonsource (Forsberg et al. 1993). In addition, the identifiable components in a fish’sstomach may actually represent the least digestible elements in its diet. Finally,stomach contents only provide a snapshot of the immediate food sources. Bycontrast, fish growth and isotope values are integrated over the lifespan of thefish. Nonetheless, qualitative observations of stomach contents of the creekchubs were indicative of stream conditions and food availability at the time ofcapture. Stomach contents of the largest chubs from Anderson Creek containeda variety of identifiable material including snails, beetle exoskeletons, earth-worms and insect appendages. In comparison, the stomachs of all the SchoolCreek chubs evaluated were either empty, or contained amorphous organicmaterial that was not identifiable. It is worth recalling the physical conditions ofSchool Creek at the time of sampling which more than likely contributed todepleted stomach contents; the stream flow was non-existent, fish were confinedin pools where the food supplies would likely have been depleted. Qualitativeanalysis of stomach contents collected during the Spring of 1997, when bothcreeks were free-flowing, indicate that the stomach contents were comparable,including adult Midge larvae (Tendipes) and adult Black Flies (Simuliidae), atthe time of study, suggesting food availability was different in the Spring of 1997than in the Fall of 1997.

CONCLUSIONS

Results from this study suggest that the open prairie stream (AndersonCreek) may be more productive than the forested stream (School Creek), asrevealed by analysis of creek chubs, a common top-level consumer. This con-


clusion is based upon significantly higher age/length and age/weight growthrates, increased survivorship, greater δ15N values, and higher quality of stomachcontents of creek chubs in Anderson Creek compared to School Creek. It is alsopossible that the periodic cessation of stream flow in School Creek contributedto reduced growth and development of fish in that stream compared to AndersonCreek, which remained free-flowing all year.

A long-term study, gathering data throughout the year would likely con-tribute to more definitive conclusions as to the structure and dynamics of thesestream ecosystems. Additional research on other components of the ecosystemincluding primary production, macroinvertebrates, and water chemistry wouldalso contribute to a better understanding of energy flow through these ecosys-tems.

ACKNOWLEDGMENTS

We would like to thank Pam Runistad and Travis Dierks for help with fieldwork and Mike Chapman for help with the stable isotope analysis. Thanks alsogo to Steve Lehtinen who introduced us to the study areas and helped with theinitial early work..

REFERENCES

Bilby, R.E., and P.A. Bisson.1992. Allochthonous versus autochthonous organicmatter contributions to the trophic support of fish populations in clear-cutand old-growth forested streams. Can. J. Fish. Aquat. Sci. 49: 540-551.

Bligh, E.G., and W. J. Dyer. 1959. A Rapid method of total lipid extraction andpurification. Can. J. Biochem. and Phys. 37: 911-917.

Busch, D.E., and S.G. Fisher. 1981. Metabolism of a desert stream. FreshwaterBiol. 11: 301-307.

Cabana, G., and Rasmussen, J.B. 1994. Modeling food chain structure and con-taminant bioaccumulation using stable nitrogen isotopes. Nature. 372:255-257.

Cummins, K.W., J.R. Sedell, F.J. Swanson, G.W. Minshall, S.G. Fisher,C.E.Cushing, R.C. Peterson, and R.L. Vannote. 1982. Organic matter bud-gets for stream ecosystems: problems in their evaluation, p. 299-353. InG.W. Minshall and J.R. Barnes (ed) Stream ecology: application and testingof general ecological theory. Plenum Press, New York, NY.

Eddy, S., and Hodson, A.C. 1982. Taxonomic keys to the common animals of theNorth Central States: exclusive of the parasitic worms, terrestrial insects,and birds. Fourth ed. Burgess, Minneapolis, MN.

Ehrlinger, J.R., and P.W. Rundel. 1989. Stable isotopes: history, units, and instru-mentation. p. 1-15 in P.W. Rundell, J.R. Ehrlinger, and K.A. Nagy (eds.).Ecological Studies 68: Stable Isotopes in Ecological Research. Springer-Verlag, New York.

Everhart, W.D. and W.D. Youngs. 1953. Principles of Fishery Science. CornellUniversity Press.Ithaca, NY.


Fisher, S.G. , and G.E. Likens. 1973. Energy flow in Bear Brook, NewHampshire: an integrative approach to stream ecosystem metabolism. Ecol.Monogr. 43: 421-439.

France, R. 1994. Critical examination of stable isotope analysis as a means fortracing carbon pathways in stream ecosystems. Can. J. Fish. Aquat. Sci. 52:651- 656.

Forsberg, B.R., Araujo-Lima, C.A.R.M., Martinell, L.A., Victoria, R.L., andBonassi, J.A. 1992. Autotrophic carbon sources for fish of the centralAmazon. Ecol. 74(3): 643-652.

Fry, B. 1991. Stable isotope diagrams of freshwater food webs. Ecology 72:2293-2297.

Fry, B., and E.B. Sherr. 1984. 13C measurements as indicators of carbon flow inmarine and freshwater ecosystems. Contributions in Marine Science 27:13-47.

Graham, M. 1929. Studies of age determination in fish. Part II. A survey of theliterature. Min. Agric. Fish and Food (U.K), Fish. Invest. Ser. II, 11(3):1- 50.

Junger, M. and D. Planas. 1994. Quantitative use of stable carbon isotope analy-sis to determine the trophic base of invertebrate communities in a boreal for-est lotic system. Can. J. Fish Aq. Sci. 51: 52-61.

Minshall, G.W. 1967. Role of allochthonous detritus in the trophic structure of awoodland spring brook community. Ecology 48: 139-149.

Minshall, G.W., 1978. Autotrophy in stream ecosystems. BioScience 28: 767-771.

Murphy, M.L., and W.R. Meehan.1991. Stream ecosystems. Am. Fish. Soc.Spec. Publ. 19:17-46.

Peterson, B.J., and B. Fry. 1987. Stable isotopes in ecosystem studies. Annu.Rev. Ecol. Syst. 18: 293-320.

Rosenfeld, J.S., and J.C. Roff. 1992. Examination of the carbon base in southernOntario streams using stable isotopes. J.N. Am. Benthol. Soc. 11:1-10.

Rounick, J.S., M.J. Winterbourn, and G.M. Lyon. 1982. Differential utilizationof allochthonous and autochthonous inputs by aquatic invertebrates in someNew Zealand streams: a stable carbon isotope study. Oikos 39: 191-198.

Triska, F.J., J.R. Sedell, and B. Buckley. 1975. The processing of conifer andhardwood leaves in two coniferous forest streams: II. Biochemical andnutrient changes. Verh. Int. Ver. Limnol. 19: 1628-1639.

Weber, P.K. 1981. Comparisons of the lower trophic levels of small streamcommunities in forest and clearcut sites, southeast Alaska. Ph.D. thesis,University of Washington, Seattle, WA.


A FIELD-ORIENTED GENERAL CHEMISTRYCOMMUNITY PROJECT:

RAPID CREEK WATER QUALITY

Martha M. Tremblay and Cathleen J. WebbDepartment of Chemistry and Chemical Engineering

South Dakota School of Mines and TechnologyRapid City, SD 57701

Rapid Creek water quality was determined during a group-oriented field

research project in freshman chemistry. Student groups selected a stretch of

Rapid Creek to map and develop a site appropriate sampling plan. Samples were

collected and analyzed for a variety of water quality parameters, including anion

and metal analysis. Course work was coordinated with aspects of the field and

lab work to encourage the linkage of chemistry to a “real” situation.

Sampling sites were selected for a variety of reasons such as impacts from

industrial waste, drainage ditch runoff, and wildlife contamination. Student

teams recommended a large number of sites for sampling. The final selection

limited the sites to fifteen. The sampling teams prepared the sample bottles by

washing them and rinsing them with distilled water. At the sample sites the bot-

tles were rinsed twice with ambient water before the samples were collected.

The pH of each sample was tested. Each field sample was prepared for metal,

anion, total suspended and total dissolved solid analysis.

The following conclusions have been made from the analysis so far com-

pleted. Urban impacts were clear, because chloride levels were clearly elevated

in urban areas. Sulfate concentrations were elevated throughout all sampling

areas. Impacts from industrial sources were obvious because almost all analyte

levels were higher where industrial inflows entered the creek, near the middle of

town. Nitrate concentrations were more ambiguous due to the date

of sample collection (November 1997). Nitrate levels, in general, are lower in

the winter and higher in the summer. The data collected, to date, has shown a

steady degradation of the water quality along the entire length of Rapid Creek,

Pactola Reservoir to the Waste Treatment Facility of Rapid City. The data also

illustrated potential impacts from the Meadowbrook Golf Course.


STRAWBERRY CLOVER AND ITS USE IN THEAMELIORATION OF CADMIUM

CONTAMINATED SITES

Peter Jauert and R. Neil ReeseDepartment of Biology/Microbiology


ABSTRACT

Cadmium is found naturally in soils throughout the Midwest as well as inold mining sites of South Dakota. Two proposed methods for cleaning up thesesites involve the use of cover crops to ameliorate the soil. These crops may beused to change the soil’s characteristics to make the Cadmium unavailable toplants. In smaller sites, cover crops which would take up high levels ofCadmium could be harvested, physically removing the Cadmium from the soil.Strawberry clover is a legume found throughout the world and is related to thered clover and the white clover, common to this area. It is a very hardy plant withthe ability to tolerate saline and alkaline soils as well as flooding. This makes ita good prospect for reclamation purposes.

An ongoing research project has been set up to look at 98 accessions ofstrawberry clover received from the Western Regional Plant Introduction Stationin Pullman, Washington. These seeds were planted in the greenhouse for com-parisons of morphological variations between accessions. Different traits will beselected for based on the type of soil remediation for which the plants will beused. Strawberry clover was also grown hydroponically to examine differencesin Cadmium uptake. Comparisons of changes in rhizosphere pH by rootsbetween accessions were made using colorometric agar plates. Future researchwill look for a correlation between differences in Cadmium uptake and the abil-ity of plants to change rhizosphere pH.


LEAD SOIL LEVELS AT FOUR ELEMENTARYSCHOOL PLAYGROUNDS IN RAPID CITY

Derek Thirstrup and Cathleen J. WebbDepartment of Chemistry and Chemical Engineering


In the Spring of 1997, students in freshman chemistry participated in agroup-oriented field research project to determine lead levels in soils at elemen-tary schools in Rapid City. Student groups were assigned an elementary schoolin the Rapid City School District as their field site. The site was mapped. Teamsof students analyzed traffic patterns at each selected school over an entire day. Asite appropriate sampling plan was then developed. Soil core samples were col-lected and analyzed for lead both as a function of location and depth, using atom-ic absorption. Students from more advanced chemistry classes assisted in theareas of sample preparation, acid digestion of the soil and performance of theactual sample analysis. Field sample collection was performed in late spring,with the final laboratory analysis completed in May.

During the first part of the semester, lecture time was devoted to building anintellectual framework for the project with, for example, a discussion of thepotential sources of lead in soils and what the background levels should be. Thework related lecture material through the context of a “real” problem. Trainingin field and sample collection procedures took place in the first part of the semes-ter. Emphasis was on quality control, establishment of a “true” background forlead levels in soils and scientific ethics (Who does the data belong to?, Whatshould be done with it?, etc.).

Lead levels varied from essentially background to 25 ppm, in solution.Higher levels of lead were observed in older schools and those near major streets,presumable reflecting the time period when leaded gasoline were the standard.Correlation with traffic patterns were partially successful.


A STUDY OF HERBICIDE RETENTION IN TWO DIFFERENT HUMIN SOIL TYPES

Walter Whiteside, Justin Kurcirek, Christine J. Guetzloff, James C. Sorenson,and Thomas F. Guetzloff, Mount Marty College, Yankton, SD 57078

INTRODUCTION

Soil is a growth medium for plants which acts as a storage site for necessarynutrients, a buffering medium for roots, and acts as an anchor for root systems.Soil is comprised of two major components, organic materials and inorganicmaterials. The inorganics are comprised of mineral matter fractions (sand, silt,clay, etc.) that define different soil types, inorganic metals, and ions. The organ-ic matter in soil is formed by the decay of plant, animal, and microbial tissues.The organic portion, also known as humus, can be classified further into humicand non-humic materials. Non-humic materials include those well-known class-es of compounds such as fatty acids, lipids, proteins, carbohydrates, etc.(Stevenson, 1994)

Humic materials can be fractionated into three groups: humic acid, fulvicacid, and humin. Humic acid is soluble in alkaline conditions, fulvic acid is sol-uble at all pHs, while the third component, humin, is insoluble at all pHs (FigureI). Whole humin is comprised of mineral matter, unbound lipids, which areremovable by organic solvents (i.e. chloroform), a bound lipid component notremovable by organic solvents, a bound humic acid component and an insolubleresidue (Rice and MacCarthy, 1990). Soxhlet-extraction of humin will removethe unbound lipids, and bromine-treatment of humin eliminates the remainingorganic material accessible to oxidation.

Herbicides are often used by farmers to terminate or prevent the growth ofunwanted plants called weeds. When designing new herbicides, researchersshould be concerned with development of herbicide site selectivity for targetweeds, and should ultimately minimize long-term resistance of their targets overtime. These two factors are important because of past experience with suchchemicals as DDT (e.g. Chiou et al., 1986). Indiscriminate use of DDT in thenot-too-distant past led to environmental effects which are still appearing inorganisms today. Some herbicides bind to various components of soil (Xie et al.,1997) which may be slightly water soluble when bound and then may be trans-ported to the water supply. It is important to elucidate these chemical interac-tions because if a substance is bound in the soil to soil organic matter and doesnot appreciably degrade over time yet is slowly transported to groundwater, thenits effect would be different than a compound that is immediately transported togroundwater reservoirs after its application (Wershaw, 1992).

The interaction of soil and soil organic matter with contaminants, pesticides,and herbicides, has received much attention in recent years. The organic com-ponents of soil (humin) are important to study in this regard herbicides may bind


to the humin. The mechanism of herbicide transport through soil via humic mate-rials warrants studies to elucidate the chemical binding mechanisms of herbi-cides to soil and soil organic matter (Chiou, 1990).

One of the physiochemical sorption mechanisms used to explore the affini-ty of organic contaminants with humic materials is partitioning. Partitioning oforganic contaminants includes the complete permeation of the nonionic organiccontaminant molecule into the humin sorbent (Chiou et al., 1983) in which thesorbed material is dissolved in an organic phase by common solution forces (e.g.van der Waals interactions). Another mechanism used to describe organic inter-action with humic materials is sorption. Sorption is commonly used to describethe adsorption of the contaminant molecule (adsorbate) on to the surface of thehumin (adsorbent).

The removal of organic components such as lipids from soil may giveinsight into the interaction of herbicides with soil. The focus of this study wasto determine the sorptive effects of the herbicides, atrazine and 2,4-D, to the soilorganic matter fraction, humin. And to study the effect of lipid removal and theremoval of organic matter on the sorption of atrazine and 2,4-D to humin isolat-ed from two different soils.

MATERIALS AND METHODS

Humin

The humin was prepared by the traditional-alkali extraction method (Riceand MacCarthy, 1992) from a Poinsett Silt Loam soil, collected from the LakePoinsett area in South Dakoa and a Nebraska Sandy Loam soil from KnoxCounty near Lewis and Clark Lake. The types of humin used in the study werewhole, Soxhlet, and brominated humin. The Soxhlet humin was extracted withchloroform for 24 hours to remove lipids from the whole humin and air dried toremove excess chloroform (Howard, 1991). The brominated humin was treatedwith bromine to oxidize the organic matter remaining in the whole humin withminimal changes to the mineral component (Mitchell and Smith, 1974; vanLangeveld et al., 1978). The humin sample weight ranges were from 0.15 to 0.34grams. The humin samples were washed with distilled water until the absorbanceat the desired wavelengths was less than 0.1 absorbance units to remove residualhumic and fulvic acid (A blank without atrazine or 2,4-D was used as a referenceblank). The percent organic matter of each humin was determined by ashing thesample at 400°C for 4-6 hours. The particle size classification was determinedusing the standard Soil Texture Triangle.

Herbicides

Atrazine and 2,4-D were the selected herbicides in this study (Figure II).Stock solutions of both herbicides were prepared for solution in the appropriateconcentration range studied (parts per million). Atrazine was studied at five, ten,


fifteen, twenty-three, and thirty parts per million in distilled deionized water (pH4.0-5.0). The 2,4-D was studied at ten, twenty, forty, sixty, and eighty parts permillion in water (pH 3.0-4.0). The various solutions were pipetted into test tubes(3-5 mL of solution per test tube) containing 0.15-0.35 g humin and shaken peri-odically for twenty-four hours at 25°C. The samples and controls were kept indarkness to minimize possible light degradation. After the twenty-four hour peri-od each tube was centrifuged for five minutes at 500 RPM to remove suspendedhumin particulate matter. The absorbance of the supernatant was collected on aPerkin-Elmer Diode Array spectrometer at 222 nm for 2,4-D and 268 nm foratrazine. The concentration of the atrazine and 2,4-D was determined by usingstandard solutions (controls) to determine the amount of atrazine or 2,4-D sorbedto humin during the twenty-four hour period.


Table I lists the % organic matter and particle size distributions (% sand, silt,and clay) for the soils and humin isolated from Nebraska (NE) and South Dakota(SD). The % organic matter of whole soil and whole humin are higher thanbrominated humin and somewhat higher than lipid-free Soxhlet humin. NE soiland humins have a greater percentage of sand than SD soil. The SD soil andhumins have a greater percentage of silt and clay than the NE soil.

The amount of atrazine sorbed to humin is shown in Figures III and IV.Whole soil and whole humin from NE displayed similar linear isotherms with Kd

values of 2.33 and 2.73, respectively (Table II). Linear adsorption isotherms areindicative to partitioning of the sorbate molecule into the sorbent. The Kd forwhole humin from SD was significantly higher than NE, even though the percentorganic matter is higher in NE than in SD. But the SD soil has a greater per-centage of % silt than NE soil. After removal of lipids (Soxhlet) and organicmatter (brominated) the uptake of atrazine by humin is a nonlinear adsorptionisotherm. Downward curvature in the isotherm plot indicates adsorption of theatrazine on to the surface of the humin and loss of the ability of the atrazine tocompletely permeate the entire volume of the humin sample. Removal of organ-ic matter (brominated) results in a greater nonlinear increase of atrazine sorptionby humin from NE than removal of lipids. Since lipids and organic matter areremoved there must now be sites open or available on the mineral surface of thehumin samples and there is now very little organic matter left in the humin to actas the sorbent medium for partitioning.

The atrazine results seem to show a correlation between lipid/organic mat-ter content and sorption of atrazine to the humins or loss of the ability of atrazineto partition into the humin. Others have noted that atrazine was absorbed on tosites in soil but they were unsure of the sites; lipids may be one of the sites ofsorption (Hamaker and Goring, 1975). Atrazine has been able to transportthrough layers of soil and through aquifers (Graber, et al., 1995). That gives rea-son for concern as to where atrazine will be eventually deposited and concen-trated after it has been applied. An important aspect of atrazine to consider is that


in soil can be degraded into smaller components that remain in soil for a verylong time (Brouwer, et al., 1990). This degradation may come into prominencewith other atrazine studies of binding to humin and/or lipids. The sorption ofatrazine on to soil clay components has been shown and discussed in previousstudies (Laird, et al., 1994). Other researchers have commented that soils mayhave different adsorption isotherms of organic compounds due to individual soilcompositions, but these differences are not really that important (Chiou, et al.,1985).

The uptake of 2,4-D for NE and SD soil and humin is shown in Figures Vand VI. The sorption of 2,4-D to SD humin shows no significant difference in theuptake of herbicide by humin even though there is a slight decrease in the amountof 2,4-D adsorbed after removal of lipids and organic matter. The Kd values are2.46, 1.74, and 1.47 for whole, Soxhlet, and brominated humins of the linearadsorption isotherms. This could be due to the exposed mineral matter which isnet negatively charged at the surface of the humin. The sorption of 2,4-D to NEhumin shows little difference between the uptake of 2,4-D for whole soil andwhole humin even though there is slight downward curvature for whole soil.Removal of lipids (Soxhlet humin) results in a larger Kd value than whole humin.And brominated humin displays nonlinear adsorption isotherms with downwardcurvature. At high equilibrium concentrations of 2,4-D there appears to be aninhibiting affect on the sorption of the 2,4-D to the humin possibly due to elec-trostatic repulsive forces.

The results show that lipids and organic matter may not play as important arole in the sorption of 2,4-D as with atrazine. This is not unexpected as otherresearchers have noted that 2,4-D may be retained in soil by electrostatic forcesalone (Madrid, et al., 1993). The mineral matter of humin is negatively chargedand 2,4-D is also negatively charged which results in less sorptive behavior.Atrazine and 2,4-D do have separate applications in the field; atrazine is usedmainly on corn, and 2,4-D is used on a greater variety of plants. 2,4-D is a syn-thetic form of auxin which itself causes plants to grow, but the synthetic hormoneis used to keep unwanted plants from growing. The actions by which 2,4-D fullyworks are not totally understood, but would involve an affect on certain growthreceptors (Raven, Evert, and Eichhorn, 1992). These differences as well as a dif-ferent composition may be why the humin didn’t absorb as much 2,4-D. The 2,4-D must be affected by other variables and one of those may be competition withphosphate as well as other negatively charged compounds (Madrid, et al., 1993).

CONCLUSIONS

In comparison of the two different soil types, NE soil was classified as aSandy Loam and SD was determined to be a Silt Loam soil. Both atrazine and2,4-D displayed linear sorption isotherms for SD soil and humin uptake. It ispossible that the organic matter of the Silt Loam soil provided a more appropri-ate medium for partitioning into the humin. Whereas the NE soil was a SandyLoam with less of the appropriate type of organic matter needed for partitioning


and the nonlinear isotherms were exhibiting downward curvature. The removalof lipids and organic matter does alter the physiochemical aspects of the huminexposing the mineral matter and forming possibly new sites for atrazine and 2,4-D when compared to whole humin and whole soil.

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Howard, K.T. 1991. M.S. Thesis. South Dakota State University.


Laird, D.A., Yen, P.Y., Koskinen, W.C., Steinheimer, T.R., and R.H. Dowdy.1994. Sorption on soil clay components. Environ. Sci. and Technol.28:1054-1061.

Madrid, L., Marillo, E., and E. Diaz-Barrientos. 1993. Fate and prediction ofenvironmental chemicals in soils, plants, and aquatic ecosystems. LewisPublishers, Boca Raton, FL.

Mitchell, B.D., and B.F.L. Smith. 1974. The removal of organic matter from soilextracts by bromine oxidation. J. Soil Science. 25, 2, 239-241.

Rice, J.A., and P. MacCarthy. 1992. Disaggregation and characterization ofhumin. The Science of the Total Environment. 117/118:83-88.

Rice, J.A., and P. MacCarthy. 1990. A model of humin. Environ. Sci. & Technol.1875-1877.

Stevenson, F.J. 1994. Humus chemistry; genesis, composition and reactions. J.Wiley and Sons, New York.

Van Langeveld, A.D., van der Gaast, S.J., and D. Eisama, D. 1978. A compari-son of the effectiveness of eight methods for the removal of organic matterfrom clay. Clays and Clay Minerals. 26, 5, 361-364.

Wershaw, R.L. 1992. Membrane-micelle model for humus in soils and sedimentsand its relation to humification. U.S. Geological Survey Open-File Report91-513.

Xie, H., Guetzloff, T.F. and J.A. Rice. 1997. Fractionation of pesticide residues


Figure I. The particle size analysis and % organic matter of two different soil and humin types.


Figure II. The structuresof atrazine and 2,4-D.

Figure III. The uptake of atrazine by humin isolated from Nebraska.


Figure IV. The uptake of atrazine by humin isolated from South Dakota.

Figure V. The uptake of 2,4-D by humin isolated from Nebraska.


Figure VI. The uptake of atrazine by humin isolated from South Dakota.


NUMERICAL TREATMENT OF AMATHEMATICAL MODEL OF TWO

COMPETITORS IN A PARTIALLYPERIODICALLY MUDDY LAKE

Alicia Vander Weyst & A. S. ElkhaderDepartment of MathematicsNorthern State University

Aberdeen, SD 57401

This work is to analyze a mathematical model that predicts the outcome oftwo microorganisms competing for a limiting resource in a periodically partiallymuddy lake. The lake environment is divided into two parts, a clear part and amuddy part. The size of the muddy part of the lake is subject to seasonalchanges. The microorganisms and nutrients flow from one region to the other.

The model consists of four ordinary differential equations of nonlinear type.The growth functions of both populations are assumed to have Michaelis-Mentenform. The system has size parameters: the washout rate, the input concentration,the maximum growth rate, the half saturation rate, the volume of the muddyregion, and the migration rate. It had been shown that coexistence is not possi-ble in either region when these parameters are constants and there is no migra-tion between the regions. However, coexistence of the two populations is possi-ble in both regions when migration between them is allowed, or when any ofthese parameters is modified to a periodic function.

In this work, we study the model when the migration rate between the tworegions is allowed to a periodic function of the volume of the muddy region andall other parameters are kept fixed. We are able to show, using the mathematicalsoftware PHASER, that coexistence is possible under these modifications. Inaddition, we will show the behavior of the model when the size of the muddylake is very small and very large.


HYDROLOGY OF GLACIAL LAKES,FORT SISSETON AREA

Perry H. RahnDepartment of Geology & Geological Engineering


ABSTRACT

The “Glacial Lakes” area in Marshall and Day Counties in northeasternSouth Dakota contain hundreds of depressions in close proximity to each other.They are glacial kettles. Many are dry most years, but some contain ephemerallakes and some contain perennial lakes up to several square miles in area. Mostof this area is internally drained during typical years.

In 1995 many depressions filled up due to above-average precipitation.From 1995 to the present time (January, 1998) the lakes in the Fort Sisseton areahave been overflowing one into another, finding an outlet at the western edge ofCattail Lake at an elevation of 1792 ft above sea level. The water then flows intoCrow Creek and thence into the James River. This has happened only one timein recorded history, during the year 1916. The unusual flooding pattern beingexperienced during the last 4 years has caused hardships to local farmers whofind their hay fields submerged. Numerous roads have been closed, and litiga-tion over responsibility for the flooding is ongoing.

INTRODUCTION

The Prairie Coteau was formed as continental glaciers repeatedlyadvanced into South Dakota and Minnesota, dividing into two lobes by adrainage divide that existed between the present James River Valley and the low-land to the East. The Dakota Lobe extended down into South Dakota, followingthe present James River Valley, and the Des Moines Lobe extended down the pre-sent Lake Traverse region into Iowa. Lateral moraines accumulated in the areaof the division of the glacier, forming the hilly land known as the Prairie Coteau(Koch, 1975; Rahn, 1975).

As the last of the glaciers melted approximately 20,000 years ago, blocks ofice left within the moraines melted, leaving depressions known as kettles. Smallkettles are locally called potholes. Today many swamps and lakes in this area fillthe depressions, occupying a single kettle or coalescing kettles. Figure 1 showsthat the drainage basin that includes Fort Sisseton has a high density of kettles.Some of the larger lakes, such as Roy Lake, are not simply one large kettle, butwere formed by constructional morainal development as well.


Inspection of topographic maps (Figure 1) indicates at first glance that mostof the Glacial Lakes area has internal drainage. The land immediately adjacentto a typical kettle slopes into it and there is seemingly no outlet. But in extreme-ly wet years, the kettles can fill up with water, and then overflow to adjacent ket-tles and lowlands. Some of the kettles probably have never filled up since theywere formed some20,000 years ago. In 1995 an unusually wet spring led to thefilling of numerous kettles in this region, and the lakes began to coalesce into agiant complex and they finally spilled over to Crow Creek and thence into theJames River and into the Missouri River.

HYDROLOGY

The average annual precipitation at Britton, the county seat of MarshallCounty, was 18.81 inches as of 1995 (NOAA, 1995). This amount of precipita-tion is normally sufficient to produce some standing water in only the deeper ket-tles.

The annual precipitation in Britton was 5.55 inches above normal in 1994,7.45 inches above normal in 1995, 0.89 inches above normal in 1996, and 0.87inches above normal in 1997 (NOAA, 1996-97). During the early summer of1995, heavy rains filled the many depressions to the point where they began tooverflow. [A tremendous volume of water is necessary to accomplish this,because the elevation of the bottom of the Cattail Lake kettle complex is approx-imately 1782 ft, based on the USGS 7.5 minute topographic maps, and the ele-vation of the water surface at its maximum depth over the Cattail Lake complexwas about 1794 ft.] The order of filling in this region was from northeast tosouthwest, so that Buffalo Lake overflowed into Red Iron Lakes, and thence toClear Lake, Roy Lake, Lost Lake, Kettle Lake, and finally Cattail Lake.Ultimately the water reached an outlet along the western shore of Cattail Lake,and on July 19, 1995, began flowing westerly off the Prairie Coteau. This out-let, formerly an unnamed slough, is now referred to as Cattail Creek. From July19, 1995 until present (January, 1998), water has discharged continuously out ofthe Cattail Lake complex, except for the period January through March, 1997,when Cattail Creek froze up (Dan Hook, pers. comm., January 2, 1998). Cattailcreek gradually increases in gradient westerly as it descends off the PrairieCoteau, ultimately descending into Crow Creek at Hickman Dam (Figure 1).

Figure 2 shows the high waters surrounding Fort Sisseton State Park.Mallard Slough is shown as a perennial lake on the USGS 1:24,000 scale topo-graphic map, but Fort Lake is shown simply as a lowland swamp. In this 1996photo, both depressions are completely full of water, and flow into and aremerged with the Kettle Lake/Cattail Lake complex. Figure 3 shows the samedepressions during the normally drier conditions which existed in 1975. Note theabsence of Fort Lake.

Figure 4 shows the County Road #5 crossing of Cattail Lake. This locationis close to the outlet of the Cattail Lake complex as it existed om 1996. There isa culvert under the road at this location, and the operation of it was a factor in a


litigation brought about by landowners who claimed their lands were being inun-dated to some degree by roads constructed by the County Highway Department(Steiner et al. vs. Marshall County, South Dakota Supreme Court, 1997). Theareas covered by water in Figure 4 are shown as hayfields in 1975 photos (Figure3).

FLOODING FREQUENCY

Figure 5 is a detailed topographic map of the Cattail Lake and surroundingareas. The ground elevation of fhe outlet of Cattail Lake is 1792 ft above sealevel. Outlined is the shoreline of the Cattail Lake complex based on the watersurface elevation of 1794 ft. Waters remained at approximately this level from1995 through 1998. Local residents do not recall the Cattail Lake complex fill-ing ever before, although it reportedly did occur in 1916, the year the U.S. SoilConservation Service (Schultz, 1975) reported as the wettest year in 54 years ofrecord.

Fort Sisseton was built in 1864, and most likely was founded at this site forthe natural protection from Indian attack that was afforded by the surroundinglakes. Thus Fort Lake, Mallard Slough, and the Kettle Lake/Cattial Lake com-plex were probably full at that time.

From the above data, the frequency of inundation can be roughly deduced.It can be assumed that the lakes were full in 1864, then dry for 52 years until1916 when it filled then dry again for the following 79 years, then full during thepast 3 years. Thus the cycle is roughly that of 2 years full, with water discharg-ing to the James River, followed by 65 years of internal drainage.

CONCLUSION

The Glacial Lakes area of the Prairie Couteau area is unique in that it is oneof the few areas of the Midwest that lacks external drainage. The surroundingareas are drained by stream systems, generally flowing off the Prairie Coteau tothe west, north, and east. To the south, the headwaters of the Big Sioux Riverextends to near the town of Waubay Lake, about 10 miles south of the drainageof the Cattail Lake complex shown in Figure 1. But the exceptionally wet yearsof 1994-98 show that the Glacial Lakes can fill up and overflow to an externaldrainage, in this case via Cattail Creek to Crow Creek and thence into the JamesRiver. In the future geologic time, perhaps as little as 10,000 years, the gradientof Cattail Creek will probably increase as it erodes a channel coming off thePrairie Coteau into Crow Creek. It would then cause the outlet of Cattail Laketo be eroded below 1792 ft, and thus eventually integrate the Cattail Lake com-plex from a quasi-internal drainage to a more normal stream/lake system con-nected to the James River.

Flood damage to eastern Soiuth Dakota during the wet years of 1995 to 1998was severe. Damage during 1996 alone reached an estimated $46 million duringthe paralyzing blizzards and spring floods (Figure 6). Watertown was one of the


communities most severely impacted by flooding as the Big Sioux River spilledover its banks. The high precipitation of 1994-98 caused hardship to the farm-ers in the Glacial Lakes area who owned hayground in the bottom of kettleswhich are usually dry. The outflow from the Cattaiil Lake complex into CrowCreek added to the misery of farmers in the James River valley who were alreadybeing flooded by the James River. The total lost productivity and road damageis difficult to assess.

REFERENCES CITED

Koch, N.C., 1975. Geology and water resources of Marshall County, SouthDakota: South Dakota Geological Survey, Bull. 233.

NOAA, 1995-97. Climatological data annual summary, South Dakota, 1995;1996; 1997. National Atmospheric and Ocean Administration.

Rahn, P.H., 1977. The origin of the Prairie Coteau, northeastern South Dakota:Proc. S.D. Acad, Sci. 56:27-33.

Schulz, L.D., 1975, Soil survey of Marshall County, South Dakota: U.S. SoilConservation Service, 116 p.


Figure 1. Topographic map of part of the USGS Milbank quadrangle, South Dakota, originally at1:250,000 scale. The drainage basin of Crow Creek above Hickman Dam is outlined, representingan area of 170 sq. miles.

Figure 2. Oblique aerial photograph of Fort Sisseton, looking northwesterly. “Fort Lake” (FL) and“Mallard Slough” (MS) are full of water at the time of this photograph, July 17, 1996.


Figure 3. Vertical aerial photograph of Fort Sisseton area. The soil unit abbreviations are explainedby Schulz (1975). “FS” is Fort Sisseton. “OL” is the outlet of the Cattail Lake complex. Fort Lakeis dry at the time of this photograph (1975), but Mallard Slough has some water in it.


Figure 4. Oblique aerial photograph of Cattail Lake, looking westerly. This photograph was takenon July 16, 1996, and shows the lake has nearly submerged County Road #5. The outlet (CattailCreek) is shown as “OL”.

Figure 5. Portion of several USGS topographic maps, originally 1:24,000 scale. The Cattail Lakecomplex is shown, based on a 1794 ft. elevation from published contour lines. “OL” is the outletfrom Cattail Lake.


Figure 6. County Road #16, approximately 3 miles west of Lake Eden. This July 17, 1996 photo-graph shows the road submerged by an arm of Cattail Lake.


SOME MODEL SELECTION PROCEDURESIN DISCRIMINANT ANALYSIS

Lu ZhangDepartment of MathematicsNorthern State University

Aberdeen, SD 57401

ABSTRACT


ABORTION IN AMERICA

Joel Vander KooiGovernment and International Affairs Department

Augustana CollegeSioux Falls, SD 57197

ABSTRACT

Abortion in America today presents real logical dilemmas which challengeour society’s most basic concepts of justice. Currently, many inconsistencies andlogical flaws exist within American public policy on the issue of abortion. Insome areas, policy allows abortion throughout pregnancy including the proce-dure termed partial birth abortion. Simultaneously, fetal p homicide laws pro-vide stiff penalties for killing the unborn from the moment of conception. Suchinconsistencies result from major misunderstandings within the AmericanJudiciary of the competing values which must be balanced in this issue as wellas the Judiciary’s unwillingness to face the issue squarely and honestly.

The proper solution to the logical problems arising from abortion may wellinclude the extension of the rights of personhood to the unborn throughConstitutional means. Such a solution would avoid the extremes of unrestrictedabortion or outlawing abortion outright while providing a clear standard bywhich cases may be decided and laws may be patterned thus delivering the legalsystem from the tangled web of current policy on the issue. However, this is onlypart of the whole solution which must also include the proper alternatives andsupport necessary to assist women through pregnancy. Those who are neverfaced directly by the issue still have a clear ethical duty to assist those who are,both mothers and unborn.


UTILIZATION OF HANDS-ON ACTIVITIES INTHE SCIENCE CLASSROOM: AN ASSESSMENT

OF STUDENT PERFORMANCE.

Nels H. Troelstrup, Jr. and Dennis ClarkDepartment of Biology and Microbiology


The objectives of this project were to (1) develop and provide hands-onenvironmental science activities for use in middle and high school science class-rooms and (2) assess changes in student performance following instruction withhands-on activities.

Math and science educators from 21 classrooms across South Dakota par-ticipated in the Environmental Classroom project during the 1996 and 1997school years. Participants were provided a workbook and instruction during two-three day workshops at the Oak Lake Field Station. In addition, participantsdeveloped a standardized student assessment tool to evaluate student perfor-mance. This assessment tool consisted of 10 multiple choice questions for eachactivity.

Teachers implemented at least two activities during the school year and eval-uated student performance using the standardized assessment tool. Studentswere evaluated immediately before and after instruction after which data weretabulated and analyzed using a sign rank test.

A total of 953 students from 13 school systems has received instruction dur-ing the 1996/97 and 1997/98 school years. Number of students per classroomranged from 10 to 114 (avg. = 33) and thirteen of 15 activities were utilized byparticipating educators. Significant improvements in student performance (avg.13%) were observed following instruction with all but one hands-on activity.Instruction with most activities resulted in improvement equivalent to one lettergrade. Changes in performance were greatest following inquiry-based lab exer-cises (avg. = 20.4 %), followed by mixed field/lab exercises (avg. 12.9%), infor-mation survey exercises (avg. 12.8%), field measurement exercises (avg.12.7%), group discussions (avg. 12.1%), Internet-based exercises (avg. 11. 2%)and library research exercises (avg. 0.7%).

Results of our analysis indicate significant improvement in student under-standing of environmental concepts following instruction with EnvironmentalClassroom activities. Furthermore, student performance appears to be enhancedfollowing those activities with inquiry-based field or laboratory components.This information supports existing education efforts sponsored by state agencieswithin South Dakota and emphasizes the need for inquiry-based instruction with-in our public school classrooms.


IMPLEMENTATION OF MULTIMEDIAINSTRUCTION IN BIOLOGY AND CHEMISTRY

COURSES: STUDENT REACTIONS

Thomas F. Guetzloff, Chemistry Departmentand

James C. Sorenson, Biology DepartmentMount Marty College, Yankton, SD 57078

“The new stress on multimedia presentations and on integrating the Internetinto instruction is a challenge to both experienced and novice teachers. Thosewho have been in the classroom for years must change their teaching styles rad-ically to accomodate these new sources of information. And new teachers haveto worry not only about mastering content and classroom teaching techniques,but also about being comfortable with several forms of technology in the teach-ing setting.” Flannery (1998) - from her review of two general biology text-books.

INTRODUCTION

Incorporating multimedia instructional materials into college science cours-es is a daunting task which has become almost a requirement for those of us whoteach such courses, as indicated by the quote above from Flannery. While sev-eral different options exist for how such implementation can occur, all approach-es face three obstacles that need to be overcome for implementation to be con-sidered successful: 1) funding constraints, 2) preparation-time constraints, and 3)acceptance by students. After a brief mention of how obstacles 1 and 2 wereovercome, we focus in this paper on our findings concerning obstacle 3: accep-tance by students.

Overcoming Obstacle 1: We were fortunate in that Mount Marty College(MMC) received a Title III grant from the U.S. Department of Education, oneaspect of which was to fund a Computer-Aided Instruction (CAI) Lab and to pur-chase software, so we were able to overcome obstacle 1 to a certain extent. OurCAI Lab has two rooms, with 10 IBM PC clones in one room and 10 PowerMacsin the other. In addition we were able to purchase many CD-ROMs and othersoftware packages in multiple copies so that student access was not a real prob-lem.

However, once past the problem of funding and obtaining the hardware andsoftware, we still faced obstacles 2 and 3.

Overcoming Obstacle 2: Putting in the “prep-time” on the multimedia materialsis an unavoidable, but necessary, requirement for successful implementation. We


all know that the Web is unregulated, so while it may be attractive to merely setstudents loose on the Web, this approach can be very time-consuming and frus-trating for students (especially for “Web-naive” students) if they begin by access-ing sites with dubious information, so instructors must provide guidance at leastto direct the students to reliable sites to start with. Or if course materials are tobe distributed through an instructor’s homepage with “reliable” links built in,there is the prep-time required to compose such a homepage, and to keep it cur-rent. And while pre-packaged materials may seem to circumvent this problem,they still need to be previewed by the instructor to evaluate accuracy, extent anddepth of coverage of the ideas, and the appropriateness of the level of presenta-tion and of the interactive quizzes that may be included. For example, we havepreviewed CDs that covered so much material in such depth that they were inap-propriate for the undergraduate level, at least for freshman level courses atMMC. We have found others that presented small inaccuracies that could poten-tially cause conceptual difficulties if unnoticed and uncorrected by the instructor.

Unfortunately, no matter which approach is taken, preview and preparationneeds to be done, but it is a task that must often be added on top of already exist-ing tasks (e.g., lab prep, course prep, textbook review, committee and depart-mental meetings, etc.). While we feel that this is a real problem that needs fur-ther discussion, we will delve into a more extensive discussion of this problemin another forum. However, for our purposes here, let us say that we were ableto overcome obstacle 2 (in addition to overcoming obstacle 1) so that we wereable to present our courses in a way that incorporated much use of multimediamaterials starting in the fall semester of 1997.

Overcoming Obstacle 3 (Acceptance by Students): In this paper we wish tofocus on student reactions to multimedia instruction, for that, we feel, is at theheart of the matter: the investment of money and time required to overcomeobstacles 1 and 2 will be for naught if the students don’t use and benefit from themultimedia materials. And while there are a number of papers in the literatureadvocating the use of multimedia in the classroom (Blystone 1993, Darensbourg1996, Dessy 1997, Graziadei and McCombs 1995, Greenhalgh 1997, Huang1991, Jones and Smith 1992, Jones and Berger 1995, Kozma 1994, Marsa 1995,Moore and Miller 1996, Smith and Stovall 1996, Waldow 1997, Watkins 1992),and a number of “how to” papers describing methods of implementing multime-dia in the classroom (Collins 1995, Hall 1996, Hayward 1996, Mounts 1996,Radice 1997), only a few references attempted to gauge student reaction to theuse of multimedia in the classroom (Amend and Furstenau 1992, Buttles 1992,Fitfield and Peifer 1994, Treadway 1996).

Here we report on student reactions (acceptance and use) to the incorpora-tion of multimedia materials in freshman-level biology and chemistry coursesbased on the responses and comments given by students to open-ended ques-tionnaires of our devising. Because of the different approaches taken in ourcourses we are able to report on the difference in acceptance and use of CD-ROMs -vs- acceptance and use of the Internet; and as a result of the differing


composition of students in our courses we are able to report on the acceptanceand use of multimedia by science majors -vs- non-science majors. Finally, basedon these results, we draw some tentative conclusions on the efficacy of incorpo-rating multimedia into the classroom.

METHODS

Two questionnaires were administered to students at the end of fall semes-ter 1997.

Questionnaire #1 was administered to both of Sorenson’s freshman-levelbiology courses: Biology 103: Principles of Biology, a course intended for sci-ence and pre-professional majors, and Biology 106: Introduction to the LifeSciences, a course intended for non-science majors and taken mostly by studentsto fulfill a General Education requirement.

Questionnaire #2 was administered to Guetzloff’s freshman-level Chemistrycourse: Chem 101, a course intended for nursing majors and other health sciencemajors, and also to Guetzloff’s sophomore-level Physics course, a courserequired of all science majors.


Sorenson’s classes emphasized the use of pre-packaged CD-ROMs to pro-vide information for review and to provide interactive quizzes, while Guetzloff’scourses emphasized the use of the Internet to provide similar features. Noticethat among the science majors and nursing majors (Bio 103, Chem 102, Physics)acceptance and use is well over 50% (Tables I, III, and IV), and the reasons mostcommonly given by students for use deal primarily with trying to enhance com-prehension of complex topics (Table I).

Sorenson’s two classes allow a comparison of the acceptance and use of CD-ROMs between science majors and non-science majors. Notice that half of thestudents in the science-majors course (Bio 103) reported using the CD-ROMs “alot”, while only one student in the non-science majors course (Bio 106) reportedusing the CD-ROMs “a lot” (Tables I and II). We believe this is indicative of thelevel of commitment and motivation of the students: science majors used theCAI materials to enhance their understanding, non-science majors were mostlyunwilling to make the time-commitment, and this is evidenced in their writtencomments. However, other possible implications that can be derived from theirresponses, and especially from their written comments, is that the non-sciencemajors are already time-limited (e.g., “I work 40 hours a week and it’s hard tofind time to get into the lab to view the CD-ROMs”) or are easily frustrated bycomputer glitches (e.g., “I tried it a couple of times and couldn’t really get it towork. Just frustrated me.”).

According to Guetzloff’s results (Tables III and IV) students feel that pre-sentations employing a digital format do improve the lecture compared to a moretraditional lecture format (Tables III and IV, questions 3 and 4). The students


seem not to like the listserv (class-list e-mail, now more commonly referred to asVirtual Office Hours) as much (Table III, question 5, 6, 7, 8). This was mainlydue to student apprehension or unwillingness to learn new computer applica-tions. There were numerous informal comments from students indicating thatthey were afraid of computers and utilizing e-mail. However, students wereforced to overcome their fear through cooperative learning when they wereforced to download assignments and messages and were assisted by other, com-puter-literate classmates. Other students, however, expressed the opinion thatthis is the best way to get information. They indicated that this approach madethem “feel like an adult” because “they can get the information on their owntime”. We were very surprised that the students did not indicate a strong appre-ciation for the internet movies and/or CD-ROM demonstrations in Guetzloff’ssurvey (Tables III and IV, question 2).

CONCLUSION

Our findings are that student reactions to the incorporation of multimediainstructional materials into courses is not automatically nor overwhelminglyfavorable. Acceptance depends on a number of factors, and perhaps key amongthem is the level of motivation of the student: science majors more readily usedthe multimedia materials than non-science majors.

Both approaches (using pre-packaged materials -vs- using the Internet) haveboth positive and negative aspects. Among the positive aspects is that either isreadily accepted and used by motivated students, and that a core group of moti-vated and computer-adept students can and will enable nearly all students to uti-lize multimedia through cooperative learning. Most of the negative aspects dealwith relative costs of implementation in terms of time and money, and theseaspects will be discussed in a different forum and warrant no further discussionhere.

Future areas of investigation will involve attempting to determine whetheror not learning is enhanced in courses that incorporate multimedia when com-pared to courses that do not incorporate such materials.

However, just as we professors feel that the task of incorporating multime-dia materials into our courses is something that has been added over-and-aboveour already existing job requirements, perhaps students see multimedia materialsin much the same way - that it doesn’t lighten their load at all, it is an addition-al requirement that has been added over-and-above the already existing courserequirements that involve reading the textbook, attending class for discussionand notes, and the requisite reviewing prior to examinations. Thus, perhaps oneimplication of our findings is that if multimedia is to be incorporated into cours-es, that a certain time-allowance be given to students to use the materials, andperhaps this is an issue that is relevant to both faculty and administrators whendeciding on the scheduling aspects of college courses.


LITERATURE CITED

Amend, John R., and Ronald P. Furstenau. 1992. Employing Computers in theNonscience-Major Chemistry Laboratory. Journal of College ScienceTeachers 22: 110-114.

Blystone, Robert V. 1993. Teaching in the Digital Republic. Academe 79:39-41.

Buttles, Sunny. 1992. “A Model for Incorporating and Evaluating Use of aComputer Laboratory Simulation in the Nonmajors Biology Course.American Biology Teacher 54: 491-494.

Collins, Michael. A.J. 1995. Using Electronic Bulletin Boards with CollegeBiology Classes. The American Biology Teacher 57:188-189.

Darensbourg, Marcetta R. 1996. Simple Techniques Can Give PleasurableResults. Journal of Chemical Education 73:A273-A274.

Dessy, Raymond E. 1997. The Size of Thoughts. Analytical Chemistry69:A615-A616.

Fitfield, Steve., and Rick Peifer. 1994. Enhancing Lecture Presentations inIntroductory Biology with Computer-Based Multimedia. Journal ofCollege Science Teachers 23:235-239.

Flannery, Maura. 1998. The Bottom-Up Approach to Biology. BioScience48:223-224.

Graziadei, William D. and Gillian M. McCombs. 1995. The 21st CenturyClassroom-Scholarship Environment: What Will It Be Like? Journal ofEducational Technology Systems 24:97-112.

Greenhalgh, J.B. 1997. Using the Internet: Concept to Classroom. TechnicalHorizons in Education Journal 24:15-19.

Hall, Donald W. 1996. Computer-Based Animations in Large-EnrollmentLectures: Visual Reinforcement of Biological Concepts. Journal of CollegeScience Teachers 25:421-425.

Hayward, Roger. 1996. New technology, new questions: Using an Internet data-base in chemistry. Australian Science Teachers Journal 42:39-42.

Huang, Samuel D. 1991. The Impact of Using Interactive Video in TeachingGeneral Biology. American Biology Teacher 53:281-284.


Jones, Loretta L., and Stanley G. Smith. 1992. Can Multimedia Instruction MeetOur Expectations? EDUCOM Review 27:39-43.

Jones, Tricia. and Carl Berger. 1995. Students’ Use of Multimedia ScienceInstruction: Designing for the MTV Generation? Journal of Educational Multimedia and Hypermedia4:305-320.

Kozma, Robert B. 1994. A Reply: Media and Methods. EducationalTechnology Research and Development 42:11-14.

Marsa, Linda. 1995. Shoptalk in Cyberspace: Virtual Labs EncourageCollaboration and Exchange. OMNI 17:14.

Moore, Randy, and Iain Miller. 1996. How the Use of Multimedia AffectsStudent Retention and Learning. 25:289-293.

Mounts, Richard D. 1996. Chemistry on the Web. Journal of ChemicalEducation 73:68-71.

Radice, Gary P. 1997. computer-Assisted Microscopy in Science Teaching andResearch. Journal of Science Teaching and Research 26:265-269.

Smith, Stanley and Iris Stovall. 1996. Networked Instructional Chemistry:Using Technology to Teach Chemistry. Journal of Chemical Education73:911-915.

Treadway, William J. Jr. 1996. The Multimedia Chemistry Laboratory:Perception and Performance. Journal of Chemical Education 73:876-878.

Waldow, Dean A. 1997. CIRRUS: A Chemistry Internet Resource for Researchby Undergraduate Students. Journal of Chemical Eduction 74:441-442.

Watkins, Beverly T. 1992. Chemistry Professors Try Technology to LureStudents Into Advanced Study. The Chronicle of Higher Education38:A17-A18.


TASTE AND ODOR AVERSION CONDITIONINGIN MUS MUSCULUS AND ASSOCIATED

CHANGES IN HEART RATE AND FLUID CONSUMPTION

Linda R. Alzieblerand

Thomas P. CoxBlack Hills State University

Spearfish, SD 57783

ABSTRACT

The chemical senses of olfaction and taste have long been recognized asimportant to survival. Taste Aversion, and to some extent odor aversion condi-tioning is a robust phenomena. We looked at affects of aversively conditionedodor or taste had on heart rates, lick rates, and water consumption of Mus mus-culus. This involved pairing either saccharine water (taste), or Amyl-Acetate(odor) with Lithium Chloride injection induced illness. Following a single con-ditioning trial mice were again exposed to taste or odor stimuli. Heart rates, lickrates, and water consumption rates were compared for both pre and post condi-tioning, and between groups. Results indicate heart rate increases as well as lickand water consumption rate decreases for taste aversion conditioned mice. Thesetendencies were also seen for odor conditioned mice, but did not reach signifi-cance at the 0.05 level.

INTRODUCTION

Olfaction and taste have long been recognized as important to most animals.Rodents are dependent on olfaction for selecting safe food, choosing appropriatemates, and for avoiding situations that could prove fatal. Rodents also dependon gustatory cues to prevent them from consuming substances that could provedeadly.

It is well recognized that animals who become ill after eating or drinking agiven substance will avoid that substance in the future (Garcia and Koelling,1965). This taste aversion condition is reproduced under laboratory conditionsby encouraging the animal to drink or eat a tasty food and then making the ani-mal ill. The animal associates the illness with the taste of what was previouslyconsumed and avoids further contact. Although taste aversion conditioning iswell established, odor aversion conditioning without the use of an associated fla-vor is difficult to obtain (Backer and Booth, 1989, Holder, 1991, Palmerino,Rusiniak, and Garcia, 1980, Panhuber, 1982, Rusiniak, Hankins, Garcia, and


Brett, 1979). Pairing of a variety of either odors combined with a variety oftastes, as well as odor only in preference studies has not yet provided clear evi-dence that single exposure conditioning for odor alone is possible (Lucas andSclafani, 1995).

Avoidance is only one reaction to possibly threatening stimuli.Physiological reactions are also associated with such stimuli. These can includechanges in heart rate, blood pressure, respiration rate, temperature, and skin con-ductance, which are all common indices of stress (Abdeen, Taylor, Youngblood,and Printz, 1995, Cocke and Thiessen, 1986, Hunt, Hess, and Campbell, 1997,Young and Leaton, 1994).

It is reasonable to assume that a cue, such as taste or odor, associated with athreatening event, such as Lithium Chloride (LiCl) induced illness, would, on re-exposure produce some of the physiological changes commonly associated withstress. We attempted to determine if mice exposed to a taste or odor stimulusassociated with LiCl induced illness would produce behavioral avoidance(decreased water consumption and decreased lick rates) as well as a physiologi-cal change associated with stress (increased heart rate).

METHODS

Subjects

Twenty-three naive male, Swiss, albino mice (Mus musculus), approximate-ly 160 days old, were housed individually in plastic cages with stainless steelcovers and maintained on a 12:12-h light:dark cycle. Mice had free access tofood and free access to water except during the eight days of experimental ses-sions and the two weeks preceding these experimental sessions.

Apparatus

A two piece aluminum electrical project box 15 x 10 x 10 cm was used as aconditioning chamber. The cover of the project box was used as a lid for the con-ditioning chamber. A standard water bottle 2.3 cm inside diameter entered thecenter of one 10 x 10 cm. side of the conditioning chamber. A .8 cm outsidediameter nozzle which accommodated standard .4 cm inside diameter plasticaquarium air hose entered at each 10 x 10 cm side of the conditioning chamber.The air hose entering the drinking spout end of the chamber supplied either odor-ized or unodorized air, while the air hose at the opposite end of the chamber rout-ed air directly to a roof mounted ventilation duct. Two plastic 1.3 cm electricalconduit clamps were mounted 2 cm apart on the bottom of the chamber. Theconduit clamps secured the mouse restraining device which consisted of a 3 cmdiameter syringe cover cut to approximately 10 cm in length. A whole was cutto approximately 1 cm in diameter in the other end of the syringe cover to accom-modate the mouse’s muzzle. A cap for the other end of the restraining device wasmade from a plastic tube just slightly larger in diameter than the syringe cover


and cut to about 4 cm in length. A hole was made in the closed end of the tubeto accommodate the mouse’s tail.

Air entering the chamber was filtered through a 100 ml flask containingeither 50 ml of d H2O, or 45 ml d H2O mixed with 5 ml of Amyl-Acetate(C7H14O2). An aquarium air pump supplied pressurized air at a volume of 1liter/minute.

Pulse rates were monitored via a light sensitive athletic pulse monitorintended for humans. The monitor was clamped to the tails of the mice to obtaintheir heart rate measures. Lick rates were measured by a lickometer consistingof a modified touch activated, capacitance switch door knob alarm. The capaci-tance switch was adjusted to close at mouse body capacitance. The switch wasattached to the stainless steel drinking spout. Contact with the drinking spoutactivated an event counter via the switch. Water consumption rates were mea-sured using a graduated drinking tube attached to the drinking spout. An elec-tronic, digital stop watch was used for timing all pre and post-conditioning trials.

Procedure

Mice were given ad lib. access to food and water for 105 days. They werethen placed on a 23 hr. water deprivation regime. The water deprivation regimebegan 7 days prior to acclimating the subjects to the apparatus.

Animals were then acclimated to the apparatus for 7 days for a period of 5minutes each day. Following acclimation trials mice were allowed ad. lib. accessto water for one hour. Acclimation trials consisted of placing mice in therestraining device and conditioning chamber with cover. The pulse rate monitorwas attached to their tails. During acclimation mice were exposed to pressurizedair supplied by an aquarium air pump at a rate of 1 liter/minute and filteredthrough a 100 ml flask of d H2O.

Subjects were then randomly assigned to one of four groups receiving oneof two treatment conditions for the purpose of collecting pre-conditioning data:Groups one and two were selected for presentation of air filtered through C7H14O2

(Amyl-Acetate). They were also allowed access to d H2O for drinking. Thesemice were placed in the restraining device inside of the conditioning chamberwith the chamber in place and with the heart rate monitor attached to their tailsfor a period of 5 minutes each day for 3 days. Pre-conditioning readings wererecorded for all three measures in the following manner: heart rates were record-ed once a minute for five minutes and averaged for per minute heart rate; lickrates were recorded once a minute for five minutes and averaged for per minutelick rate; water consumption rates were established by calculating the differencein the amount of water measured before the trial began and after the trial wascompleted. Each mouse received one hour of water each day following theirtime in the conditioning chamber. Group’s three and four were selected for pre-sentation of clean air and a 43% solution of saccharine drinking water. All otherconditions were identical to that of group’s one and two, and all readings wererecorded in the same way.


On the third day of pre-conditioning within 5 minutes of removal from theconditioning chamber, mice in group’s two and four recieved a single interperi-toneal injection of 10 ml/kg of 0.3M LiCl. Injection occured before the micewere returned to their cages for their hour access to water. All of the mice ingroup one and group three (control groups) received an interperitoneal injectionof 10ml/kg of physiological saline within 5 minutes of being removed from theconditioning chamber and before being returned to their cages and one houraccess to water.

Post-conditioning data collection began on the day immediately followingconditioning day and continued for a total of 5 days. Subjects in all groups wereplaced in the same condition as during pre-conditioning. Data was collected andrecorded for all groups in the same manner as during pre-conditioning.

RESULTS

Within group pre and post-conditioning means were compared using arepeated measures ANOVA . In addition, between mean differences were test-ed with a one way ANOVA (Table 1). All comparisons were tested at the .05 con-fidence level unless otherwise specified.

The within group comparisons for the paring of LiCl and saccharine waterreached significance as indicated by decreased post conditioning lick rates (df =1,46, SS = 13104.2, F = 5.98) (Figure 1) and decreased post-conditioning fluidconsumption rates (df = 1,46, SS = 178.8, F = 23.15) (Figure 2). However thegroup which received pairing of LiCl and Amyl-Acetate odor did not demon-strate a significant decrease in lick rate or fluid consumption. Post conditioningheart rate increases were not significant for either group.

Mice that receiving an injection of saline in conjunction with the Amyl-Acetate odor also displayed significant change in post-conditioning lick rate (df= 1,38, SS = 3971.88, F = 6.91) (Figure 1) and decreased post-conditioning fluidconsumption rate (df = 1,38, SS = 104.68, F = 13.34) (Figure 2). The group thatreceived an injection of saline in conjunction with the taste of saccharine watershowed a significant post-conditioning drop in fluid consumption (df = 1,46, SS= 189.41, F = 8.51) (Figure 2) level but post-conditioning decrease in lick ratesdid not reach significance (Figure 1). These groups did not display significantincreases in post-conditioning heart.(Figure 3).

The between groups comparisons of all groups did not reach significance butcomparisons indicate significantly lower lick rates for odor and saline than forboth taste and odor paired with saline (df = 1,53, SS = 5415.24, F = 6.44) (Figure5). Odor and saline also had a lower fluid consumption rate than taste and salinealthough this difference did not quite reach significance (df = 1,53, SS = 106.22,F = 3) (Figure 5). Lick rates for Amyl-Acetate and LiCl when compared withsaline and LiCl approached but did not reach significance (df = 1,58, SS =8242.16, F = 3.44) (Figure 6). All other comparisons for groups 2 and 4 failedto reach the .05 criterion level.


DISCUSSION

Mice exposed to a taste, (in this case saccharine water), and made ill withLithium Chloride induced illness, show a decrease in willingness to consume thattaste. Lick rates and fluid consumption rates decreased. These results mirroredthose of Garcia et al. (1965). Mice exposed to the odor of Amyl-Acetate and thenmade ill do not show the same tendency to avoid the non-tasty fluids served inconjunction with that odor. Lucas et al, (1995) speculated it may be difficult toaversively condition an odor without a corresponding taste.

What was unusual in this experiment was the lack of significant increases inheart rate measures for either the group exposed to taste and illness or the groupexposed to odor and illness. Heart rate increase is a common indicator of stress(Abdeen et al., 1995, Young et al., 1994). Forced exposure to an odor or tastethat previously caused illness should have caused a stressful situation. The factthat heart rate changes don’t appear to be a good indicator of stress in this caseprobably has something to do with experimental design. It is quite possible thata single exposure to the aversive stimuli was simply not enough to produce thedesired result.

Animals in control groups also showed a significant decrease in willingnessto consume fluids after conditioning. The pain of the saline injection alone couldhave caused this result. However saline injected control groups for other tasteconditioning studies do no support this speculation.

The animals in groups that were given saccharine water consumed morefluid than those animals in groups given distilled drinking water. This differencein lick rates and fluid consumption rates between groups had more to do withtaste than because of any other variable.

If odor conditioning is not so robust a phenomenon as taste conditioning itmay require more than a single exposure. If heart rate is a reasonable measureof stress on danger, it should have increased upon re-exposure to the aversivelyconditioned stimulus. This occurred for the saccharine and LiCl pairing unfor-tunately it was also seen for the control groups. Heart rate actually decreased forthe odor - LiCl pairing. This may reflect a requirement for repeated pairing ofodor stimuli and illness for odor aversion conditioning (figure 3).

LITERATURE CITED

Abdeen, O.A., Taylor, B.K., Youngblood, K.L., and Printz, M.P. 1995.Peripheral Beta and Adrenergic Blockade Modifies Airpuff Startle-InducedHeart Rate Responses. The Journal of Pharmacology and ExperimentalTherapeutics 272:282-289.

Baker, B.J., and Booth, D.A. 1989. Genuinely Olfactory PreferencesConditioned by Protien Repletion Appetite 13:223-227.


Cocke, R., and Thiessen, D.D. 1986. Chemocommunication Among Prey andPredator Species. Animal Learning and Behavior 14:90-92.

Garcia, J., and Koelling, R.A. 1996. Relation of Cue to Consequence inAvoidance Learning. Psychonomic Science 4:123-124.

Holder, M.D. 1991. Conditioned Preferences for the Taste and OdorComponents of Flavors: Blocking but not Overshadowing Appetite 17:29-45.

Hunt, P.S., Hess, M.F., and Campbell, B.A. 1997. Conditioned Cardiac andBehavioral Response Topography to an Olfactory CS Dissociates with AgeAnimal Learning and Behavior 25:53-61.

Lucas, F., and Sclafani, A. 1995. Carbohydrate-Conditioned Odor Preferencesin Rats. Behavioral Neuroscience 109:446-454.

Palmerino, C.C., Rusiniak, K.W., and Garcia, J. 1980. Flavor-Illness Aversions:The Peculiar Roles of Odor and Taste in Memory for Poison Science208:753-755.

Panhuber, H. 1982. Effect of Odor Quality and Intensity on Conditioned OdorAversion Learning in the Rat Physiology and Behavior 28:149-154.

Rusiniak, K.W., Hankins, W.G., Garcia, J., and Brett, L.P. 1979. Flavor-illnessAversions: Potentiation of Odor by Taste in Rats Behavioral and NeuralBiology 25:1-17.

Young, B.J., and Leaton, R.N. 1994. Fear Potentiation of Acoustic StartleStimulus- Evoked Heart Rate Changes in Rats Behavioral Neuroscience108:1065-1079.


DECREASED AGGRESSION IN GONADECTOMIZED MALE FIGHTING FISH

(BETTA SPLENDENS) SYSTEMATICALLYRELATED TO INCREASING LEVELS

OF ESTROGEN.

Michael J. Huxford and Thomas P. CoxDepartment of Psychology

Black Hills State UniversitySpearfish, SD 57783

ABSTRACT

Male Siamese fighting fish (Betta splendens) engage in a stereotypic aggres-sive threat display when they view either another male fighting fish, or their ownmirror image. Female Siamese fighting fish are less aggressive then males.Male aggressive behavior is promoted by increasing levels of 11-Keto testos-terone and/or testosterone proportionate. Males of some fish species such as thethree spined stickleback, blue gourami and sunfish remain aggressive followingcastration. Therefore, testosterone may not be required for maintenance ofaggressive behavior. We were interested in whether the hormone estrogen con-tributed to lower levels of aggression in females.

Fish were presented with their mirror image for thirty seconds and rated foraggressive display. They were observed for ten trials interspersed with five five-minute inter-trial intervals. Both male and female fish were tested.

Following an initial aggression rating male fighting fish were gonadec-tomized. This eliminated any testosterone influence. Gonadectomized fish werethen allowed to recover for five days, at which time they were injected witheither telecost solution, or telecost solution as a vehicle for one of three levels ofestrogen. Twenty-four hours were allowed for hormone absorption and the fishwere again tested for aggressive display. Comparisons of aggressive displayswere made between all levels of estrogen supplemented gonadectomized males,as well as between the aggression levels for these males and the aggression lev-els of intact female fish. Although still incomplete, the data suggests a system-atic decline in gonadectomized male aggression as estrogen levels increase, per-haps to levels mimicking those of intact female fighting fish.


OBLIGATORY WINTER DIAPAUSE IN STATOBLASTS OF PLUMATELLA CASMIANA

(BRYOZOA: PHYLACTOLAEMATA)

Melissa A. Albers, and Jonathan C. WrightDepartment of Biology, Northern State University1200 South Jay Street, Aberdeen, SD 57401-7198.

ABSTRACT

Statoblasts are asexual overwintering structures produced by colonial fresh-water invertebrates, the Bryozoa, which live attached to submerged objects.Each statoblast comprises a mass of undifferentiated cells enclosed in a bivalvedshell. When conditions favor germination, the valves separate and a white ger-minal yolk mass protrudes. Various studies have assessed whether statoblastsmust go through an obligatory dormant period (diapause), or whether they cangerminate as soon as favorable conditions arise (quiescence). To date, diapausehas not been demonstrated in statoblasts. The present study set out to explore thedormancy and germination of statoblasts in Brown Co., SD.

Statoblasts of Plumatella casmiana were field-collected in mid-winter fromshoreline debris at Moccasin Creek and maintained in the laboratory until study.Germination was studied in samples immersed in distilled water. No germina-tion was observed until March. These results demonstrate that statoblasts ofPlumatella casmiana from this population enter winter diapause. Following thebreaking of diapause, statoblasts showed a mean germination success of 22%,most germinations occurring in 7-14 days.

Statoblasts collected in the fall and brought into the laboratory could be ger-minated by immersion throughout the winter. This shows that neither seasonalchanges in photoperiod, nor desiccation, trigger diapause alone. To study possi-ble effects of chilling on the induction of diapause, statoblasts were transferredto filter paper, placed in Plexiglas tubes, and immersed in a Cole Parmer Polystatprogrammable water bath containing a 1:1 mixture of ethylene glycol and water.Cooling was studied for various periods up to 18 days. Chilling temperatures var-ied between 0 and -10_C; in all experiments, statoblasts were initially exposedto 5_C, then cooled to the experimental temperature over a 5 hour period.Results indicate that diapause is induced by brief (10h) exposure to -10_C andmore prolonged exposure to higher temperatures. Long-term viability of thesestatoblasts is currently being studied, but we can infer that cessation of germina-tion is due to diapause rather than mortality given the much more severe tem-peratures tolerated in the field. No dormancy was induced following 18 dayschilling at 0_C; statoblasts submerged throughout the winter thus probably do


not enter diapause. Effects of chilling and desiccation on post-diapause statoblasts were also

studied to determine whether they account for significant spring and summermortality. A chilling duration of 10 hours was used with various temperaturesdown to -25_C. The effect of desiccation was studied by placing statoblasts in adesiccator (anhydrous CaCl2) for 24-48 hours. These modest exposures toextreme chilling and desiccation had no significant effect on viability. The factthat such chilling regimes induce dormancy in the fall and winter suggest thatinduction of diapause can occur only within a critical developmental window, orthat it requires compound cues such as a combination of temperature and pho-toperiod.

There is a clear adaptive advantage of diapause for statoblasts in this popu-lation of P. casmiana. In the fall and early spring, sub-freezing temperatures arefrequently interspersed with brief warm periods. If these were to trigger germi-nation, the resulting colonies would not survive the winter and the overwinteringmechanism for the species would fail.


CHROMOSOMAL ANALYSIS OF DEFORMED FROGS

Angela Reister, Sara Horner and Debra CarlsonDepartment of Biology, Augustana College, Sioux Falls SD 57197

ABSTRACT

Deformed amphibians have been found with increasing incidence in NorthAmerica. Several counties in the state of Minnesota have reported an unprece-dented number of frogs with deformities, many of which involve supernumeraryor missing hind limbs. There is significant concern about the increased numbersof deformed animals since amphibians, and frogs in particular, are seen as indi-cators of environmental quality. Although several explanations (parasite infesta-tion, pesticide use and ultraviolet radiation) have been offered to explain theincreased number of abnormal animals, the evidence is still incomplete. It is pos-sible that the physical deformities are the result of environmental agents thatdamage the genetic material of the afflicted frogs. To address this question,metaphase chromosome spreads were prepared from the peripheral blood ofnorthern leopard frogs (Rana pipiens) and mink frogs (Rana septentrionalis)captured in wetland areas of affected Minnesota counties. Blood was collectedby cardiac puncture from 146 frogs and cultured in the presence of a mitogen andcolcemid. After fixation, application on slides and staining, analyzablemetaphase plates were identified from 40% of the animals. All chromosomespreads examined had the normal number of chromosomes (2N = 26 for bothspecies), and all chromosomes exhibited the normal morphology. Silver stainingof the chromosomes revealed that all complete metaphase plates had the normalnumber of nucleolar organizer regions (2 per 2N set for both species). At thelevel of resolution of this study, the chromosomes of the deformed frogs did notexhibit any abnormalities in number, morphology or nucleolar DNA. Whilesome banding procedures have been attempted, a more detailed analysis of thefrog chromosomes is not currently possible because of the technical status ofamphibian cytogenetics.


MULTIVARIATE MORPHOMETRIC VARIATIONIN THE COFFEE SNAKE, NINIA DIADEMATA

Brian E. SmithDepartment of Biology

Black Hills State University,USB #9044, Spearfish, SD 57799-9044

Traditional studies of snake systematics emphasize univariate variation in avariety of characters, and usually ignore potential multivariate differentiation.Although snakes have been described by some investigators as “heads with tubesattached”, nevertheless, it is conceivable that snakes exhibit multivariate differ-entiation, as do a variety of other taxa. In this study, I utilized discriminant func-tion analysis to analyze multivariate morphometric differentiation in head shapeamong four putative subspecies of Ninia diademata, a small semi-fossorial colu-brid snake from intermediate elevations of Central America. Linear discriminantfunction analysis is a multivariate analog of analysis of variance which derives aclassification function as well as standard measures of significance. Linear dis-criminant function analysis can only be used if covariance matrices are the sameamongst test groups. This assumption is seldom met (and also seldom tested). Ifthis assumption is not met, then quadratic discriminant function analysis can beused, however, this type of test is less powerful for detecting differences. Newtechniques which compare the structure of covariance matrices can be used toconstruct matrices constrained by commonality of matrix components, whichmay then be used in a standard linear discriminant function analysis to offer amore powerful test of an experimental hypothesis. Using these methods, I foundthat previous hypotheses of subspecific differentiation in Ninia diademata werenot supported by the data. The species appears to consist of two distinctive sub-species, with one a wide-ranging group spread throughout northern Guatemalasouthwards to northern Panama, and the other narrowly confined to the Pacificversant of southern Mexico and Guatemala, isolated by the Guatemalan high-lands. These races can be distinguished using subtle differences in ventral col-oration and some univariate measures.


SEPARATION OF MATERNAL AND EMBRYOCONTRIBUTIONS TO REPRODUCTIVE

FAILURE IN YELLOW MICE (Ay/a; C57BL/6J)

Gerald A. Dickens1, Maureen R. Diggins2, and Nels H. Granholm1

1Department of Biology & Microbiology, South Dakota State University, Brookings, SD 57007

2Department of Biology, Augustana College, Sioux Falls, SD 57197

This study was conducted to determine if reduced reproductive performancein Ay/a mice is due primarily to the maternal uterine environment not to the pres-ence of genetically yellow (Ay/a) fetuses within those uteri. Following bilateralovariectomy genetically black ovaries (a/a) were grafted into empty ovarian bur-sae of 79 experimental (Ay/a) and 54 control (a/a) females. At 120 days of agefemales were superovulated (PMS/HCG), mated to a/a males, and scored forreproductive performance. Since a/a ovaries were grafted to Ay/a females whichwere then mated to a/a males, all pregnant Ay/a as well as a/a females containedembryos of only a/a genotypes; thus, differences in reproductive performancebetween the two groups had to be due to factors other than the presence of Ay/afetuses. Uterine weights (10 days p.c.) for Ay/a (229 _ 33 mg, n=12) were notstatistically lighter than those of a/a (366 _ 82, n=13). The mean uterine weightper decidua was less (P<0.05) in Ay/a versus a/a females (46 _ 7 v. 76 _ 12 mg,respectively). Mean somites per embryo and mean normal embryos were alsosignificantly reduced in Ay/a females (P<0.01). Because of the way in which thegrafts were conducted, the genetic makeup of fetuses in both experimental (Ay/a)and control (a/a) groups was exclusively a/a. Therefore this study supports thehypothesis that Ay causes a significant decline in reproductive performance inde-pendent of its action in Ay/a embryos. Since mildly obese Ay/a females exhibit-ed reproductive problems, obesity may not be a prerequisite for Ay-inducedinfertility. Supported by research funds awarded to MRD (USDANRICGP-2245and NHG (AES089, Ehrmann, and NIHAR42757).


BOVINE AGOUTI MOLECULAR ANALYSIS

Marcus D. Johansen, Raymond R.R. Rowland, Carl A. Westby,Donald M. Marshall, and Nels H. Granholm

South Dakota State University, Brookings, SD 57007

Agouti alleles regulate pigmentation in mouse melanocytes causing the pro-duction of black and/or yellow coat color. Yellow allelic mutations result in thedysregulated or overexpression of the agouti gene and promote a number of sys-temic disorders (lethal yellow syndrome) of great interest to human health andanimal production. Objectives of this study were to 1) isolate and sequence thebovine agouti gene and 2) determine agouti expression in bovine tissues. Bovinegenomic DNA was either digested and prepared for Southern analysis or used forPolymerase Chain Reaction (PCR) to amplify specific coding regions of theagouti gene. Amplified PCR products were cloned and sequenced. Isolatedbovine total RNA was used either for Northern analyses or for reverse transcrip-tase (RT-PCR). In both Southern and Northern analyses, hybridization wasdetected using a 32P labeled murine agouti cDNA probe.

Southern hybridizations confirm that cattle possess an agouti gene.Sequence analyses of cloned PCR products revealed an agouti-specific 161 bpfragment that resembled agouti exon 2. The nucleotide sequence of bovineagouti exon 2 was 75, 82, and 88% similar to murine, porcine, and human agoutiexon 2 sequences, respectively. Furthermore the predicted amino acid sequencewas 60, 73, and 81% similar to murine, porcine, and human predicted amino acidsequences, respectively. Preliminary Northern hybridizations indicate agoutiexpression in bovine tissues. Further molecular analyses of the bovine agoutigene may enable cattle producers to select agouti alleles beneficial for rapidgrowth, high protein/low fat carcasses, enhanced reproduction, and greater vigor.Work funded by SDSU-AES(HD089), Eagles’Ehrmann Cancer Fund, and NIH(AR42757).


COMPARISON OF ADULTECHINOCOCCUS MULTILOCURARIS

EXCRETORY/SECRETORY (E/S)ANTIGENS TO THOSE OF PROTOSCOLECES

Troy L. Lackey, Margaret Perry, and Mike B. HildrethDepartment of Biology/Microbiology


ABSTRACT

Tapeworms often infect canines. Because of their low pathogenicity, mostare of little concern to veterinarians. However, there is one species of dog tape-worm that can cause a potentially lethal zoonotic disease in humans. Thisspecies is known as Echinococcus multilocularis and it causes alveolar hydatiddisease (AHD) in humans. Humans and rodents become infected with the larval,hydatid cyst stage of this species if they ingest the eggs released from adults into the feces of canines. Dogs become infected with adult worms when theyingest protoscoleces present within the cysts.

Diagnosis of E. multiloculatis infection in canines is very difficult becausethe adults are small (1.2-mm to 9.7-mm) and very hard to see in fecal samples;and the eggs are morphologically identical to eggs of other non-zoonotic tape-worms commonly present in dogs (ie. Taenia pisiformis). However, there areseveral new procedures being developed to identify E. multiloculatis infectionsin dogs. One of the most promising methods under investigation involves thedetection of coproantigens released from adult worms into the host’s intestine.These antigens can be detected with an ELISA (enzyme linked immunosorbentassay) system using antibodies generated against these antigens. Thus far, pro-teins from adult worms havebeen used to produce antibodies used in the system. The purpose of this studywas to evaluate the feasibility of using the protoscolex stage as an alternativesource of antigens.

For this study excretory and secretory (E/S) antigens were collected from theprotoscoleces and adults by first maintaining each stage in tissue culture mediaand then concentrating the antigens using an ultrafiltrate cell with a 10,000 mol-ecular weight cut off. A portion of the E/S antigens collected from the proto-scoleces were used to immunize a rabbit (3 injections, 2 week intervals).Hyperimmune serum was collected from the rabbit 3 weeks after the final injec-tion. The presence of polyclonal antibodies against protoscolex E/S antigens wasfirst detected using an Ouchterlony double diffusion assay and immuncoyto-chemistry. The protoscolex and adult E/S antigens were separated into their pro-


tein components by polyacrylamide gel electrophoresis. The proteins were eitherstained in the gels with Coomassie Blue or transferred to Immobilon-P transfermembranes (PVDF; Millipore corporation Bedford, MA). The membranes wereincubated first in Tween blocking solution for 2 hours to block non-specific sites,and then for 1 hour in the hyperimmune rabbit serum (3-ul rabbit hyperimmuneserum raised against EM per 10-mL of Phosphate Buffer Solution). Antibodiesbound on the membrane were detected using a Vectastain ABC systems kit withDAB as the substrate (Novocastra Laboratories Ltd Newcastle, UK).

Numerous protein bands were identified in the Coomassie blue-stained gelsfrom both the protoscolex and adult source of antigens. Antibodies from the anti-protoscolex serum bound to most of the protoscolex proteins in the transfermembrane. These same antibodies also bound to many of the E/S proteins recov-ered from the adult stage. Results from this study suggest that the protoscolexstage of EM can be used as a source for many of the E/S antigens released by theadult.


GANGLIOSIDOSIS IN SHEEP CAUSED BY ADEFECTIVE b GALACTOSIDASE GENE

K. Heidebrink, R. Rowland, L. Holler, T. CheesbroughDepartment of Biology/Microbiology


ABSTRACT

Gangliosidosis is a genetic disease that occurs in a variety of animals includ-ing sheep and humans. It is similar to Tay Sachs disease and Morquio B syn-drome in humans. We are using this disease in sheep as preliminary research fordeveloping tests and treatment for similar human genetic diseases.Gangliosidosis is caused by an inactive b-galactosidase gene. The normal b-galactosidase gene cleaves the terminal sugar from oligosaccharide chains allow-ing for their degradation. A buildup of these sugar chains accumulates in neu-rons and visceral organs of affected animals causing disease symptoms and even-tual death. The symptoms in affected lambs are progressive neurological degen-eration and loss of physical fitness occurring at 4-6 months with death occurringat 6-8 months. To identify mutated b-galactosidase genes, we are isolating thenormal and mutated genes from tissues and blood of normal and affected lambsrespectively. Once sequenced, the two genes can be compared to identify themutations that cause the inactivation of the mutated gene. Similar diseases inother animals have shown that mutations are mainly base pair inserts and pointmutations. The discrepancies between the two genes will be utilized to developa rapid test for in-utero identification of Gangliosidosis.


COMPARISON OF THE CELLULAR AND HUMORAL IMMUNE RESPONSE AS A PREDICTOR FOR PROTECTION

IN A TYPE II BOVINE VIRALDIARRHEA VIRUS CHALLENGE.

A.A. Ahmed1,2, A.A. Salama1, D.J. Hurley2, L.J. Braun2 and C.C.L. Chase2.

1Dept. Micro., Fac. Vet. Med., Zagazig University, Zagazig, Egypt; 2Dept.Vet.Sci., South Dakota State University, Brookings, SD 57007

ABSTRACT

Bovine viral diarrhea virus (BVDV) has emerged as the major pathogen incattle. One of the major questions involves cross protection of vaccines con-taining Type I against Type II BVDV infection and disease. A balanced study in500 # feeder calves was done with 4 controls, 4 animals vaccinated withBovishield (Type I vaccine; Pfizer) and 4 animals vaccinated with Virashield 5(Type I and II vaccine; Grand). The cellular and humoral response was moni-tored for 4 months prior to challenge with type II BVDV strain 890 and for onemonth following challenge. Both groups of vaccinated animals were protectedbut the magnitude of the protection was mirrored by the cellular response priorto challenge. The Virashield group showed no clinical signs throughout the studywhile the Bovishield had lower clinical signs than the control group. The cellu-lar response was increased in the Bovishield following challenge indicating goodimmunological memory. The cytotoxicity assay used in this study was uniqueand provided evidence of the role of cytotoxicity in protection against BVDVchallenge. The Virashield group appeared to protect against infection whileBovishield protected against disease. This study indicates that vaccines need tocontain both types of BVDV to provide adequate protection.


THE NUCLEOCAPSED PROTEIN OFPORCINE REPRODUCTIVE AND

RESPIRATORY SYNDROME VIRUS LOCALIZES TO THE NUBLEOLUS

Christopher J. Kuckleburg, Bobby L. Kervin, David A. Benfield, and Raymond R. R. Rowland.

Departments of Biology-Microbiology and Veterinary Science,South Dakota State University, Brookings, SD 57007

ABSTRACT

Porcine reproductive and respiratory syndrome virus (PRRSV) belongs tothe family, Arteriviridae, which also includes lactate dehydrogenase-elevatingvirus (LDV) of mice, equine arteritis virus (EAV), and simian hemorrhagic fevervirus (SHFV). Two distinct strains of PRRSV are represented by the EuropeanLelystad virus, and VR-2332, which was isolated in the United States in 1991.Primary porcine pulmonary macrophage cultures and MARC-145 cells, a sub-clone of monkey kidney MA-104 cells, support virus replication in vitro. Theprincipal structural components of PRRSV are the nucleocapsid (N) protein,matrix (M) and envelope (E) glycoproteins. The 15 kD N protein forms the viralcapsid and is translated from ORF7 within the smallest subgenomic mRNA. Themonoclonal antibody SDOW 17 recognizes the same conserved epitope in bothLelystad and VR-2332 N proteins. Following FITC-labeled SDOW-17 stainingof MARC-145 cells infected with the U.S. isolate, 23983, we observed a signif-icant amount of fluorescence in nucleolar-like structures within the nucleus.SDOW-17 stained mock-infected cells showed only background fluorescence.In order to demonstrate targeting of the N protein to the nucleolus we made aDNA construct consisting of ORF 7 from 23983 ligated into the multiple cloningsite of the eukaryotic expression vector, pEGFP. When transfected into mam-malian cells this vector produced a green fluorescent (GF) protein. Our fulllength ORF7 construct, named pEGFP-7-123, was transfected into MARC-145cells and intracellular localization of the N-GF fusion protein followed by fluo-rescence microscopy. The N-GF protein produced fluorescence in the nucleolus,whereas, transfection with pEGFP alone showed no nucleolar localization. Theprincipal feature of proteins that migrate from the cytoplasm to the nucleolus isthe presence of a nucleolar localization signal (NoLS) sequence, which consistsof long stretches of basic amino acids. We identified two putative NoLSsequences in the 23983 PRRSV, KQQKRKK and RGKGPGKKNKKKNPEKwhich is similar to NoLS found in other viral proteins, such as the N protein ofSemliki Forest Virus. Current work is directed at following the localization ofN-GF fusion proteins that contain deletions of the putative NoLS sequence, as


well as determining the role of the N-protein in modulating host cell protein andRNA synthesis. We also noticed that MARC-145 cells that expressed the N-GFprotein showed increased cytoplasmic granularity, became rounded, and eventu-ally detached from the plate, which is similar to the CPE observed followingPRRSV infection of MARC-145 cells. The mechanism of N protein-mediatedcell death, which resembles apoptosis, is currently under investigation.


ANALYSIS OF THE STRUCTURAL GENES OF A PRRSV STRAIN THAT GROWS

POORLY IN MACROPHAGES

K.L. Kauers1, M.S. Steffen1, D. Shah1, D.A. Benfield2, R.R.R. Rowland1

Departments of 1Biology-Microbiology and 2Veterinary Science,South Dakota State University, Brookings, SD 57007

Serial passage of porcine reproductive and respiratory virus (PRRSV) strainSDSU-23983 on MARC-145 cells results in decreased ability to replicate inporcine alveolar macrophages (PAMs). Serial passage 93 (P93) showed a 2 logdecrease in virus yield when grown on PAMs compared to wild type P6 and P83.The purpose of this study was to identify the molecular basis for macrophage-tropism by comparing amino acid sequences of P83 and P93 structural genescoded by ORFs 2-7. Amino acid sequence was derived from the DNA sequenceof both strands of RT-PCR products. This analysis revealed 4 amino acid differ-ences in ORF2, 2 in ORF3, 0 in ORF4 and 5, 2 in ORF6, and 3 in ORF 7. Theonly non-conserved changes were in ORF 3(Q-H) and ORF 7 (V-M). Theseresults suggest that inability to replicate in PAMs may be related to events down-stream from the interaction of virus and macrophage receptors since we saw nosignificant change in ORF5, which is considered the principal envelope protein.


CONGENITAL PRRSV: SITES OFVIRUS REPLICATION DURING ACUTE

AND PERSISTENT INFECTION

S.R. Lawson1, D.A. Benfield2, R.R.R. Rowland1

Departments of Biology-Microbiology1 and Veterinary Science2


Infection of late-term fetuses with porcine reproductive and respiratory syn-drome virus (PRRSV) produces enhanced pulmonary disease, increased sec-ondary infections, lymphadenopathy, and stunted growth in the offspring. Thelong term consequences of congenital PRRSV are not known. The purpose ofthis study was to characterize virus replication in a group of piglets deliveredfrom sows infected at 90 days gestation with VR-2332 or SDSU-23983. Viruswas isolated from serum and lung until 21 days post parturition (PP) and fromlymph node and tonsil for up to 132 days PP. In situ hybridization confirmed thepresence of cells supporting virus replication in lymphoid tissues. Four congen-itally infected pigs, retained for approximately 383 days, were negative forPRRSV; by serology, PCR and by in situ hybridization. These results show thatPRRSV replication, following congenital infection, is first established as a multi-organ system infection followed by long-term replication within lymphoidorgans. The mechanism of clearance is not known, but is not associated with theappearance of neutralizing antibody.


NITROGEN PARTITIONING IN A MANUREDFIELD IN SOUTHEAST SOUTH DAKOTA

H. Smeltekop, D. Malo and D. ClayPlant Science Department, South Dakota State University,

Brookings, SD 57007

ABSTRACT

Predicting the amount of nitrogen that will become available throughout agrowing season in manured fields is difficult: manure is not homogenous innutrient content and composition; manure is applied to different parts of the fieldat different times as it becomes available, and manure is not applied evenly to thefield. Landscape position and soil types also play an import role in nitrogenavailability. The objective of this study was to examine the fate of nitrogen in amanured, fertilized field planted to corn. Nitrogen transformations were moni-tored by measuring manure application, plant nitrogen uptake, nitrogen loss dueto runoff and leaching, and nitrogen mineralization.


SPATIAL AND TEMPORAL DIFFERENCES OFHERBICIDE MINERALIZATION:

RELATIONSHIP TO WEED CONTROL

Z. Liu, S. A. Clay, D. E. Clay.Plant Science Department, South Dakota State University,

Brookings, SD 57007

ABSTRACT

Non-uniform Landscapes often are found in North Central Agriculturalfields. Soil properties such as pH, N, and C content also may vary widely acrossthe field. Changes in these properties, in turn, may result in changes of herbicidebehavior in soil, such as sorption, mineralization, degradation and movement.This study investigated the spatial and temporal variability of atrazine andalachlor mineralization in the laboratory, and their relationship to weed biomassmeasured in the field. Eighteen 0 - 15 cm soil samples (15 m apart) from eachof 3 landscape positions (summit, backslope, and toeslope) were collected froma continuous corn field on May 23, August 18, and September 29, 1997. Tengrams of soil from each sample point was placed in an incubation flask and 14C-ring labeled atrazine or alachlor was added. Sodium hydroxide vials were placedin the flask as 14CO2 traps and changed weekly with fresh NaOH. Each week, analiquot of the NaOH solution was placed in scintillation vials, scintillation fluidadded, and the amount of 14CO2 trapped was determined. The experiment wasconducted at 25oC for a period of 8 weeks. Weed biomass samples were collect-ed at each of the sampled point in August by clipping weed in a 0.1m2 area nearthe sample points. The weed biomass was determined by the dry weight of thesesamples. The complete mineralization of atrazine at toeslope, backslope andsummit positions was 27.5%, 53.1% and 50.6%, respectively, for May samples,and 22.2%, 25.6% and 38.1%, respectively, for August samples, and 49.6%,41.9% and 43.1% respectively, for September samples. The complete mineral-ization of alachlor at toeslope, backslope and summit positions was 23.2%,21.8% and 26.2%, respectively, for soil sample collected in May, and 15.9%,19.9% and 22.4%. respectively, for soil samples collected in August, and 27.1%,30.3%, and 33.5%, respectively, for soil sample collected in September. Areawith higher herbicide mineralization had higher weed biomass, whereas areawith lower herbicide mineralization had lower weed biomass. These results mayprovide useful information for better agriculture management and guidelines forprecision farming practices.


ROOT MORPHOLOGY AND ENDOGENOUSCYTOKININ LEVELS OF FLOOD TOLERANT

AND FLOOD SUSCEPTIBLE SOYBEANS

Steven B. Ortmeier and R. Neil ReeseDepartment of Biology/Microbiology


ABSTRACT

Flood tolerant soybeans are plants that can withstand various amounts offlooding (from waterlogged soil to complete submergence) during a wide varietyof developmental stages with little loss of yield compared to flood susceptiblesoybeans. Flooding causes a variety of responses in these herbaceous plants.Early flood responses include ethylene production, aerenchyma tissue formation,and root growth directional changes. Later responses can include growth inhibi-tion, senescence, abscission, and wilting. This experiment focuses on the mor-phological root characteristics of aerenchyma development and adventitious rootformation in response to flooding as well as analyzing the root physiologicalcharacteristics of endogenous cytokinin production.

Using soybean lines pre-screened for flood tolerance, two-week old plantswere subjected to flooded conditions 2-3 cm above the soil line. Plants weresampled at specified times of flood duration for xylem sap extraction and lightmicroscopy sectioning. Root aerenchyma tissue development was observed andcompared between flooded and nonflooded plants using Reeves’ triple stain tech-niques and standard light microscopy. Root tips will also be compared forDNA/RNA content (relative activity) using acridine orange stain and fluores-cence microscopy.

Xylem sap samples were collected using a pressure chamber (100 kPA) forthirty minutes to force xylem exudate from the roots. The exudate was then ini-tially used for an Amaranthus cytokinin bioassay to indicate relative cytokininactivity. Xylem exudates were purified using immunoaffinity chromatographywith polyclonal antibodies specific to three main groups of cytokinins: zeatin,dihydrozeatin, and isopentenyl adenine. These purified samples were then ana-lyzed qualitatively and quantitatively for cytokinins by high performance liquidchromatography.


EFFECTS OF OUTCROSSING ON KERNELTRAITS AND GRAIN YIELD IN HYBRID MAIZE (ZEA MAYS L.)

Mack, C.T.1, Z.W. Wicks III2, P.B. Beauzay2

1Department of Agronomy, Iowa State Univ., Ames, IA2Plant Science Dept., South Dakota State Univ., Brookings, SD 57007

ABSTRACT

A six-year study was conducted to determine the effects of outcrossing ongrain yield, kernel weight, ear weight, and kernel protein concentration of maize(Zea mays L.). Both yellow and white varieties were used in crosses. The useof the white endosperm varieties allowed separation of selfed and outcrossedkernels for measurement of levels of outcrossing, as well as kernel traits for self-ed versus outcrossed kernels in the yellow varieties.

Averaged over locations, levels of outcrossing in outer rows of four-rowplots was 52.42% and 43.83% for inner rows. Grain yield associated withgreater percent outcrossing (outer rows) was 1.51 Q*Ha-1 higher than when asso-ciated with lower outcrossing levels (inner rows). This agreed with findingswhich showed outcross kernel weight exceeding selfed kernel weight. The ratioof outcrossed to selfed kernel weight was 1.071 for outer rows and 1.064 forinner rows. Ear weight did not appear to increase as levels of outcrossingincreased. Lack of response for this trait may have been due to localization ofoutcrossed kernels in the apical or basal portions of the ear which would tend tomask changes in ear weight. Finally, kernel protein concentration was shown tobe increasing for outcrossed versus selfed kernels. The differences in proteinconcentration between selfed and outcrossed kernels was greatest in inner rowswhere sink demand was lower due to fewer outcross kernels competing for lim-ited amino acid supply.


EXTRACTION AND ANALYSIS OF DNA FROMRECALCITRANT PLANT LEAF TISSUE

Galloway, E.M., and M.R. Duvall, Department of Biology and Microbiology,South Dakota State University, Brookings, SD 57007-0595

ABSTRACT

The purpose of this project was to determine the quality of DNA present inten year old samples of freeze-dried wild-rice (Zizania sp.) leaf tissue. There arefour different species of Zizania:: Z. latifolia, Z. aquatica, Z. palustris, and Z.texana. This experiment dealt with leaf tissue from Z. aquatica var. subbrevis, avariety of wild-rice endemic to southern Ontario. After extraction of DNAthrough standard protocols the DNA was found to be in very low concentrationsand highly degraded. Slight modifications to the standard procedure allowed forbetter recovery and ease in handling freeze-dried tissue. A portion of the chloro-plast genome was eventually amplified and sequenced in 300 base pair segments.These portions constituted the chloroplast gene ribulose 1,5 - bisphosphate car-boxylase/oxygenase (rbcL). The segmentation was necessary due to the amountof degradation of the DNA in the tissue. The experimental portion of the projectinvolved testing the freeze-drying process by observing the amount of DNAdegradation after a sample of fresh maize (Zea mays) tissue was freeze-driedunder identical conditions as the Zizania tissue. Maize was selected as a close-ly related, easily obtained species. rbcL was then amplified from the maize leaftissue demonstrating that the freeze-drying process was an adequate method forpreserving DNA in leaf tissue under short-term storage. After comparing tissuefrom Zizania and maize it was concluded that the subsequent treatment of theZizania was the cause of the DNA degradation not the initial freeze-drying pro-cedure. The original contribution of this project surrounds the techniques usedto successfully amplify a portion of the degraded Zizania DNA. Although rbcLcould not be amplified as one piece the sequence was easily recovered when sep-arated into smaller segments, producing strong amplification and verifiable DNAsequence.

INTRODUCTION

Extraction of DNA from tissue can often be a difficult task especially if thetissue is not fresh. Current fresh tissue methods rely on the presence of watermolecules within the tissue. Freezing these molecules over dry ice or liquidnitrogen makes them act as molecular scissors, cutting cellular membranes andcell walls, effectively leaving the internal contents of the cell free in solution.When dealing with tissue that is not fresh, the option of simply utilizing the cell’sintrinsic ‘scissors’ is not always present. In the case of herbarium samples, fos-


sils, or freeze-dried samples, the tissue has lost a great deal if not all of the wateroriginally in the tissue. This lack of water presents the problem of how to lysethe cell walls and membranes, effectively and quickly.

This method was developed for the treatment of wild-rice, (Zizania sp.).Zizania has four different species, Z. aquatica, Z. palustris, Z. texana, and Z. lat-ifolia. Within these four species there are seven varieties that are recognized (Z.aquatica var. aquatica, Z. aquatica var. subbrevis, Z. aquatica var. brevis, Z.palustris var. palustris, Z. palustris var. interior, Z. texana, Z. latifolia.). Thereis controversy among those studying Zizania as to whether or not these varietiesare truly different. Research has been done concerning the taxonomy—relatingleaf size, plant height, floral characters, and geographic distribution (1).Research has also been completed concerning isozyme differences, seed storageprotein differences, and interfertility (2, 3). Although there is a wide base ofknowledge available concerning the taxonomy of Zizania there are still questionsthat are disputed. The original goal of this project was to amplify and comparethe chloroplast gene ribulose 1,5-bisphosphate carboxylase/oxygenase (rbcL)from the different varieties of Zizania. Differences in the gene sequence couldgive some clues to the taxonomic questions that have not been answered. Thetissue used in this experiment was collected, harvested and lyophilized approxi-mately ten years prior to the date of analysis. It was hypothesized that the DNAhad degraded due to repeated freeze/thaw cycles the tissue had undergone duringthe ten year period. To test this hypothesis a secondary experiment was set up totest the effectiveness of freeze-drying as a storage technique for tissue used forDNA analysis. The main problem was how to extract and amplify the ZizaniaDNA from the tissue on hand. The method developed includes the incorporationof a so-called ‘hot CTAB’ method, a rehydration step (necessary to prepare thetissue for grinding and to limit transfer steps to conserve the small amounts ofdegraded DNA), and a modification of normal PCR procedures to accommodatethe fragmented DNA.

METHODS

Collection and Initial Treatment of Tissue

The younger the sample of leaf tissue the more active synthesis of DNA istaking place. For this reason the youngest of the collected tissue was selected.The control tissue, obtained from fresh maize seedlings, was harvested at 2weeks by cutting the plant at the base and immediately covering it in liquid nitro-gen. The tissue was kept frozen until it could be quickly transferred to the freeze-drier. To emulate the procedure used when the initial field collection of theZizania was done, the maize sample was kept frozen (-80 °C) for 48 hours. Themaize tissue was then transferred to the freeze-drier for 12 hours. The tissue wasobserved for signs of wetting. Wetting can be a potential problem initially in thefreeze-drying process; if the tissue is allowed to moisten for too long DNases canbecome active and destroy useful DNA. After the freeze-drying process was


completed the tissue was kept in an airtight container for one week to see ifdegradation would occur.

DNA Extraction

The freshly prepared maize sample and the ten year old Zizania sample (Z.aquatica var. subbrevis harvested from 2-week-old seedlings) were processedfollowing a modification of a published extraction protocol (4). An extractionbuffer containing the detergent CTAB (mixed alkyltrimethylammonium bro-mide) with 2 µl tRNA stock solution at 10 mg/ml per 10 ml extraction buffer, washeated to 60 °C and the tissue was added after it was aggressively minced withscissors. 5 mls of buffer were used per 1 g of freeze-dried tissue. Thetissue/buffer mixture was incubated for 5 min and then homogenized using anelectric cell homogenizer. This process created a small amount of foaming so anadequate-sized container was a necessity. After homogenization was completethe mixture was incubated at 60 °C for another 5 minutes, removed from heat andcombined with an equal volume of chloroform: isoamyl alcohol solution (24:1v/v). This solution was mixed by continuous gentle inversion of the tube for 2minutes. This solution was then put in a -20 °C freezer overnight. After return-ing the solution to room temperature it was centrifuged at 13,000 rpm for 5 minor until there was a translucent top layer clearly separated from the bottom. Theupper (aqueous) layer was removed and added to an equal volume of chilled iso-propanol and allowed to precipitate in the freezer for one week. The solution wasthen centrifuged at 13,000 rpm for 10 min to pellet the DNA. The supernatantabove the pellet was removed and washed with 1 ml 0.2 M sodium acetate in76% ethanol and centrifuged for 5 minutes at 13,000 rpm. After this first washthe supernatant was again removed and 1 ml 0.01 M ammonium acetate in 76%ethanol was added and the solution was centrifuged as previously. After thisfinal centrifugation was completed the pellet was dried and dissolved in sterilewater. The long incubation times were necessary for the degraded nucleic acidswhich have a tendency to stay in solution; the tRNA added to the extractionbuffer also helped pull the DNA out of solution. Before amplification the RNAwas digested from the preparation by standard Rnase digestion methods.

Amplification and Analysis

To check the quality of the DNA preparation a 5 µl aliquot was run on anagarose gel. The maize, as shown in Fig. 1, did not show much degradation. TheZizania DNA, however, showed a great deal of degradation. Most of these frag-ments were estimated to be within 250-350 base pairs in length. Using this gelan estimate was made that there should be an adequate number of 300 bp DNAfragments for amplification. Five pairs of amplification primers were thus cho-sen to produce overlapping fragments of approximately 300 base pairs (5).These primer pairs were Z1, Z346R; Z234, Z674R; S523-Z895R; Z674-Z1020R;and Z895-Z1375R. These primers are named for the sequence from which they


were derived (“Z” = rbcL from Z. mays; “S” = rbcL from spinach) and the num-ber refers to the nucleotide position of the 5’ end of the primer on the maize rbcLsequence. Two rounds of Taq-mediated amplification were performed using aPTC-100 thermal cycler (MJ Research), each beginning with a “hot start” (2 min,94 °C; 5 min, 80 °C). The first amplification protocol also included 34 cyclesof: 2 min at 94 °C; 3 min at 42 °C; and 3 min at 72 °C with a final chain exten-sion of 12 min at 72 °C. This product was run on a standard 0.8% agarose geland the bands were cut from the gel and frozen completely (-20 °C, overnight)in microcentrifuge tubes and centrifuged at 13,000 rpm. The liquid layer wasremoved from the top of the agarose. Aliquots of this product were amplified asecond time as above except with the temperature of the annealing step increasedfrom 42 °C to 48 °C. The products of this reaction were concentrated usingMicrocon-100 columns (Millipore Corp.) and run on a 0.8% agarose gel to esti-mate concentration. The concentrated products were sent together with aliquotsof the amplification primers to the DNA sequencing Facility at Iowa StateUniversity for automated DNA sequencing. Each fragment was sequencedtwice, once with each of the terminal primer sequences used in the amplification.


Both the maize and Zizania amplified well, although for the same mgamounts of tissue the maize showed a much larger estimated yield of DNA. Theamplification of DNA from all five of the approximately 300 base pair fragmentsfrom the recalcitrant leaf tissue was successful (Fig. 2). 1363 bases of readablesequence were produced from these overlapping fragments. Automatedsequencing signal strength was very high (average values were G: 541; A: 455;T: 399; C:550) indicating the quality of the template. Forward and reversesequences for each of the doubly sequenced fragments were the same in all cases.This new rbcL sequence was compared to a published rbcL sequence from Z.texana. and differed at only two nucleotide sites. These were transitions from theZ. texana. sequence at positions 687 (G to A) and 909 (A to G). Note that bothof these are third codon position mutations. The minor differences between thesesequences are as would be expected from a sequence comparison between con-generic species. When the var. subbrevis rbcL sequence was compared with thecontrol maize sequence 54 differences were found as would be expected betweentwo such distantly related grass species. The sequence from this var. subbrevistissue was thus determined to be valid and reliable and not a sequence from amaize contaminant or from another species.

Analysis of the freeze-drying process showed that it was effective for pre-serving DNA for DNA sequence analysis. However as shown here, subsequenttreatment of the tissue is important for maintenance of DNA integrity.Recalcitrant tissue is often the only source of DNA for a particular experiment.Certain features in this method that are especially important for working withfreeze-dried, degraded tissue are: 1) the initial hydrating step 2) addition oftRNA and 3) analysis of the integrity of the DNA before attempting amplifica-


tion so primers can be adjusted to optimize amplification. The initial incubationallows the buffer to hydrate the tissue making it more pliable and easier to breakapart. The tRNA, as mentioned before, increases the amount of small fragmentsof DNA that are pulled out of solution, especially if the overall amount of tissuebeing treated is very small. Lastly, breaking the amplification into several small-er segments, ie. sizes compatible with the size of the degraded tissue, yielded sin-gle amplification products that sequenced well. This process has a few moresteps than standard extraction procedures but it is a good way to utilize tissue thatbefore might have otherwise been discarded as too difficult on which to spendtime.

LITERATURE CITED

1) Terrell, E., P. M. Peterson, J. Reveal, and M. R. Duvall. 1997. Taxonomyof North American Zizania (Poaceae). SIDA 17: 533-549.

2) Duvall, M. R. 1987. A systematic evaluatoin of the genus Zizania(Poaceae). Dissertation, University of Minnesota, St. Paul.

3) Warwick, S. I. And S. G. Aiken. 1986. Electrophoretic evidence for therecognition of two species in annual wild rice (Zizania, Poaceae).Systematic Botany 11: 464-473.

4) Doyle, J. J. and J. L. Doyle. 1987. A rapid DNA isolation procedure fromsmall quantities of fresh leaf tissue. Phytochemical Bulletin 19: 11-15.

5) Zurawski G., DNAX Corporation, Palo Alto, California furnished the origi-nal primer sequences and primer stocks.


Fig. 1. DNA extracts from Zizania aquatica var. subbrevis (lane 2)and Zea mays (lane 3). The largest DNA fragments for each arenoted with arrows at approximately 25,000 base pairs (lane 3) and1000 base pairs (lane 2) although note that the bulk of the var. sub-brevis DNA is at 250-350 base pairs. Lane 1 is a DNA standard.

Fig. 2. Amplifications of five over-lapping portions of rbcL from Z. aquati-ca var. subbrevis using five different pairsof primers: Z1, Z346R (lane 2); Z234,Z674R (lane 3); S523-Z895R (lane 4);Z674-Z1020R (lane 5); and Z895-Z1375R (lane 6). Lane 1 is a positivecontrol for the amplification reaction.


MUTAGENESIS AND ANALYSIS OFAUREOBASIDIUM PULLULANS

Anna R. Oller and Carl A. WestbySouth Dakota State University, Brookings, SD 57007

Aureobasidium pullulans is a polymorphic fungus that produces a polysac-charide called pullulan. Pullulan is important in industrial applications, but dueto its high cost, using pullulan in products is not economically feasible. Pullulanelaboration on corn mash is very high, and smaller quantities are produced onsoybean media. Pullulan would be able to be grown in current ethanol and soy-bean plants, therefore causing economic growth in South Dakota agriculture.Since Aureobasidium produces such low quantities of pullulan, finding a pullu-lan gene is very important. Another gene, 1,3-beta-glucan synthase, has beenfound in the cell walls of Candida albicans and Saccharomyces cerevisiae.Since pullulan is believed to be produced in the cell wall, the glucan synthasegene may be involved in pullulan production. Our objectives are to find a pul-lulan gene and the 1,3-beta-glucan synthase gene, as well as to determine if theglucan synthase gene is involved in pullulan production.

In order to identify a pullulan gene, gamma irradiation of the wild type ofAureobasidium pullulans was performed by a cobalt source and mutants wereidentified morphologically on various media. DNA was isolated through the useof yeast DNA extraction kits. The mutants were compared to the wild typethrough gel electrophoresis after their DNA had been amplified through the poly-merase chain reaction (PCR) with known primers. In order to find the 1,3-beta-glucan synthase gene, primers were designed according to the Candida sequencepublished in a journal article. PCR was then employed with the wild type DNAand gel electrophoresis was used to determine if the gene was present.

Specific banding patterns of the wild type and the mutants have led to theidentification of a presumptive pullulan gene. The individual bands will be puri-fied and put into yeast protoplasts to determine which band, or combination ofbands, restore pullulan production, indicating a pullulan gene within thesequence. PCR results with the glucan synthase primers indicate thatAureobasidium pullulans does contain the gene. DNA obtained from Japanesescientists will be used as a labeled probe in a Southern Hybridization procedureto determine if the sequences truly are homologous.

Aureobasidium pullulans produces pullulan, which is important in commer-cial applications. A presumptive pullulan gene has been identified and furtherexperiments will reveal the sequence of the gene. A 1,3-beta-glucan synthasegene has also been found in A. pullulans, and Southern Hybridization experi-ments will demonstrate if the gene is homologous to the Candida gene.


ALLELOPATHIC POTENTIAL OF ECHINACEAANGUSTIFOLIA D.C.’S ROOT EXTRACTS

Kimberly Piechowski and R. Neil ReeseDepartment of Plant Science and Biology /Microbiology


ABSTRACT

The narrow-leafed purple coneflower, Echinacea angustifolia D.C., is aperennial native to North America. E. angustifolia is found on the dry prairiesof Texas to Saskatchewan, west to the Rocky Mountains and east into Minnesota.E. angustifolia produces many biologically active compounds that have shownan allelopathic effect on lettuce (Lactuca sativa), switchgrass (Panicum virga-tum) and prairie dropseed (Sporobolus heterolopis). Natural allelopathic chemi-cals have the potential of being used for the production of herbicides that areenvironmentally safe. These allelopathic compounds possess components thatcould be used for development of new herbicides for use in production agricul-ture. The allelopathic potential of ethanol soluble compounds from E. angusti-folia roots from five populations (South Dakota, North Dakota, Wyoming,Kansas, and Nebraska) were examined. A common allelopathy bioassay waspreformed using lettuce seeds germinated in the presence of the ethanol extractspipetted on filter paper. The extracts were brought to dryness and were resus-pended in deionized water. Analysis of the seed assay was completed by mea-suring percent germination and root elongation of 25 lettuce seeds per replicationfor extracts from each of 100 E. angustifolia plants (20 plants per population).Results from the lettuce seed germination experiment showed that germinationvaried from 100% to 0%. Variation within extract from E. angustifolia popula-tions were as significant as between populations, this suggests that environmentis not the only factor effecting the allelopathic potential of the plants. The pop-ulations do show a tend, as collection sites moved southward the allelopathicpotential decreased. Root extracts were concentrated by evaporation and ana-lyzed by reverse phase high performance liquid chromatography (HPLC) usinga C-18 column. Absorbancy was measured at 210 nm and 254 nm. HPLC analy-ses revealed eleven significant peaks in all the populations. Using multipleregression analyses, a correlation was made between germination, root elonga-tion and peak area. These results showed that compounds identified as peaknumbers three and six contribute to 72 % of the allelopathic effects of E. angus-tifolia root extracts. Purification of the first seven peaks has been completed.Since peaks three and six are of most interest they are being identified by NMR(nuclear magnetic resonance) at the University of Illinois. When the identifica-tion process is completed, another lettuce seed germination assay will be per-formed using purified compounds to determine whether allelopathic effects ofthe isolated compounds are synergistic or additive.


A STUDY OF FOSSIL AND MODERN MEMBERSOF THE PANICEAE (POACEAE) FROM

NORTH AND SOUTH AMERICA

J.J. Smith and M.L. GabelDepartment of Biology, Black Hills State University, Spearfish, SD 57799

ABSTRACT

While fossil grasses have been reported numerous times from Miocene stra-ta in North America, this is the first report of three dimensional opalized grassfossils from South America. Fossil members of the genus Setaria (Poaceae) havebeen reported only one time prior to this study, from the Miocene Ash HollowFormation of Lincoln County, Nebraska. We are reporting the second collectionof fossil Setaria from the province of Catamarca, Argentina. The MioceneAndalhuala Formation of Catamarca is composed of high percentages of sandand ash, similar in nature to the Ash Hollow Formation in North America. Acomparison was made between the fossils from North America and SouthAmerica, as well as to modern specimens of the South American Paniceae.Preservation of fossils from both North America and South America are exquis-ite, with epidermal cells intact. Cuticle was removed in both fossil and modernspecimens prior to analyses. Characteristics of the paleas and lemmas includingepidermal cell dimensions, shapes, and ornamentation were used for detailedanalyses. Results from cluster analyses and principal component analyses indi-cate that the fossils from South America are most similar to S. tenassisima and S.scandens. Fossils from North America were most similar to S. sulcata.


A STUDY OF LYCOPODIUM DENDROIDEUM INTHE BLACK HILLS OF SOUTH DAKOTA

Terri Hildebrand1, Mark Gabel2, Cecilie Steib2, and Shane Sarver2

1Hildebrand Contracting, Sturgis, SD 577852Department of Biology, Black Hills State University, Spearfish, SD 57799

ABSTRACT

Lycopodium dendroideum has a circumboreal distribution, but is listed as asensitive species in South Dakota by the U.S. Forest Service and the SouthDakota Department of Game, Fish and Parks. We are in the third year of a multi-year study of the Lycopodium populations in the Black Hills. We have locatednumerous new populations, and with the cooperation of the U.S. Forest Service,have begun intensive studies on the effects of logging on three populations. Weare also determining population genetics of several of the Black Hills popula-tions and making comparisons with populations within the Black Hills and fromtheir nearest neighbors outside the Black Hills in Idaho, Manitoba, andMinnesota.

Protein analysis using isozyme electrophoresis has shown no variation inphosphoglucoisomerase, glutamate oxaloacetate transaminase, esterase, glucose-6-phosphate dehyrogenase, leucine amino peptidase, phosphoglucomutase, ormalate dehydrogenase from populations in the Black Hills or among the nearestoutlying populations. These results do not correlate with previous studies ofother species of Lycopodium.


DYNAMICS OF SOURCE-SINK LANDSCAPESRELATIVE TO WATERFOWL POPULATIONS IN

THE NORTHERN GREAT PLAINS

Kenneth F. Higgins1, Rex R. Johnson2, David E. Naugle2, Matthew M. Holland2, Thomas R. Cooper2, and Michael E. Estey2

1 South Dakota Cooperative Fish and Wildlife Research Unit, USGS/BRD,Box 2140B, South Dakota State University, Brookings, SD 57007

2 Wildlife and Fisheries Sciences Department, Box 2140B, South Dakota State University, Brookings, SD 57007

ABSTRACT

Migratory waterfowl populations annually occupy habitats with variablesuitability within broader landscapes or regions. Current theory regarding sourceand sink populations indicates that landscape variables such as land use compo-sition and local wetland complex features may be used to identify geographicsource and sink areas. Although models that incorporate landscape variables areuseful in predicting potential areas of use, land managers also must consider tem-poral variability and local conditions that may cause source areas to shift locallywithin broadly defined regions. In this presentation, we identify areas previous-ly considered potential sinks for breeding waterfowl that periodically function assource areas and contribute to waterfowl populations during wet hydroperiods.Our evidence of temporal shifts in habitat suitability have direct importance tospatial strategies for wetlands protection and acquisition, to long-term manage-ment plans for waterfowl, and to conceptual logistics relative to source and sinkhabitat theory.


GOLDEYE RECRUITMENT AND GROWTH INTWO MISSOURI RIVER BACKWATERS

Daniel N. Moon, Shannon J. Fisher, and David W. WillisDepartment of Wildlife and Fisheries Sciences


ABSTRACT

Goldeye Hiodon alosoides was found to be an abundant fish species in theMissouri River system and interactions with the lotic community at all levelslikely occur. Therefore, understanding the population dynamics (recruitment,growth, and mortality) of goldeye in this system has the potential to be impor-tant. Goldeye were sampled in two backwater habitats in western North Dakotawith experimental gill nets during August and September of 1997 and scaleswere collected. From the scales, we determined age structure and back-calculat-ed length at age. Mean back-calculated length at age was found to be 121-, 189-, 234-, 276-, 298-, 305-, and 314-mm total length at ages 1-7, respectively. Theage-frequency histogram indicated the presence of goldeye in each age groupthrough age 7 and the stair-step decline in goldeye frequency from ages 2through 7 indicated that recruitment of goldeye in this system was relatively con-sistent. A total mortality rate of 47% for age 2-6 goldeye was determined usingthe Chapman-Robson method. The population dynamics of goldeye in upperMissouri River backwaters were similar to goldeye studies in other MissouriRiver segments and both lotic and lentic populations in Canada.

INTRODUCTION

Goldeye Hiodon alosoides is an adundant fish species in many northernwaters and is typically classified as slow growing and long-lived (Kennedy andSprules 1967). Miller (1970) found that growth patterns of goldeye at variouslocations in Garrison Reservoir, North Dakota, were statistically different, whichwas attributed to differences in turbidity and temperature. Goldeye had maxi-mum growth increments during the first year of life in Lake Oahe, South Dakota,decreasing only slightly in the second year, with marginal increments in latteryears of life (Miller and Nelson 1974). Goldeye recruitment has been found tobe quite consistent in the Missouri River system (Miller and Nelson 1974); how-ever, mortality will respond to environmental disturbances and the species is sus-ceptible to exploitation (Kennedy and Sprules 1967). However, Elrod andHassler (1971) found that in natural conditions, annual survival rates typicallyexceed 50%.


Although the species is common in the Missouri River system, assessmentsof goldeye population dynamics are sparse or unavailable. Because goldeye area primary community member in the potentially critical habitats of the upperMissouri River, understanding their biology is important. Therefore the objec-tives of this study were to 1) draw inferences about goldeye recruitment from theage structure, 2) determine the rate of mortality from age-frequency data, and 3)assess incremental and annual growth from the back-calculated mean length atage analysis.

STUDY SITE AND METHODS

This study was conducted in two Missouri River backwaters locatedbetween river kilometers 2510 (mile 1569) and 2538 (mile 1574) in northwest-ern North Dakota. We defined a backwater as an off-channel habitat that con-tained water with limited or absent flow and was connected to the main riverchannel during all or a portion of the year. Goldeye were sampled from thesebackwaters during August and September of 1997 with experimental gill nets(38-m long, 1.8-m deep, and 7.6-m panels of 19-, 25-, 38-, and 76-mm bar-mea-sure mesh). Scales were collected from below the lateral line and directly behindthe pectoral fin from 179 goldeye. Otoliths were removed from 12 goldeye andaged with a dissecting scope. Scales were aged by two readers. Scales for whichthe readers differed in age assignments were pressed on acetate slides and re-examined by both readers, discussed, and an age was assigned. Age structureand back-calculated length at age of adult goldeye from the Missouri River back-waters were determined from the scales.

Scale measurements were digitized using the DISBCAL computer softwareprogram (Frie 1982), and back calculated length-at-age was determined. We cal-culated the y-intercept for the body length-scale radius relationship from our dataset (50 mm). Recruitment was qualitatively assessed through visual inspectionof the age-structure histogram. Because we used multiple-size meshes on ourgill nets we assumed that our sample was representative of the goldeye popula-tion. Thus, missing or substantially reduced numbers of goldeye from given yearclasses would be indicative of inconsistent recruitment. From the age-frequencydata, we calculated an estimated mortality rate using the Chapman-Robsonmethod (Everhart et al. 1975).


Scales have traditionally been used to determine ages of goldeye (Donaldand Kooyman 1977a) and are useful indicators to at least three years of age(Donald and Kooyman 1977b). Craig et al. (1989) and Donald et al. (1992)found that bony and calcified structures are preferable for age determination inolder goldeye. Removal of otoliths from larger goldeye proved to be difficultand inefficient; thus, we only report here the results of our scale aging and analy-sis. The two readers only had an initial agreement percentage of 61, indicatingthat precision of assigned ages was somewhat low.


We concluded, based on the age-frequency histogram, that goldeye rangingin age from 1 to 7 were present in our sample (Figure 1). The 1995 (age 2) gold-eye year class was the most abundant and abundance of the 1994 (age 3) yearclass appeared to be somewhat low. The low frequency of age-1 goldeye waslikely the result of gear bias ( i.e, we ineffectively sampled goldeye of that size).In general, the presence of goldeye in representable numbers for year classesfrom 1990 through 1996 is indicative of relatively consistent recruitment.However, Donald (1997) reported that dominant goldeye year classes resultedfrom warm and calm weather during early life stages.

The total annual mortality rate for age 2 through age 6 was 47%. Age-1goldeye were excluded from the analysis because they had not recruited to a sizeefficiently captured by our gill nets. The age-7 goldeye were also excludedbecause age groups with fewer than five fish should not be included in the analy-sis (Everhart et al. 1975). Total annual mortality rates of 40% were reported byMiller and Nelson (1974) for goldeye in Lake Oahe, South Dakota.

Mean back-calculated lengths for goldeye from the upper Missouri Riverbackwaters were comparable to those reported by Miller and Nelson (1974) forthe Lake Oahe system. Carlander (1969) reported mean back-calculated lengthsfrom several Canadian waters that were also comparable through age 4, butgrowth for older goldeye was faster (Table 1). In our study the greatest growthincrement occurred during the first year of life, declined by approximately 50%in the second year of life, and then remained nearly constant from ages 4-7 (Table2).

In summary, we inferred from inspection of the age-frequency histogramthat goldeye in the upper Missouri River backwaters had relatively consistentrecruitment and mortality was comparable to that reported in other waters. Meanlength at age for goldeye from this area was also comparable to other studies.Thus, goldeye might be a relatively stable component of the Missouri River fishcommunity. Consistent recruitment, moderate mortality, and relatively stablegrowth increments in older individuals all suggest that the population may notsubstantially fluctuate during differential hydrograph cycles. Thus, goldeye maybe quite adaptable to changes in habitats and food resources.

ACKNOWLEDGEMENTS

We would like to thank the Ecological Services and Fisheries AssistanceOffices of the U.S. Fish and Wildlife Service, South Dakota State University,North Dakota State Game and Fish Department, and the South DakotaCooperative Fish and Wildlife Research Unit for their essential contributions tothe completion of this study. This manuscript was approved for publication bythe South Dakota Agricultural Experiment Station as Journal Series No. 3066.


LITERATURE CITED

Carlander, K. D. 1969. Handbook of freshwater fishery biology. Iowa StateUniversity Press, Ames.

Craig, J. F., K. Smiley, and J. A. Babaluk 1989. Changes in the body composi-tion with age of goldeye, Hiodon alosoides. Canadian Journal of Fisheriesand Aquatic Sciences 46:853-858.

Donald, D. B. 1997. Relationship between year-class strength for goldeyes andselected environmental variables during the first year of life. Transactions ofthe American Fisheries Society 126:361-368.

Donald, D. B., J. A. Babaluk, J. F. Craig, and W. A. Musker. 1992. Evaluationof the scale and operculum methods to determine ages of adult goldeyeswith special reference to a dominant year class. Transactions of theAmerican Fisheries Society 121:792-796.

Donald, D. B., and A. H. Kooyman. 1977a. Migration and population dynamicsof of the Peace-Athabasca delta goldeye population. Canadian WildlifeService Occasional Paper 31.

Donald, D. B., and A. H. Kooyman. 1977b. Food, feeding habits, and growthof goldeye, (Hiodon alosoides) (Rafinesque), in waters of the Peace-Athabasca Delta. Canadian Journal of Zoology 55:1038-1047.

Elrod, J. H., and T. J.Hassler. 1971. Vital statistic of seven fish species in LakeSharpe, South Dakota 1964-69. Pages 27-40 in G.E. Hall, editor. Reservoirfisheries and limnology. American Fisheries Society, Special PubicationNo. 8, Bethesda, Maryland.

Everhart, W. H., A.W. Eipper, and W. D. Youngs. 1975. Principles of fishery sci-ence. Cornell University Press, Ithaca, New York.

Frie, R.V. 1982. Measurement of scales and back-calculation of body lengthusing a digitizing pad and microcomputer. Fisheries 7(5):5-8.

Kennedy, W. A. and W. M. Spules. 1967. Goldeye in Canada. Fisheries ResearchBoard of Canada, Bulletin 161, Ottawa.

Miller, D.E. 1970. A comparison of biological characteristics of goldeye, Hiodonalosoides (Rafinesque), in Garrison Reservoir. M.S. Thesis. University ofNorth Dakota, Grand Forks.

Miller, G. L., and W. R. Nelson. 1974. Goldeye, Hiodon alosoides, in LakeOahe: abundance, age, growth, maturity, food, and the fishery, 1963-69.U.S. Fish and Wildlife Service, Technical Paper 79, Washington, D.C.


SPRING MIGRATION OF RAPTORS IN MOODYCOUNTY SOUTH DAKOTA

Andy E. Gabbert andNatoma A. Schneider

Department of Wildlife and Fisheries SciencesBox 2140B, South Dakota State University

Brookings, SD 57007

ABSTRACT

The number of raptors in the northern Great Plains increases dramaticallyduring the spring and fall migration periods. However, little published informa-tion can be found on the timing, abundance, and composition of these migrationsin South Dakota. We used 4 road routes to monitor the timing of migration,species diversity, and annual variations in the relative abundance of raptors ineastern South Dakota during spring and early summer of 1995 and 1996.Surveys were conducted from 31 March 1995 to 2 June 1995 and 1 January 1996to 5 June 1996. Observation points were marked approximately every 805 malong each route. We observed 139 raptors representing 7 species during ourstudy. Red-tailed hawks (Buteo jamaicensis) (n = 56) and American kestrel(Falco sparverius) (n = 36) were the most abundant species observed. The peakrelative abundance of raptors occurred the first week of April and steadilydeclined from the third week of April until June.

INTRODUCTION

Spring and fall migrations of raptors (i.e., hawks, eagles, falcons) is oftennoted by the public. Local bird enthusiast look forward to the raptor migrationsto view species which can only be seen in South Dakota during these times.However, little published information can be found on the timing, abundance andcomposition of these raptor migrations in South Dakota (L. D. Flake pers. com-mun.). The spring and fall raptor migrations provide excellent opportunities toobserve these birds because of the high concentrations and variety of speciesmigrating to breeding or wintering grounds. Long-term surveys of migratingraptors could provide important information relating to population changes andconservation status of raptor species common to South Dakota and the northernprairies.

The staple diet of many raptors includes birds and mammals (Martz andKochert 1995, Norelius 1984) and migration patterns of raptors may be influ-enced by the availability of food (Niles et al. 1996). In addition to a generalinterest in observing these raptors, there is also an interest in their potential influ-


ence on the survival of gamebirds. Previous researchers (Leif 1996, Snyder1985, Weigand 1980) have indicated that the spring influx of avian predators wasa principal factor affecting the spring survival of ring-necked pheasants(Phasianus colchicus) and grey partridge (Perdix perdix). The highest densitiesof ring-necked pheasants occurs in eastern South Dakota (Trautman 1982) andthe peak period of non-hunting mortality of adult birds occurs during the spring(Leif 1996). Ground dwelling birds (e.g., Phasianidae, ?) of eastern SouthDakota may be particularly vulnerable to avian predation in spring due to sparseprotective ground cover, a corresponding increase in raptors during spring migra-tion, and the presence of trees for use as perching sites (Leif 1996, Petersen 1979,Snyder 1985).

This study was supplementary to studies specific to the survival and repro-duction of ring-necked pheasants in eastern South Dakota (Gabbert 1997, Purviset al. unpub., data). It was initiated to evaluate the specific period between win-ter and summer when avian predation could negatively affect the survival andsubsequent reproduction of ring-necked pheasants. However, the informationpresented could be valuable to future investigative studies conducted in easternSouth Dakota, where the possible effects of avian predators to the population ofinterest is unknown. The information presented provides a basic knowledge ofthe timing, relative abundance, and composition of raptors during the springmigration in eastern South Dakota.

METHODS

We used 4 road routes to monitor the timing of migration, species diversity,and annual variations in the relative abundance of raptors in Moody County,South Dakota during spring and early summer 1995 and 1996. Surveys wereconducted weekly from 31 March 1995 to 2 June 1995 and 1 January 1996 to 5June 1996. All surveys were performed by a single observer within a season toreduce possible variations in observer bias; however, the same observer was notused in both years. In 1995, we used 2-16 km (10 mile) road routes, with eachroute conducted on the same day. For 1996, we added 2-16 km routes to moni-tor a broader geographic area. We conducted 2 of 4 road routes (paired) on thesame day each week with paired routes conducted every other week. Surveysbegan mid-day (1200 - 1400 hours) when sky’s were clear to partly cloudy. Nosurveys were conducted during rain, snow, or other weather events that reducedobserver visibility to < 400 m.

Observation points were marked approximately every 805 m (0.5 mile) fromthe start to to the end of each route. The observer drove to the start of a route,recorded the start time, and then proceeded to look for raptors. At each obser-vation point, the observer exited the vehicle and scanned the area in a circularpattern for 1 minute without the aid of binoculars. If a raptor was sighted, binoc-ulars were used to aid species identification. For the 1996 survey, a 60x spottingscope was also used to help identify perched raptors. Date, time, species, andbehavior (perched or flying) was recorded for each raptor sighting. Only raptors


observed at observation points were recorded. After the completion of the firstroute, the ending time was noted and the observer proceeded to the next route.We alternated the starting route for each successive survey. Raptor relative abun-dance was calculated by dividing the total number of raptors observed per weekby the total road route length (32.18 km).


We observed 139 raptors representing 7 species during our study (Table 1).We identified 117 individuals to species, while 22 were classified as unknown orunknown Buteo species. Species identified include red-tailed hawk (Buteojamaicensis), rough-legged hawk (B. lagopus), Swainson’s hawk (B. swainsoni),bald eagle (Haliaeetus leucocephalus), northern harrier (Circus cyaneus),American kestrel (Falco sparverius), and peregrine falcon (F. peregrinus). Red-tailed hawks and American kestrels were the most common raptors observed.Two species (i.e., bald eagle, peregrine falcon) were observed exclusively duringthe 1996 survey. Excluding American kestrel observations (due to their erraticbehavior), 66% of 116 raptors were observed in flight while 34% were observedon a perch site. The peak relative abundance for all species combined, occurredduring the first week of April and steadily decline into June both years (Fig. 1).However, the 1995 survey did not begin until 31 March, and the first raptorrecorded during the 1996 survey was observed in early March.

Red-tailed hawk and northern harrier were the initial species recorded dur-ing both the 1995 (31 March) and 1996 (4 March) surveys. In addition, aSwainson’s hawk was observed on 31 March 1995. This was one of the earliestreported sightings of this species in South Dakota (SDOU 1991). The peak peri-od for Swainson’s hawk observations in South Dakota normally occurs duringthe last week of April (SDOU 1991) and is typically 3 weeks behind the red-tailed hawk migration (Janes 1994). The timing of all remaining raptor observa-tions recorded during our study occurred within the expected range for SouthDakota (SDOU 1991).

We observed an apparent greater relative abundance of raptors during the1995 survey versus the 1996 survey. However, there was little variation in thetiming of migration pulses between years (Fig. 1). Red-tailed hawk relativeabundance peaked the last week of March and again on the second week of Aprilfor both survey years (Fig. 2). American kestrel relative abundance peaked thelast week of March in 1995 and the first week of April in 1996 (Fig. 3). Northernharrier relative abundance peaked the second week of April in 1995, but werefound from the first week of March to the last week of April in 1996 (Fig. 4).Apparent differences in relative abundance between years may be due to actualdifferences in population abundance, or differences in observer abilities.Although relative abundance was lower in 1996, species richness was greater.We surveyed a larger geographic area in 1996. Therefore, the likelihood ofencountering more diverse habitats increased and consequently the diversity ofspecies observed increased (e.g., the association of riparian zones and baldeagles).


Our observations on spring raptor migrations in eastern South Dakota sug-gests that the best time to view raptors occurs between the last week of Marchand the second week of April. Greater than one third of raptors recorded in ourstudy were observed on a perch. The perching behavior by migrating raptorssuggests that the areas resources were used for feeding and resting purposes(Niles et al. 1996). Thus, there is a potential for increased predation by migrantraptors from March to June, with the greatest potential occurring in April.

LITERATURE CITED

Gabbert, A. E. 1997. Food plot effects on winter home range and survival ofradio-marked pheasant hens in east-central South Dakota. M. S. Thesis,South Dakota State University, Brookings. 76pp.

Janes, S. W. 1994. Partial loss of red-tailed hawk territories to Swainson’shawks: relations to habitat. Condor 96:52-57.

Leif, A. P. 1996. Survival and reproductive chronology of female ring-neckedpheasants in South Dakota. Prairie Nat. 28:191-200.

Marti, C. D., and M. N. Kochert. 1995. Are red-tailed hawks and great-hornedowls diurnal-nocturnal dietary counterparts. Wilson Bull. 107:615-628.

Niles, L. J., J. Burger, and K. E. Clark. 1996. The influence of weather, geog-raphy, and habitat on migrating raptors on Cape May Peninsula. Condor98:382-394.

Norelius, S. E. 1984. Use of eastern South Dakota shelterbelts by nesting birdsof prey. M. S. Thesis, South Dakota State University, Brookings. 48pp.

Petersen, L. !979. Ecology of great horned owls and red-tailed hawks in south-eastern Wisconsin. Wis. Dep. Nat. Resour. Tech. Bull. 111. 63pp.

Snyder, W. D. 1985. Survival of radio-marked hen ring-necked pheasants inColorado. J. Wildl. Manage. 49:1044-1050.

South Dakota Ornithological Union. 1991. Birds...........

Trautman, C. G. 1982. History, ecology and management of the ring-neckedpheasant in South Dakota. South Dakota Dep. Game, Fish and Parks Wildl.Res. Bull. 7. 118pp.

Weigand, J. P. 1980. Ecology of the Hungarian partridge in north-centralMontana. Wildl. Monogr. 74. 106pp.


Figure 1. Relative abundance (raptors/32.18km) of 7 raptor species observed in MoodyCounty, South Dakota, between 27 Februaryand 5 June, 1995 and 1996.


Figure 2. Relative abundance (raptors/32.18 km)of red-tailed hawks (Buteo jamaicensis) observedin Moody County, South Dakota, between 27February and 5 June, 1995 and 1996.

Figure 3. Relative abundance(raptors/32.18 km) of Americankestrels (Falco sparverius)observed in Moody County,South Dakota, between 27February and 5 June, 1995 and1996.


WHITE CRAPPIE BIOLOGY IN AN UPPERMISSOURI RIVER BACKWATER

Randy A. Sheik, Shannon J. Fisher, and David W. WillisDepartment of Wildlife and Fisheries Sciences


ABSTRACT

Research conducted in Erickson Island Slough (EIS), a large Missouri Riverbackwater in western North Dakota, revealed the presence of a substantial whitecrappie Pomoxis annularis population. The function of centrarchids in northern-latitude riverine backwaters is not well understood. Thus, we collected whitecrappie in May, July, and September to assess the structure and dynamics of thispopulation. Mean back-calculated length at age was 77-, 163-, 228-, 245-, 277-, and 299-mm total length (TL) at ages 1-6, respectively. White crappie season-al and annual growth in EIS was similar to other regional populations. Sizestructure varied seasonally, ranging from a proportional stock density (PSD) highof 64 in May to a low of 28 in July. Incremental relative stock density (RSD)values indicated that the population was dominated by white crappies between130- and 250-mm TL. Assessment of age structure indicated that the 1995 yearclass (age 2) was especially abundant. White crappie body condition values werehighest for all length groups in July, with mean relative weight (Wr) values of106 to 122. Mean Wr values were lowest in September, ranging from 78 to 99.The food habits of white crappies in EIS varied with crappie length; however, allcrappies had diets dominated by Cladocera, Corixidae, and fish eggs in May.White crappies less than 200-mm TL in September primarily consumed zoo-plankton, whereas crappies greater than 200 mm preyed upon fishes andCorixidae.

INTRODUCTION

White crappie Pomoxis annularis have been extensively studied and supportimportant sport fisheries in both reservoirs and natural lakes (Hammers andMiranda 1991). Although this species is native to the Missouri River system, ourunderstanding of lotic crappie Pomoxis spp. populations, particularly in northernlatitudes, remains limited. Centrarchid growth and year-class strength are influ-enced by biotic and abiotic resource availability during various life stages (Ziglerand Jennings 1993). Habitat availability in unmodified large riverine systemstends to be diverse and includes backwater habitats with submergent and emer-gent macrophytes that are important for fish production (Dewey and Jennings1992).


By gaining insight into white crappie population dynamics, populationstructure, and community interactions in the potentially productive backwaterhabitats of the upper Missouri River, we can better assess the role of this cen-trarchid in the local aquatic community. Thus, the objectives of this study were(1) to assess the structure and dynamics of a white crappie population in aMissouri River backwater and its potential as a sport fishery, and (2) to initiatean assessment of the role of white crappies in the aquatic community of the upperMissouri River.

STUDY SITE

Research conducted in 1997 indicated that Erickson Island Slough (EIS)contained a white crappie population that was suitable for an initial biologicalstudy of the species. EIS is a backwater located along the Missouri River on thecounty line between Williams and McKenzie counties in northwestern NorthDakota. The backwater has a surface area of approximately 1100 ha; however,extreme seasonal fluctuations in surface area and mean depth coincide with theMissouri River hydrograph. The hydrograph of this Missouri River stretch tendsto be quite dynamic due to its location below the confluence with theYellowstone River, the longest free-flowing river in the United States.

METHODS

White crappies were collected during the months of May, July, andSeptember of 1997 from EIS using trap nets. We expended 18 net nights of effortper sample period (nine nights with nets having 0.9- x 1.8-m frames, 9.5-mm barmesh, and 16- x 0.9-m leads; nine nights with nets having 0.6- x 0.9-m frames,9.5-mm bar mesh on the frames, 3-mm bar mesh on the hoops, and 7- x 0.6-mleads).

White crappie density was indexed using catch per unit effort (CPUE),which was defined as the number caught per trap net night. The total number ofwhite crappies from each net was documented and catch results from both largeand small frame trap nets were combined. We separated the white crappies intoadult and juvenile components, with the 13-cm minimum stock length(Gabelhouse 1984) as the division. Fish > 13-cm total length (TL) were consid-ered adults and those < 13 cm were categorized as juveniles. CPUE was thenseparately calculated for adult and juvenile white crappies.

Length data were collected from the white crappies and summarized usinglength-frequency histograms. The length frequencies were then quantified usingproportional stock density (PSD) and incremental relative stock density (RSD;Willis et al. 1993). PSD and RSD are defined as:

PSD = Number of fish Æ quality length x 100, andNumber of fish Æ stock length

Incremental RSD = Number of fish in a length group x 100. Number of fish Æ stock length


The minimum total lengths for white crappie stock, quality, preferred, memo-rable, and trophy length categories are 13, 20, 25, 30, and 38 cm (Gabelhouse1984). Confidence intervals (95%) were determined for the stock density indices(Gustafson 1988).

Scales were removed from white crappies below the lateral line and direct-ly behind the pectoral fin (DeVries and Frie 1996), except for the known age-0fish captured in September (i.e., crappie < 65-mm TL). To better assess whitecrappie age structure and to compare ages derived from different aging struc-tures, otoliths were also collected. Scales and otoliths were aged by two readers,discrepancies were discussed, and an age was assigned. Percent agreementbetween readers and between aging structures was calculated.

Mean back-calculated length at age for EIS white crappies was determinedusing DISBCAL software (Frie 1982). We used a standard y-intercept value forthe body length to scale radius relationship of 35 mm (Carlander 1982).Seasonal growth was calculated by determining the mean TL at the time of cap-ture for age-1 and age-2 white crappies from each sample month.

We used relative weight (Wr) to index white crappie condition. Wr is cal-culated as follows:

Wr = W/Ws * 100,where W is the weight of a fish in g and Ws is the length-specific standard weightfor a fish of that length. The Ws equation for white crappie is

log10Ws = -5.642 + 3.332(log10TL), where Ws is the standard weight in g and TL is the total length in mm (Neumannand Murphy 1991). Mean condition values were calculated by length category.

An annual mortality rate for white crappies collected in 1997 from EIS wasestimated using age-structure data. We used two methods to estimate the annu-al mortality rate: 1) the Chapman-Robson method and 2) the Ricker catch curvetechnique, which have both been suggested to provide reasonable total annualmortality rate estimates (Everhart et al. 1975).

Stomachs from the captured white crappies were removed and preserved in10% formalin. Stomach contents were quantified and reported as percent bynumber. To better identify dietary shifts based upon crappie size, we separatedthe crappies into four length categories [65- to 129-mm TL, 130- to 199-mm TL,200- to 249-mm TL, and > 250-mm TL]. The mean percent by number for eachfood item and the standard error of the mean were determined for each lengthgroup on each sample date.


Relative Abundance

Few white crappie juveniles were captured prior to the September sample.The juvenile white crappie CPUE was thus near 0 in both May and July; how-ever, the age-0 crappies recruited to our nets by September and their presencewas reflected by the sharp increase in juvenile CPUE (Table 1). The CPUE of


adult white crappie was higher in May and September than in July (Figure 1).This seasonal pattern is commonly found by biologists who sample crappie pop-ulations in lentic populations (Pope and Willis 1996) because greater numbers ofcrappie are likely captured in the spring and fall due to seasonal changes in habi-tat use. White crappies in lentic waters spawn in nearshore areas, disperse todeeper waters during the summer, and then return to littoral habitats in the fall(Guy et al. 1994). During the summer, CPUE tends to decline in the near-shorehabitat sampled with conventional trap net techniques.

Size Structure

The length-frequency histograms indicated that the spawning population ofwhite crappie in EIS contained many fish between 150- and 250-mm TL (Figure1). The number of adult crappies sampled in July and September varied asexpected, with the sample consisting of fewer large fish in July with greater num-bers present in September.

PSD values reflect expected seasonal trends typically found in lentic waters(Pope and Willis 1996), with higher values in spring and fall and lower values inmid-summer (Table 1). The May PSD of 64 indicates that this population con-tained a substantial proportion of fish longer than 20-cm TL. However, incre-mental RSD analysis indicated that, regardless of season, the crappie populationwas dominated by fish between stock and preferred lengths (i.e., 13- to 25-cmTL; Table 1). Memorable-length fish (> 300-cm TL) were only captured in May,which was likely during the spawning period.

Age Structure

We collected white crappies as old as age 6 (Figure 2). Young fish domi-nated this population, with an apparently strong year class occurring in 1995 (age2). We believe that the assigned ages are reliable. The agreement on scale-assigned ages between the two readers was 84.8%. The agreement between read-ers on otolith-assigned ages was 89.7%. The level of agreement between scaleand otolith ages was also high (92.7%). Kruse et al. (1993) reported greater than94% agreement between scales and otoliths for black crappies Pomoxis nigro-maculatus collected from two South Dakota waters.

Growth

Mean back-calculated lengths for EIS white crappie revealed comparablegrowth patterns with those found in other northern waters (Table 2). EIS meanlength at age values were slightly higher than the mean for northern watersreported by Carlander (1977). Seasonal growth trends (Figure 3) were similar tothose found for white crappies in Lake Goldsmith, South Dakota, where mostgrowth for young crappies occurred in mid- to late summer (Guy and Willis1995).


Condition

White crappie Wr values indicated that the crappies in EIS were in goodbody condition in most sampling periods, but that some seasonal variability didoccur (Table 1). The Wr of white crappies in EIS often was considerably betterthan those found in many other populations. Gabelhouse (1991) found that in aKansas reservoir with a white crappie population having desirable size structure,mean Wr rarely exceeded 95 for fish between stock and preferred lengths, andthat condition tended to increase for larger crappies. Declines in EIS white crap-pie condition values occurred in the September sample; however, no visible pat-terns in Wr among length categories could be detected.

Mortality

The two methods of estimating total annual mortality provided somewhatdifferent rates. The Chapman-Robson method indicated that the total annualmortality for age-1 and older white crappie in EIS was 51%. The catch-curvetechnique was more conservative and indicated a total annual mortality of 40%.In a turbid Oklahoma reservoir, total annual mortality for white crappies wasfound to be 54%, most of which occurred prior to attaining age 3 (Muoneke etal. 1992).

Food Habits

Small white crappies (< 200-mm TL) primarily consumed zooplankton,while larger crappies consumed fishes and corixids (Table 3). These results arequite similar to those reported for white crappies in Lake Goldsmith, SouthDakota (Guy and Willis 1993), except that larger crappies in South Dakota tend-ed to utilize more zooplankton and dipterans. In addition, white crappies in EISutilized substantially more corixids than did the South Dakota fish. Bowen(1996) suggested that fishes captured and held in trap nets may have stomachcontents that are not fully representative of their natural diets, although trap netswere used to collect crappies in both our study and the South Dakota study.

IMPLICATIONS

Both CPUE and size structure were highest in spring and fall, and lowest inmid-summer indicating seasonal movements by adult white crappies. Whitecrappie size structure in EIS during May included substantial numbers of 23-cmand longer fish. Age-structure analysis indicated that year classes were producedannually from 1991 to 1997, although recruitment was apparently variableamong years. White crappie reached 228 mm at age 3, slightly above the meanlength at age 3 reported by Carlander (1977) for northern waters. Total annualmortality estimates ranged from 40 to 51%, which are typical values for bothcrappie species across their range. Mean Wr values varied by season and length


category but peaked at 122, which is in the upper 4% for white crappies acrosstheir range. Small (<200 mm) white crappies primarily consumed zooplankton,while 200-mm and longer fish primarily consumed corixids and fishes. Giventhe density and population structure and dynamics of this white crappie popula-tion, there is potential for a sport fishery. The food habits and relative abun-dance of this population also indicate that white crappies may help shape andregulate the aquatic community in this Missouri River backwater habitat.

ACKNOWLEDGMENTS

We would like to thank Charles Pyle, the Ecological Services and FisheriesAssistance Offices of the U.S. Fish and Wildlife Service, South Dakota StateUniversity, North Dakota State Game and Fish Department, and the SouthDakota Cooperative Fish and Wildlife Research Unit for their essential contribu-tions to the completion of this study. This manuscript was approved for publi-cation by the state of South Dakota Agricultural Experiment Station as JournalSeries No. 3065.

LITERATURE CITED

Bowen, S. H. 1996. Quantitative description of the diet. Pages 513-529 in B.R. Murphy and D. W. Willis, editors. Fisheries techniques, second edition.American Fisheries Society, Bethesda, Maryland.

Carlander, K. D. 1977. Handbook of freshwater fishery biology. Iowa StateUniversity Press, Ames.

Carlander, K. D. 1982. Standard intercepts for calculating lengths from scalemeasurements for some centrarchid and percid fishes. Transactions of theAmerican Fisheries Society 111:332-336.

DeVries, D. R., and R. V. Frie. 1996. Determination of age and growth. Pages483-508 in B. R. Murphy and D. W. Willis, editors. Fisheries techniques,second edition. American Fisheries Society, Bethesda, Maryland.

Dewey, M. R., and C. A. Jennings. 1992. Habitat use by larval fishes in a back-water lake of the upper Mississippi River. Journal of Freshwater Ecology7:363-372.

Everhart, W. H., A. W. Eipper, and W. D. Youngs. 1975. Principles of fisheryscience. Cornell University Press, Ithaca, New York.

Frie, R. V. 1982. Measurement of fish scales and back-calculation of bodylengths using a digitizing pad and microcomputer. Fisheries 7(5):5-8.

Gabelhouse, D. W., Jr. 1984. A length-categorization system to assess fishstocks. North American Journal of Fisheries Management 4:273-285.

Gabelhouse, D. W., Jr. 1991. Seasonal changes in body condition of white crap-pies and relations to length and growth in Melvern Reservoir, Kansas. NorthAmerican Journal of Fisheries Management 11:50-56.


Gustafson, K. A. 1988. Approximating confidence intervals for indices of fishpopulation size structure. North American Journal of Fisheries Management8:139-141.

Guy, C. S., and D. W. Willis. 1993. Food habits of white crappies in LakeGoldsmith, South Dakota. Proceedings of the South Dakota Academy ofScience 72:51-60.

Guy, C. S., and D. W. Willis. 1995. Growth of crappies in South Dakota waters.Journal of Freshwater Ecology 10:151-161.

Guy, C. S., J. J. Jackson, and D. W. Willis. 1994. Biotelemetry of white crap-pies in a South Dakota glacial lake. Transactions of the American FisheriesSociety 123:63-70.

Hammers, B. E., and L. E. Miranda. 1991. Comparison of methods for esti-mating age, growth, and related population characteristics of white crappies.North American Journal of Fisheries Management 11:492-498.

Kruse, C. G., C. S. Guy, and D. W. Willis. 1993. Comparison of otolith andscale age characteristics for black crappies collected from South Dakotawaters. North American Journal of Fisheries Management 13:856-858.

Muoneke, M. I., C. C. Henry, and O. E. Maughan. 1992. Population structureand food habits of white crappie Pomoxis annularis Rafinesque in a turbidOklahoma reservoir. Journal of Fish Biology 41:647-654.

Neumann, R. M., and B. R. Murphy. 1991. Evaluation of the relative weight(Wr) index for assessment of white crappie and black crappie populations.North American Journal of Fisheries Management 11:243-251.

Pope, K. L., and D. W. Willis. 1996. Seasonal influences on freshwater fisheriessampling data. Reviews in Fisheries Science 4:57-73.

Willis, D. W., B. R. Murphy, and C. S. Guy. 1993. Stock density indices: devel-opment, use, and limitations. Reviews in Fisheries Science 1:203-222.

Zigler, S. J., and C. A. Jennings. 1993. Growth and mortality of larval sunfishin backwaters of the upper Mississippi River. Transactions of the AmericanFisheries Society 122:1080- 1087.


RESOURCE OVERLAP AND PARTITIONINGBETWEEN LARVAL YELLOW PERCH AND ADULT FATHEAD MINNOWS IN A PRAIRIE

GLACIAL LAKE

Shannon J. Fisher, Gene F. Galinat, and David W. WillisDepartment of Wildlife and Fisheries Sciences


ABSTRACT

Fathead minnow Pimephalas promelas population densities can becomeextremely high in shallow prairie lakes and wetlands. Adult fathead minnowsmay compete with other fishes and waterfowl for limited food resources. Inmany of the same lakes where high density fathead minnow populations some-times occur, inconsistent recruitment of yellow perch Perca flavescens is com-mon. Although adult yellow perch feed on large macroinvertebrates and benthos,larval perch may require food resources that are also utilized by fathead min-nows. We collected fathead minnows and larval yellow perch during May andJune of 1995 with an ichthyoplankton surface trawl from Pelican Lake, SouthDakota. Stomach contents were examined and dietary selection determined.Selection of food items was determined using a linear electivity index that con-trasted the proportion of each consumed taxon with the proportion available inthe environment. We also quantified the stomach contents for each species usingfrequency of occurrence and percent composition by number. Fathead minnowsand larval yellow perch both selected calanoid copepods as their primary dietitem, while secondary prey taxa were somewhat partitioned between the twospecies. Overall, the diets overlapped substantially. However, fathead minnowstended to select larger copepods and yellow perch the smaller individuals. Dietoverlap was greatest after larval yellow perch attained lengths greater than 14mm. At those lengths, given the dietary selection and resource overlap of fatheadminnows and larval yellow perch, competition could result if calanoid copepodabundance declined to densities where prey availability became a limiting factorfor growth and survival.

INTRODUCTION

Yellow perch Perca flavescens are a popular panfish in South Dakota; how-ever, populations of this species tend to be highly dynamic and sport fisheries areinconsistent. Mills et al. (1989) found greater than 50-fold differences in yellowperch year-class strengths in Oneida Lake, New York. Lott (1991) identified two


yellow perch population types in eastern South Dakota glacial lakes, one ofwhich commonly produced weak or absent year classes. Year-class strength isdetermined by the proportion of offspring that hatch and then reach a length atwhich they contribute to the adult population (e.g, maturation and spawning).During the recruitment process, there are many possible factors that influence thesurvival and growth of yellow perch larvae, including the availability of foodresources. Clady (1976) and Whiteside et al. (1985) observed cases where lim-ited food resources were a substantial factor in larval yellow perch mortalityrates. However, several researchers (e.g., Hansen and Wahl 1981; Mathias andLi 1982) have suggested that yellow perch are highly selective feeders; therefore,size structure and species composition were more important than overall zoo-plankton density.

The availability of food resources can contribute to larval fish growth andsurvival, especially when prey densities are low and intra- and interspecific com-petition exists (Welker et al. 1994). May (1974) suggested that competition dur-ing larval fish stages may be critical because fish are more susceptible to starva-tion during this period. Although adult yellow perch primarily feed on largemacroinvertebrates in eastern South Dakota lakes (Lott et al. 1996), larval yel-low perch typically consume copepod adults and nauplii (Fisher and Willis1997). Held and Peterka (1974), Price et al. (1991), and Hambright and Hall(1992) all found that fathead minnows Pimephales promelas preferred cladocer-ans, chironomid larvae, and amphipods when available; however, copepods wereconsumed and selected in the absence of other prey species. Duffy (1996) foundthat fathead minnow densities can reach levels at which they affect zooplanktondensities through predation pressure, resulting in a decline of zooplankton abun-dance. Thus, fathead minnows can alter zooplankton community structure andspecies composition, reducing resource availability to other zooplanktivores andeven ducklings (Bouffard and Hanson 1997).

Many shallow eastern South Dakota lakes have high densities of fatheadminnows. Mean fathead minnow densities in South Dakota wetlands have beenrecorded up to 47,620/ha (Carlson and Berry 1990) and fathead minnow pro-duction can exceed 17.5 kg/ha per year (Payer and Scalet 1978). High densitiesof fathead minnows were observed in Pelican Lake, South Dakota - a lake that has also exhibited inconsistent yellow perch recruitment(Fisher 1996). Thus, the objectives of this study were to assess and contrast adultfathead minnow and larval yellow perch 1) stomach contents, 2) prey selection,and 3) resource overlap and prey partitioning.

STUDY SITE AND METHODS

We completed this study on Pelican Lake, South Dakota, in May and Juneof 1995 after observing large numbers of fathead minnows in our larval yellowperch samples. Pelican Lake is a warmwater semi-permanent highly eutrophiclake (Stueven and Stewart 1996). The lake has a surface area of 1,131 ha and arelatively shallow mean depth of 1.7 m. We collected fathead minnows and lar-


val yellow perch on four dates using a 0.76-m diameter ichthyoplankton trawl(500-um bar mesh) towed slightly subsurface at 1-2 m/sec for 2-5 min. Six towswere conducted randomly on each sample date between 1100-1500 hours. Aflow meter, mounted in the mouth ofthe trawl, allowed for estimation of water volume filtered and number of fish-es/m3. We also estimated the number of fathead minnows and larval yellowperch per ha by applying the density per unit volume estimates to the estimatedlake volume (surface area x mean depth). All collected fishes were preserved in5% formalin and returned to the laboratory for analysis. Zooplankton and otherlimnetic organisms were sampled with a 2-m tube sampler (DeVries and Stein1991) at each trawl location on each sample date. Samples were filtered througha 63-um net and preserved in 4% sucrose-formalin (Haney and Hall 1973) forlater analysis. Zooplankton densities were expressed as number/L and the pro-portion of each potential prey item was determined for use in the electivity analy-sis. The proportion of calanoid copepods that were <0.80, 0.80-0.89, 0.90-0.99,1.00-1.09, and >1.09 mm for each sample date was determined from a subsam-ple of total lengths collected from 180 (10 per sample) calanoid copepods. Wecompleted an electivity analysis on copepod length separately to help determinethe fathead minnow and larval yellow perch preferred copepod size.

Stomach contents of up to 15 fathead minnow adults and 25 yellow perchlarvae per cm length group per sample date were assessed and lengths of up to10 calanoid copepods per stomach were recorded. Stomach contents were quan-tified into percent composition by number, and frequency of occurrence was alsocalculated. Prey selection by larval yellow perch and fathead minnow adults wasdetermined with a Linear Electivity Index (LEI; Strauss 1979). The degree ofelectivity was determined with the following formula:

LEI = ri - pi,where ri equals the relative abundance of prey taxon ‘i’ in the diet and pi equalsthe relative abundance of prey taxon ‘i’ in the environment. This index has arange of -1 to 1, where a value of -1 indicates complete avoidance and 1 repre-sents complete selection.

To better assess dietary partitioning and overlap, we incorporated thePercent Resource Overlap Index (PROI; Schoener 1971). The degree of overlapwas determined with the formula:

PROI = 100(1-0.5í pxi - pyi),where pxi is the mean proportion of food category ‘i’ in the diet of species x andpyi is the mean proportion of food category ‘i’ in the diet of species y. PROI val-ues range from 0 to 100, with a 0 indicating no overlap and 100 representative ofcomplete overlap between species x and y. Wallace (1981) suggested that PROIvalues >60 have the potential to be biologically significant.


Fathead minnow density estimates ranged from 0.56 to 1.23/m3 (Figure 1),which were then converted to population estimates of 9,520 to 20,910/ha. Larval


yellow perch densities were considerably lower, peaking at 0.43/m3 on 30 May(Figure 1), a density that converted to a population estimate of 7,310/ha. Peakfathead minnow density coincided with the peak yellow perch density, afterwhich abundance of both species in our trawl samples declined. As fathead min-now and larval yellow perch abundance declined over time, zooplankton densi-ties tended to increase across sample dates (Table 1).

Adult fathead minnows and larval yellow perch both had diets dominated bycalanoid copepods (Table 2). The LEI indicated that both species strongly select-ed calanoid copepods, meaning that the prey item composed a greater proportionof the diets than what was present in the environment. Fathead minnows alsoselected Bosmina sp. and Keratella spp., whereas secondary prey items for lar-val yellow perch included Diaphanosoma birgei and cyclopoid copepods (Table2). Although the Keratella sp. made up a substantial portion of the fathead min-now diets, the prey category was extremely abundant in the environment and notproportionally consumed. Both fathead minnows and yellow perch maintainednearly neutral selection for all other diet items, although there was some vari-ability by sample date.

The strong selection for the shared calanoid copepod resource was indica-tive of considerable dietary overlap between these two species. The PROIassessment indicated that biologically significant (PROI > 60) resource overlapexisted between adult fathead minnows and larval yellow perch on most sampledates. The PROI values ranged from 56 to 82 (Figure 2), indicating that if foodresources were to become limited, competition could likely result. However,secondary prey resources did not substantially overlap, allowing us to infer thatsome prey resource partitioning also occurred.

We suspected that the calanoid copepod resource may have been partitionedby copepod size. The LEI results for fathead minnow electivity of copepod sizesindicated that the species had the greatest preference for calanoid copepods>0.99 mm on all sample dates (Table 3). Larval yellow perch selected calanoidcopepods <0.90 mm on the first two sample dates; however, perch selected larg-er copepods as the perch grew (Table 3). Yellow perch larvae and adult fatheadminnows did not overlap in their copepod size selection until the last sample datein June, when both species selected calanoid copepods >0.89 mm in June, eventhough the mean copepod size available in the environment had declined to 0.87mm.

Calanoid copepods appear to be an important prey item for both fatheadminnows and yellow perch. Food resource availability during the early life his-tory stages of yellow perch is widely believed to be critical to growth and sur-vival. Our results suggest that, in Pelican Lake during 1995, substantial parti-tioning of prey taxon and size likely occurred - even though dietary overlap wasindicated by the PROI assessment. However, as the yellow perch grew, theytended to select prey items also preferred by fathead minnows. During periodsof limited food resources, interspecific competition between adult fathead min-nows and larval yellow perch could result, particularly for the calanoid copepodresource. Based on our observations, competitive interaction between these two


species would not likely occur until the late larval or juvenile growth stages,when considerable dietary overlap, declining calanoid size structure, and increas-ingly similar prey selection occur.

ACKNOWLEDGMENTS

Field assistance for this study was provided by R. Hanten, S. Rustin, and J.Ackerson. A special thank you to C. Pyle for completing the stomach contentassessments. Partial funding for this study was provided by the South DakotaDepartment of Game, Fish and Parks through Federal Aid in SportfishRestoration Project F-15-R, Study 1555. This manuscript was approved for pub-lication by the South Dakota Agricultural Experiment Station as Journal SeriesNo. 3061.

LITERATURE CITED

Bouffard, S.H., and M.A. Hanson. 1997. Fish in waterfowl marshes: waterfowlmanagers perspective. Wildl. Soc. Bull. 25:146-157.

Carlson, B.N., and C.R. Berry, Jr. 1990. Population size and economic value ofaquatic bait species in palustrine wetlands of eastern South Dakota. PrairieNat. 22:119-128.

Clady, M.D. 1976. Influence of temperature and wind on the survival of earlystages of yellow perch. J. Fish. Res. Bd. Can. 33:1887-1893.

DeVries, D.R., and R. Stein. 1991. Comparison of three zooplankton samplers:a taxon specific assessment. J. Plank. Res. 13:53-59.

Duffy, W.G. 1996. Population dynamics, production and prey consumption offathead minnows in prairie lakes and wetlands. South Dakota Departmentof Game, Fish, and Parks, Fisheries Division Completion Report 96-6,Pierre.

Fisher, S.J. 1996. Early life history of yellow perch in eastern South Dakotalakes. M.S. Thesis. South Dakota State University, Brookings.

Fisher, S.J. and D.W. Willis. 1997. Early life history of yellow perch in twoSouth Dakota glacial lakes. J. Fresh. Ecol. 12:421-429.

Hambright, K.D., and R.O. Hall. 1992. Differential zooplankton feeding behav-iors, selectivities, and community impacts of two planktivorous fishes. Env.Biol. Fishes 35:401-411.

Haney, J.F., and D.J. Hall. 1973. Sugar-coated Daphnia: a preservation tech-nique for Cladocera. Limn. and Ocean. 18:331-333.

Hansen, M.J., and D.H. Wahl. 1981. Selection of small Daphnia pulex by yel-low perch fry in Oneida Lake, New York. Trans. Amer. Fish. Soc. 110:64-71.

Held, J.W., and J.J. Peterka. 1974. Age, growth, and food habits of the fatheadminnow in North Dakota saline lakes. Trans. Amer. Fish. Soc. 103:743-756.

Lott, J.P. 1991. Food habits of yellow perch in eastern South Dakota glaciallakes. M.S. Thesis. South Dakota State University, Brookings.


Lott, J.P., D.W. Willis, and D.O. Lucchesi. 1996. Relationship of food habits toyellow perch growth and population structure in South Dakota lakes. J.Fresh. Ecol. 11:27-37.

Mathias, J.A., and S. Li. 1982. Feeding habits of walleye larvae and juveniles:comparative laboratory and field studies. Trans. Amer. Fish. Soc. 111:722-735.

May, R.C. 1974. Larval mortality in marine fishes and the critical period con-cept. Pages 3-19 in J.H.S. Blaxter, editor. The early life history of fish.Springer-Verlag, New York.

Mills, E.L., R. Sherman, and D.S. Dobson. 1989. Effect of zooplankton abun-dance and body size on growth of age-0 yellow perch in Oneida Lake, NewYork, 1975-1986. Can. J. Fish. Aquat. Res. 46:880-886.

Payer, R.D., and C.G. Scalet. 1978. Population and production estimates of fat-head minnows in a South Dakota prairie wetland. Prog. Fish-Cult. 40:63-66.

Price, C.J., W.M. Tonn, and C.A. Paszkowski. 1991. Intraspecific patterns ofresource use by fathead minnows in a small boreal lake. Can. J. Zool.69:2109-2115.

Schoener, T.W. 1971. Theory of feeding strategies. Annual Review of Ecologyand Systematics 2:369-404.

Strauss, R.E. 1979. Reliability estimates for Ivlev’s electivity index, the forageratio, and a proposed linear index of food selection. Trans. Amer. Fish. Soc.108:344-352.

Stueven, E., and W.C. Stewart. 1996. 1995 South Dakota lake assessments.South Dakota Department of Environment and Natural Resources, FinalReport, Pierre.

Wallace, R.K. 1981. An assessment of diet overlap indexes. Trans. Amer. Fish.Soc. 110:72-76.

Welker, M.T., C.L. Pierce, and D.H. Wahl. 1994. Growth and survival of larvalfishes: roles of competition and zooplankton abundance. Trans. Amer. Fish.Soc. 123:703-717.

Whiteside, M.C., C.M. Swindoll, and W.L. Doolittle. 1985. Factors effectingthe early life history of yellow perch. Environ. Biol. Fishes 12:47-56.


QUATERNARY AMMONIUM RESEARCH

Dianna WineingerAugustana College, Sioux Falls, SD 57197

ABSTRACT

Reactions for the formation of quaternary ammonium compounds from ter-tiary arnines using dimethyl carbonate, DMC, as the alkalating agent were stud-ied. During this study it was discovered that little data was available on themethyl carbonate anion; therefore, reactions were run using sodium methoxideand dimethyl carbonate in order to isolate the methyl carbonate anion. The prod-ucts of these reactions were studied spectrally using FT-IR spectroscopy andmass spectrometry. Once identified, the methyl carbonate anion was observed toconvert to the hydrogen carbonate anion in the presence of water.

The anion of methyl carbonate quaternaries can be replaced with the anionsof acids. A number of different reactions involving this conversion of the methylcarbonate anion were conducted using various acids.


QUATERNARY AMMONIUM SYNTHESIS

Michael HansonAugustana College

Sioux Falls, SD 57197

ABSTRACT

Industrially and domestically, the importance of quaternary ammoniumcompounds can not be overestimated. Their uses range from hair care productsto additives in asphalt.

The synthesis of quaternary ammonium compounds presently entails usingmethyl chloride or dimethyl sulfate as the methylating agent in conjunction witha secondary or tertiary amine. Though efficient as methylating agents, methylchloride and dimethyl sulfate are both highly toxic and pose a significant healthrisk. In addition the quaternary ammonium compound formed in this way arevery slowly biodegradable. An alternative methylating agent is clearly needed.

An alternative has been found in the compound dimethyl carbonate. Theadvantages of using dimethyl carbonate are, its low toxicity and its biodegrad-ability. Dimethyl carbonate’s ability to methylate tertiary amines is poor how-ever, and fairly extreme conditions are needed to cause alkylation. Theseextreme conditions unfortunately place this reaction beyond the reach of indus-trial reactors. Less extreme conditions must be found, while at the same timekeeping the reaction time to a minimum.

Using quantitative nucleur magnetic resonance spectroscopy the half-life ofa reaction could be determined. From this rudimentary kinetic information thebest conditions for running the reaction could be determined by manipulatingparameters such as: pressure in the reaction vessel, volume, ratios of reactantsand solvent, and temperature. These parameters were manipulated not only togive the best rate of reaction but also to keep the pressure and temperature with-in reach of industrial capabilities.


STABILITIES OF NON-G•U MISMATCHES ININTERNAL LOOPS OF RNA HAIRPINS

Peter deLannoy, LaHoma Easterwood, and Stephanie Price Department of Chemistry

Black Hills State UniversitySpearfish, SD 57783

ABSTRACT

Four RNA hairpins containing a symmetrical internal loop consisting of twonon-G•U mismatches were synthesized by T7 transcription and their thermal sta-bilities were determined. The model hairpin (U61) containing an A•C mismatchwas derived from the naturally occurring Caenorhabditis elegans U6 spliceoso-mal snRNA. Three additional stem loops were designed including one withoutthe internal loop (U62), one containing an A•G mismatch (U63), and one con-taining an U•U mismatch (U64). The main findings are: (i) the Tms varied withionic strength; (ii) the wild-type (U61) internal loop destabilizes the hairpin by4 kcal/mole; and (iii) the A•G and U•U mismatches have a stabilizing effect rel-ative to the wild-type A•C mismatch. These data should be useful in predictingthe stability of other hairpins and will serve to enhance and improve the para-meters currently understood for secondary structure prediction of RNA.

INTRODUCTION

RNA is required for a variety of biological functions. Several RNAs arerequired for protein translation including the rRNAs, tRNAs, and mRNAs(Trachsel, 1990). Recently the Escherichia coli 23S rRNA was implicated in thepeptidyl transferase activity during translation (Noller et al. 1992). Formation ofthe spliceosome and removal of the intron from pre-mRNAs requires a combi-nation of five different snRNAs and splicing factors (Nilson, 1994). Numerouscatalytic RNAs have now been characterized that function on a variety of differ-ent substrates (Gestland and Atkins, 1993). The functions of these various RNAsdepend on their abilities to form three-dimensional structures (Chastain andTinoco, 1991). An RNA’s three-dimensional structure is composed of a primarysequence of nucleotides which interact to form secondary structural elementswhich, in turn, interact to form a tertiary structure. The tertiary interactions thatpromote formation of the three-dimensional structure are generally assumed tobe weaker than the sum of interactions that form the secondary structures (Turneret al. 1988). Thus, the total free energy of formation for a three-dimensionalRNA can be approximated by the sum of free energies of formation for its sec-ondary structures.


Secondary structure is formed by the matching of palindromic sequenceswithin the primary sequence, usually purine—pyrimidine base pairs, thoughother, non-Watson-Crick base pairs have been noted (Saenger, 1984; Morse andDraper, 1995). Secondary structural elements can include double helices, exter-nal loops, internal loops, and bulges (Chastain and Tinoco, 1991; Tinoco et al.1987).

There are several approaches to predicting RNA secondary structure, includ-ing phylogenetic studies, structure mapping, and thermodynamic stability(Turner et al. 1988; Serra et al. 1994). The best approach for defining RNA sec-ondary structure is by using a combination of all three methodologies. The mainlimitation of the thermodynamic approach is the lack of data for secondary ele-ments other than helices and small external loop sequences (Serra et al. 1994).

External loop stability is dependent on the size of the loop but may not bedependent on its composition (Groebe and Uhlenbeck, 1988). The stability of anexternal loop is also dependent on the closing base pair of the loop and the firstmismatch in the loop (Serra et al. 1994; Serra et al. 1993). Bulge nucleotideshave also been found to affect hairpin stability (Groebe and Uhlenbech, 1989).

In contrast to external loops in RNA secondary structure, much less is cur-rently known about internal loops and their contribution to overall thermody-namic stability of secondary structure motifs. Most notably, Santa Lucia et al.(1991) have shown that an internal loop stabilizes the duplex when the mis-matches are G•A, U•U, or C•C+, and destabilizes the duplex when the mis-matches are G•G, C•A, C•U, A•A, or C•C. It is interesting to speculate whetherthese trends exist in the internal loops of naturally occurring RNAs. In order toapproach this problem we have designed a model hairpin derived from theCaenorhabditis elegans U6 spliceosomal RNA containing an internal loop withtwo consecutive mismatches (Thomas et al. 1990; Figure 1a). In this paper wecharacterized the thermodynamic stability of four hairpins: U61 containing thewild-type internal loop with no modifications (Fig. 1b), U62 lacking the internalloop (Fig. 1c), as well as U63 containing a A•G mismatch and U64 containing aU•U mismatch (Figs. 1d and 1e, respectively).

MATERIALS AND METHODS

Oligonucleotide Synthesis. RNA hairpins were synthesized by the in vitrotranscription of single-stranded DNA templates by T7 RNA polymerase(Milligan et al. 1987; Ambion, Inc.). The resulting oligomers were purified bydenaturing gel electrophoresis in 20% polyacrylamide -7 M urea using standardprocedures (Maniatis, 1982) and quantified by UV-vis spectroscopy usingextinction coefficients described previously (Puglisi and Tinoco, 1989). Nativegel electrophoresis of stem loops was by conditions previously described(Jacques and Susskind, 1991).

Melting Curves. The conditions used for the melting studies were by Puglisiand Tinoco (1989) with the following modifications. The buffer used for ther-modynamic studies was 10 mM sodium cacodylate pH 7 with NaCl as specified


below. In experiments where RNA hairpin concentration was varied, NaCl wasmaintained at 1M. Alternatively, the salt dependency of the hairpins was deter-mined by varying the concentration of NaCl between 0.01M NaCl-1M NaCl.For these experiments RNA concentration was maintained at 3µM. For experi-ments comparing U61, U62, U63, and U64, the RNA concentration was main-tained at 3µM with a salt concentration of 0.01M.

Samples were mixed as described previously (Puglisi and Tinoco, 1989) andplaced in a quartz cuvette (0.5 cm path length) for spectral measurements. Priorto each study the sample was heated to high temperature and allowed to subse-quently reach equilibrium at the appropriate starting temperature. Absorbancevs. temperature melting curves were measured at 260nm with a heating rate of0.2˚C per minute on a Hewlett-Packard 8452-A spectrophotometer interfacedwith a Hewlett-Packard Peltier temperature controller. Absorbances were takenover a particular temperature range with data acquisition every 0.2˚C. After eachrun, samples were allowed to cool back to the starting temperature, the A260 wasrecorded, and if the value differed from the starting A260 by more than 1%, thedata were not used. A minimum of four runs was obtained for each desired con-centration of NaCl or RNA hairpin tested.

Data Analysis. Upon completion of each experiment, the results were veri-fied graphically on the UV-vis spectrophotometer. Absorbance melting curveswere fit to a two-state model with sloping baselines as described previously(Puglisi and Tinoco, 1989; Petersheim and Turner, 1983). For the melting tran-sitions of the hairpins, the program was adapted for a unimolecular transition.The melting temperature of the hairpins and resulting thermodynamic parameterswere concentration independent and obtained from an average of the fits of indi-vidual melting curves.

The thermodynamic parameters for each hairpin were determined throughthe use of the van’t Hoff relation and standard thermodynamic equationsdescribed previously (Puglisi and Tinoco, 1989). Once the thermodynamicparameters were obtained, the standard deviations for the experiments weredetermined. The data were accepted only if the standard deviations were withinpublished experimental error values (Santa Lucia et al. 1991).

RESULTS

The model RNA hairpin was selected from the U6 spliceosomal RNA of thenematode Caenorhabditis elegans. (Figure 1a). This model was chosen becauseof its natural occurrence, its known secondary structure, and its inclusion of asymmetrical internal loop with non-G•U mismatches. In order to synthesize thehairpins used in these studies, we designed unique single-stranded DNA tem-plates and used T7 RNA polymerase (Milligan et al. 1987). The templates con-tained two permutations including the addition of an A•U base pair at the base ofthe RNA stem and the addition of three G residues creating a 5’ overhang. Theaddition of the A•U base pair was to provide additional thermodynamic stabilityto the terminal portion of the RNA stem thus enhancing the probability of a


smooth melting transition (Turner et al. 1988). Although the additional Gresidues provided no benefit with regard to the analysis of the hairpin, single-stranded DNA templates containing C residues in the +1 and +2 positionsincrease the overall yields of transcription (Milligan et al. 1987). Additionally,T7 RNA polymerase adds one non-templated G during transcription to the 5’-endof the in vitro RNA. The resulting U61 hairpin containing the additional base-pair and the 5’ overhang is shown in Figure 1b.

As a first step in characterizing the thermodynamic stability of U61 RNA,melting experiments were performed over a ten-fold range in hairpin concentra-tion under conditions of constant ionic strength. This experiment was necessaryin order to demonstrate that the model hairpin was preferentially forming the pre-dicted hairpin structure versus a bimolecular dimer. The Tm for the unimolecularmelting transition would be independent of hairpin concentration whereas thethermal melting for the bimolecular transition is concentration dependent and theTm will increase with increasing concentration of single strands. As shown inTable 1, the Tm was constant within experimental error over a ten-fold range inconcentration of U61 RNA. The Tm data in Table 1 may vary + 0.09˚C and the∆G˚ may vary + 0.01kcal/mol at 37˚C. The demonstration of the independenceof the Tm on the RNA concentration shows that the melting transition for the U61model RNA hairpin was unimolecular. Furthermore, native gel electrophoresisconfirmed that U61 and the stem loops mentioned below were hairpins and notbimolecular duplexes (Jacques and Susskind, 1991; data not shown).

In order to determine the contribution of the internal loop to the thermody-namic stability of U61, three additional hairpins were synthesized by T7 tran-scription. The U62 hairpin lacks the internal loop (1c), U63 contains an A•Gmismatch (1d), and U64 contains an U•U mismatch (1e). Initial experiments todetermine the relevant thermodynamic parameters for U62 at 1M NaCl wereunsuccessful since it melts at 88˚C (data not shown). However, using the rulesdescribed by Serra and Turner (1995) and mfold (Walter et al. 1994), the esti-mated theoretical Tm for U62 was 88˚C. Thus, in order to compare the thermo-dynamic stability of the four hairpins experimentally, melting studies were per-formed at 0.01M NaCl as shown in Table 2. In all the RNAs tested, the meltingtransitions exhibited an upper melting temperature that was concentration inde-pendent, indicating that the hairpins were behaving as monomeric species (datanot shown). However, a low temperature transition was also observed and thedata presented here and below was taken using base lines defined at the top ofthe transition. A low temperature transition was recently reported by another lab-oratory analyzing the melting behavior of the U1 snRNA stem/loop II sequence(Hall, 1994). In that case, the lower transition was sensitive to the sequence ofthe ten nucleotide external loop and the resulting thermodynamic parameters ofthe stem loop were determined from the upper melting transition. Moreover,non-Watson-Crick base pairs probably form to shorten large external loopswhich may explain the low temperature transition (reviewed in Wyatt andTinoco, 1993). Because the melt associated with the stem was independent ofthe low temperature transition it was not considered further in this manuscript.

Our data show that the internal loop containing the A•C mismatch destabi-


lizes U61 by a ∆∆G˚ of 4.17kcal/mol at 37˚C, and the Tm is decreased by approx-imately 17.2˚C relative to U62. Moreover, the internal loop containing the A•Gmismatch destabilized U63 by a ∆∆G˚ of 3.7kcal/mol at 37˚C, and the Tm wasdecreased by approximately 13.9˚C. Concurrently, the internal loop containingthe U•U mismatch destabilized U64 by a ∆∆G˚ of 4.1kcal/mol at 37˚C, and theTm in this case was decreased by approximately 14.4˚C. Both hairpins (U63 andU64) were more stable than U61 which was consistent with the data previouslyreported for consecutive and tandem mismatches in duplexes where A•G andU•U mismatches were more stable than A•C mismatches (Santa Lucia et al.1991; Wu et al. 1995).

In order to further characterize U61 and its biophysical behavior under avariety of salt conditions, melting transitions were performed over a hundred-fold range in NaCl concentration using all four hairpins. The resulting Tm dataare shown in Table 3. The Tm data may vary + 0.9˚C and the largest standarddeviation was + 16.4cal/mol for ∆S˚, + 5.3kcal/mol for ∆H˚, and + 0.6kcal/molfor ∆G˚ at 37˚C. The data show that the Tm increases about 20˚C over the hun-dred-fold increase in salt concentration with an approximate increase of 9.45˚Cover each ten-fold increment demonstrating a linear dependence of the Tm on theincrease in ionic strength. These data were consistent with the results shown byothers using similar systems (Record, 1967; Puglisi and Tinoco, 1989).

DISCUSSION

We have designed a model system derived from the naturally occurring U6spliceosomal RNA from Caenorhabditis elegans. The U61 RNA hairpin consistsof a symmetrical internal loop containing non-G•U base pairs and an externalloop of 11 nucleotides. Melting studies were performed to characterize themodel RNA hairpin and three other stem loops under a variety of conditions.Our data shows that the RNA hairpins undergoe unimolecular melting transitionsover a ten-fold range in RNA concentration. The folded state of these moleculeswere further analyzed by native gel electrophoresis. Our results confirmed themelting studies suggesting that the RNA was forming unimolecular stem loopsand not bimolecular duplexes. We have also determined the melting profiles andthermodynamic parameters for all four hairpins. Our data demonstrated that theU61 (A•C mismatch) wild-type internal loop destabilizes the stem loop byapproximately 4.17kcal/mol at 37˚C. By comparison, the U63 (A•G mismatch)and U64 (U•U mismatch) stabilize the stem loop relative to the wild-type A•Cmismatch. Moreover, the A•G mismatch stabilized the hairpin to a greaterdegree than the U•U mismatch. These results and the trends seen in terms of therelative stability of a particular mismatch were consistent with the data publishedfor tandem mismatches in RNA duplexes (Santa Lucia et al. 1991; Wu et al.1995). In those studies tandem mismatch stability was ranked whereA•G>U•U>A•C for small RNA duplexes. Thus, these results further extend thepreviously published data that showed the stability of tandem mismatches


depends on the mismatch sequence in small RNA duplexes.We have also characterized the stem loops mentioned above for their bio-

physical behavior over a 100-fold range in salt concentration. As shown in Table3, our data indicated that for each ten-fold increase in salt concentration the Tm

increased by approximately 9.45˚C. Thus, the Tm for each of the stem loops test-ed was a linear function of ionic strength and was in agreement with results pre-viously reported for polynucleotide double helices (Record, 1967) and RNA hair-pins (Groebe and Uhlenbeck, 1989; Hellendorn et al. 1996). Using these resultswe more accurately estimated the Tm for U62 at 1M NaCl. Each of the four hair-pins tested demonstrated a strong correlation between salt concentration and theeffect on the subsequent Tm. Thus, using the average ∆Tm = 9.45˚C per 10-foldincrease in salt concentration, we determined the Tm for U62 to be 84˚C. Asmight be expected this value was within 4˚C of the predicted value. Using thesame logic we also calculated the ∆G˚ to be approximately -10.6 kcal/mol at37˚C which is within 1.7 kcal/mole of the theoretical value determined by mfoldand was similar in magnitude to data previously determined for a hairpin with asimilar sequence and composition (Groebe and Uhlenbeck, 1989).

Ultimately the goal of this laboratory was to determine the thermodynamicstability of the internal loop as the function of the mismatch and adjacent basepairs. Therefore, more mismatches will be analyzed including G•A and C•C+mismatches which should also stabilize the hairpin (Santa Lucia et al. 1991; Wuet al. 1995). Mismatches that have been previously shown to destabilize duplex-es will also be tested including G•G, C•A, A•A, C•U, U•C, and C•C. The result-ing data should lead to a better understanding of the parameters used for the pre-diction of RNA secondary structure and ultimately provide a clearer understand-ing of how RNAs fold into their functional three-dimensional form.

ACKNOWLEDGMENTS

We would like to thank Greg Parker for his technical assistance in providingthe native electrophoresis of the stem loops. This work was funded in part by theCenter for Innovation and Economic Development and a grant from the BHSUCommittee for Faculty Research.

REFERENCES

Chastain, M. and Tinoco, I., Jr. (1991). Structural elements in RNA. Prog. Nucl.Acid Res. Mol. Biol. 41, 131-177.

Ehresmann, C., Baudin, F., Mougel, M., Romby, P., Ebel, J.P., and Ehresmann,B. (1987). Probing the structure of RNAs in solution. Nucleic AcidsResearch 15, 9109-9129.

Gesteland, R. F. and Atkins, J. F. (1993). The RNA World (Cold Spring Harbor,


New York: Cold Spring Harbor Laboratory Press).

Groebe, D.R. and Uhlenbeck, O.C. (1988). Thermal stability of RNA hairpinscontaining a four-membered loop and a bulge nucleotide. Biochemistry 28,742-747.

Groebe, D.R. and Uhlenbeck, O.C. (1989). Characterization of RNA hairpin loopstability. Nucleic Acids Research 16, 11725-11735.

Hellendorn, K., Michiels, P.J.A., Buitenhuis, R., & Pleij, C.W.A. (1996).Protonatable hairpins are conserved in the 5’-untranslated region oftymovirus RNAs. Nucleic Acids Research 24, 4910-4917.

Jacques, J-P. and Susskind, M.M. (1991). Use of electrophoretic mobility todetermine the secondary structure of a small antisense RNA. Nucleic AcidsResearch 19, 2971-2977.

Maniatis, T., Fritsch, E. F., Sambrook, J. (1982). Molecular Cloning - ALaboratory Manual (Cold Spring Harbor, New York: Cold Spring HarborLaboratory Press).

Milligan, J.F., Groebe, D.R., Witherell, G.W. and Uhlenbeck, O.C. (1987).Oligoribonucleotide synthesis using T7 RNA polymerase and syntheticDNA templates. Nucleic Acids Research 15, 8783.

Morse, S.E. and Draper, D.E. (1995). Purine-purine mismatches in RNA helices:evidence for protonated G-A pairs and next-nearest neighbor effects.Nucleic Acids Research 23, 302-306.

Nilson, T. W. (1994). RNA-RNA interactions in the spliceosome: unraveling theties that bind. Cell 78, 1-4.

Noller, H. F., Hoffarth, V. and Zimniak, L. (1992). Unusual resistance of pep-tidyl transferase to protein extraction procedures. Science 256, 1416-1419.

Petersheim, M. and Turner, D.H. (1983). Biochemistry 22, 256-263.

Puglisi, J.D. and Tinoco, I., Jr. (1989). Absorbance melting curves of RNA.Methods in Enzymology 180, 304-325.

Saenger, W. (1984). Principles of Nucleic Acid Structure. Springer-Verlag, NewYork.

Santa Lucia, Jr., J., Kierzek, R. and Turner, D.H. (1991). Stabilities of consecu-


tive A-C, C-C, G-G, U-C, and U-U mismatches in RNA internal loops: evi-dence for stable hydrogen-bonded U-U and C-C+ pairs. Biochemistry 30,8242-8251.

Serra, M.J. and Turner, D.H. (1995). Predicting thermodynamic properties ofRNA. Methods in Enzymology 259, 242-261.

Serra, M. J., Axenson, T. J. and Turner, D. H. (1994). A model for the stabilitiesof RNA hairpins based on a study of the sequence dependence of stabilityfor hairpins of six nucleotides. Biochemistry 33, 14289-14296.

Serra, M. J., Lyttle, M. H., Axenson, T. J., Schadt, C. A. and Turner, D. H. (1993).Nucleic Acids Research 21, 3845-3849.

Record, Jr., M.T. (1967). Electrostatic effects on polynucleotide transitions. I.Behavior at neutral pH. Biopolymers 5, 975-992.

Thomas, J., Lea, K., Zucker-Aprison, E. and Blumenthal, T. (1990). The spliceo-somal snRNAs of Caenorhabditis elegans. Nucleic Acids Research 18,2633-2642.

Tinoco, I., Jr., Davis, P.W., Hardin, C.C., Puglisi, J.D., Walker, G.T. and Wyatt,J. (1987). RNA structure from A to Z. Cold Spring Harbor Symposia onQuantitative Biology LII, 135-146.

Trachsel, H. (1991). Translation in Eukaryotes (Boston, Massachusetts, CRCPress, Inc.)

Turner, D.H., Sugimoto, N. and Freier, S.M. (1988). RNA structure prediction.Ann. Rev. Biophys. Biophys. Chem. 17, 167-192.

Walter, A.E., Turner, D.H., Kim, J., Lyttle, M.H., Müller, P., Mathews, D.H.,Zuker, M. (1994). Coaxial stacking of helices enhances binding of oligori-bonucleotides and improves predictions of RNA folding. Proc. Natl. Acad.Sci. U.S.A. 91, 9218-9222.

Wu, M., McDowell, J.A. and Turner, D.H. (1995). A periodic Table of symmet-ric tandem mismatches in RNA. Biochemistry 34, 3204-3211.


EFFECT OF LIGAND DEUTERATION ON THE.EU3+ (5D0) LIFETIME IN

TRIS (2,2,6,6-TETRAMETHYL-3-5-HEPTANEDIONATO)EUROPIUNI(III)

Todd C. Schwendemann, Paul S. May, and Mary T. BerryDepartment of Chemistry

University of South Dakota, Vermillion, SD 57069

The rate of decay of the 5D0 excited electronic state of Eu3+ in the complexEu(2,2,6,6-tetramethyl-3,5-heptanedionato)3 has been studied using timeresolved luminescence. The contributions to the rate from radiative and variousnon-radiative mechanisms have been accessed separately. Multiphonon (non-radiative) contributions to the rate, which can be attributed to energy transferfrom the metal to C-H stretching vibrations, have been determined by comparingdecay rates in the perdeutero complex in which the hydrogens of the ligand havebeen replaced by deuterium. The presence of a low-lying ligand-to-metalcharge-transfer state is invoked to explain both the strong temperature depen-dence of the rate and the unusually high radiative rate of decay.


PRODUCT ANALYSIS FOR THE PHOTOCHEMICAL ADDITION OF ETHANOL

TO MALEIC ACID AND ITS ESTERS

M. Robert Stoner, Thomas Tran, and Roy C. PrebbleDepartment of Chemistry

University of South DakotaVermillion, SD 57069

ABSTRACT

Previous work showed that the irradiation of maleic acid and benzophenonein ethanol as solvent yields tetrahydro-2-methyl-5-oxo-3-furancarboxylic acid(R= H), but no information was given about the stereochemical nature of theproduct. We have

studied the stereochemical distribution of the products produced in the photo-chemical reaction. We have extended the study to the reactions of dimethyl (R= CH3) and diethyl maleate (R = CH2CH3) as well. Irradiations were carried outin borosilicate glass containers using mercury vapor lamps as the light source.The products were separated by the use of flash chromatography. Identificationof the lactone products was accomplished by the use of gaschromatography/mass spectrometry and IR and 1H-NMR spectroscopy. Thecompositions of the photochemical reaction mixtures were determined by gaschromatography. We have found that both the cis and trans isomers of the lac-tone acid and esters are formed, but that the trans-isomer predominates in allcases.

The photochemical reaction occurs by a free radical addition mechanism inwhich the triplet state of the benzophenone functions as the radical initiator. Tlestereochemistry of the product is apparently determined at the time that thehydroxyethyl radical adds to the carbon-carbon double bond of the acid or ester.The preference for the trans-isomer in the product allows us to speculate aboutthe preferred orientation of the hydroxyethyl radical as it approaches the doubly-bonded carbon of the maleic acid or maleate ester.


ISOMERIZATION OF TRIMETHYL PHOSPHITE

Drew J. Paulson and Arlen VisteDepartment of Chemistry, Augustana College


ABSTRACT

The purpose of this research was to investigate the interaction of trimethylphosphite, (CH3O)3P, with sulfur trioxide (SO3) and with chlorosulfonic acid(ClSO3H), using gas chromatography/mass spectrometry as the principal analyt-ical tool to examine the distribution of product mixtures in methanol.

Particular attention was given to the possible formation of sulfur-phospho-rus bonds, but none were observed by this analytical method. However it wasfound that both sulfur trioxide and chlorosulfonic acid lead to isomerization oftrimethyl phosphite to methyl methylphosphonate.

(CH3O)3P = CH3P(O)(OCH3)2

Dimethyl sulfate, (CH3O)2SO2, was also observed in the GC/MS. This ratherhazardous material is a byproduct in methanol solutions, and may be a reactiveintermediate as well.Some conversion of trimethyl phosphite to dimethyl phosphonate was alsoobserved.

(CH3O)3P + H2O = HP(O)(OCH3)2 + CH3OHThe water may arise indirectly from methanol.In addition, some trimethyl phosphate, OP(OCH3)3, was formed by oxidation ofthe trimethyl phosphite.

Similar observations were made with triethyl phosphite. In this case ethanolsolutions were used in the GC/MS analysis, and some diethyl sulfate,(C2H5O)2SO2, was also observed as a byproduct or reaction intermediate.

(C2H5O)3P = C2H5P(O)(OC2H5)2

(C2H5O)3P + H2O = HP(O)(OC2H5)2 + C2H5OH

ACKNOWLEDGMENTS

This research was carried out as part of the Summer Science Institute, dur-ing Summer 1997, at Augustana College.


FTIR SPECTRA OF POLYMERS INREFLECTANCE AND TRANSMISSION

John D. Gilbertson and Arlen VisteDepartment of Chemistry, Augustana College


ABSTRACT

The purpose of this research was to investigate the FTIR (Fourier TransformInfrared) spectra of polymers, using both transmittance and reflectance methods,and to augment the available spectral library.

Methods used included Specular Reflectance, Attenuated Total Reflectance(ATR), and transmission. The FTIR spectrometer was IBM Instruments IR/32,now somewhat elderly but still working dependably. The Specular Reflectanceaccessory was Buck Scientific Model 56. The ATR was IBM InstrumentsVariable Angle ATR, with KRS-5 (thallium bromide/thallium iodide) crystal.The most common sample form was the thin polymer film, such as polyethylene.Beads of several polymers were hot pressed into thin films. Samples represent-ing the standard recycling codes 1-6 were included. Specular reflectanceobserved from sufficiently thin polymer films sometimes included some contri-bution from transmission through the film, reflection from the mirror in back,and transmission through the film a second time, in addition to true reflectance.Specular reflectance with shiny commercial black bakelite (phenol-formalde-hyde resin) indicated that most of the infrared radiation was absorbed. Thisprobably represented electronic excitation through small or zero electronic bandgaps. ATR required careful adjustment of mirror angles, and good contact withthe polymer sample, but with care was successful. One other reflectance method,Cylindrical Internal Reflectance (Circle cell), using a ZnSe rod, can be appliedto solutions, including aqueous solutions which are difficult to handle by othermethods.

Library search was carried out successfully, using both transmission andreflectance samples.

ACKNOWLEDGMENTS

Most of this research was carried out as part of the Summer ScienceInstitute, during Summer 1997, at Augustana College.


FOURIER IMAGE ANALYSIS

John P. Berdahl and Karel Vander Lugt, Augustana College, Sioux Falls, SD 57197

ABSTRACT

Fourier image analysis uses the mathematical properties of an image tomanipulate the image in a desired manner. The procedure involves having atwo-dimensional image and taking the Fourier Transform of it. The image isnow in the Fourier transform plane. While the image is in the Fourier trans-form plane it is easy to manipulate it with different computer techniques, main-ly filtering processes. Then the inverse Fourier transform is done and a similar,but enhanced, image is created.


AN INEXPENSIVE DEVICE FORMEASUREMENT AND PRESENTATION OF

ATOMIC SPECTRA

James L. LeffertsAssociate Professor of Chemistry and Physics

Dakota Wesleyan UniversityMitchell, SD 57301

ABSTRACT

An inexpensive device for viewing of atomic spectra can be made using aholographic grating together with a video camera and monitor. This system canbe adapted for both classroom presentation and laboratory experiments. In theGeneral Chemistry laboratory this has been used to measure wavelengths of thelines in the visible spectra of gaseous elements normally present in the atmos-phere. The video monitor was used to display the spectrum. Optimal magnifi-cation of the spectrum was obtained by placing the grating 90 centimeters fromthe source lamp. The camera was located 75 centimeters from the grating.Calibration of a wavelength scale is accomplished by measuring the position ofthe lines in the mercury spectrum, and plotting their wavelengths versus position.Student’s values for wavelengths of the hydrogen spectrum are obtained from thecalibration line. Errors (%) ranged from 0.2% to 1.9% over two lab sectionstotaling sixteen students. Only one calibration line was established for each labsection. Least squares analysis of student data gives results no better than thoseobtained by free-hand graphing.


ELECTRONIC STATES IN SEMIMAGNETICQUANTUM WELLS

James Niggemann and A. G. PetukhovPhysics Department, South Dakota School of Mines and Technology,

Rapid City, SD 57701-3995

ABSTRACT

There are several experiments that reveal very interesting properties of semi-magnetic, semimetal or semiconductor quantum wells, in particular, ErAs andGaMnAs [1-2]. The results included the manifestation of spin-dependent reso-nant tunneling through hole states, along with the dependence of transport prop-erties on the relative orientation of the magnetic field to the interface. The resultsof first-principle calculations (obtained from the linear muffin-tin orbitalmethod) are available for the bulk electronic structure of these materials [3-4].However, to investigate the properties of quantum wells we need a more intuitivedescription. The description of the hole states in terms of the Kohn-LuttingerHamiltonian, with the exchange field taken into account, is an excellent way todo it.

In ErAs, the exchange field comes from the magnetization of 4f core elec-trons and the strong exchange coupling of valence electrons with the 4f spinmagnetic moments. For the holes, there is both exchange splitting at the G point(center of the Brillouin zone) and differences in the effective mass for spin “up”and spin “down” in the absence of spinorbit coupling.The Kohn-Luttinger Hamiltonian, including the contribution from the exchangefield and spin-orbit coupling, was set up and applied to the quantum wells for dif-ferent orientations of the magnetic field and interface. The valence band split-ting was most pronounced for magnetic fields perpendicular to the interface,while for in-plane magnetic fields it was close to zero.This striking anisotropy of exchange splitting may have serious implications onthe electronic and optical properties of a novel class of semiconductor structuressuch as semimagnetic quantum wells and spin-dependent resonant tunnelingdiodes.

REFERENCES

[1] D.E. Brehmer, Kai Zhang, Ch. J. Schwarz, S.-P. Chau, S.J. Allen, J.P.Ibbetson, J.P. Zhang, A.G. Petukhov, C.J. Palmstrom, and B. Wilkens, Solid-State Electronics 40, 241 (1996)

[2] H. Ohno et al., to be published in Appl. Phys. Lett. (1998)


[3] A.G. Petukhov, W.R.L. Lambrecht, and B. Segall, Phys. Rev. B 53, 3646(1996)

[4] W.R.L. Lambrecht, B. Segall, Ria Bogaerts, and A.G. Petukhov, Phys. Rev.B 55, 9239 (1997)


TEMPERATURE CONTROLLER FOR AMICROCALORIMETER CONTROL SYSTEM

L. W. Watson and T. AshworthDepartment of Physics, South Dakota School of Mines and Technology,

Rapid City SD 57701

The purpose of the project was to design a computer control system for aSetaram BT 215 low temperature, microcalorimeter. A program was written inthe C language which controlled the temperature of the microcalorimeter and theduration of the experiment through real time measurement of voltage and resis-tance. Interface between the software and hardware was achieved with an IEEE488 interface circuit card installed in the computer. The hardware included thecalorimeter and various items of test equipment. An integral part of the systemhardware was a temperature controller for the microcalorimeter. The tempera-ture controller featured an Omega CN6082 ramp and soak temperature controller.It was powered by stepped-down, fused, 120 VAC current and had an on-off tog-gle switch. The ramp and soak cycles were determined by the experimenter whocould program any desired temperature and time sequences. The software con-verted resistance measurements from the microcalorimeter to temperature read-ings. When the appropriate temperature was achieved, and the required time hadpassed, the temperature control system adjusted the temperature of themicrocalorimeter accordingly. As a result, a very precise computer control sys-tem was constructed to replace an older, outdated control system for which repairparts were no longer available.


SELF-CONSISTENT PERTURBATION THEORYCALCULATIONS FOR TWO-DIMENSIONAL

SUPERCONDUCTORS

John J. DeiszDivision of Mathematics, Science, and Technology

Black Hills State UniversitySpearfish, SD 57799-9051

Certain materials, called superconductors, conduct electricity without elec-trical resistance at low temperature. In the temperature region where a materialswitches from being a superconductor to a normal conductor, thermodynamicproperties are often well described by BCS theory, a scheme which approximateselectron-electron interactions with a self-consistently determined, static field.

There are at least two reasons to believe that BCS theory is insufficient forcharacterizing copper-oxide superconductors, a class of materials for whichsuperconductivity persists to relatively high temperatures. First, the crystalstructure is composed of well-separated conducting planes for which it is under-stood that strong dynamical superconducting fluctuations will appear, a fact thatis inconsistent with the assumption of an average, static field. Second, for tem-peratures above the superconducting transition temperature, these materials donot behave like normal metals. In contrast, BCS theory presumes a normalmetallic state above the superconducting transition temperature.

I describe a scheme, the fluctuation exchange approximation (FEA), forincluding the effects of the superconducting fluctuations that are omitted in BCStheory. Although the FEA is substantially more complex mathematically thanBCS theory, an algorithm developed for computations on parallel supercomput-ers has made numerical solution of the FEA feasible for many cases. In particu-lar, we show that these calculations demonstrate that the FEA correctly producesa superconducting transition in planar structures without relying on the qualita-tively incorrect assumption of a static effective field as is done in BCS theory.Further, superconducting fluctuations included in the FEA produce large devia-tions from normal metallic behavior, as is observed in the cooper oxide super-conductors.


CLIMATOLOGICAL TRENDS IN THE BLACK HILLS

Stephen D. Trimarchi, James R. Miller, Jr., and L. Ronald JohnsonInstitute of Atmospheric Sciences

South Dakota School of Mines and TechnologyRapid City, SD 57701-3995

ABSTRACT

This paper represents parts of an M.S. thesis entitled “A Climatology of theBlack Hills Region”, written by the first author. The purpose of this study was toexamine climatological trends for the Black Hills through much of the 20th cen-tury, and determine how elevation affects temperature, precipitation, and snow-fall at locations in and around the Black Hills. A comparison between climato-logical results in this study was made with climatological results from the 1949study by Harley Johnson. Trends over the 1910 to 1994 period indicated a slightincrease of 0.4°F in temperature and an increase of 0.3 inches in annual precipi-tation in the Black Hills. Elevation was found to be a significant forcing factorfor temperature and snowfall in the region. Comparisons were made with the1949 study to see how the averages and extremes had changed over the interme-diate 45-year time span. It was found that mean annual temperatures did notappear to have changed, but annual precipitation had increased up to 20% insome areas, while annual snowfall had generally increased in all areas fromabout 5% to over 100% over the last 50 years.

INTRODUCTION

Weather in the northern Great Plains is quite variable with rapid and extremechanges in temperature and calm-to-storm transition. The Black Hills riseprominently from the adjacent plains of western South Dakota and easternWyoming and add an extra element to the weather in this area. Temperature andprecipitation totals are often quite different between the surrounding plains andhigher elevations of the Black Hills. The orographic effects of the Black Hillsallow for large differences in weather in small distances.

The Black Hills are dome shaped with the major axis oriented 340 degreesby 160 degrees (Thornbury, 1954). They rise from about 900 to 1200 metersabove the surrounding plains. The area of the Black Hills is approximately 200kilometers long from north to south and 80 kilometers wide from east to west.

The Black Hills region has its own micro-climate compared to the northernGreat Plains region surrounding it. If not for the Black Hills, this area would bethe ideal type of continental climate with high daytime temperatures, low night-time temperatures, extreme alterations of hot and cold, and generally dry with awet season in the spring and early summer. In the Black Hills, the continental


type climate clashes with a mountain type climate, characterized by an increasein rainfall up to elevations of 1500 to 1800 meters, a rapid decrease of humid-ity with altitude, and wind movements altered (Johnson, 1949).

DATA SET

The instrumented sites which were chosen for this study were allDepartment of Commerce sites consisting of either National Weather Servicesites or cooperative station sites, where people volunteer to take observations ofmaximum and minimum temperatures and/or precipitation readings on a dailybasis (usually taken in the morning). Not every station was located in the centerof town and their location from the center of town is designated after the town’sname (see Appendix A). For example, the station Hermosa 3SSW was locatedabout 3 miles south–southwest of the center of Hermosa.

The data for this study was taken from the National Climatic Data Centers’(NCDC) Summary of the Day CD-ROM, distributed by EarthInfo, Inc. (1996),which contains temperature, precipitation, snowfall, and where applicable, evap-oration values for every NWS station and cooperative station going back to 1948,with a few going back even further.

The sites that were chosen for the study needed to fulfill two requirementsin order to be used. One was that the site needed to be located in or within about20 kilometers of either the South Dakota or Wyoming Black Hills. BelleFourche and Newell are a little beyond the 20 km perimeter, but are consideredimportant locations that can be used for comparisons to locations in the BlackHills in the analysis. The other consideration was that the sites needed to haveat least 30 years of continuous records and include the years of 1965 through1994.

There were 17 stations with temperature data and 21 sites with precipitationdata that satisfied all criteria (see Appendix A). Figure 1 shows the locations ofthe stations presented in the study. All these sites had temperature, precipitation,and snowfall data, except Buffalo Gap, Buskala Ranch, Hermosa, and Hill City,at which just precipitation and snowfall data were available.

CLIMATOLOGICAL ANALYSIS

Temperature and Precipitation Trends in the Black Hills

When looking at temperature trends, there are different ways to developthem. One general way is a linear trend model (Figure 2), which develops astraight line trend of how temperature has changed over the preceding years.Non-linear trends are better treated by a quadratic trend model (Figure 3), whichcan show increasing or decreasing trends for the same period of record overshorter periods of time.

Trends of annual mean temperatures were evaluated using averaged values


for four of the station sites: Hot Springs, Lead, Rapid City, and Spearfish. Thesesites give a good representation of the Black Hills, in general, over the 1910 -1994 period. Annual mean temperatures ranged over 7°F (4°C) difference on ayearly basis (from ~43°F to 50°F). The linear trend indicated an increase ofabout 0.4°F (0.2°C) over the 1910 to 1994 period (Fig. 2). The quadratic trend(Fig. 3) shows an increase in annual mean temperature of 0.65°F (0.35°C) till theearly 1960’s and then a decrease through the rest of the period of about 0.25°F(0.14°C).

The linear R2 value indicates a poor correlation to the yearly temperaturevalues, as would be expected considering the wide variability of the yearly tem-peratures. The t-test significance of the linear trend slope coefficient showed a tvalue of 0.67, which is low. The significance of the slope was about α = 0.25;thus no significant trend was found.

As with the temperature trends, precipitation trends were looked at using thelinear trend model (Figure 4) and the quadratic trend model (Figure 5). The year-ly mean values were based on mean annual precipitation using the same four sta-tions (Hot Springs, Lead, Rapid City, and Spearfish). The annual precipitation atthese four sites ranged from just over 10 inches (250 mm) in 1936 to over 33inches (840 mm) in 1945. An increase of about 0.35 inches (8.8 mm) was shownover the 1910 - 1994 period. The quadratic model showed most of the increasetaking place in the earlier half of the century and the trend leveling off at the endof the period. There is large variability in annual precipitation, even on a year toyear basis.

In these plots, the correlation of the yearly precipitation values to the trendline was even poorer than for the temperature trends. The t-statistic value of thelinear trend slope coefficient was only 0.21, which was significant at α = 0.42;thus, no significant trend was found.

Overlapping the temperature and precipitation trend figures, it could be seenthat dry years were typically associated with warmer than average temperatures,such as took place in the 1930’s and the 1950’s. However, the opposite was notnecessarily true with wetter than average years, although it is often cooler thanaverage or near average in those years. A scatter plot of temperature vs. precipi-tation is shown in Fig. 6. The best fit line had a correlation coefficient, r, of 0.38.This relationship suggested that lower than average annual temperature yearswere accompanied by higher than average annual precipitation years at an α of< 0.001.

Temperature, Precipitation, and Snowfall as a Function of Elevation

In Figure 7, the scatter plot of annual mean temperature as a function of eleva-tion shows a rather uniform decrease in temperature with height. Each point andnumber represents a station listed in the legend to the right, and its correspond-ing annual mean temperature. The fitted linear regression equation, using theelevation and annual mean temperatures, were calculated using the Minitab ver-sion, a statistical software package.


The linear regression equation, above the graph, represents the expected pre-cipitation values, based on elevation, from the values used to derive the equation.This line appears to be a good representation of the data. The average decreaseof temperature with height was about 0.7∞F (0.4∞C) per 100 meters which wassmaller than that expected.

Above the plot, the linear regression equation, along with the R2 value, anda t-statistic value are given. In this temperature analysis, the observed t-statisticvalue for the slope coefficient was –4.07. The given value for α = 0.05 is 1.75.The absolute value A1–4.07A1 was greater than 1.75; thus, elevation can be con-sidered a significant variable at α = 0.001.

The plot also suggested the possibility that the temperature decrease withelevation could be nonlinear at higher elevations. Deerfield’s annual mean tem-perature was much lower than the fitted regression’s expected value at that ele-vation. However, there were not enough high elevation stations to evaluate atrend.

The two stations that deviated the furthest from the fitted regression linewere Mt. Rushmore (higher) and Deerfield (lower). Mt. Rushmore’s tempera-ture was higher than would be expected likely because of the location of itsrecording site. The National Weather Service office in Rapid City, responsiblefor the maintenance of the stations, reported that this site’s location was near therocky base of the monument mountain. The granite rocks release heat at nightback into the atmosphere, keeping night-time temperatures 3∞F to 5∞F (1.5∞Cto 3∞C) warmer than would be expected at that elevation. Deerfield’s tempera-ture was lower than would be expected at its elevation possibly because it is oneof the highest locations in the Black Hills, where the lower water vapor contentof the air absorbs less of the outgoing infrared radiation (IR) resulting in greaterradiational cooling.

Figure 8 shows mean annual precipitation as a function of elevation for theBlack Hills. Each point and number represents a station and its mean annual pre-cipitation value. The middle solid line is the fitted linear regression line for theentire Black Hills. Closer examination of the scatter plot of the precipitation val-ues reveals that many of the southern Black Hills locations are to the left of theBlack Hills regression line and many of the northern Black Hills locations are tothe right of the regression line. Therefore, it appeared appropriate to split theBlack Hills into north and south sections and do a regression for each. The BlackHills data were thus split in half approximately perpendicular to the prevailingNNW winds of the region. In the figure, each station was designated as a south-ern Black Hills (S.H.) or northern Black Hills (N.H.) location in the legend. Thefitted regression lines for the southern Black Hills (dashed line) and the northernBlack Hills (dash-dot-dash line) are shown in Fig. 8.

The Black Hills regression line slope showed that precipitation increasedapproximately 0.71 in. (18 mm) per 100 m of increased elevation. The southernBlack Hills had a smaller rate of increase, about 0.5 in. (13 mm) per 100 m, whilethe northern Black Hills had a slightly larger rate of increase of 0.75 in. (19 mm)


per 100 m. The snowfall as a function of elevation plot (Figure 9) shows a quite diverse

distribution of mean annual snowfall with increase in elevation. Each point andnumber represent a station listed in the legend and its snowfall value. The samemethods and regressions were performed with mean annual snowfall as with themean annual precipitation.

The snowfall regressions show a much stronger correlation to the data thanthe precipitation regressions, especially when the stations are split into the north-ern and southern Black Hills. The significance tests at a = 0.05 show a strongcorrelation for locations in the southern Black Hills and even stronger in thenorthern Black Hills. All were significant to alpha values of less than 0.005. Theregression equation for the entire Black Hills shows an average increase of 9.8in. (25 cm) of snowfall per 100 meters rise in elevation, while the southern BlackHills shows an increase of only about 3 inches (7.5 cm) per 100 meters and thenorthern Black Hills shows an increase of over 12 inches (30 cm) per 100 meters.

Elevation was found to be significant in determining precipitation valueswhen using the entire Black Hills data set. Elevation was not shown to be asstrongly related to precipitation as it was to temperature and snowfall.

Comparison of this Climatological Study with the 1949 Study

The last detailed climatological study of the Black Hills region was com-pleted by Harley N. Johnson of the then-United States Weather Bureau in 1949.This study compared means from the lead authors’ thesis (Trimarchi, 1998)study, “A Climatology of the Black Hills Region”, covering the 1965 - 1994 peri-od for selected sites, to the analysis from Johnson’s study completed almost 50years earlier. An important note about Johnson’s 1949 study was that the meansin the following tables (Tables 1 - 3) were based on varying years of record, rang-ing from 20 to 61 years, and not all were continuous records.

Looking at the temperature comparisons between the two studies in Table 1,there appear to be no significant differences, except in a couple of northern loca-tions, Belle Fourche and Sundance, WY. At these locations there has been anannual mean temperature increase of 1.2∞F (0.7˚C) at Belle Fourche and 2.7˚F(1.5˚C) at Sundance.

At Rapid City, there has been an increase in annual mean temperature of1.0˚F (0.6˚C). This increase could be a small urban heat island effect due togrowth in the city, which would have contributed to a small rise in temperatureover the years. The temperature differences took place throughout the year andwere not seasonal.

When comparing mean growing seasons (Table 1, last column), it was a lit-tle surprising to find that the mean growing season appears to be of shorter dura-tion than those in the past. The difference ranges from a few days up to a cou-ple of weeks.

Many people have thought that a general increase in annual precipitation hasoccurred in the Black Hills and surrounding region. The precipitation trend


showed such an increase. Though there were periods of years, such as the 1920’sand 1960’s (Figure 4), when precipitation was often higher than average, therewere other periods, such as the 1930’s, 1950’s, and 1980’s (Figure 4), where itwas lower than average.

Comparing modern with the historic precipitation data, Table 2, there did notappear to be a clear-cut trend. Some stations experienced an increase in meanannual precipitation, some a decrease, while others showed no change.Deadwood’s precipitation was lower by 2.5 inches (64 mm) while Lead’sincreased by over 4 inches (100 mm). These could be due to changes in measur-ing techniques and/or gage location, and/or period of record. It was found thatDeadwood did not record precipitation between 1923 and 1942.

Comparison of number of days with precipitation of 0.01” or more showedthat all the stations in this study experienced a greater number of days with pre-cipitation than the 1949 study. Some sites were slightly greater, while otherswere more so. Even at locations where current mean annual precipitation wentdown, the number of days with measurable precipitation was higher. Theincreased frequency of precipitation days could have been due to a difference inhow the observations were taken by the observers in the first half of the century.

Mean annual snowfall amounts (Table 3) were greater in every location inthis study as compared to the 1949 study. Most were small differences explainedby low annual snowfall amounts in the 1920’s and 1930’s (Johnson, 1949) andannual snowfall increases in this part of the century, especially the last 10 yearsin the northern Black Hills. But a couple of sites, Lead and Deerfield, were muchhigher now than in the Johnson study. In the Johnson study period, Deadwoodreceived more mean snowfall than Lead. These changes are hard to explain sincethe difference in annual snowfall in Deadwood is not any different than at theother station, percentage wise. The apparent increase in Lead’s snowfall totalslikely involves several factors, including measuring accuracy at the station, pre-cipitation increases in the colder months, and possible changes in location of theobserving site within the town over time. Small changes in location could resultin significant differences in snowfall amounts on an annual basis.

In general, it appears temperatures were nearly constant, precipitation wentup at some locations, down in others, and snowfall increased from the first halfto the second half of the century. Overall, these two studies supported anincrease in precipitation over the Black Hills in the wintertime in the second halfof this century. The trend analysis also supported a small increase in precipita-tion over the same time period.

SUMMARY

The Black Hills region is a very diverse area weatherwise. Despite its smallsize, it plays a major role in the weather of the area. In the span of only a fewmiles, the weather can be dramatically different at any time of the year.Temperatures between observation stations differ by as much as 10˚F (6˚C) fromone location to another, annual precipitation doubled from the surrounding plains


to some higher northern Black Hills locations, and snowfall increased even morein those same locations. These wide variations spread to other climatic variables,such as heating and cooling degree days, length of growing season, and numberof days with certain temperature thresholds.

The temperature and precipitation trends (Figures 2 - 5) both indicate smallincreases over the 1910 to 1994 period. Temperatures have increased by about0.4°F (0.2°C) over that period and precipitation has increased by about 0.3 inch-es (8 mm). These trends have leveled off over recent years, however. In theBlack Hills, temperature, precipitation, and snowfall appear to be strongly relat-ed to elevation. In particular, elevation is a very significant variable to tempera-ture and snowfall in the region.

When comparing the results of this study to the earlier study done on theBlack Hills in 1949, some differences do show up between the two time periods.While temperatures stayed fairly constant, precipitation increases in some areas,decreases in others. Snowfall generally increased everywhere since the earlierpart of the century. These results generally support a wintertime increase in pre-cipitation over the years of up to 20% in some locations.

Future topics of research related to this study could involve looking at pos-sible microscale weather processes, which appear to be occurring in some areas,such as the very localized, heavy snowfall events in portions of the northernBlack Hills, around the Lead area. Another possibility is to look in-depth at thesynoptic and mesoscale weather patterns which affect the weather in the BlackHills region and what the specific effects are for different parts of the Black Hills.Another study could involve looking at the historic weather records in the BlackHills, and examining closely the differences between the past and present aver-ages and why the differences are occurring.

ACKNOWLEDGMENTS

The authors thank Ms. Connie Crandall for the fine job of preparing themanuscript. This research was sponsored by the State of South Dakota.

REFERENCES

Earthinfo, Inc.. 1996. NCDC Summary of the Day, West2 CD2. EarthInfo, Inc.,Boulder, CO. CD-Rom.

Johnson, N. H. 1949. A climatological survey of the Black Hills. Black HillsEngineer, S. D. School of Mines and Technology. 29: 3-35.

Thornbury, W.D., 1954: Principles of Geomorphology. Wiley and Sons, NewYork.

Trimarchi, S.D.. 1998. A Climatology of the Black Hills Region. M.S. Thesis,Department of Atmospheric Science, South Dakota School of Mines and


SENEGAL’S THEORY OF ATOMIC PARTICLES

Claffey J. Senegal, Jr.c/o Telecare Choices

3851 Rosecreans StreetSan Diego, CA 92110`

The electron and proton are electro-magnetic energy bubbles. The neutronsare electrons and protons together and the atoms are electro-magnetic energybubbles. The electron does not orbit the nucleus. But, the electron sits on anelectro-magnetic energy shield or “bubbles.” They form bubbles because elec-tro-magnetic energy has surface tension, just like water has. On a large wave-length of radiowave, it bends around corners because there is less surface tensionover that wavelength. And, on a small wavelength of electo-magnetic spectron,like microwave and higher frequencies, the wave travels in a straight linebecause of the large surface tension over that wavelength.

The nucleus of an atom consists of proton and neutrons. The neutrons arebipolar and they stick to the proton like foam of bubbles. And, on an atom orbubbles around angstrom which consist of secondary bubbles, third and fourth,fifth and sixth, and so on sit electrons. They sit still and do not radiate energy.The particle made from cyclotron like anti-proton, anti-electron, Quartz etc., areelectro-magnetic energy bubbles: and, when they slowly pop like bubbles, theyform a wave. When the gamma rays come close to an proton, they form bubbles.Atoms stick together like suds.


PATTERNS IN AVIAN COMMUNITYSTRUCTURE AND NON-POINT SOURCE

DISTURBANCE POTENTIAL ALONG THELAND-WATER INTERFACE OF

A PRAIRIE POTHOLE LAKE

Kristel K. Bakker1 and Nels H. Troelstrup, Jr.2

Department of Biology and MicrobiologySouth Dakota State University

Brookings, SD 57007

ABSTRACT

This study was conducted to test the hypothesis that bird community struc-ture varies significantly between areas which are prone versus not prone to dis-turbance within the land-water interface of a prairie lake. On-site assessment ofland cover and a Geographic Information System (GIS) with coverages of slope,soil erodability, soil hydrologic group and drainages were used to delineate sitesprone and not prone to non-point source disturbance. Habitat and communityattributes were examined within these site classes during the summer of 1995.While habitat differences between classes (prone and not prone) were inconclu-sive, results of avian community analyses indicated that sites prone to distur-bance had significantly higher average densities (69 individuals/ha vs. 56),species richness (10.9 vs. 8.1 species) and diversity (H’=2.05 vs. 1.72) than sitesnot prone to disturbance. These results are consistent with observations of otherecological communities exposed to intermediate levels of disturbance.Furthermore, these results suggest a relationship between GIS generated non-point source disturbance potential and ecological communities within the land-water interface.

INTRODUCTION

Pickett and White (1985) state “A disturbance is any relatively discrete eventin time that disrupts ecosystem, community or population structure, and changesresources, availability of substratum, or the physical environment”. Disturbanceis also viewed as any irregular, erratic or unusual event that forces organismsfrom a static, near-equilibrium condition (Pickett and White 1985). These devi-ations from the norm, whether positive or negative, may lead to elimination ofill-adapted species within a community (Odum et al. 1979).

Ecological monitoring programs are often implemented to detect thesechanges and understand factors that influence them. Biomonitoring uses thecharacteristics of organisms to assess the environment (Hunsaker and Carpenter


1990). The use of organisms has several advantages over physical and chemicalmeasurements, including integration of environmental changes through time,simultaneous consideration of multiple trophic levels and consideration of organ-ism health (Furness and Greenwood 1993).

Standing freshwater makes up less than 4-5% of the Earth=s surface, yet asmany as 11-23% of all bird species are dependent on these waters and their mar-gins and many more species use these areas during their life cycle (Furness andGreenwood 1993). In eastern South Dakota alone, 100 bird species depend onwetlands (Johnson et al. 1997). Freshwaters, like all ecosystems, are subject tonatural and anthropogenic changes which may have many biological repercus-sions. Prairie wetlands have been altered by drainage, siltation and changingagricultural practices leading to marked changes in vegetation (Kantrud 1990).These alterations in habitat may lead to dramatic changes in resident animalspecies.

This research was conducted to determine if habitat characteristics and aviancommunity structure differ between areas of high and low disturbance potentialfrom non-point source pollution.

METHODS AND MATERIALS

Research efforts were conducted along the shoreline of Oak Lake,Brookings County, SD (T112N, R48W, Section 12) (Figure 1). Oak Lake has anarea of 163 ha and perimeter of 9.8 Km. Habitats around the lake include shal-low and deep water wetlands, oak forest, upland mixed grass prairie and agroe-cosystems.

Eight sampling sites were chosen along the perimeter of Oak Lake based ontheir disturbance potential (Figure 1, Table 1). Shoreline patch attributes alongthe perimeter of the Oak Lake basin (erodability, hydrologic soil group, slope,land use and drainages) were scored individually and summed to provide anoverall site disturbance potential score following Sivertun et al. (1988) (Table 1).A detailed description of GIS scoring procedures is provided in Foley (1997).Drainages were the focal point of sites with high disturbance potential as they arelinked to the hydrology of the basin through runoff events, making them criticalareas for study. Each of the study sites encompassed an area of 6000 squaremeters. Four sites each were selected representing low and high disturbancepotential.

Habitat sampling was conducted in July, after avian counts were completedso that nesting habitat would not be disrupted (Ralph et al. 1993). Vegetationcomposition and density were measured using randomly placed, nested quad-rants along five random transects at each site (Higgins et al. 1994).Measurements included grass, forb, cattail and woody stem species occurrenceand density. In addition, tree species, diameter at breast height, visual obstruc-tion and canopy cover measures were taken within a 10x10m quadrant.

A fixed-width transect survey (Mikol 1980) was used to determine avianspecies richness, avian density and presence/absence of species. Counts were


initiated on 16 May 1995 and ended 30 June 1995. These dates were chosen socounts would not be biased by migrants or young birds (Ralph et al. 1993 andMikol 1980). Transects were walked between one half hour before and three tofour hours after sunrise (Emlen 1971; Ralph et al. 1993). Sites were rotated aftereach round of counts to eliminate bias due to time of day. Counts were not com-pleted when 1) there was heavy to moderate rain, 2) heavy fog, 3) strong winds(> 10 km per hour), or 4) extreme temperatures (<7 or > 24 C) (Ralph et al.1993).

Birds leaving the transect during approach were counted, if they were with-in the 100m length and 60 m fixed width (Mikol 1980). Transects were walkedat an approximate speed of 0.5 to 1.5 mph (Ralph et al. 1993). Species and thenumber of each occurring within the transect width were counted and recorded(Mikol 1980). Short stops were allowed to observe detected birds (Mikol 1980,Ralph et al. 1993). Both visual and auditory detections were counted.

Avian species richness, density, and Shannon-Wiener diversity index(Shannon and Weaver 1963; Washington 1984) were analyzed for site class anddate effects using a two-way analysis of variance with date as a blocking vari-able (Steel and Torrie 1980). Habitat differences between site classes were ana-lyzed using a one-way analysis of variance. Means comparisons among siteclasses and dates were analyzed using the Tukey=s mean comparison test.

Runs and Wilk-Shapiro tests were used on all data to determine if the datawere normally and independently distributed. Data not normally distributedwere ranked and analyses were performed on the ranks (Conover and Iman1981).

RESULTS

Typha glauca, grass, forb and shrub stem densities were higher and morevariable at sites prone to disturbance (Table 2). In addition, measured canopycover was zero at prone sites due to small numbers of scattered, large trees.However, no statistical differences could be detected for vegetation attributesbetween site classes (ANOVA, p > 0.05).

Fifty-six avian species were recorded from Oak Lake study sites, 50 ofwhich were observed at sites prone to disturbance and 43 of which were observedat sites not prone to disturbance (Table 3). Red-winged blackbirds were the mostabundant species observed at all eight sites, accounting for 32% of the total indi-viduals observed (Table 3). Common yellowthroats, yellow warblers, song spar-rows and marsh wrens were also abundant, accounting for 30% of all observedindividuals. Species occurring on more than one date and observed only in sitesnot prone to disturbance included the white-breasted nuthatch, house wren, rose-breasted grosbeak and belted kingfisher (Table 3).

Bird densities were higher at sites prone to disturbance relative to not pronesites (69 individuals/ha versus 56 individuals/ha; 2-way ANOVA, p = 0.001,n=4)(Table 4). Lower densities were observed late in the counting session (2-way ANOVA, p = 0.005, n=12) but no significant treatment by date interactionwas observed (p = 0.362).


Sites prone to disturbance had a mean richness of 10.9 (n=48) species whilesites not prone had a mean richness of 8.1 (n=48) species (2-way ANOVA,p<0.001, n=4) (Table 4). Lower richness values were observed toward the endof the counting period (2-way ANOVA, p<0.049, n=12). No treatment by dateinteraction was observed (p=0.668).

Shannon-Wiener diversity was significantly higher at sites prone to distur-bance with a mean index value of 2.05 (n=48) while not prone sites had a meanindex value of 1.72 (n=48)(2-way ANOVA, p<0.001, n=4) (Table 4). No signif-icant date (p=0.378, n=12) or interaction (p=0.870) effects were observed.

DISCUSSION

Avian density, richness and diversity were all significantly higher in patch-es which were prone to disturbance. Prone sites had higher mean stem densitiesand greater variance between replicates for Typha glauca, forbs and grasses. Inaddition, scattered trees and shrubs increased habitat heterogeneity relative to notprone sites. Marsh wrens, yellow-headed blackbirds and sedge wrens all requirewetland vegetation types for nesting habitat (Ehrlich et al. 1988). These specieswere two to three times more abundant at prone sites. In addition, several edgespecies, including common grackles, American goldfinches and brown-headedcowbirds, were three to ten times more abundant at prone sites.

Sites not prone to disturbance exhibited lower habitat heterogeneity and sup-ported greater numbers of mature trees and shrubs. Three avian species occur-ring only in sites not prone to disturbance, the white-breasted nuthatch, housewren and rose-breasted grosbeak, depend on mature trees. White-breastednuthatches and house wrens are cavity nesters while rose-breasted grosbeaks pre-fer deciduous woodlands and nest 5 to 15 feet off the ground (Ehrlich et al.1988). In addition, belted kingfishers were observed foraging only from thecanopy of not prone sites.

The presence of more heterogeneous habitat at sites prone to disturbance(i.e., some trees and shrubs and higher densities of grasses, forbs and cattails)provided greater habitat diversity for avian species. Greater heterogeneity facil-itates nonrandom use of habitat and food resources by resident animals andimproves foraging efficiency (Pickett and White 1985).

The results of this study may provide support to the IntermediateDisturbance Hypothesis (Connell 1978). This hypothesis contends that distur-bances occur at various scales, intensities and frequencies (Connell 1978).Greater habitat diversity and reduced competitive interactions result from inter-mediate frequencies and intensities of disturbance. Our results suggest that dis-turbances to littoral patches around Oak Lake maintain greater habitat hetero-geneity at prone sites, leading to greater avian abundance, species richness and species diversity.

Reice (1994) suggests that biological systems are normally in some stage ofrecovery from the last disturbance. Successional processes follow these distur-bance events, leading toward habitat homogeneity and strong competitive inter-


actions. Oak Lake patches not prone to disturbance exhibit characteristics ofmore mature successional stages with lower habitat heterogeneity, avian numbersand species. These results are consistent with those of others who have foundhigher avian species richness and density during intermediate stages of succes-sion (Odum 1950; Johnston and Odum 1956; Karr 1968; Shugart and James1973; Meslow 1978; Morrison and Meslow 1984; Raphael et al. 1987; Meyersand Odum 1991).

Data collected from Oak Lake indicate differences in avian communitystructure between sites with high and low disturbance potential. Avian abun-dance, species richness and species diversity were all higher at sites prone to dis-turbance. These differences appear to be related to maintenance of high habitatheterogeneity at sites prone to disturbance and are consistent with observationsof other ecological communities exposed to intermediate levels of disturbance.Furthermore, these results suggest a relationship between GIS generated distur-bance classification and ecological attributes of pothole habitats.

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MINUTES OF THE EIGHTY-THIRD ANNUAL MEETING OF THE SOUTH DAKOTA

ACADEMY OF SCIENCE

The Eighty-third Annual Meeting of the South Dakota Academy of Sciencewas held April 17,1998 at Black Hills State University in Spearfish, SouthDakota.

The Executive Council met at 7:00 pm Thursday, April 16, for a final checkof plans for the annual meeting to follow.

OPENING GENERAL SESSION

President Sharon Clay called the Academy to Opening Session at 9:00 am.Dr. Thomas Flickema, President of Black Hills State University, welcomed theAcademy to the campus. In his address at the opening session President-ElectRoyce Engstrom emphasized the importance and significance of contributionsmade by rural Americans in his address titled: Science and Rural America.

Papers for the Senior Academy began at 10:00 am and continued through theday. There were fifty-six papers presented. The Junior Academy did not con-duct an annual meeting this year. Ninety six members (47 associate and 49 reg-ular) registered for the annual meeting.

ANNUAL BUSINESS MEETING

The Academy was called to its Annual Business Meeting at 12:45 pm byPresident Sharon Clay. President Clay extended the Academy’s sincere gratitudeto Audrey and Mark Grabel for making the local arrangements for the AnnualMeeting.

Secretary-Treasurer Bill Soeffing asked and reminded the membership tosubmit email addresses and institutional addresses to him for future mailings. Healso reported that the membership dues increase made in 1994 has had a stabi-lizing affect on cash flow for the academy. He also indicated that some monieswere available for investment and that he would be seeking support for invest-ment of those monies later during the meeting. A list of new regular and associ-ate members was presented for the Academy’s approval.

Editor Emil Knapp extended his report through the Secretary-Treasurer. Hesuggested that additional assistance was needed to produce the Proceedings ofthe South Dakota Academy of Science in a more timely fashion. He also indi-cated an interest in resigning his position as the editor, but concomitantlyexpressed a commitment to helping a new editor.

President Clay pronounced Dr. Kenneth Higgins as the first Fellow of theSouth Dakota Academy of Science. Dr. Higgins was to arrive later to receive theaward. Following the announcement, the Application for the FellowsNomination was reviewed and the procedures for nomination and election reaf-firmed.


Dakota State University has invited the Academy to hold its 84th AnnualMeeting in Madison, South Dakota on April 16-17, 1999. Drs. DonnaHazelwood and Dale Droge will be co-chairs for the local arrangements. Long-range meeting arrangements were discussed and announcements made for theTri-State (MN, ND and SD) Meeting in 2000 at Moorhead, MN. The 2001Annual Meeting will be in conjunction with Space Day at the Discovery Centerin Pierre, South Dakota.

The Junior Academy of Science was discussed. The Senior Academy reaf-firmed its commitment to the Junior Academy and discussed plans for increasingparticipation. President Clay suggested that President-Elect Engstrom appointthe First and Second Vice Presidents to work collaboratively to support theJunior Academy. A rotation system would naturally evolve from such arrange-ments and provide the continuity that might be needed to support such anendeavor. Scholarships to various public and private colleges and universitieswere announced as motivation for many of the student participants.

MOTION (Tatina/Granholm): The SDAS will invite the regional sci-ence fair winners to attend the Junior Academy with travel andlodging being subsidized by the Senior Academy.

Motion Carried.

RESOLUTION COMMITTEE

President Clay announced that US Representatives John Thune and TomDaschle had responded to the Academy’s Resolution pertaining to paleontologi-cal artifacts and fossils.

Drs. Gary Larson and Robert Tatina presented the following resolutions onbehalf of the committee.

REPORT OF THE RESOLUTIONS COMMITTEE

The membership of the South Dakota Academy of Science thanks Dr.Thomas Flickerma, President of Black Hills State University, for making thecampus facilities available and for hosting the 1998 annual meeting of theAcademy.

The Academy extends its thanks to the local planning committee - Audreyand Mark Gabel - for making the excellent arrangements for the meeting.

Thanks to President-Elect Royce Engstrom for his address, AScience andRural America@. His presentation provided valuable insight into South Dakota’sstatus in the scientific community and how funding of scientific research mightbe improved in the state.

We also wish to thank Dr. Norman Myers of Oxford University for his fas-cinating address, AMass Extinction of Species: What Can We Do about It?@

President Sharon Clay is commended for the leadership he has provided theAcademy this year.


We are appreciative of the time, effort, and perservance of the AcademySecretary-Treasurer, Bill Soeffing, and the Editor of the Proceedings, EmilKnapp.

The Academy congratulates Ronald Campbell, Arlington High School, whowas recognized as the Outstanding Physical Science Teacher of the Year. Wealso extend our congratulations to Donald Decker, Lincoln High School, SiouxFalls, recipient of the Outstanding Biology Teacher of the Year award.

Respectfully submitted,Nels H. Troelstrup, Jr. and Gary E. Larson

MOTION (Reese/Leifferts): Accept the General Resolutions.Motion Carried.

DISTANCE EDUCATION

Whereas, distance learning technologies are gaining ever greater popularityas means to deliver courses to remote audiences, and

Whereas, South Dakota institutions of higher education are being encour-aged to compete in the preparation and delivery of Internet courses, includingscience courses, and

Whereas, many science courses entail indispensable hands-on laboratoryand field experiences, many of which are impossible to simulate on the comput-er, and

Whereas, science educators and professionals are the proper authorities whoshould determine course content and mode of delivery, and

Whereas, no foolproof method has yet been devised to verify learning out-comes by testing over the Internet, and

Whereas, the learning outcomes of Internet courses should be comparable tothe learning outcomes of traditional courses,

Therefore, be it resolved the South Dakota Academy of Science affirms theauthority of science faculty in determining what should and should not be appro-priately delivered via the Internet.

Discussion followed pertains to the type of courses being currently offeredby South Dakota’s institutions of higher education and the transferability ofcourses outside of the State.

MOTION (Tatina/Engstrom): Accept the Distance Education Resolutions.Motion Carried.

South Dakota Academy of Science recognizes the years of experience, wis-dom and collective passion of retiring public educators throughout South Dakota.These dedicated educators will be replaced, but in fact they are not replaceable.The academy wishes to recognize the following educators for their contributiontoward the education of our young students. We wish them happiness and pros-perity in their new endeavors.


Mr. Marv SelnesMr. John LandegentMr. Ron CampbellMr. Vernon EggertMr. Jay HenniesMr. Joe AustinPatrick Henry School, Sioux FallsSioux Falls Lincoln High School, Sioux FallsArlington High School, ArlingtonHoward High School, HowardVermillion High School, VermillionMadison High School, Madison

Respectfully submitted,Nels H. Troelstrup, Jr., Gary L. Larson and Bob Tatina, Resolutions Committee

MOTION (Hawk/Reese): Accept the Proclamation of RetiringSecondary Teachers contingent upon completing the listing to rep-resent the entire State.

Motion Carried.

TEACHER CERTIFICATION

Whereas, it is the responsibility of the South Dakota Academy of Science topreserve the integrity of science (Resolution. 1982. Proc. Acad. Sci. 61:11-12),and

Whereas, science is a systematic method of investigation based on continu-ous experimentation, observation and measurement leading to explanations ofnatural phenomena, such explanations being open to further testing and modifi-cation (Resolution. 1982. Proc. Acad. Sci. 61: 11-12), and

Whereas, the theory of descent with modification by natural selection fullysatisfies these criteria, and is the central theory upon which all of biology isbased (Resolution. 1982. Proc. Acad. Sci. 61: 11-12), and

Whereas, Chapter 24:16:08:16, entitled 7-12 Science Education Program, ofthe proposed Administrative Rules of South Dakota governing teacher certifica-tion omits evolution as a topic required in the preparation of secondary schoolteachers, and

Whereas, these same rules omit the philosophy and methods of science as arequired topic in the preparation of secondary school teachers, and

Whereas, the proposed South Dakota Content Standards (South DakotaScience Standards Draft II, March 1988) lack a direct reference to the theory ofdescent with modification,

Therefore, be it resolved the South Dakota Academy of Science, through itsPresident, urge the South Dakota Board of Education to add evolution and phi-losophy and methods of science as required teacher education topics to the


Administrative Rules, and evolution (as the theory of descent with modificationby natural selection) to Section 24:16:08:16 of the Science Content Standards.

MOTION (Larson/Haertel): Accept the Teacher CertificationResolutions

Motion Carried.

AUDITING COMMITTEE

Charles Estee announced the Auditing Committee found the books andrecords of the SDAS in good order and financial standing.

MOTION (Estee/Reese): Accept the Auditing Committees action.Motion Carried.

President Clay announced the Outstanding Biology Teacher of South Dakotawas Mr. Don Decker and the Outstanding Physical Science Teacher of SouthDakota was Mr. Ronald Campbell. She also announced that Gateway 2000 Inc.has been approached to sponsor a three-year cycle of awards for an OutstandingComputer Science or Mathematics Teacher of South Dakota.

NOMINATING COMMITTEE

The Nominating Committee presented a slate of candidates and electionswere held. The officers for the 1998-99 South Dakota Academy of Science areas follows:

President Royce Engstrom, USDPresident-Elect R. Neil Reese, SDSU1st Vice-President Lenore Koczon, NSU2nd Vice-President Charles Lamb, BHSUSecretary-Treasurer William Soeffing, USFProceedings Editor Emil Knapp, Augustana College1st Past President Sharon Clay, SDSU2nd Past President John Thomas, USDMembers-at-Large Audrey Gabel, BHSU

Donna Hazelwood, DSUGary Larson, SDSUJim Sorenson, Mount Marty College

BIODIVERSITY COMMITTEE

Nels Granhom has sent out a “working document” to the committee mem-bers in an attempt to present South Dakota’s biodiversity at a comprehensivelevel.


TREASURER’S REPORTStatement of Receipts, Disbursements and Changes

in Cash Balances for Fiscal Year 1997

Cash Balance on 1 January 1997Certificate of Deposit $5161.47Savings Account $9.15Checking Account $4788.25

Total Beginning Cash $9958.87

ReceiptsMembership Dues

(Life-34 / Regular-98+2 / Associate-16) $2080.00Warrant Replacement $2330.00Gifts to Jr. Academy $120.00Jr. Academy Annual Meeting Registration $19.00Reprint Charges $365.00Page Charges $5344.50Annual Meeting Registration $600.00Banquet Tickets $215.00Interest on Investments $279.01

Total Receipts $11352.51

DisbursementsCorporate Fee (SD Secretary of State) $40.00NAAS Dues $$67.00Jr. AcademySupplies and Materials $369.34Substitute Teacher Pay $122.00Research Grants $600.00SDAS Proceedings Publication $4207.00Stale Warrants $2411.50Annual Meeting $763.27Postage $81.32Duplication $232.11

Total DisbursementS $8893.54

Cash Balance on 31 December 1997Certificate of Deposit $5500.00Savings Account $9.15Checking Account $6908.69

Total Ending Cash $12417.84


NEW BUSINESS

1st Vice President R. Neil Reese discussed a Internet/World Wide Web Sitedevelopment program for the SDAS. Additional information will distributed tothe membership.

MOTION (Soeffing/Larson): Direct the Secretary-Treasurer to invest$3000.00 into a certificate of deposit as per the discretion of theExecutive Council at the time of investment.

Motion Carried

At the close of the meeting, President Sharon Clay expressed her pleasure inpassing the gavel to President-Elect Royce Engstrom and wished him the best inthe 1998-99 presidency. At 2:00 pm, President Engstrom declared the SouthDakota Academy of Science adjourned until the April 16-17, 1999 AnnualMeeting at Dakota State University in Madison, South Dakota.

Respectfully submitted,Bill Soeffling

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