bacteriological assessment of spoon river water qualityspoon river in the vicinity of havana was...

APPLIED MICROBIOLOGY, Aug. 1974, p. 288-297Copyright i 1974 American Society for Microbiology

Vcl. 28, No. 2Printed in U.S.A.

Bacteriological Assessment of Spoon River Water QualitySHUNDAR LIN, RALPH L. EVANS, AND DAVIS B. BEUSCHER

Water Quality Section, Illinois State Water Survey, Peoria, Illinois 61601

Received for publication 24 April 1974

Data from a study of five stations on the Spoon River, Ill., during June 1971through May 1973 were analyzed for compliance with Illinois Pollution ControlBoard's water quality standards of a geometric mean limitation of 200 fecalcoliforms per 100 ml. This bacterial limit was achieved about 20% of the timeduring June 1971 through May 1972, and was never achieved during June 1972through May 1973. Ratios of fecal coliform to total coliform are presented. Byusing fecal coliform-to-fecal streptococcus ratios to sort out fecal pollutionorigins, it was evident that a concern must be expressed not only for municipalwastewater effluents to the receiving stream, but also for nonpoint sources ofpollution in assessing the bacterial quality of a stream.

Pathogenic bacteria indicators are used todetermine the presence of disease-causing orga-nisms originating from fecal pollution. Indica-tors such as total coliform (TC), fecal coliform(FC), and fecal streptococcus (FS) are usedbecause of the laborious technique and equallyexpensive equipment required to isolate patho-genic bacteria and viruses from water.The use of coliform bacteria as a measure of

the fecal contamination of lakes and streamshas been in practice for many years. Thecoliform group, however, includes'a heterogene-ous mixture of bacteria, many of which havelittle in common with each other except the factthat they are always present in the intestinaltract of humans and other warm-blooded ani-mals. Thus the occurrence and densities of theTC group have been useful in assessing thesanitary conditions of water even though it iswell known that some bacteria in the group haveorigins other than fecal material.More recent developments have shown that

the use of FC as an indicator of pollution fromwarm-blooded animal feces is a more precisebacteriological tool for assessing water quality,and the Illinois Pollution Control Board (10)(IPCB) has adopted rules requiring adherenceto certain limitations on bacterial quality inwaters of the state measured by the occurrenceand density of fecal coliforms.One of the weaknesses of the FC test is its

inability to distinguish between whether or notthe contributing sources are human or nonhu-man warm-blooded animals. Geldreich andKenner (2, 4) have reported upon the use oftests for FC correlated with tests for FS as a

reliable procedure for differentiating sources ofbacterial pollution. Their findings showed thatFS densities were significantly higher than FCdensities in all warm-blooded animals fecesexamined except that of humans. The applica-tion of these findings, within limits, should beuseful in determining the comparative relation-ship of urban and rural areas as sources of fecalpollution in Illinois waters.A 2-year water quality study of the Spoon

River commenced on 1 June 1971. The stream'swaters were sampled at weekly intervals forbacteria and plankton enumerations as well asfor major chemical constituents until 29 May1973. The objectives of the study were (i) todetermine the bacteria densities at selectedsites on the main stem of the Spoon River and(ii) to ascertain if meaningful relationships existbetween the densities and distribution of TC,FC, and FS.

MATERIALS AND METHODSThe Spoon River, with a drainage area of about

1,800 square miles, (4,660 sq. km) extends approxi-mately 161 miles (260 km) in length from its conflu-ence with the Illinois River in the vicinity of Havana,Ill. From its two sources, the East Fork and West Forkoriginating in Bureau and Henry counties, respec-tively, the Spoon River flows in a southernly directionthrough the counties of Stark, Peoria, Knox, andFulton. In addition to its origin, the principal tribu-taries of the Spoon River are Indian, Walnut, Court,Cedar, and Big Creeks. The slope of the main stem ofthe river ranges from 4 feet (about 1.21 m) per mile(about 1.6 km) near its mouth (0.012 to 0.076%). Theextent of the basin is shown in Fig. 1.

There are 16 municipalities served by wastewater288

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BACTERIOLOGICAL ASSESSMENT OF SPOON RIVER

FIG. 1. Spoon River Basin showing sampling stations.

treatment plants with effluents that discharge in thebasin. The largest communities are Kewanee andCanton with populations of 15,800 and 14,200 persons,respectively. Of the other 14 communities, seven havepopulations equal to or less than 1,500 persons, andthe other seven have populations greater than 1,500persons. None of these 14 communities have popula-tions in excess of 3,200 persons. The sewered popula-tion in the basin is approximately 53,800 persons.

For the purpose of the study, five sampling stationswere established (Table 1). Their respective locationsare shown in Fig. 1.

During periods of sampling, stream flows as mea-sured at Seville varied from 18 fts/s (about 5.4 m3/s)on 18 September 1971 to 16,000 ft'/s (about 480 ma/s)on 24 April 1973. These extreme flow ranges suggestthe sensitivity of the Spoon River to runoff. Watertemperatures varied from 0 to 29 C and ice cover

prevailed during January and February. The temper-

TABLE 1. Sampling stations used

Station Mile points Drainage area

Station ~~~~~~~(sq.mi.)"Havana 2.5 (1.6) 1,805 (704.0)Seville 37.7 (23.4) 1,600 (624)London Mills 68.4 (42.4) 1,070 (417.3)Elmore 113.7 (70.5) 569 (221.9)Modena 141.2 (87.5) 154 (60.1)a River miles above mouth. Numbers in parentheses

indicate kilometers."Numbers in parentheses indicate square kilome-

ters.

ature of the stream water, in general, increased as thewater progressed downstream.

Although tests for TC and FC bacteria were per-formed at weekly intervals during the 2 years of

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LIN, EVANS, AND BEUSCHER

sampling, tests for FS were limited to the second year.All samples for bacterial examination were collectedseveral inches below the surface of the water atmidchannel, in sterile 250-ml glass bottles, and placedon ice immediately. Bacterial examinations wereperformed on the morning after the day of collection.Membrane filter techniques for TC, FC, and FS

were performed in accordance with StandardMethods (1). The previous studies (5, 6) suggest thatthe membrane filter procedure is comparable to themultiple-tube method for TC, FC, and FS determina-tions in the river waters. TC counts were made withthe M-Endo agar LES two-step procedure. M-FCbroth and M-Enterococcus agar were used for FC andFS enumerations, respectively. Three duplications foreach sample were filtered through 0.45-jim membranefilters for each test.

RESULTS AND DISCUSSIONBacteria density. From June 1971 through

May 1973, total coliform densities varied ran-domly from a minimum of 160/100 ml to amaximum of 13,000,000/100 ml. Both of theseextremes occurred at Seville. FC densitiesranged from 16/100 ml at London Mills to

160,000/100 ml at Modena, and FS countsreached a minimum of 17/100 ml at Havana anda maximum of 42,000/100 ml at Modena.The mathematical distribution of the bacte-

rial density data was determined by plotting thedata on log probability paper. Figure 2 shows atypical pattern of distribution for the first year'sdata at two stations. Since a straight line can bedrawn with some confidence through each set ofplotted points, the data reflect the characteris-tics of a log-normal distribution with the geo-metric mean at the 50% quantile. The centraltendencies and dispersion of log-normal distrib-uted data can best be expressed in geometricterms, i.e., geometric mean and geometric stan-dard deviation. The slope of the line reflects thevariability of the data, and the geometric stan-dard deviation (variability) is larger for theHavana data (Fig. 2).The yearly ranges, geometric means, and

geometric standard deviations of the bacterialdensities observed at each of the sampling sitesare summarized in Table 2. An inspection of thegeometric means for FC and FS in the table

1100/100 ml0viation (a6 7 5.01

906 0 8 0 5 9 99.

PROBABILITY OF NOT EXCEEDINGFIG. 2. Log-probability plot of fecal coliforms densities in the Spoon River, June 1971 through May 1972.

290 APPL. MICROBIOL.


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generally suggests a higher concentration oforganisms in the upper reaches, with lowerconcentrations in the downstream sectors. How-ever, after applying statistical tests to the data,it was determined that there are no significantdifferences between the geometric means for thestations at Modena and Elmore, a distance ofapproximately 31 miles (50 km); similarly,there was no difference between the geometricmeans at London Mills and Seville, a distanceof about 28 miles (45 km). Since it is unlikelythat FC and FS experience regrowth in thestream, and considering the fact that the orga-nisms do die off, it is probable that the lack ofdifferences in bacterial densities between thestations is due to bacterial loadings originatingfrom nonpoint rural sources of pollution alongthe intervening distances between stations.Monthly geometric means of TC and FS bac-

teria densities are shown in Fig. 3 and 4,respectively. It is apparent from Fig. 3 andTable 2 that TC densities for the first year of thestudy were much lower than that for the secondyear. Stream flows were considerably higherduring the second year. There was not a generalpattern, suggesting seasonal variations in bacte-ria densities (TC and FS) at any of the stationssampled. Generally, however, there were de-creases in TC densities with downstream move-ment (Table 2).An exception to this general rule occurred

during January through April 1973 (Fig. 3) forthe Havana station. An examination of theIllinois River recorded pool stages for thatperiod suggests that the sampling station on theSpoon River in the vicinity of Havana wassignificantly influenced by the floodwaters ofthe Illinois River. It is quite probable duringthis period of high water that the Illinois Riverwas receiving inadequately treated sewages up-stream of the Havana area to the extent thathigher than normal TC densities were found inits backwaters in the Spoon River.Comparison of FC standards. The general

rule for bacterial quality, adopted by the IllinoisPollution Control Board (10) and applicable tothe most Illinois streams including the SpoonRiver, is Rule 203(g), which states:

"Based on a minimum of five samplestaken over not more than a 30-dayperiod, fecal coliforms shall not exceeda geometric mean of 200/100 ml, norshall more than 10% of the samplesduring any 30-day period, exceed400/100 ml."

The FC densities, recorded for the Spoon

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J J A S 0 N D J F M A M J J A S 0 N D J F M A M1971 1972 1973

FIG. 3. Monthly geometric means of total coliforms densities in the Spoon River, 1971 through 1973.

uJ 310

210JUN JUL AUG SEP OCT NOV UEC JAN FEB MAR APR MAY

1972 1973FIG. 4. Monthly geometric means of fecal streptococci densities in the Spoon River, 1972 through 1973.

River at five locations from June 1, 1971 to May29, 1973 were evaluated in terms of this ruleusing a programmable Wang 720 calculator(Fig. 5 and 6). There were some 30-day periodsfor which five samples were not available forevaluation. These omissions are indicated onthe abscissa of the figures. Also depicted on the

figures are the FC limits adopted by the IPCB,i.e., the geometric mean of 200/100 ml thatmust not be exceeded.

It appears that only during the latter part ofOctober and the month of November 1971 wasacceptable bacterial quality achieved at allstations (Fig. 5). Acceptable bacterial quality,


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The geometric mean is plotted at the first sampling date ofa 30-day period. Plotted points on the abscissa indicateless than five samples taken over a 30-day period and showthe sampling dates.

10JJi JOJL AUd SEP OCT N0V DEC JAN FEb MAR APR MAY

1971 1972

FIG. 5. Geometric means of fecal coliforms densities in the Spoon River, 1971 through 1972.

JU'4 JUL 0G SEP OUT 'Jo. DJC JAI, FEB CAR APR MAY1972 1973

FIG. 6. Geometric means of fecal coliforms densities in the Spoon River, 1972 through 1973.

as measured by the geometric mean, occurred atsome stations during the period December 1971through March 1972. Ice cover prevailed duringthe months of January and February.

It is apparent that satisfactory bacterial qual-ity was not achieved at any of the five stationsduring the 12-month period, June 1972 to May1973 (Fig. 6). As mentioned earlier, the bacte-rial quality observed during the second year ofthe study was substantially worse than thatfound for the first year of the study. This isprobably due to the above-normal precipitationevents resulting in excessive rural and urbanrunoff during 1972 through 1973, compounded

by the necessity to bypass sewage treatmentfacilities at frequent intervals.A comparison of the observed FC densities

with water quality requirements is summarizedin Table 3. There were periods of compliancewith the IPCB rule (Fig. 5). However, on anoverall basis (Table 3), only in about one ofevery five periods of 30 days was a geometricmean of 200/100 ml or less achieved; and onlyabout one period in 12 periods of 30 days metthe conditions of the rule pertaining to FCdensities not exceeding 400/100 ml for morethan 10% of the samples.

It would appear that the portion of the rule

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TABLE 3. Evaluation of fecal coliforms densities of the Spoon River during June 1971 May 1972with IPCB Rule 203(g)

Observed data Data adjusted for ommissions

Compliance with Compliance with 10% Compliance with IPCBSamping Total no. of geometric mean of samples during Rule 2

station (MP) 30-day periods (<200/100 ml) 30 days ( <400/100 ml) Total no. of Rule 203g)evaluated 30-day periods.

No. of Percentage No. of Percentage evaluated No. of Percentage30-day of 30-day 30-day of 30-day 30-day of 30-dayperiods periods periods periods periods periods

141.2 44 4 9.1 1 2.2 49 1 2.0113.7 48 4 8.3 3 6.3 49 3 6.168.4 45 11 24.4 8 17.8 49 8 16.337.7 44 13 29.5 2 4.5 49 2 4.12.5 39 10 25.6 4 10.3 49 6 12.2

Overall 220 42 19.1 18 8.2 245 20 8.2

limiting geometric mean densities is not thegoverning factor in assessing the bacterial qual-ity of the Spoon River. Rather the limitingfactor is that portion of the requirementwhereby no more than 10% of the samples shallexceed 400/100 ml. Whether or not this is thecase for other Illinois streams remains to bedetermined.FC/TC ratio. The historical record of bacte-

rial examinations in Illinois streams is com-

posed principally of total coliform data. Itseemed worthwhile, therefore, to determine FC/TC ratios with the thought in mind that a FCbaseline record might, at sometime, be derivedfrom the TC record of previous years.

We used the arithmetic mean in evaluatingthe FC/TC ratios. The ratios developed for thefive stations during the period June 1971 to May1973 are summarized in Table 4. The ratiosvaried considerably, ranging from 0.0001 to0.567 with a 2-year overall average of 0.095. Inother words, 9.5% of the TC consisted of FC.This overall average for the Spoon River ishigher than that observed on the Upper IllinoisWaterway (7) (8.8%) and lower than that re-

ported for the Ohio River (9) (14.0%). The range

of the ratios for the Spoon River was greaterthan that of the Upper Illinois Waterway (0.002to 0.38) and the Ohio River (0.004 to 0.45).The monthly average ratios for each of the

five sampling stations are depicted in Fig. 7.The period of relatively low ratios occurredduring January through March 1972 and No-vember 1972 through May 1973. To produce lowratios, either the FC densities must decrease or

the TC densities must increase. During the twoperiods of low ratios, the FC densities did notsignificantly decrease (Fig. 5 and 6). On theother hand, TC densities did increase signifi-cantly during these periods (Fig. 3). It can be

TABLE 4. Statistical results of FC/TC values of theSpoon River

(SMP Study period Range M. b

141.2 6/1971-5/1972 0.002-0.530 0.115 0.1306/1972-5/1973 0.001-0.300 0.064 0.063Two-year 0.093 0.109

113.7 6/1971-5/1972 0.004-0.391 0.113 0.1036/1972-5/1973 0.001-0.473 0.054 0.065Two-year 0.084 0.088

68.4 6/1971-5/1972 0.003-0.567 0.143 0.1396/1972-5/1973 0.001-0.451 0.071 0.086Two-year 0.108 0.118

37.7 6/1971-5/1972 0.001-0.400 0.131 0.1176/1972-5/1973 0.0001-0.295 0.060 0.073Two-year 0.096 0.104

2.5 6/1971-5/1972 0.002-0.429 0.110 0.1066/1972-5/1973 0.001-0.188 0.057 0.055Two-year 0.083 0.089

a M, Arithmetic mean.b a, Standard deviation.

concluded that the lower ratios on the SpoonRiver are due to increasing TC densities ratherthan decreasing FC densities.

Strobel (11) reported that the relationshipbetween FC and TC varied with the sources ofpollution, level of sewage treatment, character-istics of the receiving waters, and precipitationon the watershed. Since so many factors mayinfluence the overall value of FC/TC ratios, itwould seem unwise to rely on overall averagevalues based upon a year or more of observa-tion. In fact it would be preferable to limitjudgment to only those ratio values obtainedduring stable stream flow conditions. However,on the basis of the results shown in Fig. 7, themonthly or seasonal mean might be useful.The ORSANCO Water Users Committee (9)

suggests that higher FC/TC ratios might indi-



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J J A S O N D J F M A M J J A S O N D J F M A M1971 1972 1973

FIG. 7. Monthly average of FC/TC values in the Spoon River.

cate the proximity of inefficient wastewatertreatment operations or conditions where treat-ment facilities are being bypassed during stormrunoff. Low ratios (<0.20) are most likelycaused by aftergrowths of Aerobacter aerogenesresulting in abnormally high TC densities (9).In the Spoon River, 437 samples of 494 (88.5%)produced FC/TC ratios less than 0.20. This isindicative of A. aerogenes aftergrowths in thestream.FCFS ratio. The use of FS in conjunction

with FC was first suggested by Geldreich et al.(10). They felt the use of a ratio, FC/FS, wouldbe a more valuable informational tool for assess-ing pollution sources than relying solely on FCdensities. In applying the FC/FS ratio to anatural stream system, best results are obtainedif the stream samples are collected within a 24-hstream flow time of a pollution source.From a series of studies (2, 4), it was deter-

mined that ratios greater than 4 were indicativeof a pollution source primarily of human originsuch as domestic wastewater, whereas ratiosless than 0.7 suggest the likelihood of thepollution source to be that of waste from warm-blooded animals other than humans, i.e., live-stock and poultry wastes. Table 5 summarizesthe relationship of a range of FC/FS ratio valuesto their respective probable origins (2, 4, 8).

Determinations for FS densities were limitedto the second year of the study which coveredthe period June 1972 through May 1973. Thiswas a very wet year with excessive variability instream flows and the computed ratios reflectedthis variability. The low value of 0.18 occurredat the Havana station; a high of 27.8 occurred atthe Modena station. Both of these extremesoccurred on 27 December 1972.There was no distinguishable pattern in aver-

TABLE 5. Relationship between FC/FS values andpollution sources

FC/FS = X Indicative source of pollutionrange

X > 4 Human wastes4 > X > 2 Predominance of human wastes in

mixed pollution2 > X > 1 Uncertain of interpretation

1 > X > 0.7 Predominance of animal wastes inmixed pollution

0.7 . X Livestock or poultry wastes

age monthly values and thus the yearly averagevalues were meaningless. In an effort to evalu-ate the FC/FS ratios for comparison with therelationships set forth in Table 5, a cumulativefrequency diagram was prepared (Fig. 8). Areview of the figure suggests that the percent-age of fecal bacteria originating from animalwastes other than human increased with down-stream movements. This increase in occurrenceof fecal bacteria from animal wastes is apparentfrom the values set forth in Table 6, if the sum-mation of columns 1 and 2 are compared withthe summation of columns 4 and 5. It is quiteapparent from Table 6 that fecal bacteria in theupper reaches of the Spoon River originatedprimarily from humans during the period ofstudy; from London Mills downstream, the shiftin origin is. toward other animal wastes with asignificant change of bacterial origin occurringin the stretch of the stream between Seville andHavana. The data emphasize the importance ofdefining nonpoint sources of pollution in addi-tion to point sources when assessing the bacte-rial quality of streams.We draw the following conclusions from our

data.

VOL. 28, 1974 295

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FIG. 8. Cumulative frequency for FC/F'

TABLE 6. Percentage of time fecal bactvarious sources in the Spoon River ox

Station IndeterPredonHuman nantly - nantli(MP) huan inate anmhuman animal

141.2 17 25 35 8113.7 9 30 35 1768.4 8 23 39 1437.7 9 18 39 182.5 6 27 24 14

(i) Bacteria densities, although va

domly in time, exhibited a log-normation at each station permitting ti

tendencies and dispersion of the dexpressed in geometric terms.

(ii) Bacteria densities, in general,with downstream movement.

(iii) During periods of high Illixstage, the bacterial quality of the loiRiver was influenced by the Illinois I

(iv) From comparing fecal coliform.with the bacteria requirements of lacceptable bacterial quality in the SIwas achieved about 8.3% of the time di1971 through May 1972. During the pa1972 through May 1973, satisfactoryquality was not achieved at any o

sampling stations during periods of s

(v) Since compliance with the I

achieved about 20% of the time dtthrough 1972 under that portion o:

lp-- referring to a geometric mean limitation of 200FC/100 ml, and only 8.3% of the time whenconsidering the conditions of the rule pertainingto not exceeding bacterial densities of 400FC/100 ml for more than 10% of the samples, itwould appear that the latter condition is thegoverning factor in assessing the bacterial qual-ity of the Spoon River.

(vi) To derive meaningful baseline fecal coli-form data from historical records of total coli-form measurements by current FC/TC ratios, itwould be preferable to limit the data used to

1.2 that obtained from stable stream flow condi-8.4 tions. However, monthly or seasonal values may2. 5 - be useful with proper judgment.

(vii) Although bacterial densities decreasedwith downstream movement, the decrease in

4 0 total coliform was less dramatic because of theoccurrence of A. aerogenes aftergrowths in the

S values, stream.(viii) The use of FC/FS ratios indicated that

fecal bacteria in the upper reaches of the Spooneria from River originated primarily from human wastes;ccurred from London Mills downstream the originnim shifted to animal wastes with a significant

y Animal change occurring between Seville and Havana.al (ix) The FC/FS ratios emphasizes the need to-

5consider nonpoint as well as point sources of

19 pollution in assessing the bacterial quality of

16 streams.1629

ACKNOWLEDGMENTS

We express our appreciation to all the staff of the WaterQuality Section for their advice and assistance during this

trying ran-phase of the Spoon River study, and especially to Dennis

rying ran- Dooley of the Natural History Survey and Jack Williams who1 distribu- performed much of the sampling, Pamella Martin for herhe central assistance in bacterial determinations, and John Brother, Jr.,lata to be who prepared the illustrations.

decreased LITERATURE CITED

1. American Public Health Association et al. 1971. Stan-no0s River dard methods for the examination of water and waste-wer Spoon water, 13th ed. American Public Health Association,River. Inc., New York.densities 2. Geldreich, E. E. 1967. Fecal coliform concents in streampollution. Water Sewage Works 114:R98-R109.

the IPCB, 3. Geldreich, E. E., H. F. Clark, and C. B. Huff. 1964. Apoon River study of pollution indicators in a waste stabilizationuring June pond. J. Water Poll. Cont. Fed. 36:1372-1379.eriod June 4. Geldreich, E. E., and B. A. Kenner. 1969. Concept of fecalstreptococci in stream pollution. J. Water Poll. Cont.bacterial Fed. 41:R336-R352.

of the five 5. Lin, S. D. 1973. Evaluation of coliform tests for chlorin-,ampling. ated secondary effluents. J. Water Poll. Cont. Fed.[PCB was 6. 45:498-506.

6 Lin, S. D. 1974. Evaluation of fecal streptococci tests foriring 1971 chlorinated secondary sewage effluents. J. Environ.f the rule Eng. Div. Amer. Soc. Civil Eng. 100:253-267.

A MP 14o MP 11A MP 6D MP 3* MP



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VOL. 28, 1974 BACTERIOLOGICAL ASSESSMENT OF SPOON RIVER 297

7. Lin, S. D., and R. L. Evans. 1974. An analysis of coliform bacterial quality. J. Water Poll. Cont. Fed. 43:630-640.bacteria in the Upper Illinois Waterway. Water Res. 10. State of Illinois, Environmental Protection Agency. 1972.Bull., vol. 10. Water pollution regulations of Illinois. State of Illinois,

8. Millipore Corporation. 1972. Biological analysis of water Springfield.and wastewater. Application manual AM 302. Mil- 11. Strobel, G. A. 1968. Coliform-fecal coliform bacteria inlipore Corp., Bedford, Mass. tidal waters. J. Sanit. Eng. Div. Amer. Soc. Civil Eng.

9. ORSANCO Water Users Committee. 1971. Total coli- 94:641-656.form: fecal coliform ratio for evaluation of raw water


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bacteriological assessment of spoon river water qualityspoon river in the vicinity of havana was...

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