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MMS 97-0024

Chronic Toxicological Effects of Produced Water on Reproduction and Development in Marine Organisms

Final Technical Summary Final Study Report

U.S. Department of the Interior Minerals Management Service Pacific OCS Region

Chronic Toxicological Effects of Produced Water on Reproduction and Development in Marine Organisms

Final Technical Summary Final Technical Report

Authors

Gary N. Cherr Teresa W-M. Fan Principal Investigors

Prepared under MMS Cooperative Agreement No. 14-35-0001-30471 by Southern California Educational Intiative Marine Science Institute University of California Santa Barbara, CA 93106

U.S. Department of the Interior Minerals Management Service Camarillo Pacific OCS Region January 1997

Disclaimer

This report has been reviewed by the Pacific Outer Continental Shelf Region, Minerals Management Service, U.S. Department of the Interior and approved for publication. The opinions, findings, conclusions, or recommendations in this report are those of the author, and do not necessarily reflect the views and policies of the Minerals Management Service. Mention of trade names or commercial products does not constitute an endorsement or recommendation for use. This report has not been edited for conformity with Minerals Management Service editorial standards.

Availability of Report

Extra copies of the report may be obtained from: U.S. Dept. of the Interior

Minerals Management Service Pacific OCS Region 770 Paseo Camarillo

Camarillo, CA 93010 phone: 805-389-7621

A PDF file of this report is available at:

http://128.111.226.115/SCEI/

Suggested Citation

The suggested citation for this report is: G.N. Cherr and T. W-M. Fan. Chronic Toxicological Effects of Produced Water on Reproduction and Development in Marine Organisms. MMS OCS Study 97-0024. Coastal Research Center, Marine Science Institute, University of California, Santa Barbara, California. MMS Cooperative Agreement Number 14-35-0001-30471. 71 pages.

Table of Contents

FINAL TECHNICAL SUMMARY ....................................................................... 1 FINAL TECHNICAL REPORT............................................................................ 5 Background .............................................................................................................. 5 Objectives ................................................................................................................. 5 Effects of produced water and its toxic constituents on developing systems...... 5 Barium uptake by mussels in the field and laboratory......................................... 8 References ................................................................................................................ 9 Appendix I Inhibition of Cellular Events During Early Algal Gametophyte Development: Effects of Select Metals and an Aqueous Petroleum Waste.......................... 11 Appendix II Nuclear Events During Early Development in Gametophytes of Macrocystis pyrifera, and the Temoporal Effects of a Marine Contaminant .................... 31 Appendix III Developmental Effects of Barium Exposure in a Marine Bivalve (Mytilus californianus) .................................................................................. 41 Appendix IV Developmental Abnormalities and DNA-Protein Crosslinks in Sea Urchin Embryos Exposed to Three Metals................................................................ 49 Appendix V Barium Concentrations in Mussel Shells and Correlation with Distance from Outfall and Performance....................................................................... 71

Final Technical Summary – Cherr and Fan

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FINAL TECHNICAL SUMMARY

STUDY TITLE: Chronic Toxicological Effects of Produced Water on Reproduction and Development in Marine Organisms

REPORT TITLE: Chronic Toxicological Effects of Produced Water on Reproduction and Development in Marine Organisms - Final Report.

CONTRACT NUMBER: 14-35-0001-30471

SPONSORING OCS REGION: Pacific

APPLICABLE PLANNING AREA: Southern and Central California

FISCAL YEAR(S) OF PROJECT FUNDING: FY92, FY93

COMPLETION DATE OF THE REPORT: December 1996

COST(S): FY 1992-1993: $53,000; FY 1993-1994: $82,242

CUMULATIVE PROJECT COST: $135,242

PROGRAM MANAGER: Russell J. Schmitt

PRINCIPAL INVESTIGATORS: 1G.N. Cherr and 2T. W.-M. Fan

AFFILIATION: 1Bodega Marine Laboratory and 2Dept. Land, Air & Water Resources

ADDRESS: 1P.O. Box 247, Bodega Bay, CA 94923 and 2University of California at Davis, Davis, CA 95616

KEY WORDS: Santa Barbara Channel; produced water; chronic toxicity; embryo development; marine organsisms; barium; biomarkers; contaminant effects; toxic mechanisms; invertebrate toxicology; macroalgal toxicology

BACKGROUND: The largest discharge associated with offshore oil and gas production is produced water. In the Santa Barbara Channel, produced water is discharged into the coastal environment in 10-15 meters of water. As long-term effects on reproduction and development of marine organisms can often be subtle, it is difficult to assign cause-and-effect relationships amid the background of natural variability, particularly in an environment such as the Santa Barbara Channel where multiple complicating factors are present. Such high level of variations limits our ability to detect subtle impacts of contaminants. However, if the mechanism of action of toxicants under controlled laboratory conditions are better understood, the observed pattern of bioeffects in the natural environment can be more directly linked with specific contaminant(s). Such a marriage of laboratory and field studies must take place in order to achieve scientifically valid information that is also environmentally relevant. Among the myriad of effects that can be measured, gametogenesis and embryonic development are catagories that deserve special attention because of their relevance to effects at the population level. A mechanistic understanding of contaminant effects on cellular functioning during gametogenesis and embryonic development are generally unclear; understanding these is required for linking laboratory bieffects with field observations. Biological field surveys alone may not detect the cause of population changes in polluted environments due to the presence

Chronic Toxicological Effects of Produced Water

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of other complicating factors. It has been suggested that the most promising approach would be to relate molecular and cellular alterations to individual-level physiological changes that lead to perturbations of reproduction and embryonic development. These effects are then relatable to potential impacts at the population level.

Previously we have shown that the divalent cation fraction of PW is responsible for the toxicity to developing mollusc embryos and larvae (Higashi et al., 1992). This fraction primarily contains barium and strontium, and based on concentrations found, that barium is likely the cause of the toxicity. In addition, preliminary data suggested that microtubule-based processes in cells (which can be perturbed by divalent cations) may be impacted by PW.

OBJECTIVES: There were two major objectives in the proposed project: l) define the effects of PW and its constituents on early embryonic development in several coastal marine organisms with the purpose of exploring a common cellular mechanism for these effects; 2) determine, in parallel, the effects of PW on adult mussels (including (gametogenesis and subsequent development of embryos from adult mussels) outplanted at the PW discharge site and from adult mussels exposed to PW and its toxic constituents in the laboratory.

DESCRIPTION: The effect of both PW and the PW water-soluble fraction (WSF) have been investigated (Garman et al., 1994a), and were assessed for their ability to affect gametophytic development. Neither PW or PW-WSF exhibited any significant effect on germination. Germ tube growth was only slightly effected by both, with the PW-WSF being somewhat less effective. However, both PW and PW-WSF inhibited nuclear translocation in developing gametophytes, indicating that the major effect of PW and its toxic constituents are on the microtubule-dependent developmental events. It should be noted that kelp gametophytes are not particularly sensitive to PW, however, the response observed (inhibition of nuclear translocation) has not been reported for other toxicants; in fact, germination and germ tube growth are the common endpoints used to assess toxicity, and are usually perturbed by both simple and complex toxicants investigated to date. We also found that arsenic, a metal constituent common to a number of different PWs also inhibited nuclear events. Further investigations have shown that not only does arsenic specifically inhibit the microtubule-mediated nuclear division and translocation, but it also affects DNA synthesis prior to division (Garman et al., 1994b). The latter was determined quantitatively using single-cell microspectrofluorometry. Experiments were conducted to determine if PW perturbs mussel (Mytilus californianus) embryo development in a stage-specific manner (Spangenberg and Cherr, 1996). PW does not appear to have any toxic effect on embryos prior to hatching (15 hrs.), but does perturb later development at relatively low concentrations. This was determined by pulse-chase experiments in which zygotes were "pulsed" with PW and cultured for 15 hrs., and then "chased" by washing in fresh seawater. The embryos exposed continually exhibited abnormalities between the 1 % and 2% concentrations. This would suggest that early cleavage stages are not affected, while gastrulation and cell differentiation are. The basis for stage-specific toxicant effects was examined using sea urchin embryos exposed to select metals, and the basis may include the induction of persistent DNA-protein crosslinks which result in delayed effects during later stages of differentiation (Garman et al., 1996).

Final Technical Summary – Cherr and Fan

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In a mussel outplant study at Carpinteria, we found that the barium load in mussel tissues (an average of 10 individuals) was correlated with the distance from the outfall of PW. The mussels outplanted at the 1 m site contained a much higher level of Ba in their tissues than those from other sites farther away from the outfall (see inset of Figure 6). It is interesting to note that the growth and reproductive performance of the 1-5 m site mussels was also poorer. These findings suggest that barium in the PW may be accumulated in mussel tissues, thereby affecting their growth and reproductive development. It should also be noted that the barium accumulation dropped off quickly with distance from the outfall. The barium effects observed in adult mussels in the field, together with the presence of barium in the toxic fraction of PW (Higashi et al. 1993) predicate our interest to further examine the relationship between barium and PW bioeffects. It is essential to account for the apparent discrepancy that barium concentration in natural seawater is kept low (i.e. at ppb level) due to precipitation by sulfate. Therefore, a direct uptake from seawater by mussels may not be a dominant mechanism. However, it is possible that when barium concentration is below certain values (e.g. 1-200 ppb), it may remain soluble or suspended even in the presence of high sulfate concentrations. There is an alternative route to passive uptake into mussels, and that is via marine phytoplankton. It is generally recognized that toxicants accumulation can be amplified through the food chain and barium has been shown to be accumulated in phytoplankton (Martin and Knauer, 1973). Laboratory experiments exposed mussels to barium through a common coastal phytoplankton Skeletonema costatum and through seawater only. At the end of exposure, we measured barium load in these mussels and compared this information with assessment on gonadal (Fan et al., 1993; Fan and Lane, 1992) and embryo development. We also investigated how barium impacts coastal primary production as it is both an environmentally important question and directly relatable to the mussel bioeffects. We have developed a convenient algal microassay to test the effect of barium on phytoplankton growth. It is interesting to note that barium effect on S. costatum growth was complex; it was slightly stimulatory at 28 ppb while somewhat inhibitory at 113 ppb. It should also be noted that this range of barium concentration was comparable to that found at the Carpinteria site. At higher concentrations, the effect was diminished, similar in trend to that observed for the mussel embryo, perhaps as a result of barium aggregation or precipitation.

STUDY PRODUCTS: Cherr, G.N., G.D. Garman, R.M. Higashi, M.C. Pillai, and T.W-M. Fan. 1993. Effects of

produced water on reproduction and development in marine organisms. Proc. of 14th annual Society of Environmental Toxicology and Chemistry Meeting. p127.

Garman, G.D., Cherr, G.N., and Pillai, M.C. 1992. Toxicant inhibition of gametophyte development in the giant kelp, Macrocystis pyrifera. Proc. of 6th Annual Research Symposium, UC Toxic Substances Research and Teaching Program, pp. 23.

Garman, G.D., Cherr, G.N., and Pillai, M.C. 1993. Toxicant inhibition of gametophyte development in the giant kelp, Macrocystis pyrifera. 73rd Annual Meeting of the Western Society of Naturalists. p. 16.


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Garman, G.D., M.C. Pillai, and G.N. Cherr. 1994a. Inhibition of cellular events during algal gametophyte development: Effects of select metals and an aqueous petroleum waste. Aquatic Toxicology 28:127-144.

Garman, G.D., M.C. Pillai, L.J. Goff, and G.N. Cherr. 1994b. Nuclear events during early development in Macrocystis pyrifera gametophytes and the temporal effects of a marine contaminant. Marine Biology 121:355-362.

Garman, G.D., Cherr, G.N., and S.L. Anderson. 1994. Induction of DNA-protein crosslinks in developing embryos of the purple sea urchin, Strongylocentrotus purpuratus. Proc. of 15th annual Society of Environmental Toxicology and Chemistry Meeting, Denver, CO, p.69.

Garman, G.D., S.L. Anderson, and G.N. Cherr. 1997. Developmental abnormalities and DNA-protein crosslink induction in sea urchin embryos exposed to metals. Aquatic Toxicology 39:247-265.

Final Study Report – Cherr and Fan

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FINAL STUDY REPORT

BACKGROUND

The largest discharge associated with offshore oil and gas production is produced water. In the Santa Barbara Channel, produced water is discharged into the coastal environment in 10-15 meters of water. As long-term effects on reproduction and development of marine organisms can often be subtle, it is difficult to assign cause-and-effect relationships amid the background of natural variability, particularly in an environment such as the Santa Barbara Channel where multiple complicating factors are present. Such high level of variations limits our ability to detect subtle impacts of contaminants. However, if the mechanism of action of toxicants under controlled laboratory conditions are better understood, the observed pattern of bioeffects in the natural environment can be more directly linked with specific contaminant(s). Such a marriage of laboratory and field studies must take place in order to achieve scientifically valid information that is also environmentally relevant. Among the myriad of effects that can be measured, gametogenesis and embryonic development are categories that deserve special attention because of their relevance to effects at the population level. A mechanistic understanding of contaminant effects on cellular functioning during gametogenesis and embryonic development are generally unclear; understanding these is required for linking laboratory bioeffects with field observations. Biological field surveys alone may not detect the cause of population changes in polluted environments due to the presence of other complicating factors. It has been suggested that the most promising approach would be to relate molecular and cellular alterations to individual-level physiological changes that lead to perturbations of reproduction and embryonic development. These effects are then relatable to potential impacts at the population level. Previously we have shown that the divalent cation fraction of PW is responsible for the toxicity to developing mollusc embryos and larvae (Higashi et al., 1992). This fraction primarily contains barium and strontium, and based on concentrations found, that barium is likely the cause of the toxicity. In addition, preliminary data suggested that microtubule-based processes in cells (which can be perturbed by divalent cations) may be impacted by PW. OBJECTIVES

There were two major objectives in the proposed project:') define the effects of PW and its constituents on early embryonic development in several coastal marine organisms with the purpose of exploring a common cellular mechanism for these effects; 2) determine, in parallel, the effects of PW on adult mussels (including (gametogenesis and subsequent development of embryos from adult mussels) outplanted at the PW discharge site and from adult mussels exposed to PW and its toxic constituents in the laboratory. Effects of PW and its Toxic Constituents on Developing Systems We have been investigating the stage-specific effects of PW on kelp gametophyte development and mussel embryo development. M. pyrifera reproduction involves the release of haploid zoospores from the adult plant, and these motile cells settle onto a substrate, initiate germination within 4-5 hours, and produce a 12-15 pin long germ tube; these events have been


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recently described by us as part of a previous SCEI supported project (Pillai et al., 1992). Coincident with germ tube formation, there is a cytoplasmic streaming and organelle translocation along the tube. Shortly after completion of the germ tube growth, the zoospore nucleus undergoes a division and, one of the daughter nuclei is then immediately translocated to the distal end of the germ tube. Following this nuclear translocation, the first gametophytic cross wall is formed and, subsequently, the daughter nucleus remaining in the original zoospore body undergoes repositioning, assuming a position within the germ tube near the cross wall. Developmental events in M. pyrifera, which occur prior to differentiation into male or female gametophytes, are temporally and mechanistically distinct (Pillai et al., 1992). During germination of these microscopic gametophytes, actin dynamics are critical for normal germ tube growth, while microtubules are involved in subsequent nuclear division and translocation. It appears that intracellular allWinization occurs in the germ tube during actin-mediated germination based on singlecell intracellular pH measurements, while acidification occurs during microtubule-based nuclear events. The effect of both PW and the PW water-soluble fraction (WSF) have been investigated (Garman et al., 1994a; see Appendix 1), and were assessed for their ability to affect gametophytic development. Neither PW or PW-WSF exhibited any significant effect on germination. Germ tube growth was only slightly effected by both, with the PW-WSF being somewhat less effective. However, both PW and PW-WSF inhibited nuclear translocation in developing gametophytes, indicating that the major effect of PW and its toxic constituents are on the microtubule-dependent developmental events. It should be noted that kelp gametophytes are not particularly sensitive to PW, however, the response observed (inhibition of nuclear translocation) has not been reported for other toxicants; in fact, germination and germ tube growth are the common endpoints used to assess toxicity, and are usually perturbed by both simple and complex toxicants investigated to date (Anderson and Hunt, 1988). The time course of DNA synthesis in developing haploid gametophytes of the giant kelp Macrocystis pyrifera was determined, and the effects of arsenic (As) on the temporally distinct nuclear events, DNA synthesis and subsequent nuclear division/translocation, were investigated to establish which of these specific events may be disrupted by this contaminant (Garman et al., 1994b; see Appendix 2). Experiments were carried out on material collected from kelp beds near Santa Barbara, California from 1993-1994. Timing of DNA synthesis was determined during development by use of the fluorochrome, DAPI (4',6-diamidino-2-phenylindole), and single-cell microspectrofluorometry. Zoospores, which result from meiosis, had already undergone two rounds of DNA synthesis at the time of release. The developing gametophytes underwent 16 rounds of replication of DNA by 16 h of development, and following the first nuclear division/translocation, the gametophyte contained 8 times the minimum DNA level throughout subsequent development. Both DNA synthesis and nuclear division/translocation, were found to be inhibited by As. Phosphate enrichment reduced the inhibitory effects of As on division/translocation of the nucleus, supporting the hypothesis that As interferes with phosphorylation. Gametophytes were more severely affected by As under light conditions, as opposed to dark, suggesting that photosynthesis may be more sensitive than dark metabolism.


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Experiments were conducted to determine if PW perturbs mussel (Mytilus califomianus) embryo development in a stage-specific manner (Spangenberg and Cherr, 1996; see Appendix 3). Produced waters contain variable mixtures of organic and inorganic chemicals, and are often high in the divalent cations barium (Ba) and strontium (Sr). In order to assess toxicity of Ba and Sr in sea water, mussel embryos (Mytilus californianus) were exposed to barium agate and strontium chloride by static exposure from fertilization through development of veliger larvae. Only Ba exhibited bioactivity at environmentally relevant levels. Ba exhibited teratogenicity between 200-900 pg/L (ppb), with higher concentrations associated with decreased toxicity and apparent precipitation of Ba from sea water. Barium concentrations between 100 and 900 pg/L were stable in filtered sea water over two weeks based on analysis of the soluble Ba concentration by inductively coupled plasma emission spectroscopy. Adverse effects on embryos included abnormal shell calcification and embryo morphology. Exposure of embryos to Ba in stage-specific, pulsechase experiments demonstrated that although specific developmental stages exhibited similar types of abnormalities, they were differentially affected. Gastrula were the most sensitive, while exposure of blastula and trochophore larvae to 500 pg/L Ba did not result in obvious abnormalities. Teratogenic effects observed in embryos exposed during the gastrula stage were not reversible despite washing and completion of culture through the veliger stage in clean sea water. These findings are the first to demonstrate that 200-900 pg/L Ba in seawater can be teratogenic, and is of potential concern in the marine environment. Furthermore, these results corroborate findings of previous studies which indicated that Ba is primarily responsible for produced water toxicity to developing marine embryos. The basis for stage-specific toxicant effects was examined using sea urchin embryos exposed to select metals, and the basis may include the induction of persistent DNA-protein crosslinks which result in delayed effects during later stages of differentiation (Garman et al., 1996; see appendix 4). Two sublethal responses were used to investigate the effects of genotoxic metals on embryos of the purple sea urchin, Strongylocentrotus purpuratus. In addition to the standard measurement of developmental success, we used a novel genotoxic response, DNA-protein crosslink (DPC) induction, to assess the effects of embryo exposure to pentavalent arsenate (As), nickel (Ni) and hexavalent chromate (Cr). The procedure for sea urchin embryo DPC measurement was adapted from a mammalian cell assay using potassium-SDS precipitation and a DNA fluorochrome to quantify relative amounts of free and protein-bound DNA. Developmental abnormality and DPCs increased in a dose-dependent manner after a 48-h exposure to each of the three metals. Lowest observable effect concentrations (LOECs) for development were 0.011 mg/L As, 0.40 mg/L Ni, and 2.5 mg/1- Cr. LOECs calculated for the DPC response to these same three metals were 0.023 mg/L, 8.0 mg/L, and 10.0 mg/L, respectively. DPCs were transiently high in control embryos through the blastula stage, which is prior to the dramatic increase in transcription of the embryonic genome. By the gastrula and prism stages (subsequent to embryo gene transcription) there was a significant decrease in DPCs. Ni-exposed embryos exhibited the greatest magnitude of adverse effect in embryos exposed through the blastula stage, as compared to those exposed from blastula through late gastrula stage. We hypothesize that stage-sensitivity to Ni in sea urchin embryos may be related to the induction of persistent DPCs, and the prevention of normal transcription of the embryonic genome.


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Initial studies on the cellular effects of PW have focused on kelp gametophyte development. This system is excellent for dissecting mechanisms of toxicity due to the distinct cellular events which occur during development (Pillai et al., 1992). In addition, the determination of cellular and subcellular effects of PW, and its constituents, on mollusc embryos required the stage-specific determinations described above prior to initiation of these studies. Initial studies with kelp gametophytes have shown that the PW and PW-WSF both effect nuclear translocation, a microtubule-mediated developmental event (Pillai et al., 1992). This response, and the lack of response for germination and germ tube growth, are similar as those observed for the microtubule-specific inhibitors colchicine and amiprophys methyl (Pillai et al., 1992). The distribution of microtubules in control and PW-exposed gametophytes at the time of nuclear translocation was investigated using a monoclonal antibody to tubulin. In controls, microtubules were observed along the entire germ tube as translocation of the daughter nucleus occurred. These microtubules form the leading edge of the daughter nucleus. In PW exposed gametophytes, the only staining of microtubules was observed at the base of the original zoospore body; microtubules were not observed within the germ tube or at the leading edge of the nuclei. Barium Uptake by Mussels in the Field and Laboratory In a mussel outplant study at Carpinteria (in collaboration with C. Osenberg), we found that the barium load in mussel shell (an average of 10 individuals) was correlated with the distance from the outfall of PW (Appendix 5). The mussels outplanted at the 1 m site contained a much higher level of Ba in their tissues than those from other sites farther away from the outfall. It is interesting to note that the growth and reproductive performance of the 1-5 m site mussels was also poorer. These findings suggest that barium in the PW may be accumulated in mussel tissues, thereby affecting their growth and reproductive development. It should also be noted that the barium accumulation dropped off quickly with distance from the outfall. The barium effects observed in adult mussels in the field, together with the presence of barium in the toxic fraction of PW (Higashi et al., 1992) predicate our interest to further examine the relationship between barium and PW bioeffects. It is essential to account for the apparent discrepancy that barium concentration in natural seawater is kept low (i.e., at ppb level) due to precipitation by sulfate. Therefore, a direct uptake from seawater by mussels may not be a dominant mechanism. However, it is possible that when barium concentration is below certain values (e.g. 1-200 ppb), it may remain soluble or suspended even in the presence of high sulfate concentrations. There is an alternative route to passive uptake into mussels, and that is via marine phytoplankton. It is generally recognized that toxicants accumulation can be amplified through the food chain and barium has been shown to be accumulated in phytoplankton (Martin and Knauer, 1973). Laboratory experiments exposed mussels to barium through a common coastal phytoplankton Skeletonema costatum and through seawater only. At the end of exposure, we measured barium load in these mussels and compared this information with assessment on gonadal (Fan et al., 1993; Fan and Lane, 1992) and embryo development. We also investigated how barium


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impacts coastal primary production as it is both an environmentally important question and directly relatable to the mussel bioeffects. We have developed a convenient algal microassay to test the effect of barium on phytoplankton growth. It is interesting to note that barium effect on S. costatum growth was complex; it was slightly stimulatory at 28 ppb while somewhat inhibitory at 113 ppb. It should also be noted that this range of barium concentration was comparable to that found at the Carpinteria site. At higher concentrations, the effect was diminished, similar in trend to that observed for the mussel embryo, perhaps as a result of barium aggregation or precipitation. These results are summarized in Appendix 6. REFERENCES: Anderson, B.S. and Hunt, J.W. 1988. Bioassay methods for evaluating the toxicity of heavy metals, biocides and

sewage effluent using microscopic stages of giant kelp Macrocystis pyrifera (Agardh): A preliminary report. Mar. Environ. Res. 26:113134.

Cherr, G.N., G.D. Garman, R.M. Higashi, M.C. Pillai, and T.W-M. Fan. 1993. Effects of produced water on reproduction and development in marine organisms. Proc. of 14th annual Society of Environmental Toxicology and Chemistry Meeting. p 127.

Garman, G.D., M.C. Pillai, and G.N. Cherr. 1994a. Inhibition of cellular events during algal gametophyte development: Effects of select metals and an aqueous petroleum waste. Aquatic Toxicology 28:127-144.

Garman, G.D., M.C. Pillai, L.J. Goff, and G.N. Cherr. 1994b. Nuclear events during early development in Macrocystis pyrifera gametophytes and the temporal effects of a marine contaminant. Marine Biology 121:355-362.

Garman, G.D., S.L. Anderson, and G.N. Cherr. 1997. Developmental abnormalities and DNA-protein crosslink induction in sea urchin embryos exposed to metals. Aquatic Toxicology 39:247-265

Higashi, R.M., G.N. Cherr, C.A. Bergens, T.W-M. Fan, and D.G. Crosby. 1992. Toxicant isolation from a produced water source in the Santa Barbara Channel. Pages 223-233. In: Produced Water: Technological/Environmental Issues and Solutions, J.P. Ray, ed., Plenum Publishing, New York.

Krause, P. R. 1994. Effects of an oil production effluent on gametogenesis and gamete performance in the Purple sea urchin (Strongylocentrotus purpuratus Stimpson). Environ. Toxicol. Chem., 13(7):1153-1161.

Neff, J.M. 1993. Barium in produced water: Is it a toxicological hazard to the marine environment? Proceedings, 14th annual SETAC meeting, Houston, TX, USA, 11/93, p. 131.

Osenberg, C.W., Schmitt, R.J., Holbrook, S.J., 1992. Spatial scale of ecological effects associated with an open coast discharge of produced water. Pages 387-402. In: Produced Water: Technological / Environmentallssues and Solutions, J.P. Ray, ed., Plenum Publishing, New York.

Pillai, M.C. , Baldwin, J.D. and Cherr, G.N. 1992a. Early development in an algal gametophyte:regulation of germination and nuclear events by cytoskeletal elements. Protoplasma 170:34-45.


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APPENDIX I:

Inhibition of Cellular Events During Early Algal Gametophyte Development: Effects of Select Metals and an Aqueous Petroleum Waste.

G.D. Garman, M.C. Pillai, and G.N. Cherr. 1994. Aquatic Toxicology 28:127-144.


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APPENDIX II:

Nuclear Events During Early Development in Gametophytes of Macrocystis pyrifera, and the Temoporal Effects of a Marine Contaminant.

G.D. Garman, M.C. Pillai, L.J. Goff, and G.N. Cherr. 1994. Marine Biology 121:355-362.


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APPENDIX III:

Developmental Effects of Barium Exposure in a Marine Bivalve (Mytilus californianus).

J.V. Spangenberg and G.N. Cherr. 1996. Environmental Toxicology and Chemistry 15:1769-1774.


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APPENDIX IV:

Developmental Abnormalities and DNA-Protein Crosslinks in Sea Urchin Embryos Exposed to Three Metals.

G.D. Garman, S.L. Anderson, and G.N. Cherr. 1997. Aquatic Toxicology 39:247-265.


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Appendix V: Barium Concentrations in Mussel Shells and Correlation with Distance from Outfall and Performance


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The Department of the Interior Mission As the Nation's principal conservation agency, the Department of the Interior has responsibility for most of our nationally owned public lands and natural resources. This includes fostering sound use of our land and water resources; protecting our fish, wildlife, and biological diversity; preserving the environmental and cultural values of our national parks and historical places; and providing for the enjoyment of life through outdoor recreation. The Department assesses our energy and mineral resources and works to ensure that their development is in the best interests of all our people by encouraging stewardship and citizen participation in their care. The Department also has a major responsibility for American Indian reservation communities and for people who live in island territories under U.S. administration.

The Minerals Management Service Mission As a bureau of the Department of the Interior, the Minerals Management Service's (MMS) primary responsibilities are to manage the mineral resources located on the Nation's Outer Continental Shelf (OCS), collect revenue from the Federal OCS and onshore Federal and Indian lands, and distribute those revenues.

Moreover, in working to meet its responsibilities, the Offshore Minerals Management Program administers the OCS competitive leasing program and oversees the safe and environmentally sound exploration and production of our Nation's offshore natural gas, oil and other mineral resources. The MMS Royalty Management Program meets its responsibilities by ensuring the efficient, timely and accurate collection and disbursement of revenue from mineral leasing and production due to Indian tribes and allottees, States and the U.S. Treasury.

The MMS strives to fulfill its responsibilities through the general guiding principles of: (1) being responsive to the public's concerns and interests by maintaining a dialogue with all potentially affected parties and (2) carrying out its programs with an emphasis on working to enhance the quality of life for all Americans by lending MMS assistance and expertise to economic development and environmental protection.

ocs study mms 97-0024 chronic toxicological effects of ......mms ocs study 97-0024. coastal research...

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