OCS Study 

•

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 

1 

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: 1

G.N. Cherr and 2T. W.-M. Fan 

AFFILIATION: 1

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

ADDRESS: 1

P.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 

2 

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 

3 

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. 
Chronic Toxicological Effects of Produced Water 

4 

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 

5 

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 
Chronic Toxicological Effects of Produced Water 

6 

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. 
Final Study Report – Cherr and Fan 

7 

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. 
Chronic Toxicological Effects of Produced Water 

8 

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 
Final Study Report – Cherr and Fan 

9 

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. 
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 

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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.