Technical Reports and Proceedings Citations and Abstracts

The formation of halo-organic compounds by chlorination of estuarine waters has been investigated under both laboratory and field conditions. Haloforms are readily generated in the laboratory with chlorine doses of 1 to 10 mg/l, the range employed by many coastal power plants. At salinities above 1 g/kg, Br is the principal halogen in the haloform products. On a molar basis, more than 4% of the chlorine was converted to haloforms in some tests. Ozone in the laboratory also generated haloforms in estuarine water; the yields were similar to those obtained from chlorine. However, only traces of haloforms were found in a power plant field site, where apparently haloform-bypassing reactions consume free chlorine much faster than in the laboratory. Identification of these reactions is uncertain, but they may involve formation of stable halogenated macromolecules. A large sewage treatment plant served as a volatile halocarbon source to study the fate of these compounds. The major loss mechanism appears to be volatilization to the atmosphere. Rates for this process are estimated. However, there appears to be some loss under winter ice cover, perhaps because of chemical or biological degradation.

Neff, Jerry M. 1981. Fate and Biological Effects of Oil Well Drilling Fluids in the Marine Environment: A Literature Review. EPA-600/3-82-064. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 150 p. (Avail. from NTIS, Springfield, VA: PB82-240391)

The major environmental concerns about discharge of used drilling muds to the ocean are that they may be acutely toxic or cause deleterious sublethal effects in sensitive organisms and ecosystems and that heavy metals associated with drilling muds may be accumulated by marine organisms to dangerous concentrations. A majority of major drilling mud ingredients are biologically inert or have a very low order of acute toxicity. Of the major drilling mud ingredients, only chrome- and ferrochrome-lignosulfonates can be considered at all toxic. Their toxicity is quite low to all but a few sensitive species (e.g., some corals). Minor ingredients of some environmental concern include sodium phosphate salts, detergents, biocides (chlorinated phenols no longer are permitted for offshore disposal), chromate salts and asphalt/oil-based ingredients. Ordinarily, these materials are not used in large enough quantities to cause concern. Their concentrations should be kept low in drilling muds destined for ocean disposal. Where possible, less toxic substitutes should be used. To date, the acute toxicity and sublethal biological effects of more than 20 used offshore-type drilling muds have been evaluated with more than 60 species of marine animals from the Atlantic, Pacific, Gulf of Mexico and Beaufort Sea. Representatives of five major animal phyla have been tested, including Chordata, Arthropoda, Mollusca, Annelida and Echinodermata. Larvae and other early life stages, and oceanic species (considered to be more sensitive than adults and estuarine species to pollutant stress) were included. In all but a few cases, acute toxicity, usually measured as 96-hr. LC50, was 10,000 ppm or higher drilling mud added. The lowest acute LC50 value was 500 ppm for stage I larvae of dock shrimp Pandalus danae exposed to a high density ferrochrome lignosulfonate drilling mud from Cook Inlet, Alaska. Chronic or sublethal responses were observed in a few cases at concentrations as low as 50 ppm.

Schimmel, Steven C. 1981. Results: Interlaboratory Comparison--Acute Toxicity Tests Using Estuarine Animals. EPA-600/4-81-003. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 14 p.

Environmental Protection Agency's Office of Pesticide and Toxic Substance (OPTS) is required by law to supply industry with recommended test standards to determine the toxicity of new chemicals. One proposed series of test standards is the acute toxicity test, flow-through and static, using estuarine animals. In proposing a test standard, OPTS requires knowledge of the precision of the data generated in these tests i.e., The expected variability in test results, given the same standard, test chemical and test species. This final report summarizes the results of static and flow-through toxicity tests conducted by six laboratories using the chemicals endosulfan and silver nitrate. Saltwater species tested were the copepod, Acartia tonsa, the mysid shrimp, Mysidopsis bahia and the sheepshead minnow, Cyprinodon variegatus. Results of the M. bahia and C. variegatus tests indicated that the mean ratio of the highest LC50 to the lowest LC50 generated for each chemical and test type was 4.0 (i.e. the greatest variability in the LC50 values expected was a factor of 4.0). Results of A. tonsa studies were not conclusive because different practices were used in the culture, age selection and testing of the species by the participating laboratories.

Ahearn, D.G., W.L. Cook and S.A. Crow. 1981. Effects of Pollutants on Microbial Activities in Estuarine Surface Films. EPA-600/4-81-009. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 20 p. (Avail. from NTIS, Springfield, VA: PB81-171183)

Samples of inshore surface films from Escambia Bay, Florida and from sites in the North Sea yielded populations of aerobic, heterotrophic microorganisms up to 108 ml-1 or 106 cm-2. Hydrocarbonoclastic organisms occurred in relatively low populations. A comparison of species of yeasts prevalent in North Sea waters before and after oil production activities indicated a shift to a more widespread distribution of hydrocarbonoclastic forms with possible inhibition of a non-hydrocarbon utilizing species. Examination of various hydrocarbons and chlorinated compounds with the potential of being sequestered in natural films indicated that 66% could potentially alter microbial metabolic processes in the slick. In microcosm studies of estuarine systems, representative compounds demonstrated a selective effect for microfungi.

Carpenter, James H., Carroll A. Smith and Rodney J. Zika. 1981. Reaction Products from the Chlorination of Seawater: Final Report. EPA-600/4-81-010. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 52 p. (Avail. from NTIS, Springfield, VA: PB81-172280)

Current methods underestimate the residual oxidants in chlorinated seawater by as much as 70% depending upon the details of the procedures. Chlorination of seawater in the presence of light produces bromate ions which can influence standard analytical procedures and represent an unknown factor in estuarine and coastal waters. Toxicity of bromate ion and persistence in coastal waters has not been determined. The copper complexing capacity of Biscayne Bay, Florida, water was reduced with the addition of chlorine. Analysis was by anodic stripping voltammetry on water samples after successive additions of copper sulfate solutions. Chlorination of seawater may produce toxicity and growth reduction through the indirect mechanism of copper release and/or reduced binding capacity. Laboratory chlorination of water from the intake of the Port Everglades, Florida, power plant produces bromoform levels comparable to that found in the plant. Chloroform extracts of chlorinated Biscayne Bay water are found to contain halogenated compounds which are new and different, and which pose unusual analytical problems. Studies usng GC/EC, GC/MS, HPLC, 1H NMR, differential pulse polarography and other techniques on natural extracts and synthesized compounds are reported.

Schimmel, Steven C. and Richard L. Garnas. 1981. Results: Interlaboratory Comparison-Bioconcentration Tests Using the Eastern Oyster. EPA-600/4-81-011. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 199 p. (Avail. from NTIS, Springfield, VA: PB82-194689)

This report summarizes the results of an interlaboratory comparison for bioconcentration (BCF) testing using the eastern oyster (Crassostrea virginica) and the organic chemicals p,p'-DDE, 1,2,4-trichlorobenzene, and pentachlorophenol. The mean BCF's and high to low BCF ratios for p,p'-DDE, 1,2,4 trichlorobenzene, and pentachlorophenol were 52,600 (3.4), 168 (2.3), and 64 (2.4), respectively. The test method (ASTM Draft 9) used for all three chemicals and four participating laboratories resulted in data with a mean to low BCF ratio of 2.7, which represented a 'worse case' situation for an estimate of variability. Considering the varying degrees of experience between participants in conducting BCF tests, the wide geographic distribution between participants, and the different dilution water characteristic, the results were remarkably reproducible.

Klecka, G.M. and D.T. Gibson. 1981. Bacterial Degradation of Dibenzo-p-dioxin and Chlorinated Dibenzo-p-dioxins. EPA-600/4-81-016. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 74 p. (Avail. from NTIS, Springfield, VA: PB81-171639)

Pseudomonas sp. N.C.I.B. 9816, strain 11, when grown on salicylate in the presence of dibenzo-p-dioxin, accumulated cis-1,2-dihydroxy-1,2-dihydrodibenzo-p-dioxin and 2-hydroxydibenzo-p-dioxin in the culture medium. Crude cell extracts prepared from the parental strain grown with naphthalene oxidized cis-1,2-dixhydroxy-1,2-dihydrodibenzo-p-dioxin to 1,2-dihydroxydibenzo-p-dioxin.Further degradation of the metabolite was not detected. Whole cells of the parent strain of Beijerinckia, grown with succinate and biphenyl, oxidized dibenzo-p-dioxin and several chlorinated dioxins. A mutant strain (B8/36) of Beijerinckia oxidized dibenzo-p-dioxin to cis-1,2-dihydroxy-1,2-dihydrodibenzo-p-dioxin. The mutant organism also oxidized two monochlorinated dibenzo-p-dioxins to cis-dihydrodiols. No metabolites were detected from two dichlorinated dibenzo-p-dioxins. Growth of the parent strain of Beijerinckia on succinate was inhibited after four hours when 0.05% Dibenzo-p-dioxin was present in the culture medium. Resting cell suspensions of the parent organism oxidized dibenzo-p-dioxin to a compound identified as 1,2-dihydroxydibenzo-p-dioxin. Further degradation of this metabolite was not detected, as the compound was found to be a potent mixed-type inhibitor of two ring-fission oxygenases present in this organism.

Snedaker, Samuel C. and Melvin S. Brown. 1981. Water Quality and Mangrove Ecosystem Dynamics. EPA-600/4-81-022. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 80 p. (Avail. from NTIS, Springfield, VA: PB81-204109)

Field studies were made to determine the relationship between general water quality parameters, with emphasis on pesticides and metal pollutants, and the functioning of the halophytic mangroves. It was concluded, from a broad range of ecological sample analyses, that mangroves are relatively insensitive to toxic materials in the parts per million range and lower. Further, they do not significantly concentrate synthetic organics or metals to levels which could be considered harmful to detritus feeders, although it is not known whether further biological concentration occurs during decomposition enrichment. Observed large differences in mangrove production and structure do appear to be related to: (1) terrestrial runoff and its entrained nutrients, (2) periodic inundation by runoff and/or tidal activity, and (3) deposits of organic matter in the substrate.

Ferrante, John G. 1981. Fate and Effects of Whole Drilling Fluids and Fluid Components in Terrestrial and Freshwater Ecosystems: A Literature Review. EPA-600/4-81-031. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 28p. (Avail. from NTIS, Springfield, VA: PB81-197766)

Drilling fluids represent an important aspect of offshore and land based drilling operations. The fluids perform a multiplicity of functions, ranging from lubricating to prevention of blowouts when encountering high pressure. Periodically, the fluids must be changed or they become old and the spent fluids are disposed of in on-land facilities. Introduction into the environment of the chemically complex fluids has prompted effects research addressing terrestrial and freshwater habitats and their respective biological components. Studies with terrestrial plants in laboratory and field experiments show that the fluids and some fluid components exhibit phytotoxicity properties reducing seed germination, growth and yield. Phytotoxicity in whole drilling fluids is attributed to soluble salt concentrations. Preference/avoidance reactions were observed in experiments with whole drilling fluids are also collated and discussed. The range of lethal concentrations of fluid components in toxicity studies was from less than 1 to 75,000 mg/l and that for whole drilling fluids from 0.29 to 85% by volume. Various reasons for observed toxicity are discussed and recommendations made for future freshwater and terrestrial research with drilling fluids.

Schmidt-Collerus, J.J., N.L. Couse, J. King and L. Leffler. 1981. Investigation of Enzymatic Screening Tests for Mutagens in Environmental Pollutants from Synfuel Operations. EPA-600/4-81-038. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 77 p. (Avail. from NTIS, Springfield, VA: PB81-209579)

The objective of this research program was to develop an enzymatic screen for chemical carcinogens based on the selective in vitro stimulation of microsomal biphenyl-2-hydroxylase by known chemical carcinogens. An attempt was made to repeat published work using a spectrophotofluorometric assay for biphenyl metabolites. It was found that this assay system is not valid for use with complex mixtures, and that metabolites must be separated from interfering compounds prior to quantitation. A high pressure liquid chromatography method was developed which permitted rapid separation of metabolites. Nanogram quantities of metabolites were detectable using this chromatographic separation in conjunction with a spectrophotofluorometric detector. Using this method, it was not possible to demonstrate in vitro stimulation of biphenyl-2-hydroxylase by chemical carcinogens. Alternative assays were also examined. Terphenyl is metabolized to at least three different compounds by hamster microsomes. Further work is necessary to validate the utility of this substrate in an enzymatic screen for carcinogens. A marine protozoan, Parauronema acutum metabolizes biphenyl in vivo to 2- and 4-hydroxybiphenyl. This organism may provide a reliable, inexpensive source of biphenyl hydroxylase for an in vitro enzymatic assay system.

Ehrlich, Kenneth, George Cawley and Elias Klein. 1981. Methods for the Recovery of Organic Carcinogens from Water. EPA-600/4-81-039. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 33 p. (Avail. from NTIS, Springfield, VA: PB81-209694)

The organic carcinogens benzo(a)pyrene, dieldrin, and N-acetyl-2-aminofluorene were recovered on XAD-2 macroreticular resin in yields of 90% or more from distilled water or seawater in yields of 40% or more from Lake Pontchartrain water containing a high concentration of organic material. The original solutions contained less than 500 parts per trillion of carcinogen. These results show that XAD-2 provides an efficient means for recovering nonpolar organic carcinogens from dilute solutions. More polar carcinogens such as dimethylnitrosamine were not effectively recovered on XAD-2 columns. Since XAD-2 binding would not be selective for carcinogens, we investigated methods which might bind carcinogens selectively from a mixture of organic compounds. We tested the ability of the above carcinogens to bind to nucleic acid using direct binding, equilibrium dialysis, nuclei binding, and binding to DNA-cellulose. Radio-labeled carcinogens were used to quantify the amount bound. Either rat liver nuclei (0.1 mg DNA) or DNA-cellulose (1 mg DNA) bound 18% of the acetylaminofluorene and up to 66% of the dieldrin from solutions containing 150 to 280 nmoles of compound. Up to 30% of the benzo(a)pyrene from solutions containing as much as 320 pmoles was bound. Ten-fold or lower recoveries were found when direct-binding or equilibrium-binding methods were used.

Burton, Dennis T. and Leonard B. Richardson. 1981. Investigation of the Chemistry and Toxicity of Ozone-Produced Oxidants and Bromate to Selected Estuarine Species. EPA-600/4-81-040. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 88 p. (Avail. from NTIS, Springfield, VA: PB82-116351)

This research program was initiated to determine the chemical and biological effects of ozone in estuarine systems and to evaluate its suitability as an alternative to chlorine. Chemical studies were performed to determine oxidant decay rates and bromate formation in both natural and artificial estuarine water. Ozonation of estuarine or marine waters can produce significant amounts of bromate if the natural levels of ammonia-nitrogen and other competing compounds are low and the initial residual oxidant and bromide concentrations are high. Toxicity studies showed that the concentrations of bromate which theoretically could be formed in an ozonated discharge were not toxic to the early life stages of striped bass (Morone saxatilis) and juvenile spot (Leiostomus xanthurus). Acute toxicity studies of ozone-produced oxidants (OPO) showed that the early larval stages of the American oyster (Crassostrea virginica) and recently hatched striped bass ichthyoplankton were most sensitive to OPO; this sensitivity decreased with progressive development. Juvenile blue crabs (Callinectes sapidus), juvenile Atlantic menhaden (Brevoortia tyrannus) and adult white perch (Morone americana) were more resistant than molluscan and ichthyoplankton larval stages. Hematological and gill histopathological sublethal studies of adult white perch showed that OPO caused death by hypoxia as a result of excessive gill damage. Moderately damaged gill tissue repaired itself within a few days when fish were moved to clean waters. A comparison of the ozone data obtained in this study with similar chlorine data from the literature showed that the toxicity of ozonated and chlorinated estuarine water is similar. Thus, ozone appears to offer few, if any, advantages over chlorine in reducing toxicity to estuarine organisms.

Environmental Research Laboratory, Gulf Breeze, FL. 1981. Acephate, Aldicarb, Carbophenothion, DEF, EPN, Ethoprop, Methyl Parathion, and Phorate: Their Acute and Chronic Toxicity, Bioconcentration Potential, and Persistence as Related to Marine Environments. EPA-600/4-81-041. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 255 p. (Avail. from NTIS, Springfield, VA: PB81-244477)

The toxicity, bioconcentration, and persistence of the pesticides acephate, aldicarb, carbophenothion, DEF, EPN, ethoprop, methyl parathion, and phorate were determined for estuarine environments. Static acute toxicity tests were conducted to determine the 96-h EC50 values for algae, 48-h EC50 values for oyster larvae, and 96-h LC50 values for at least two crustacean and fish species. Flow-through acute toxicity tests, based on measured concentrations, were conducted to determine the 96-h LC50 values of the pesticides for at least two crustacean and fish species. In addition, Maximum Acceptable Toxicant Concentrations (MATC) were determined in life-cycle toxicity tests with mysid shrimp (Mysidopsis bahia) and sheepshead minnows (Cyprinodon variegatus), or in partial life-cycle tests with grass shrimp (Palaemonetes pugio). MATCs were estimated from embryo-juvenile toxicity tests with sheepshead minnows. Persistence studies on carbophenothion, DEF, EPN, methyl parathion, and phorate investigated processes in marine systems that contribute to those pesticides' disappearance. The relative importance of biological and nonbiological processes (including biodegradation, photolysis, hydrolysis, sediment/water partitioning, and volatility) were examined. Bioconcentration factors for fish or mollusks exposed to carbophenothion, EPN, ethoprop, and phorate were determined at steady state or after greater than or equal to 28-day exposures.

Krone, Michael A. 1981. Sublethal Metabolic Responses of the Hermatypic Coral Madracis decactis Exposed to Drilling Mud Enriched with Ferrochrome Lignosulfonate: Final Report. EPA-600/4-81-049. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 67 p. (Avail. from NTIS, Springfield, VA: PB81-231623)

Madracis decatis corals were exposed for 17 days in laboratory aquaria to suspensions of 100 ppm drilling mud spiked with 0, 3, and 10 ppm ferrochrome lingosulfonate (FCLS). During the first week of exposure, these corals increased their oxygen consumption and ammonium excretion, relative to uncontaminated controls. Those corals exposed to the highest enrichments of FCLS demonstrated the greatest increases in respiration and excretion and also the largest variations in respiration and excretion between individual experimental animals. Corals reached their highest average rates of respiration and excretion by the end of the first week of continuous exposure. Rates then decreased during the next week and, after a secondary increase in excretion and respiration between days 10-13 which was most pronounced in those corals exposed to FCLS enrichment, leveled off at near-initial rates by the end of the second week. Two corals, one exposed to 100 ppm drill mud plus 3 ppm FCLS and the other to 100 ppm drill mud plus 10 ppm FCLS, became noticeably moribund as the experiment entered its second week. These were the two corals which showed the most rapid and most consistent increases in excretion and respiration during the first week of exposure to FCLS. By week two, polyp expansion in both of these corals was dramatically reduced, and each was removed from the experiment when areas of bare corallum suggested the onset of polyp death. All corals exposed to FCLS reacted by reducing their polyp expansion behavior, although only the two cited above showed mass polyp mortality. When exposure to drill mud plus FCLS was discontinued, respiration and excretion of surviving corals remained low and stable while their polyp activity returned to normal levels within 48 hours.

Borthwick, Patrick W. and Gerald E. Walsh. 1981. Initial Toxicological Assessment of Ambush, Bolero, Bux, Dursban, Fentrifanil, Larvin, and Pydrin: Static Acute Toxicity Tests with Selected Estuarine Algae, Invertebrates, and Fish. EPA-600/4-81-076. U.S. Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL. 9 p. (Avail. from NTIS, Springfield, VA: PB82-137654)

Selected static toxicity tests were conducted with Ambush, Bolero, Bux, Dursban, Fentrifanil, Larvin, and Pydrin to determine the sensitivity of species representing four major phyla. Algal bioassays were conducted with marine algae to determine the concentration of pesticide that would inhibit population growth by 50% in 96 h. Static toxicity tests with mollusk larvae estimated the concentration of pesticide that would cause 50% of the exposed larvae to develop abnormally in 48 h. Static acute lethality tests with crustaceans and fish determined the concentration of pesticide that is lethal to 50% of the test organisms during a 96-h exposure