Petroleum hydrocarbons in the marine bivalve Venus verrucosa: accumulation and cellular responses

Cellular and subcellular responses of the marine burrowing bivalve Venus verrucosa collected from the north-eastern coastline of Malta from January to June 1985, after exposure to petroleum hydrocarbons (PHC) were investigated. After long-term exposure to 100 gl^-1 of water-accomodated fractions (WAF) of crude oil, PHC were found to accumulate most rapidly in the digestive gland and then in the gills, with saturation levels being reached within 100 d of exposure in both cases. PHC accumulation, both in the mantle and muscle tissues, was more gradual and consistent throughout the whole exposure period. After 150 d of exposure, the digestive cells of the digestive gland were significantly reduced in height (atrophy) and exhibited reduced lysosomal membrane stability. After 144 h of exposure to higher concentration of PHC (820 and 420 gl^-1), several cytological effects were recorded, including an increase in cell volume and activity of gill mucocytes as well as in the number of haemocytes in gill blood sinuses. There was also evidence of damage to the epithelial lining of the foot, stomach and style sac and marked atrophy of the digestive cells of the digestive gland. The significance of such responses is discussed.     

Accumulation of 14C-1-naphthalene by an oceanic and an estuarine copepod during long-term exposure to low-level concentrations

Accumulation of the bi-cyclic aromatic hydrocarbon ¹⁴C-1-naphthalene in adult female Calanus helgolandicus Claus and adult female Eurytemora affinis Poppe in sea water concentrations of hydrocarbon ranging from 0.2 to 992 g/l was studied during exposure periods of up to 15 days as part of an investigation of the possible effects on marine zooplankton of persistent exposure to low levels of petroleum hydrocarbons. With both species the body levels of radioactivity increased rapidly during the first few days of the exposure period, but after exposure for 7 to 8 days to sea water containing 50 g hydrocarbon/l an equilibrium condition was approached; in some experiments where E. affinis was exposed to 1.0 and 10 g hydrocarbon/l for 15 days there was no further increase in body levels of radioactivity after 7 to 8 days. Using a low concentration of hydrocarbon (1 g/l), the quantity of radioactivity accumulated after 10 days was found to be nearly fifty times greater in the smaller species, E. affinis, than in C. helgolandicus, when expressed in terms of body weight. After they had been exposed to the hydrocarbon for several days the copepods contained a considerable proportion of radioactivity that was no longer identifiable as naphthalene and was presumably present as metabolites. Radioactivity accumulated in the copepods after several days was rapidly lost after they were transferred to uncontaminated sea water: e.g. C. helgolandicus lost nearly 90% of its body level of radioactivity in 24 h. Thereafter the rate of loss was greatly reduced, and 5% of the original body level of radioactivity still remained in the copepods at the end of 11 days. Experiments on the breakdown of naphthalene added at low concentrations to sea water samples containing natural microbial populations indicated degradation rates of 0.1 to 0.2 g/l/24 h in oceanic water, and 2.6 g/l/24 h in inshore water samples. The results are discussed in terms of the possible transfer of hydrocarbon to a higher trophic level in areas subjected to constant low-level inputs of petroleum hydrocarbons.     

The persistence of chronically accumulated hydrocarbons in the hard shell clam Mercenaria mercenaria

Hard shell clams, Mercenaria mercenaria, from the chronically polluted environment of the Providence River, USA, were transferred to a clean laboratory system located in lower Narragansett Bay, Rhode Island. The hydrocarbon contents of these transplanted clams were monitored for 120 days after transfer. After this period only slight depuration had occurred. A 41.9 g g wet weight^-1 average initial hydrocarbon burden decreased to 29.3 g g^-1, a decrease that became statistically significant (P<0.1) only after 120 days. At most, only 30% of the clams' hydrocarbons were lost. These findings indicate that the duration of the exposure period and the chemical nature of the source hydrocarbons are both instrumental in determining the persistence of these compounds in filter-feeding bivalves. Chronically accumulated petroleum hydrocarbons are strongly retained by the organisms and are only very slowly depurated.     

Effects of copper on the latency of lysosomal hexosaminidase in the digestive cells of Mytilus edulis

Mytilus edulis collected from Tomales Bay, California, USA, during mid-winter 1979 were exposed to increased concentrations of dissolved copper under controlled laboratory conditions. A dose-dependent reduction in the latency of lysosomal hexosaminidase activity in digestive cells was induced after a 30 d exposure to copper. The half-time of the hexosaminidase staining reaction in sections of digestive gland from control mussels was 15.5 min; for mussels exposed to 25, 50, and 75 g Cu l^-1 it was 11.8, 8.5 and 5.5 min, respectively. In addition, the dyecoupled reaction product was seen earlier in sections from individuals exposed to 50 and 75 g Cu l^-1 (after 30 s) and 25 g Cu l^-1 (1 min) than in sections from control individuals (2.5 min). Copper accumulations were demonstrated histochemically to have the same distribution as the hexosaminidase reaction product, indicating that copper is sequestered in lysosomes. Copper concentrations in digestive gland tissue, were related to the concentrations of copper in the water to which the mussels were exposed.     

Fate of petroleum hydrocarbons taken up from food and water by the blue crab Callinectes sapidus

Radiolabeled paraffinic and aromatic hydrocarbons, including benzopyrene, fluorene, naphthalene, methylnaphthalene, methylcholanthrene, hexadecane, heptadecane and dotriacontane, were taken up from food and water by the blue crab Callinectes sapidus. In 2 days, approximately 10% of the benzopyrene and fluorene were taken up from the water when their concentrations were 2.5 and 30 g/l, respectively. When given food with radiolabeled hydrocarbons, 2 to 10% of the hydrocarbons were assimilated by the carbs, with the remainder excreted. After uptake of hydrocarbon from water or food, a major pathway for the elimination of hydrocarbon and metabolites was through fecal material. All hydrocarbons used in the study were metabolized, with similar rates for both paraffinic and aromatic hydrocarbons. More than 50% of the radioactivity assimilated by the crabs was in the hepatopancreas, suggesting that the hepatopancreas was the site of hydrocarbon metabolism. Twenty-five days aftex exposure to radiolabeled hydrocarbons, radioactivity was found only in the hepatopancreas. The hepatopancreas contained highly polar hydrocarbon metabolites, including dihydroxy-compounds and their conjugates, while blood contained both monohydroxy-and dihydroxy-compounds. No evidence was found of storage of hydrocarbons by any of the crab tissues.     

Lysosomal responses to experimentally injected anthracene in the digestive cells of Mytilus edulis

Experimentally injected anthracene (10 to 100 g mussel^-1) has been shown to induce dose-dependent lysosomal destabilisation and release of hexosaminidase in the digestive cells of Mytilus edulis after 24 h. This destabilisation was accompanied by cytological evidence of cytolysis of the digestive cells. The destabilising effect of 100 g of injected anthracene persisted for 96 h with a return to the control condition by 168 h. These results are discussed in the context of environmental contamination by polycyclic aromatic hydrocarbons.     

Hydrocarbon uptake and loss by the mussel Mytilus edulis

The dynamics of accumulation and elimination of hydrocarbons by the blue mussel Mytilus edulis were studied in a continuous-flow system. Mussels were exposed for as long as 41 days to 200 – 400 g/l of diesel fuel adsorbed on kaolin particles. Hydrocarbons were accumulated in the tissues in excess of 1000 times the exposure levels. Upon termination of dosing, the mussels exhibited a rather rapid loss of hydrocarbons for the first 15 to 20 days (biological half-life=2.7 to 3.5 days). Subsequently, however, elimination was reduced to a minimum and a considerable fraction of the hydrocarbons could be recovered from the tissues after as long as 32 days of depuration. The mussels exhibited definite signs of physiological stress due to chronic exposure to diesel fuel, although recovery was rapid upon termination of dosing. It is concluded that mussels could be utilized as a test organism for monitoring long-term hydrocarbon pollution in marine waters. The implications for the mussel culture industry are discussed.     

Effect of environmental parameters on lysosomal marker enzymes in the tropical blood clam Anadara granosa

The lysosomal marker enzymes, arylsulfatase and acid phosphatase, in a tropical burrowing arcid clam Anadara granosa L. have been found to exhibit seasonal variations. The activity of both enzymes decreased with increase in ambient temperature and fell with increase in salinity. Lysosomal latency for these enzymes, however, was not significantly affected by environmental parameters, including salinity, temperature, nutritional status, breeding season, etc. The physico-chemical characteristics of the habitat were found to induce reversible changes in the lysosomal latency and enzyme activity. Exposure to elevated temperature (T=7° to 8°C), towards the upper limit of its physiological tolerance, was found to labilize the lysosomes. The biochemical methods optimized for the demonstration of lysosomal stability under exposure to environmental stressors could also be extended to quantify the impact of various pollutants, including heavy metals, radionuclides, etc. at the subcellular level.     

Alkanes and alkenes in marine benthic algae

Saturated and olefinic hydrocarbons were determined in additional species of benthic marine algae from the Cape Cod (Massachusetts, USA) area (see: Youngblood et al., 1971). The distribution of homologous and isomeric olefins was studied in plants of different age and in morphologically different parts of the same specimen. With two minor exceptions, only normal alkanes and alkenes are present. The methylene-interrupted C₁₉- and C₂₁-polyolefins are particularly abundant; ¹-heneicosahexaene and the corresponding pentaene are common to all brown algae, while the corresponding ³-isomers occur in green algae. The hydrocarbon concentration, the alkene-to-alkane ratio and the polyolefin content are highest in young plants or in rapidly growing tissues of older plants. This suggests a deeper involvement in cell biochemistry of straight-chain hydrocarbons than previously considered. The biosynthesis of the plant polyolefins remains to be explored; no immediately obvious precursors of the ¹-polyolefins were found among the algal fatty acids. The hydrocarbon composition of these benthic algae differs greatly from that of fossil fuels in its simplicity and predominately unsaturated nature. The separation of the isomers by gas chromatography and their structural elucidation by mass spectrometry, alone and in combination with hydrogenation and ozonolysis, are discussed.Contribution No. 3155 of the Woods Hole Oceanographic Institution.     

Behavioural responses of the marine bivalve Macomona liliana exposed to copper- and chlordane-dosed sediments

Sublethal behavioural responses including avoidance and burial rate were compared with 10 d acute morbidity and mortality in bioassays using juvenile Macomona liliana 1 to 3 mm long. The bivalves showed significant movement away from all copper-dosed sediments, with maximal movement after 96 h at 25 mg Cu kg (dry wt)^-1. Increasing copper concentrations slowed the rate of burial, and above 15 mg Cu kg (dry wt)^-1, most shellfish failed to bury after 90 min. After 10 d exposure, morbidity (defined as inability to rebury) occurred at 15 mg Cu kg (dry wt)^-1 and mortality at 30 mg Cu kg (dry wt)^-1. In avoidance trials, chlordane had a significant effect on overall movement from 20 g kg (dry wt)^-1, but increasing concentrations reduced movement away from dosed sediment. Chlordance appeared to have no effect on burial rate. Survival was affected by 10 d exposure to 400 g chlordane kg (dry wt)^-1, with increased morbidity and death. The relative sensitivities of the responses shown by M. liliana to copper were avoidance > burial/morbidity > mortality, with a 6-fold difference in the response threshold between avoidance and mortality. With chlordance, an avoidance response was detected at a concentration 20-fold lower than that causing morbidity. Behavioural responses of M. liliana have the potential to provide a sensitive bioassay.     

Physiological evaluation of naphthalene intoxication in the tropical acrid clam Anadara granosa

Changes in vital physiological activities — shell movements and burrowing behaviour, uptake and depuration, filtration and respiration — on exposure to sublethal concentrations of naphthalene (Nap) were investigated in the tropical arcid blood clam Anadara granosa L. from the Bombay coast in 1986. On exposure to ambient concentrations exceeding 5g Nap ml^-1, shell valves opened widely within the first hour of exposure. The compressed muscular foot was stretched vertically upwards with copious secretion of mucus, and did not exhibit any evidence of burrowing behaviour. Those exposed to 5g Nap ml^-1 regained their normal physiological activities on transfer to stressor-free medium, whereas those exposed to higher levels became intoxicated and narcotized. Bioaccumulation of Nap was dependent upon environmental concentration, increasing exponentially with time over a 9h exposure period. Further exposure, up to 96h, however, did not increase tissue levels substantially. About 65% of the total body burden of Nap was depurated within 3h of transfer to Nap-free medium. Rates of filtration and oxygen consumption were significantly reduced (ca 70%, p<0.001) compared to control clams. Percent inhibition in these physiological activities was dependent upon tissue and ambient concentrations of Nap. On transfer to pollutant-free medium, clams exhibited remarkable recovery. Rates of both filtration and oxygen consumption were gradually increased and restored to normal levels, as observed in controls. However, clams exposed to upper limits lost their ability to burrow back into the sedimentary bed, leaving them susceptable to predators. Furthermore, induction of anaerobiosis and disruption of osmotic balance on exposure to Nap together with aerial exposure at low-water periods and salinity changes, acted synergistically and proved detrimental. The rates of growth and mortality observed in the natural population of blood clams, harvested from the coastal waters off Bombay, have been explained in terms of the impact of petroleum and allied waste products released from petrochemical industries.     

Biochemical evaluation of naphthalene intoxication in the tropical acrid blood clam Anadara granosa

Exposure of the tropical arcid blood clam Anadara granosa L., collected from Bombay waters in 1986, to sublethal concentrations of naphthalene (Nap) (5, 10 and 15g Nap ml^-1), induced various changes in vital biochemical systems. The condition index (CI) remained largely within the control range, although tissue water content after 96h exposure increased significantly (p<0.001). Glycogen levels appreciably decreased, both in muscles (p<0.001) and digestive glands (p<0.01). AMP activity remained unaltered irrespective of the ambient Nap concentration. ADP levels decreased by more than 20% and ATP levels also decreased significantly (p0.001) by more than 60% with increases in ambient Nap levels from 5 to 15 g ml^-1. Lysosomal marker enzymes-arylsulphatase and acid phosphatase-showed various changes. Levels of the former were 10 to 17 times higher than the latter. Total and free arylsulphatase activity in hepatic cells decreased on exposure of clams to 5 and 10g Nap ml^-1, but not significantly. However, it was significantly inhibited (p<0.001) towards the upper limits of exposure concentration. The same was also true of acid phosphatase activity. Nicotinamide adenine dinucleotide (NADH₂) oxidation of lipochrome showed a dramatic decrease in activity in the range of 37 to 88% over controls with increasing Nap from 5 to 15 g ml^-1. Fatty acids also exhibited significant changes in composition with a decrease in polyunsaturated acids and an increase in certain saturated fatty acids. Protein and carbohydrate levels increased significantly (p<0.001) towards upper limits of exposure concentration. Pyruvic acid levels increased significantly by 2 to 6 times over controls on exposure to 5 to 15 g Nap ml^-1. Explanation of these changes is attempted in terms of the glycolysis pathway involving pyruvic acid oxidation and oxidative phosphorylation in the electron transport system.     

Stable lead isotope profiles in smelter and general urban communities: a comparison of environmental and blood measures

High-precision lead isotope ratios and lead concentrations have been compared statistically and graphically in women of child-bearing age (n = 77) from two smelter communities and one general urban community to evaluate the relative contributions to blood lead of tissue lead stores and lead from the contemporaneous environment (soil, floor dust, indoor airborne dust, water, food). Blood lead (PbB) contents were generally low (e.g. <10 g dL^–1). Statistically significant isotopic differences in blood and environmental samples were observed between the three cities although isotopic differences in blood for individual subjects living in close proximity (200 m radius) was as large as the differences within a city. No single environmental measure dominated the biological isotope profile and in many cases the low levels of blood lead meant that their isotopic profiles could be easily perturbed by relatively small changes of environmental exposure. Apportioning of sources using lead isotopes is possibly not feasible, nor cost effective, when blood lead levels are <5 g dL^–1. Interpretations based on statistical analyses of city-wide data do not give the same conclusions as when the houses are considered individually. Aggregating data from multiple subjects in a study such as this obscures potentially useful information. Most of the measures employed in this study, and many other similar studies, are markers of only short-to-medium integration of lead exposure. Serial sampling of blood and longer sampling times, especially for household variables, should provide more meaningful information.     

Effects of copper and cadmium on growth,swimming and predator avoidance in Eurytemora affinis (Copepoda)

In this study we demonstrate the sensitivity of swimming behavior and predator-escape responses of nauplii of the estuarine copepod Eurytemora affinis to sublethal doses of Cu and Cd. Behavior was generally altered at metal doses below those affecting growth rates or survival of the copepods. Swimming velocities of Cu-dosed nauplii were different from controls at all concentrations of Cu tested (10–50 g l^-1 total Cu) after 24- to 48-h exposure, whereas development rate of nauplii was significantly reduced only after 96 h at 25 g l^-1. The 96 h LC50 for Cu was approximately 30 g l^-1 Cu. Naupliar swimming velocity was also affected by Cd. Swimming speeds were reduced after 24 h at 130 g l^-1, and development was slowed after 48 h at 116 g Cd l^-1. The 96-h LC50 was >120 g l^-1. Little is known of the adaptive role of specific motile behaviors in the success of larval copepods. We investigated the relationship of swimming speed to predator — prey interactions of the nauplii using both real and simulated predators. Nauplii exposed to Cu for 24 h were observed to be generally hyperactive, a condition which could increase their encounter frequency with predators. Reduced numbers of escape responses of nauplii to a simulated predator, another indication of increased vulnerability to predation, were observed only after 48-h exposure to Cu. Nevertheless, feeding rates of non-dosed larval striped bass on dosed nauplii (24 h at 25 g Cu l^-1) were significantly higher than on control nauplii. Feeding rates of larval mysid shrimp, however, were not higher on similarly dosed nauplii; 24 h exposure of nauplii to >30 g Cu l^-1 did result in increased predation by mysids.Contribution No. 272 of the US EPA Environmental Research Laboratory, Narragansett, Rhode Island 02882, USA     

An assessment of lead absorption from soil affected by smelter emissions

The purpose of this study was to assess the oral bioavailability of lead in soil collected from a former smelter site in Sandy, Utah, USA. Sprague-Dawley rats (approximately 4 weeks of age, 5 of each sex in group) were given either soil lead or lead acetate mixed in a purified diet (AIN-93G ) at four different concentrations for 31 consecutive days. Food consumption measurements were used to compute mean daily lead exposures for the soil lead and lead acetate groups. The lead acetate treatment yielded higher concentrations of lead in the blood and bone than the soil lead treatment. Mean blood lead values ranged from below the detection limit (3 g dL^–1) to 27.25 g lead dL^–1 for the lead acetate groups at dose levels of 0.10–2.91 mg lead kg body weight^–1 and from below the detection limit to 8.8 g lead dL^–1 for the soil lead groups at doses of 0.11–3.43 mg lead kg body weight^–1. At these same doses, mean bone values ranged from 0.52 to 26.92 g lead g^–1 for the lead acetate groups and from 0.64 to 13.1 g lead g^–1 for the soil lead groups. Relative per cent bioavailability was estimated by modelling the dose-blood concentration curves for the lead acetate treatment and the dosed soil lead treatment, and then comparing doses that produce an equivalent blood lead concentration. The ratio of the doses of lead acetate and soil lead that produced the same tissue response (i.e., concentration) provided an index of relative bioavailability. For lead, the bioavailability of soil lead relative to lead acetate was 41% at a blood concentration of 6 g lead dL^–1.     

Sensitivity of marine fishes to toxins from the red-tide dinoflagellate Gonyaulax excavata and implications for fish kills

Marine fishes (Atlantic herring, American pollock, winter flounder, Atlantic salmon, and cod) were dosed orally and intraperitoneally (i.p.) with paralytic shellfish toxins extracted from Bay of Fundy Gonyaulax excavata (tamarensis) cells. The toxins are lethal to these fishes in low oral doses, and in extremely low i.p. doses. Symptoms are the same among these fishes, both for oral and i.p. administrations, including loss of equilibrium within 5 to 15 min, followed by immobilization and shallow, arrhythmic breathing. Death generally occurs within 20 to 60 min of toxin administration. Dose responses are also similar among these fishes. Oral LD50 values are 400 to 750 g saxitoxin (STX) equivalent kg^-1 body weight. Intraperitoneal LD50 values are 4 to 12 g STX equivalent kg^-1. Toxins are undetectable in fish muscle tissue following lethal oral doses. The similarity of symptoms and dose responses suggest that fish as a group are sensitive to G. excavata toxins. Results, in combination with reports implicating these toxins in herring, sand lance, and menhaden kills, show the plausibility that the nearly worldwide blooms and red tides of G. excavata and its relatives may cause kills of a variety of fishes.     

Effects of pressure,temperature and oxygen on the oxygen consumption rate of the Midwater copepod Gaussia princeps

In this study we assessed the amount of lead accumulated in the body of a grazing mollusc by transfer from its algal food in laboratory experiments, and compared these results with the amounts found in naturally occurring molluscs and seaweed. Near La Jolla, California (USA), where the concentration of lead in seawater is probably less than 0.08 g l^-1, most of the naturally occurring Egregia laevigata contains less than 0.4 g Pb g^-1 wet weight. The total body masses, without shells, of juvenile Haliotis rufescens fed on this seaweed for 3 to 6 months showed similar concentrations. When, however, E. laevigata is placed for 1 to 6 days in seawater to which lead has been added (0.1 or 1.0 mg l^-1) both the seaweed and the abalone subsequently fed with it accumulate proportionally larger amounts of lead. After 6 months, young abalone fed on E. laevigata pretreated with 1.0 mg Pb l^-1 accumulated up to 21 g Pb g^-1 wet weight. This amount of lead had no apparent consequences on the growth or activity of the molluscs. Analyses of 6 different organs from adult abalone showed that the lead was selectively concentrated in the digestive gland. In the foot (muscle tissue), which is the part normally consumed by humans, only negligible amounts were found.     

Cadmium effects on the carbon and energy balance of mudsnails

The present study examined the ability of shortterm physiological measurements to predict growth rates of a gastropod, Hydrobia ulvae, in response to pollutant exposure. Whereas growth rates of snails collected from Munkebo Harbor, Funen, Denmark in 1989 were substantially reduced by exposure to cadmium after ca. 3 wk, we could detect no change in total carbon balance measured in short-term radioisotope experiments. At 23 S, the reduction in percent growth after 3 wk was 186.4 for snails exposed to 100 g Cdl^-1 and 225.7 for snails exposed to 200 g Cdl^-1. At 13 S, the reduction in percent growth was 102.6 at 100 g Cdl^-1 and 106.4 at 200 g Cdl^-1. We found no effect of cadmium on sediment egestion rate over 18 d of exposure. Nor could we detect effects on carbon absorption efficiency or metabolic loss of absorbed carbon during the first 24 h following its ingestion. The fractional loss of ¹⁴C from uniformly-labeled snails showed a decrease in CO₂ and an increase in dissolved carbon in snails exposed to 200 g Cdl^-1. Such changes in the partitioning of carbon loss from body stores suggest a partial shift from aerobic to anaerobic metabolic pathways which could reconcile the seeming disparity between growth and carbon balance in response to cadmium exposure.     

Effects of chronic copper exposure on the green mussel Perna viridis

The effects of chronic copper exposure on growth and physiological responses of the green mussel Perna viridis were investigated by exposing the mussels to 50 μg l⁻¹ Cu for 3 mo at 17 and 25 °C. These temperatures represent, respectively, the winter and summer seawater temperatures in Hong Kong. Differences in the level of response between mussels exposed for 3 mo to 50 μg Cu l⁻¹ generally increased with duration of exposure. The tissue concentration of copper had increased by 280 and 450% after 3 mo exposure at 17 and 25 °C, and growth performances were reduced, with the 25 °C sets suffering from larger negative impact of copper in most responses. The inhibitory effects of copper on production of the various body components generally followed the order linear shell growth (greatest) > tissue production > byssus production > shell production. There were also decreases in the condition index (43 and 35% reductions at 17 and 25 °C), clearance rates (10.3 and 18.5%), faeces production (11 and 16.3%), assimilation efficiency (6.8 and 9.2%) and oxygen consumption rate (12.8 and 24.8%). In contrast, the organic content of the faeces (9.2 and 13.2% increases at 17 and 25 °C) and rate of ammonia excretion (21 and 28.6%), increased upon chronic copper exposure. Many of the responses (e.g. changes in tissue copper content, body dry wt, shell organic content, clearance rate and oxygen consumption rate) exhibited fluctuating levels of impact during prolonged copper exposure, while others (e.g. faecal production rate, assimilation efficiency, tissue organic content) demonstrated steady decreasing trends with increasing exposure time. Received: 17 September 1999     

Effects of kelthane on the estuarine shrimp Crangon tranciscorum

The chlorinated hydrocarbon pesticide Kelthane was assayed for effects on food consumption, molting rate, cannibalism, respiration, mortality and behavior of the estuarine shrimp Crangon franciscorum Stimpson. The test system was a single-pass pulse-flow apparatus employing a modified Mount-Brungs style diluter. Treatment levels were 0, 14, 33 and 81 g l^-1 Kelthane at 16.6°C and 19 S over an exposure period of 317 h. Exposure to Kelthane reduced food consumption, molting rate, cannibalism, and respiration. Behavior also was abnormal in exposed shrimp. They spent more time swimming, were less coordinated and had feeble escape reactions. Kelthane was very toxic to C. franciscorum with LT₅₀ values of 163, 196 and 264 h for shrimp exposed to 81, 33 and 14 g l^-1 respectively. The incipient lethal level of Kelthane is probably below 14 g l^-1.