Elucidating cellular and behavioural effects of contaminant impact (polycyclic aromatic hydrocarbons,PAHs) in both laboratory-exposed and field-collected shore crabs,Carcinus maenas (Crustacea: Decapoda)

Monitoring effects of contaminants at lower levels of biological organisation (e.g. biochemical and cellular) allows for mechanistic evaluation of effects of contaminant exposure through laboratory exposures. However, higher level organism effects (e.g. physiological and behavioural) are deemed more ecologically-relevant. In the present study, cellular (cell viability and immune function), physiological (cardiac activity) and behavioural (foraging behaviour) responses were evaluated in field-collected shore crabs Carcinus maenas from three estuaries [a ‘relatively’ (polycyclic aromatic hydrocarbon) PAH-contaminated site (Plym Estuary) and two ‘comparatively clean’ field sites (Avon and Yealm Estuaries)] and compared with responses of crabs exposed in the laboratory to a model organic contaminant (PAH) pyrene (200 μg l⁻¹ for 28 days). The hypothesis that behavioural indicators may be more sensitive than other more traditional methods was tested. No significant impacts were observed at the cellular or physiological level in Plym-collected crabs (the ‘relatively’ contaminated site), but foraging behaviour was significantly altered (increased prey handling time) compared to individuals collected from the ‘relatively’ uncontaminated sites (Avon and Yealm). When given a cockle as a prey item, both Plym-collected and laboratory-exposed crabs took longer to handle and break into cockle shells. Therefore, ecologically-relevant behavioural observations may serve as valuableindicators of environmental quality.     

Seasonal differences in the physiology of Carcinus maenas (Crustacea: Decapoda) from estuaries with varying levels of anthropogenic contamination

This study reports the seasonal variability in aspects of the physiology of the shore crab Carcinus maenas from three estuaries in South-west England, each with varying anthropogenic inputs: Avon Estuary (‘relatively low’ impact), Yealm Estuary (‘intermediate’ impact) and Plym Estuary (‘relatively high’ impact). Crabs collected over 12 months from the Avon had a significantly ‘lower’ physiological condition in winter and spring compared to summer and autumn; in particular, haemocyte phagocytic capability (a general indicator of immune function) was significantly higher in winter and spring compared to summer and autumn, and total haemolymph antioxidant status (an indicator of oxidative stress) was significantly lower in winter compared to the remainder of the year. Potentially, shore crabs may be more susceptible to the effects of contaminant exposure, such as increased immunotoxicity (thus, reduction of immune function) and/or oxyradicals (or reactive oxygen species) exposure) especially in seasons of increased susceptibility i.e. summer/autumn (lower phagocytic capability) and winter (lowest antioxidant function). As the Avon was taken to represent the ‘reference’ site, this pattern is considered to reflect the ‘normal’ seasonal variability in shore crab physiology. Shore crab physiological condition from the ‘relatively high’ impact estuary (Plym) revealed increased cellular viability and antioxidant status in autumn and winter compared with that of the ‘standard’ pattern (Avon) However, crabs from the intermediate impact estuary (Yealm) only demonstrated significant physiological differences in summer as shown by a lower cellular viability. All crabs had been exposed to PAHs (confirmed by the presence of PAH metabolites in their urine) which may account for the observed differences in shore crab physiology. In conclusion, to aid understanding of the potential contaminant impacts on biota it is imperative that the ’normal’ seasonal variability of physiological condition be established. Biological effects-based monitoring studies should therefore be employed seasonally to potentially highlight ‘windows of sensitivity’ to contaminant impact.     

Population ecology of fiddler crab, Uca lactea annulipes (Decapoda: Ocypodidae) in Sirik mangrove estuary,Iran

Fiddler crabs are known as the most conspicuous bioturbating animals in mangrove forest, which by their burrowing activities promote nutrient cycling in ecosystems. Mangroves, crabs and bacteria are connected in the form of positive feedback loop, whereby an increase in the activity of any participant will tend to increase the activity of all others as well. Population structure, growth, mortality, recruitment, biomass and production of fiddler crabs as dominant species of deposit-feeder crabs have been investigated from May 2005 to March 2006 in the Sirik mangrove estuary in the eastern part of Strait of Hormuz. Ten 1 m² squares of the substrate were randomly sampled during low tide. There was a significant change in organic content in the sampling area during study period. The sex ratio was significantly deviated from 1:1 ratio in large sizes, where males were dominated year around. Fiddler crabs showed seasonal growth pattern during a year, which the maximum growth was observed during autumn and early summer and ceased during winter by low temperature. Growth parameters were estimated by Appeldoorn's method and fitted with seasonalized von Bertalanffy growth models. The Kimura likelihood ratio test revealed significant differences in the overall growth curves between males and females, originated from the differences in k$k$ and L_∞$L_{\mathrm{\infty }}$ between sexes. Males were found to exhibit higher growth rate and larger size than females, this could be attributed to the higher investment of energy for reproduction in females. Reproduction was restricted to spring and early summer, while recruitment occurs at the end of summer. Total annual production of males and females was 37.90 and 10.05 g dry weight/10 m² respectively and major production took place during early summer. Because of impact of temperature on the activity of fiddler crabs in subtropical regions, density of crabs has significant positive correlation with temperature (P < 0.01), while growth rate shows significant negative correlation with organic matter of sediment (P < 0.05). In conclusion these crabs showed lower mortality and production rate with the seasonal growth and recruitment in comparison with tropical species.     

Heavy metal stress induced hyperglycemia in blue swimmer crab, Portunus pelagicus

The study was to find out the effect of cadmium and mercury on levels of hemolymph glucose, crustacean hyperglycemic hormone (CHH) and hepatopancreas glycogen in the blue swimmer crab Portunus pelagicus. The experiments were performed in both intact and eyestalk ablated crabs. Quantification of CHH was done by the indirect ELISA with the aid of primary anti-Carcinus maenas-CHH antibody. Higher glucose concentration was observed on exposure to 8×10^-6 of cadmium ((825.6±5.42) μg/mL) and 6×10^-6 of mercury ((90.5±6.25) μg/mL) after 48 h and 24 h respectively. Higher level of hemolymph glucose was observed in eyestalk intact crabs on exposure to cadmium and mercury than eyestalk ablated crabs. Decrease in the levels of CHH was observed in both eyestalk intact and ablated crabs on heavy metal exposure. Decline of the hepatopancreas glycogen level was also witnessed with the exposure to heavy metal, which validated its utilization in the production of glucose. Thus this study brings to light, the variations in hemolymph glucose, CHH and hepatopancreas glycogen on heavy metal stress. These carbohydrate metabolites can be used as biomarkers in assessing heavy metal contamination in water bodies.     

Application of the biomarker responses in scallop (Chlamys farreri) to assess metals and PAHs pollution in Jiaozhou Bay,China

Scallops (Chlamys farreri) were collected at three sites in the Jiaozhou Bay, China in April and July 2007. The responses of biomarkers were evaluated for assessment of physiological status of scallops and pollution in the areas. Compared with site S1, the activity of aryl hydrocarbon hydroxylase was induced notably (P < 0.05) and glutathione S-transferase activity was inhibited significantly (P < 0.05) in S2 and S3. The levels of DNA alkaline unwinding, DNA protein crosslinks, protein carbonyl and lipid peroxidation were tested and indicated the oxidative stress situation of scallops. There was significant difference of biomarker levels between sampling seasons (P < 0.05), and between digestive gland and gill (P < 0.05). The results provided the reference data for multiple-pollution assessment in the marine environment and indicated that tissue type and seasons affect biomarkers, therefore these factors should be taken into consideration when biomarkers are used for environmental assessment purposes.     

Phase I and II biotransformation enzymes and polycyclic aromatic hydrocarbons in the Mediterranean mussel (Mytilus galloprovincialis,Lamarck, 1819) collected in front of an oil refinery

The aim of the present study was to investigate the responses of phase I and II biotransformation enzymes and levels of PAHs in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected from three sites at different distance from an oil refinery. Phase I enzyme activities as NAD(P)H-cyt c red, NADH ferry red, B(a)PMO and phase II as UDPGT, GST were measured in digestive gland while 16 PAHs (US-EPA) in whole soft tissue. An added value to the data obtained in the present study rely on the RDA analysis which showed close correlations between PAHs levels and phase I enzyme activities in mussels collected in front of the refinery. And again a significant spatial correlation between B(a)P levels and NADPH-cyt c red activities was observed using linear models. No differences among sites for B(a)PMO and phase II GST activities were observed, while the application of UDPGT as biomarkers requires further investigation.     

Seasonal variation of antioxidant and biotransformation enzymes in barnacle, Balanus balanoides, and their relation with polyaromatic hydrocarbons

Seasonal variations in the antioxidant enzymes (catalase, superoxide dismutase [SOD], NADH-DT diaphorase), biotransformation enzyme, glutathione-S-transferase (GST) and microsomal lipid peroxidation in digestive tissue of barnacle, Balanus balanoides, from polluted and non-polluted populations have been evaluated. Relationships with accumulated polyaromatic hydrocarbon (PAH) concentration in barnacle tissues and environmental parameters (water temperature, salinity, dissolved oxygen concentration, water pH) were determined. As a general trend, maximum antioxidant enzyme and GST activities were detected in the pre-monsoon period or summer (March-June) followed by a gradual decrease during the monsoon (July October) with a minimum in the post-monsoon period or winter (November February). This pattern was similar to tissue concentrations of PAHs, resulting in a significant positive correlation with antioxidant enzymes, mainly catalase and SOD. Microsomal lipid peroxidation exhibited an almost reverse trend of seasonal variation to that of antioxidant enzyme activities indicating an enhanced susceptibility of barnacle tissues to oxidative stress. Among the environmental parameters, only water temperature seemed to have a significant effect on observed variations of antioxidant enzymes and GST activities. The barnacles from polluted and non-polluted populations exhibited seasonal differences in the activities of all the enzymes studied, particularly catalase, SOD and GST, suggesting the possibility of some biochemical adaptation in organisms from a chronically polluted environment. The results indicated that antioxidant defense components, catalase and SOD, are sensitive parameters that could be useful biomarkers for the evaluation of contaminated aquatic ecosystems. The results also suggested the potentiality of barnacle, B. balanoides, as a bioindicator organism against organic pollution.     

Seasonal changes of antioxidant and oxidative parameters in the coral Pocillopora capitata on the Pacific coast of Mexico

The physiological responses of the coral Pocillopora capitata to environmental conditions common in winter and summer were studied in 2007 during February–March (winter) and June–July (summer) at La Boquita reef (Manzanillo, Colima, Mexico). Shallow and deep sampling stations were established at different distances from a small jetty built next to the Juluapan Lagoon. We analyzed superoxide radicals () and lipid peroxidants (TBARS); the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione‐S‐transferase (GST); chlorophyll a (Chl a), zooxanthellae density (ZD); and mycosporine‐like amino acids (MAAs). Our results showed that the , TBARS, CAT, GST, MAAs, and Chl a, levels were significantly higher in summer (P < 0.05); no seasonal difference was found for GPx, GR or ZD. We found significant differences (P < 0.05) in winter only for Chl a and ZD at shallow sites and, in contrast, for at deeper sites. The results of this study indicate that increasing temperature and radiation associated with seasonal changes (from winter to summer), the efficiency of the enzymes GST, CR and GPX, and the production of MAAs together form a powerful mechanism for P. capitata to offset the detrimental effects of environmental change.     

Toxic effects of certain heavy metals (Hg,Cd, Pb,As and Se) on the intertidal crab scylla serrata

The toxic effects of certain heavy metals (Hg, Cd, Se, As and Pb) on Scylla serrata, the common edible Indian marine crab, are evaluated. Mercuric chloride, phenyl mercuric acetate (PMA), cadmium chloride, selenium dioxide, arsenic trioxide and lead nitrate were tested individually in laboratory static bioassays using artificial seawater to determine the 24 to 96 h LC₅₀ values. As indicated by 96 h LC₅₀ values PMA was found to be the most toxic and lead nitrate the least toxic. CdCl₂ showed a 6·6 fold increase in toxicity with increase in the time of exposure from 24 to 96 h. Marked histopathological changes were noticed in the hepatopancreas and gill of Scylla serrata exposed to acute 96 h LC₅₀ values of these metal compounds. 30 days' long-term exposure to sublethal concentrations of mercury and cadmium brought about degenerative changes in the hepatopancreas and gill of exposed animals. Extensive cellular proliferation (hyperplasia) of secondary lamellae accompanied by cyst formation and necrotic regions were often seen in the gills of cadmium treated crabs. Interestingly enough, the free amino acid pool from muscle and hepatopancreas of treated animals, either after acute or long-term exposure, did not show any reduction in number or quantity     

Soldier crab (Dotilla myctiroides) distribution,food resources and subsequent role in organic matter fate in Ao Tang Khen,Phuket, Thailand

This study aimed to elucidate Dotilla myctiroides (H. Milne-Edwards) food resources and how the soldier crab influences the organic matter composition on the intertidal flat. A preliminary assessment of D. myctiroides size, sex, and population distribution was conducted to clarify the characteristics of the inhabiting population. Overall, D. myctiroides size and abundance was normal across the tidal flat area. Comparison of different sediment types from the soldier crab habitat (surface sediments, igloos, burrow lining, burrow pellets, feeding pellets, and feces) using the fatty acid (FA) analysis technique revealed D. myctiroides removes organic matter via the significantly lower contributions of polyunsaturated FAs, essential FAs, and dinoflagellates in the feeding pellets. Dotilla myctiroides tissue analysis of the hepatopancreas and hindgut in individuals that had been starved for 24 h confirmed not only the consumption of dinoflagellates, but diatoms and macroalgae as well. In addition, the FA metabolism within the digestive system implied the ability of the soldier crab to synthesize highly unsaturated FAs (HUFAs) such as 20:4n–6, 20:5n–3 and 22:6n–3.     

Population Abundances, Tidal Movement, Burrowing Ability and Oxygen Uptake of Emerita analoga (Stimpson) (Crustacea, Anomura) on a Sandy Beach of South-Central Chile

Abstract. Field sampling and other experiments were carried out during February 2001 to determine whether different morphodynamic characteristics occurring within an intermediate sandy beach of southern Chile (ca. 39°S) convey differences in population abundance, tidal movement, burrowing ability and oxygen uptake of the anomuran crab Emerita analoga (Stimpson, 1857). Crabs were collected along transects extended between the lowest swash levels and the retention zone above the effluent line of the south and north end of the beach. Burrowing times of nearly 70 crabs collected at each study site were measured in saturated sands collected from the lowest swash level of each site. Oxygen uptake of crabs was measured in incubation glass bottles. The intertidal zone of the north end of the beach was wider (56 m) and flatter (1/14) than that of the south end (45 m and 1/9, respectively). In general, the swash zone of the north end was significantly wider than the south end throughout the sampling period. The frequency of swashes and number of swash crossings above the effluent line, plus up‐wash speed, were usually higher at the steeper south end of the beach. The mean population abundance of E. analoga per linear metre of beach was significantly higher at the north end, whereas density per square metre was significantly higher at the south end. No differences were found in biomass figures. Although the highest abundance of crabs at the north end was usually observed at the lowest swash levels, similar population abundances occurred along all the tidal levels sampled at the south end. Burrowing times of crabs collected from both ends of the beach increased significantly with increasing carapace length and body mass. The mean burrowing time of crabs collected at the south end of the beach was shorter than that of those collected at the north end. Oxygen uptake of E. analoga was positively and exponentially correlated with the size of individuals collected from both ends of the beach. Results of ANCOVA showed no significant difference between the regression lines obtained for the oxygen uptake of crabs collected at both ends of the beach. It is concluded that physical features of each end of the beach seem not to differ enough to produce differences in oxygen uptake of E. analoga, or in the biomass, population structure and body size of crabs, within a single beach of south‐central Chile.     

Population biology of the fiddler crab Uca annulipes (Brachyura: Ocypodidae) in a tropical East African mangrove (Mozambique)

The community structure of the tropical fiddler crab Uca annulipes (H. Milne Edwards) has been studied in Mozambique for the first time. Ten 0.25 m² squares were randomly sampled on a monthly basis during low tide periods from January to December 2002 at Costa do Sol Mangrove, Maputo Bay, southern Mozambique. A total of 981 crabs were captured, of which 494 were males (50.36%), 231 non-ovigerous females (23.85%) and 253 ovigerous females (25.79%). The overall size frequency distribution was unimodal, with males being larger than both ovigerous and non-ovigerous females. Females were more abundant in the smallest size classes while males outnumbered females in the largest ones. The overall sex ratio (1:0.99) did not differ from the expected 1:1 proportion, but significant deviations were observed in January, April, August and November. Results indicate that the size frequency distributions of this species can also be determined through direct analysis of burrow openings. Breeding took place year-round with two peaks of spawning in summer (January and December). Recruits were present throughout the year, with high abundance in summer probably due to the high reproductive activity observed in this season. The present results may allow us to suggest that U. annulipes follows a rapid breeding cycle accompanied by a rapid larval development and settlement in the study area.     

Recruitment of flatfish species to an estuarine nursery habitat (Lima estuary,NW Iberian Peninsula)

One of the present concerns of fish biologists involves defining and identifying nursery habitats in the context of conservation and resource management strategies. Fish nursery studies usually report upon nursery occupation during the latter juvenile stages, despite the fact that recruitment to nurseries can start early in life, during the larval phase. Here we investigated the use of a temperate estuarine nursery area, the Lima estuary (NW Portugal), by initial development stages of flatfish species before and after metamorphosis, integrating the larval and juvenile phases. The Lima estuarine flatfish community comprised twelve taxa, seven of which were present as pelagic larvae, six as juveniles and three as adults. There was a general trend of increasing spring–summer abundance of both larvae and juveniles, followed by a sharp winter decrease, mainly of larval flatfishes. The Lima estuary was used by Solea senegalensis, Platichthys flesus and Solea solea as a nursery area, with direct settlement for the two first species. In contrast, indirect settlement was suggested for S. solea, with metamorphosis occurring outside the estuarine area. Estuarine recruitment of S. senegalensis varied between years, with young larvae occurring in the estuary throughout a prolonged period that lasted 6–9 months, corroborating the protracted spawning season. P. flesus, the second most abundant species, exhibited a typical spring estuarine recruitment, without inter-annual variations. Developed larvae arrived in the estuary during spring, whereas the 0-group juveniles emerged in the following summer period. The present study contributes new insight to our understanding of the economically important S. senegalensis, and highlights the importance of integrating the planktonic larval phase into traditional flatfish nursery studies.     

Health status of red mullets from polluted areas of the Spanish Mediterranean coast,with special reference to Portmán (SE Spain)

A suite of general physiological indicators and hepatic biomarker responses were determined in red mullet (Mullus barbatus) from priority pollution areas of W Mediterranean Sea, including the highly metal polluted area of Portmán (Cartagena, SE Spain). Concentrations of metals and persistent organic pollutants in fish muscle tissue and sediment samples were also analysed. Our results showed that fish from Portmán accumulated the highest concentrations of mercury, lead and arsenic and also of some polycyclic aromatic hydrocarbons and polychlorinated biphenyls congeners. In addition, they had significantly lower condition factor, muscle lipid content and gonadosomatic index, as well as the lowest levels of DNA integrity and the highest ethoxyresorufin-O-deethylase (EROD) activity in liver of the areas investigated. Contaminant body burden in fish only partly corresponded to chemical characteristics of the sediments in the areas in which they were collected. Our findings indicate that red mullets from Portmán had suboptimal health status that warrant further study.     

Expression and activity of glutathione S-transferases and catalase in the shrimp Litopenaeus vannamei inoculated with a toxic Microcystis aeruginosa strain

Microcystin (MC) produced during cyanobacteria blooms is notably toxic to human and wildlife. Conjugation with reduced glutathione (GSH) by glutathione S-transferase (GST) and the antioxidant enzymes defenses (e.g. catalase, CAT) are important biochemical defense mechanisms against MCs toxicity. We investigated the enzymatic activity of CAT and GST and the gene expression levels of CAT and eight GST isoforms in the hepatopancreas of the globally farmed shrimp Litopenaeus vannamei 48-h after injection with a sub-lethal dose of 100 μg kg⁻¹ of a toxic Microcystis aeruginosa extract. MCs caused up-regulation for GST Ω, μ and a MAPEG isoform, by 12-, 2.8- and 1.8-fold, respectively, and increases in the total GST enzyme activity and CAT enzyme activity. The study points to the importance of further characterization for the L. vannamei GST isoforms and GST/CAT post-translational regulation processes to better understand the key mechanisms involved in the shrimp’s defense against MC exposure.     

Biomarkers of biochemical and cellular stress in Carcinus maenas: an in situ field study

Biomarkers are useful diagnostic tools for identifying exposure to physical, chemical, and environmental stresses. In this study a suite of techniques—measurements of metallothionein, lysosomal integrity, and osmoregulatory ability—have been used in an attempt to provide an in situ assessment of environmental quality. Five estuarine sites were studied, two clean sites, two urban sites, and one intermediate. Shore crabs, Carcinus maenas, were collected and haemolymph taken for the lysosomal and osmoregulatory assays. Animals were killed and tissue preserved for metallothionein analysis. Lysosomal integrity was significantly different in crabs from clean sites and the urban sites (n = 40, p < 0.05). The potential value of the biomarker approach is that it could provide data for impact assessment and is applicable in the field.     

Implementing geostatistical analysis to study spatio-temporal distribution patterns of swimming crabs(Portunus trituberculatus)

This study attempted to compare the performance of local polynomial interpolation,inverse distance weighted interpolation,and ordinary kriging in studying distribution patterns of swimming crabs.Cross-validation was used to select the optimum method to get distribution results,and kriging was used for making spatial variability analysis.Data were collected from 87 sampling stations in November of 2015(autumn) and February(winter),May(spring) and August(summer) of 2016.Results indicate that swimming crabs widely distributed in autumn and summer:in the summer,they were more spatially independent,and resources in each sampling station varied a lot;in the winter and spring,the abundance of crabs was much lower,but the individual crab size was bigger,and they showed the patchy and more concentrative distribution pattern,which means they were more spatially dependent.Distribution patterns were in accordance with ecological migration features of swimming crabs,which were affected by the changing marine environment.This study could infer that it is applicable to study crab fishery or even other crustacean species using geostatistical analysis.It not only helps practitioners have a better understanding of how swimming crabs migrate from season to season,but also assists researchers in carrying out a more comprehensive assessment of the fishery.Therefore,it may facilitate advancing the implementation in the pilot quota management program of swimming crabs in northern Zhejiang fishing grounds.     

Comparing quality of estuarine and nearshore intertidal habitats for Carcinus maenas

Estuarine and nearshore marine areas are vital habitats for several fish and benthic invertebrates. The shore crab Carcinus maenas (Crustacea: Brachyura: Portunidae) inhabits a variety of coastal, estuarine and lagoon habitats. At low tide, habitat structural complexity may be most important for crabs in the intertidal, providing refuge from predation and desiccation. The quality of different vegetated and nonvegetated estuarine and rocky shore habitats in SW Portugal and SW England was evaluated for intertidal C. maenas populations. We estimated population density, size–structure, and potential growth (RNA/DNA ratios) to investigate habitat quality. Vegetated estuarine habitats supported higher crab densities, than nonvegetated estuarine and rocky shore habitats. Investigation of population size–structure revealed that all habitats seem important recruitment and nursery areas although estuarine habitats in SW Portugal appeared to support higher densities of new recruits than equivalent habitats in SW England. Significant variation was found in RNA/DNA ratios among habitats. Ratios were highest in the rocky shore suggesting a high quality habitat where growth potential is high. We speculate that competition from other top-predators (Pachygrapsus spp.) rather than low habitat quality may limit the occurrence of C. maenas in intertidal rocky shore habitats in SW Portugal. In estuarine environments RNA/DNA ratios were significantly higher in the vegetated than in the nonvegetated estuarine habitats in SW Portugal but not in SW England, suggesting geographic differences in the extent to which highly structure habitats represent high quality. Our results challenge the current paradigm that structured habitats are necessarily those of higher quality for C. maenas.     

Marine invertebrate glutathione-S-transferases: purification, characterization and induction

High glutathione-S-transferase (GST) activity was found in hepatopancreas and gill cytosol of the blue crab (Callinectes sapidus) and the digestive gland cytosol of two marine gastropods (Nassarius obsoletus and Cerithium floridanum).Purification of GST from crab hepatopancreas by Sephadex G-200, DEAE-Sephacel and chromofocusing resulted in the isolation of two isoenzymes with isoelectric points of 5·9 and 5·7 (GST 5·9 and GST 5·7). Antibodies were prepared to these two isoenzymes and the two forms cross-reacted immunologically. The two transferases had similar molecular weights, amino acid compositions, substrate specifities and kinetic parameters.Crab gill cytosol showed one isoenzyme which reacted with antibodies to GST 5·9 and GST 5·7. The major isoenzyme of N. obsoletus was a basic form while C. floridanum showed a homodimer acidic form. The gastropod GST forms did not react with antibodies to crab GST. The presence of the phenolic antioxidant, butylated hydroxytoluene, in the diet of blue crab or shrimp (Penaeus aztecus) resulted in high hepatic GST activity.