Physiological responses of juvenile and adult shore crabs Carcinus maenas (Crustacea: Decapoda) to pyrene exposure

This study measured aspects of the physiology of juvenile [(<35mm carapace width (CW)] and adult (>60mm CW) Carcinus maenas to test the hypothesis that these different life-history stages exhibit different sensitivities to environmental contamination. Newly-collected juveniles had significantly (P<0.05) lower immunocompetence (phagocytosis and cellular integrity), lower metabolic energy (haemolymph glucose) and increased scope for growth compared with adults. Seven day exposure to a sub-lethal concentration (200microgL(-1)) of pyrene significantly (P<0.01) reduced immunocompetence, elevated basal heart rate and decreased respiration (at rest) for juveniles but had no overall impact on adult crab physiology. Results confirm that physiological differences exist between juvenile and adult shore crabs, and cause juveniles to be more susceptible to the effects of pyrene exposure than adults. Such differences in sensitivity to contamination between life-cycle stages of the same species have to be taken into account during the risk assessment process.     

Cypermethrin induces glutathione S-transferase activity in the shore crab,Carcinus maenas

Cypermethrin is a synthetic pyrethroid that is particularly toxic to crustacea. It is therefore applied as a chemotherapeutant in the salmonid aquaculture industry for the treatment of sea lice infestations. After use, cypermethrin is released directly into the marine environment, to be diluted by fresh seawater. The shore crab, Carcinus maenas is found in the vicinity of fish farms, and may come into contact with released cypermethrin. The detoxification enzyme, glutathione S-transferase (GST) has been implicated in cypermethrin metabolism in terrestrial arthropods, but this has not yet been demonstrated in crustacea. In this paper we investigate the response of GST activity in Carcinus to cypermethrin exposure, and also the time course of the induction process. GST activity is significantly increased in Carcinus exposed to nominal concentrations of 50 and 500 ng/l of water-borne cypermethrin. Carcinus demonstrate a significant elevation in GST activity following intra-cephalothoracic injection with 10 ng of cypermethrin. GST activity returns to basal levels after 36 h. The potential application of GST activity in Carcinus as a biomarker of cypermethrin exposure is discussed.     

A non-destructive assessment of the exposure of crabs to PAH using ELISA analyses of their urine and haemolymph

Urine and haemolymph can be repeatedly sampled from crabs with no (or limited) damage to the organism. Their analysis offers a measure of the animals' exposure to biologically available contaminants. Shore crabs (Carcinus maenas) were exposed to the PAHs phenanthrene and pyrene at concentrations ranging from 20 to 200 microg l(-1). After 48 h, urine and haemolymph samples were taken and analysed using ELISA and UV-fluorescence spectrophotometry. High correlations were recorded between the two sets of results from the urine analyses (r2 = 0.83 for phenanthrene and r2 = 0.88 for pyrene). Contaminant concentrations were much lower in haemolymph than in the urine. Analyses of urine taken from crabs collected from clean and contaminated sites confirm the suitability of these analyses for environmentally exposed organisms. Again, a good correlation was recorded between the ELISA and spectrofluorimetric analysis (r2 = 0.83). In this instance, difficulties were experienced with haemolymph analyses owing to a lack of sensitivity.     

Temperature-dependent physiological response of Carcinus maenas exposed to copper

The effect of temperature on the heart rate (fH) of Carcinus maenas exposed to a sublethal nominal concentration (0.5 mg l(-1)) of copper is reported. Adult, intermoult males (4 cm carapace width) were collected from Stavanger Fjord (Norway) in August (seawater TEMPERATURE=17 degrees C) and maintained in the laboratory (fed ad libitum) at 5, 15 and 25 degrees C for 7 days. Following this holding period, crabs were exposed to waterborne copper at the same temperature. After 3 days of exposure, individual fH was measured using the non-invasive Computer Aided Physiological MONitoring system (CAPMON) method. Copper-exposed individuals demonstrated significantly increased fH compared with controls at 5 and 25 degrees C (P<0.01). It is inferred that physiological function in C. maenas may be more vulnerable to copper contamination at seasonal temperature extremes (5 and 25 degrees C) than at the 'standard' test temperature (15 degrees C). Additionally, cardiac arrest observed at 5 degrees C is interpreted as a physiological response to limit copper uptake while the erratic fH in the 25 degrees C group reflected enhanced copper toxicity.     

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.     

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.     

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.     

Evaluation of fixed wavelength fluorescence and synchronous fluorescence spectrophotometry as a biomonitoring tool of environmental contamination

A field study was carried out to investigate whether polyaromatic hydrocarbon (PAH) contamination in shore crabs (Carcinus maenas) could be measured using fluorescence spectrometry. Crabs were collected as part of an integrated study to assess PAH contamination in Southampton Water, UK. Urine was collected from crabs via a non-destructive technique and analysed using fixed wavelength fluorescence (FF) and synchronous fluorescence spectrophotometry (SFS) in order to determine the presence of PAH metabolites, indicative of PAH exposure. By scanning at wavelength pairs specific to a variety of PAH groups the fluorescence of metabolites was analysed and attributed to the following PAH groups; naphthalenes, pyrenes and benzo[a]pyrene (BAP). Naphthalene-type, pyrene-type and BAP-type metabolites were detected using FF and SFS techniques and two distinct parameters were measured; Peak Intensity and Peak Area. Results showed that crabs obtained near a petroleum refinery, a point source of PAH discharge, exhibited significantly higher PAH metabolites for all PAH groups, than at sites sampled along the estuary (ANOVA, p<0.05). This study highlights the effectiveness of using FF and SFS as a rapid and cost-effective biomonitoring tool for detecting PAH contamination.     

Copper complexation by fulvic acid affects copper toxicity to the larvae of the polychaete Hydroides elegans

Copper toxicity is influenced by a variety of environmental factors including dissolved organic matter (DOM). We examined the complexation of copper by fulvic acid (FA), one of the major components of DOM, by measuring the decline in labile copper by anodic stripping voltammetrically (ASV). The data were described using a one-site ligand binding model, with a ligand concentration of 0.19micromol site mg(-1) C, and a logK' of 6.2. The model was used to predict labile copper concentration in a bioassay designed to quantify the extent to which Cu-FA complexation affected copper toxicity to the larvae of marine polychaete Hydroides elegans. The toxicity data, when expressed as labile copper concentration causing abnormal development, were independent of FA concentration and could be modeled as a logistic function, with a 48-h EC(50) of 58.9microgl(-1). However, when the data were expressed as a function of total copper concentration, the toxicity was dependent on FA concentration, with a 48-h EC(50) ranging from 55.6microgl(-1) in the no-FA control to 137.4microgl(-1) in the 20mgl(-1) FA treatment. Thus, FA was protective against copper toxicity to the larvae, and such an effect was caused by the reduction in labile copper due to Cu-FA complexation. Our results demonstrate the potential of ASV as a useful tool for predicting metal toxicity to the larvae in coastal environment where DOM plays an important role in complexing metal ions.     

Genetic heterogeneity in populations of the Mediterranean shore crab, Carcinus aestuarii (Decapoda,Portunidae), from the Venice Lagoon

Heterogeneity in genetic composition among recruits, mostly due to a large variance in reproductive success mediated by oceanographic processes, has been reported for marine species but is less understood in coastal lagoons' organisms. Temporal genetic variation in natural populations of the Mediterranean shore crab Carcinus aestuarii was quantified over a multi-year sample. A total of 486 adult crabs were collected at eight different sites of the Venice Lagoon during the period 2005–2007 and screened for genetic variation using 11 microsatellite loci. Two additional samples (N = 115) from neighbouring sites, located approximately 100 km North and South to the Venice Lagoon, were included for the sake of comparison. Our results show significant differences in allelic frequencies at the micro-geographic scale of the Venice Lagoon, observed between sites of collection, typologies of habitat, and areas with different class of ecological risk or pattern of hemocyanin expression. However, this pattern was not constant between years, with significant differences observed mainly in 2005 and 2006, but not in 2007. Our results indicate significant temporal differences suggesting the existence of dynamic processes that act on the genetic pool of this species. Although natural selection and gene flow might play a role, we suggest that genetic drift linked to large variation in the reproductive success of individuals is the most probable scenario to explain the local genetic patterns of differentiation in the Mediterranean shore crab. Our study, by providing the first evidence for the existence of genetic differences in this species at the micro-geographic scale, suggests that a better comprehension of the link between reproduction, recruitment and oceanography is critical to understand how colonization and maintenance of genetic variation is achieved in ephemeral and vulnerable environments such as coastal lagoons.     

Purification of two crab-paralysing polypeptides from the sea anemone Bolocera tuediae

Two toxic polypeptides BTTX I and BTTX II were isolated from the sea anemone Bolocera tuediae. Toxin isolation was achieved by alcoholic extraction of the homogenized tentacles and filaments, batchwise adsorption onto ion-exchangers (QAE-Sephadex A25, CM-Cellulose, Se-Cellulose, Sp-Sephadex C25), gel filtration on Sephadex G50 and G25 and ion-exchange chromatography on SP-Sephadex C25 and QAE-Sephadex A25. The toxins were tested on the shore crab Carcinus maenas by intramuscular injection. LD₁₀₀ for BTTX I is 500 μg kg⁻¹ C. maenas, whereas the BTTX II it is 40 μg kg⁻¹. The molecular weights of both toxins are estimated to be 5000 Dalton.     

Physical forcing of onshore transport of crab megalopae in the northern Portuguese upwelling system

Wind-driven circulation is a mechanism that may be involved in the shoreward transport of invertebrate larvae. This study investigated the temporal variability of the supply of crab megalopae to an estuary located in the northwest coast of Portugal, which is affected by seasonal upwelling, and related variations in supply to several upwelling indices. Megalopae of the European common shore crab Carcinus maenas (L.). were collected daily, using artificial settlement substrates, during a 3-month period, from March 15 to June 15, 2000. Two settlement episodes were detected, which were consistent with the variation in abundance and size of juvenile crabs. Variations in subtidal sea level and sea surface temperature anomaly were correlated with the strength of along-shore wind stress and consistent with upwelling theory. Contingency analyses showed positive significant associations between increasing intensity of southerly winds and megalopal supply, for time lags of 0–6 d, indicating that shoreward transport occurred during relaxation or reversal of upwelling favourable winds. Associations between megalopal supply and tidal range or the duration of nocturnal flood were not detected. Compared to other studies of the supply of crab megalopae to coastal habitats that used a similar methodology, very low numbers of megalopae were collected during this study. The paucity of C. maenas megalopae in the Ria de Aveiro is in agreement with the low densities of this larval stage in the plankton, and indicates that maintenance of the population is achieved despite a low supply of megalopae.     

Dominance of blue mussels versus consumer-mediated enhancement of benthic diversity

In the shallow subtidal of Kiel Fjord (western Baltic Sea), the blue mussel Mytilus edulis is the dominant competitor for space. Vertically suspended settlement substrata in the upper 6 m of the water column almost invariably become dominated by M. edulis within a few summer months. However, not all naturally available hard substrata bear mussel monocultures.In three in situ experiments we investigated the dominance of mussels and the influence of local consumers on establishment and dynamics of a benthic community: (a) the natural course of succession in the absence of benthic consumers was followed on vertically suspended settlement substrata, (b) settling plates were exposed to natural recruitment being either accessible or inaccessible to all benthic consumers, and (c) the three major local consumer species, viz. the shore crab Carcinus maenas, the starfish Asterias rubens and the periwinkle Littorina littorea, were enclosed separately in cages containing a settling panel to assess species-specific consumer effects on recruitment.The results illustrate that in this region of the Baltic Sea mussels do have the potential to dominate ungrazed substrata within a few weeks and that top-down effects (predation and grazing) may control community structure. While some species - mussels in particular - were suppressed by consumption, others seemed to benefit from the presence of consumers. Thus, barnacles and algae thrived when consumers were present. Blue mussels being the locally dominant competitor, the beneficial effects of consumers on barnacles and algae were presumably indirect ones through consumer-caused release from asymmetrical competition.The isolated effects of C. maenas, A. rubens and L. littorea, on recruitment differed in quality (positive vs. negative), quantity (strength of effect) and specificity. Barnacle recruitment was significantly reduced in the presence of both starfish and shore crabs. Diatom recruitment was significantly reduced by snails and shore crabs. Scyphozoan recruitment was significantly reduced by crabs only, but was significantly increased by snails. The influence of the shore crabs was most remarkable: recruitment by any potential coloniser species was impeded to a point where in the presence of single individuals of C. maenas the plates stayed perfectly clean macroscopically.     

4-tert-octylphenol and 17beta-estradiol applied by feeding to flounder Platichthys flesus: induction of vitellogenin and accumulation in tissues

In order to simulate a possible natural administration route of xenoestrogens male flounder Platichthys flesus were exposed via the diet to the alkylphenol 4-tert-octylphenol. Treatment with 4-tert-octylphenol (10, 50 and 100 mg OP kg(-1) BW) or 17beta-estradiol (0.05 mg kg(-1) BW) every second day during a period of 11 days resulted in a significant increase in plasma vitellogenin (Vtg) concentrations. The induction of Vtg was greatest in the fish receiving 50 mg OP kg-' BW. A significant accumulation of 4-tert-octylphenol was found in liver and muscle tissue of the OP treated groups. The tissue concentrations of 4-tert-octylphenol and the plasma vitellogenin concentration were positively correlated. The results show that 4-tert-octylphenol accumulates in liver and muscles of flounder P. flesus, and exerts estrogenic effects such as vitellogenin induction.     

Marine invasive aliens on South African shores: implications for community structure and tropillc functioning

Up to 12 marine mollusc and three crab species are thought to have been deliberately or accidentally introduced to South Africa. Of these, only the Mediterranean mussel Mytilus galloprovincialis and the European shore crab Carcinus meanas have become invasive. M. galloprovincialis, probably introduced in the late 1970s, is already the dominant mussel throughout the Cape west coast. As a result, indigenous intertidal mussels Aulacomya ater have been largely displaced, mussel standing stock has increased and the upshore limit of mussel beds has become considerably elevated. Implications include competitive exclusion of large limpets but enhanced recruitment of juvenile limpets, increased habitat availability for mussel infauna, reduction of habitat for algal infauna and enhanced food availability for predators, particularly terrestrial species. C. maenas was first recorded from Table Bay Docks in 1983 and currently ranges from Camps Bay to Saldanha Bay. Although it is a voracious predator, it appears unable to colonize wave-swept shores, so is unlikely to displace indigenous crabs, or to severely impact prey species on the open coast. Valuable conservation areas and mariculture sites in sheltered lagoonal areas are, however, threatened by it.     

Horizon scanning for alien predatory crabs: insights from South Africa

Due to the presence of few dominant predators on South African rocky shores, this coastline could be vulnerable to invasion by predatory crabs. This study applied horizon scanning to create an ordered watch list of alien crab species that could establish along this coastline under present-day and future temperature scenarios. This was done by: (i) identifying the species with both an invasion history and a possible pathway to South Africa; (ii) comparing the temperature ranges of the species’ native and introduced distributions to those of each of the four South African ecoregions; and (iii) ranking the species based on their potential ecological impacts. Of the 56 alien predatory crab species known worldwide, 28 species have pathways to South Africa. Incompatible temperature ranges excluded only two species from each ecoregion. Negative ecological impacts in their invaded ranges placed Japanese shore crab Hemigrapsus sanguineus, brush-clawed shore crab H. takanoi and Chinese mitten crab Eriocheir sinensis at the top of the watch list. This study highlights that many alien crab species have the potential to reach South Africa, with most likely to survive. This watch list should be used to support targeted monitoring and so facilitate early detection of these species, should they reach South Africa.     

Transport and accumulation of organochlorines in the ovaries of Atlantic croaker (Micropogonias undulatus)

The role of vitellogenin in the transport of organochlorines was investigated in Atlantic croaker (Micropogonias undulatus) by exposing them through the diet to o,p′-DDT at a concentration of 10.8 μg/100 g fish/day for 14 days or Aroclor 1254 (PCB) at a concentration of 0.5 mg/100 g fish/day for 30 days during gonadal recrudescence. Tissue samples were taken from the fish at various times after initial exposure, and o,p′-DDT and PCBs were extracted in acetonitrile and analyzed by gas chromatography. Analysis of the ovarian tissue collected 2 and 3 weeks from the start of exposure revealed that the o,p′-DDT concentration increases as the gonadosomatic index (GSI) increases (r² = 0.63), with accumulation ranging from less than 1% to as much as 8% of the total dosage. Interestingly, o,p′-DDT did not accumulate in the testes during the same exposure period. Accumulation of PCBs was found to be 40 times higher in the ovaries than in the testes. Gel filtration of plasma from exposed females showed that o,p′-DDT elutes in the low density lipoprotein and vitellogenin fractions. Control plasma incubated with o,p′-DDT at 4 °C for 16 h followed by chromatography on Sepharose 6B gave similar results with an o,p′-DDT concentration of 0.6μg/mg protein in the vitellogenin fraction. Furthermore, both o,p′-DDT and PCBs were found to bind to purified croaker vitellogenin. These results suggest that lipoproteins, including vitellogenin, are involved in the transport and accumulation of organochlorines in the ovaries of exposed fish.     

Kinetics of vitellogenin protein and mRNA induction and depuration in fish following laboratory and environmental exposure to oestrogens

Plaice, flounder and sand goby were exposed to ethynylestradiol (EE2) for 21 days and then followed for up to 31 days after removal of the oestrogen. Plasma vitellogenin (VTG) and hepatic VTG mRNA were determined in groups of fish sampled during the induction and post-exposure phases. VTG mRNA increased slightly earlier than plasma protein, but both reached maxima by 21 days. In contrast, VTG mRNA decayed much more rapidly than protein after EE2 exposure was terminated (typical values t(1/2) mRNA 3 days, protein 15-30 days). Vitellogenin and VTG mRNA thus measure different temporal events and this is illustrated by field data of male flounder in which both parameters have been determined. Few fish show co-ordinate increased VTG mRNA and vitellogenin but rather more fish have increased vitellogenin. Low level increases of VTG mRNA (5 X) is observed in some fish without increased vitellogenin and this may represent polymorphic differences between individual fish.     

Can starvation influence cellular and biochemical parameters in the crab Carcinus aestuarii?

Crustacea experience periods of starvation during moulting or when limited food availability occurs. The effects of starvation on Crustacea physiological responses have been demonstrated, whereas the effects of starvation on Crustacea immune parameters remain to be more fully studied. In the present study the effects of starvation on immune parameters and antioxidant enzyme activities of the crab Carcinus aestuarii were evaluated for the first time. Treated crabs were starved for 7 days, whereas control crabs were fed daily with mussels. Total haemocyte count (THC), haemocyte diameter and volume, haemocyte proliferation, cell-free haemolymph (CFH) glucose and total protein levels, and phenoloxidase (PO) activity in both haemocyte lysate (HL) and CFH were measured in crabs. In addition, superoxide dismutase (SOD) and catalase (CAT) activities were evaluated in both gills and digestive gland from crabs, in order to evaluate whether starvation induced oxidative stress in C. aestuarii. THC increased significantly in starved crabs, with respect to controls, whereas no significant variations were observed in haemocyte diameter, volume and proliferation. In CFH of starved animals glucose concentration significantly increased, whereas total protein concentration significantly reduced. A significantly higher PO activity was recorded in HL from starved crabs, than in control crabs. Conversely, PO activity did not vary significantly in CFH. Starvation did not cause significant alterations in antioxidant enzyme activities in both gills and digestive gland. Results obtained demonstrated that starvation influenced crab immune parameters, but did not induce oxidative stress. Results also indicated that C. aestuarii can modulate its cellular and biochemical parameters in order to cope with starvation.     

Effects of temperature on cellular and biochemical parameters in the crab Carcinus aestuarii (Crustacea, Decapoda)

The effects of temperature on cellular and biochemical parameters of the crab Carcinus aestuarii were evaluated. Crabs were kept for 7 days at 4, 17 (reference value) and 30 °C (salinity of 35 psu), and total haemocyte count (THC), haemocyte volume, haemocyte proliferation, phenoloxidase (PO) activity in both haemocyte lysate (HL) and cell-free haemolymph (CFH), CFH total protein and glucose levels, superoxide dismutase (SOD) and catalase (CAT) activities in both gills and digestive gland were evaluated. The lowest and the highest temperature significantly decreased THC, whereas haemocyte volume and haemolymph glucose concentration did not differ significantly among experimental conditions. Haemolymph protein concentration significantly reduced in crabs maintained at 30 °C, when compared with that of animals kept at 4 and 17 °C. Haemocyte proliferation increased significantly in crabs kept at 4 and 30 °C, when compared with that of crabs held at 17 °C. Likewise, a significantly higher PO activity was recorded in CFH from crabs kept at 4 and 30 °C, than in control crabs. Conversely, PO activity did not vary significantly in HL. With regard to antioxidant enzyme activities, a significant decrease in CAT activity was observed in gills from crabs kept at 4 °C, when compared to that of crabs kept at 17 °C and 30 °C. Results obtained demonstrated that the highest and lowest temperature tested influenced crab biological responses, and indicated that C. aestuarii modulated its cellular and biochemical parameters (mainly haemocyte proliferation, CFH protein concentrations and CFH PO activity) in order to cope with temperature.