Expression analysis of vasa in Asian paddle crab (Charybdis japónica) exposed to Bisphenol A

Background: Bisphenol A (BPA) is an endocrine-disrupting chemical (EDC) with a weak estrogen-like activity in fish that is found ubiquitously in aquatic environments. However, there has been little study about BPA on the endocrine disrupting effects of crab. In the present study, cDNA of vasa was cloned and characterized in the Charybdis japonica. Histological structures of testis and expression patterns of vasa gene in the testis of C. japonica after treatment with BPA were investigated. Results: The cDNA of vasa is composed of 3051 bp with a 2166 bp open reading frame encoding 721 AA. The deduced amino acid sequence contained eight conserved domains of the DEAD-box protein family. The tissue distribution showed that vasa mRNA was specifically expressed in ovary and testis. Histologically, the sperm cells were decreased in number and an acellular zone was seen in the testis. The transcript level of vasa gradually increased with a significant difference between the experimental and control groups. After BPA exposure with 0.50 and 1.00 mg/L for 1,3, 6 and 9 d, the expression levels of vasa increased. Conclusion: These findings suggest that BPA can increase the expression level of vasa mRNA and influence the development of the testis in C. japonica.

Changes in oxygen consumption and heart rate of the blue swimming crab, Portunus pelagicus (Linnaeus, 1766) following exposure to sublethal concentrations of copper.

Weight specific oxygen consumption and heart rates were investigated in blue swimming crab, Portunus pelagicus to indicate the stress in response to increasing salinities (50, 75, 100 and 125%SW) and increasing copper concentration (0, 0.75, 1.5, 3 and 6 ppm copper) at temperature 25 degrees C. Oxygen consumption (micromol.g(-1).h(-1) was measured using closed system respirometer (Strathkelvin Instruments oxygen meter model 781). Heart rates (beat.min(-1)) were recorded using impedance techniques. It has been found that oxygen consumption and heart rate increased with decreasing salinities and increasing copper concentration. The statistic showed weight specific oxygen consumption of P. pelagicus after 96 hr exposure to 6 ppm copper in all salinities were significantly different (P < 0.05) from 0 ppm copper. Mean heart rate of P. pelagicus in 50%SW after 96 hr exposure to 6 ppm copper were significantly different (P < 0.05) from 0 ppm copper.

Chronic toxicity bioassay and effect of endosulfan, HCH, copper and zinc on the survival and intermoult duration of larvae of mangrove crab Macrophthalmus erato De Man.

The effects of insecticides (endosulfan, HCH) and heavy metals (copper and zinc) were studied on survival and duration of the moult cycle of larvae and megalopa of a common mangrove crab, Macrophthalmus erato, that inhabits Pitchavaram mangroves, southeast coast of India. In chronic toxicity test, percentage of survival from hatchling to megalopa stage at different concentrations of endosulfan (0.048 to 0.36 microg/L), HCH (0.32 to 2.4 microg/L), copper (9.0 to 67.5 microg/L) and zinc (15.2 to 139.0 microg/L) were 6, 12, 8 and 12 respectively. Endosulfan, HCH, copper and zinc considerably increased the total period in development from first zoea to megalopa stage from 18 days in the control to 23, 21, 20 and 22 days in the treated specimens. It is concluded that concentrations well below the observed chronic concentrations, may prove to be more deleterious to larva of this species in natural environment leading to reduction in the success of recruitment to the adult population.

Identification of hemolytic and neuroactive fractions in the venom of the sea anemone Bunodosoma cangicum

Sea anemones are a rich source of biologically active substances. In crayfish muscle fibers, Bunodosoma cangicum whole venom selectively blocks the I K(Ca) currents. In the present study, we report for the first time powerful hemolytic and neuroactive effects present in two different fractions obtained by gel-filtration chromatography from whole venom of B. cangicum. A cytolytic fraction (Bcg-2) with components of molecular mass ranging from 8 to 18 kDa elicited hemolysis of mouse erythrocytes with an EC50 = 14 æg/ml and a maximum dose of 22 æg/ml. The effects of the neuroactive fraction, Bcg-3 (2 to 5 kDa), were studied on isolated crab nerves. This fraction prolonged the compound action potentials by increasing their duration and rise time in a dose-dependent manner. This effect was evident after the washout of the preparation, suggesting the existence of a reversible substance that was initially masking the effects of an irreversible one. In order to elucidate the target of Bcg-3 action, the fraction was applied to a tetraethylammonium-pretreated preparation. An additional increase in action potential duration was observed, suggesting a blockade of a different population of K+ channels or of tetraethylammonium-insensitive channels. Also, tetrodotoxin could not block the action potentials in a Bcg-3-pretreated preparation, suggesting a possible interaction of Bcg-3 with Na+ channels. The present data suggest that B. cangicum venom contains at least two bioactive fractions whose activity on cell membranes seems to differ from the I K(Ca) blockade described previously

Acute and chronic effects of cadmium on blood homeostasis of an estuarine crab, Chasmagnathus granulata, and the modifying effect of salinity

Whole body oxygen consumption and some hemolymph parameters such as pH, partial pressure of gases, level of ions and lactate were measured in the estuarine crab Chasmagnathus granulata after both acute (96 h) and chronic (2 weeks) exposure to cadmium at concentrations ranging from 0.4 to 6.3 mg/l. In all instances, the crabs developed hemolymph acidosis, but no respiratory (increased PCO2) or lactate increases were evident. Hemolymph levels of sodium and calcium were always increased by cadmium exposure. The chronic toxicity of cadmium was enhanced at 12 0/00 salinity, even causing a significantly higher mortality in comparison with the higher salinity (30 0/00) used. A general metabolic arrest took place at 12 0/00 salinity in the crabs chronically exposed to cadmium, as indicated by decreases of oxygen consumption and PCO2, an increase of PO2, along with no changes in lactate levels. These imbalances were associated with severe necrosis and telangiectasia in the respiratory gills, probably leading to respiratory impairment and finally histotoxic hypoxia and death of the animals

Zinc accumulation in phosphate granules of Ucides cordatus hepatopancreas

Amorphous phosphate granules are present in vertebrate and invertebrate organisms. The functions attributed to these structures depend on their mineral contents and organic matrix composition. In the present study we have determined zinc concentrations in the hepatopancreas of the crab Ucides cordatus from regions contaminated with zinc, and the elemental composition of hepatopancreal phosphate granules. Organisms were collected from the contaminated areas of Sepetiba Bay (SB) and Guanabara Bay (GB), and from a non-contaminated area, Ribeira Bay (RB). The first two sites are located near the metropolitan region of Rio de Janeiro city, Brazil. Atomic absorption spectroscopy (AAS) showed a significant difference (P<0.05) for zinc concentration in the hepatopancreas from organisms collected at the contaminated sites GB (210 + or - 20 µg/g dry weight) and SB (181 + or - 16 µg/g dry weight) compared to the non-contaminated site RB (76 + or - 14 µg/g dry weight). Phosphate granules isolated from hepatopancreatic tissue were studied by electron diffraction (ED), energy dispersive X-ray analysis (EDX) and electron spectroscopic imaging (ESI). ED of granules presented no diffraction spots, indicating that these structures are in an amorphous state, while EDX of granules isolated from a contaminated area contained P, Ca and Zn. Mg, Cl and Fe were also found in some of the spectra. ESI showed that O, P and Ca were colocalized in the mineralized layers of most granules observed. The correlation between the results obtained by AAS and those obtained by microanalytical techniques suggests that the hepatopancreatic granules of U. cordatus may be related to the phenomenon of heavy metal retention

Occurrence of tetrodotoxin and paralytic shellfish toxins in Phallusia nigra (Tunicata, Ascidiacea) from the brazilian coast

We have already shown the presence of guanidine neurotoxins in calcareous red algae and mussels collected in the Säo Sebastiäo channel State of Säo Paulo,Brazil). It is known that these neurotoxins comprise more than 25 analogues such as tetrodotoxin (TTX) and derivatives plus the paralytic shellfish toxins (PST) found in a variety of marine, freshwater and amphibious species. Filter feeding animals generally possess large amounts of these neurotoxins. The tunicates are sessil marine animals with a high rate of sea water filtration. The tunics and siphons of 50 specimens of Phallusia nigra were dissected and the visceral organs were immersed in methanol containing acetic acid 0.02 N ph 5.0. The extract was prepared by homogenization, filtration and the methanolic phase was concentrated under reduced pressure and defatted with chloroform. The polar phase was evaporated and the residue dissolved in deionized water for further purification in ionic-exchange resin column (Bio-Gel P-2) and HPLC analysis. The extract showed paralytic effects on mouse assay (26.9 MU/100mg) and on crustacean isolated nerve preparations. The chemical analysis for TTX and PST revealed toxins with retention times similar to gonyautoxins, saxitoxins and TTX. These findings are important to explain future toxin envenoming outbreaks on the Brazilian coast.

Respiratory responses and blood sugar level of the crab, Barytelphusa cunicularis (Westwood), exposed to mercury, copper and zinc.

Barytelphusa cunicularis (Westwood), a freshwater crab was exposed to mercuric chloride, copper sulphate and zinc sulphate from 0 to 12 hr and the oxygen consumption of the animal was measured in order to study the stress caused by these heavy metals. Normal oxygen consumption was affected by the three heavy metals. Similarly, crabs exposed to sublethal concentrations of the same pollutants for 24, 48, 72 and 96 hr showed an elevation in the blood sugar level with a maximum increase at 48 hr. The results indicate a switch towards glycolysis in order to overcome the anaerobic stress caused by the heavy metals.