Changes in the Level of Proteins,Free Amino Acids and Protease Activities of Clarias batrachus in Response to Sodium meta‐arsenite Intoxication

Increased hazard of arsenic toxicity at many provinces of India causing extensive contamination of their wetlands. To illustrate the consequences of the arsenic menace on the aquatic fauna, a nutritionally important freshwater catfish Clarias batrachus (Linn.) was subjected to the stress of sublethal concentration (1 mg/L; 5% of 96‐h LC₅₀ value) of sodium meta‐arsenite. The alterations induced by the arsenic salt on certain biomolecules (proteins, free amino acids (FAA), and protease activities) in five tissue components (muscle, liver, brain skin, and gills) of the fish using biochemical techniques were estimated. Significant (p < 0.01) decrease in the concentration of proteins in all the tissues (19.74–44.84% in muscles; 3.17–49.39% in liver; 10.05–36.16% in brain; 12.62–52.01% in skin and 7.34–61.69% in gills), and significant (p < 0.01) increase in FAA (6.84–269.89% in muscles; 125.89–358.03% in liver; 1.07–152.90% in brain; 29.31–401.62% in skin and 38.53–165.07% in gills), and protease activities (43.59–183.33% in muscles; 19.52–131.90% in liver; 8.85–46.72% in brain; 33.33–261.53% in skin and 25.39–50.79% in gills) of the exposed fish were observed. This increase in amino acids and enzyme activity point out toward breakdown of protein leading to their depletion.     

Acute and Chronic Toxic Effects of Cadmium on some Haematological,Biochemical, and Enzymological Parameters in the Fresh Water Teleost Fish Channa punctatus Akute und chronisch toxische Wirkungen von Cadmium auf einige hämatologische,biochemische und enzymologische Parameter im Süßwasserfisch Channa punctatus

The haematological, biochemical, and enzymological alterations produced on exposure of Channa punctatus to LC₅₀ (11.2 mg/L) for 96 h and to a sublethal concentration (1.12 mg/L) of cadmium (Cd²⁺) for 15, 30, 60, and 120 days have been studied. The fish were hypoglycemic and hypolactemic. The pyruvate content of blood and liver decreased in acute and all stages of chronic exposure except for 30 days where significant increase was recorded. Depletion was noted in the total protein and glycogen content of liver and muscle, and the level of lactate in liver in the two types of exposure. The levels of muscle lactate and pyruvate increased in both exposures. The activities of hexokinase, glucose-6-phosphatase, and lactate dehydrogenase decreased in liver after 96 h and all stages of chronic exposure except 30 days. The activities of hexokinase and lactate dehydrogenase in muscle and glutamate-oxalacetate transaminase and glutamate-pyruvate transaminase in serum, liver, and muscles increased in all exposures. The activities of succinate and malate dehydrogenases in liver and muscles decreased after acute and all stages of chronic exposure. The above results suggest that glycolysis and gluconeogenesis were impaired in liver but glycolysis prevailed in muscle. Aerobic oxidation is adversely affected in metal exposed fish, while proteolysis was increased.     

Effect of Parthenium Extract on the Metabolism of Young Colisa Fasciatus (BL. and SCHN.) (Pisces,Perciformes)

Parthenium hysterophorus seed extract possesses the potentiality to alter the fish metabolism. Lower concentrations (sublethal) of the seed extract stimulate the respiratory muscles, resulting in an increased rate of O₂ uptake, whereas higher concentrations (lethal) exhibit a neuro-toxic effect causing paralysis of the respiratory muscles manifested by a decreased rate of O₂ uptake, cessation of the respiratory pumps and casualty by asphyxiation. The seed extract also inhibits the activity of the enzymes SDH and carbonic anhydrase at the gas exchange sites. The log/log plotting of the O₂ uptake (mg/h) and the exposure period (2, 6, 12, 24, 72 and 168 h) showed slope values of 0.0744, 0.0778, 0.0322 and 0.0164, respectively, in four concentrations (10, 20, 25 and 30 %) of Parthenium extract.     

Tissue enzymes as biomonitors of PCP intoxication in Notopterus notopterus

In this paper, toxicity effects of pentachlorophenol have been studied using enzymes, phosphatases, transaminases and hydrogenases as indicators for toxicity. The sublethal concentrations used were 0.019, 0.0063 and 0.0038 mg/l (1/5th, 1/15th and 1/25th fractions of LC₅₀) and the experimental fishes were exposed for the periods of 15, 30, 45 and 60 days. Most of the significant alterations in enzymes were observed at 1/5th and 1/10th fractions except at few places in the liver where a significant alteration in the enzyme acid phosphatase occurs at 1/15th fraction. Glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) and lactic dehydrogenase (LDH) showed a significant increase in different tissues in all the fractions of treatment.     

Acute toxicity of platinum to coho salmon (Oncorhynchus kisutch)

The effects of short-term exposure to tetravalent platinum on survival, opercular movement and post-treatment growth of coho salmon fry (Oncorhynchus kisutch) was investigated. Employing a static water acute toxicity bioassay with platinum as PtCl₄2HCl·6 H₂O, at 8.5±0.2°C, and a water hardness of 55.9±3.5 mg l.^?1 (as CaCo₃), the 24, 48, and 96-h LC₅₀ values were 15.5, 5.2, and 2.5 mg Pt⁴⁺ l.^?1 respectively.Rates of opercular movement for fish exposed to platinum increased with increasing concentrations to a level of 1.0 mg l.^?1. No further significant increases were evident above this level. Hypoactivity of fish exposed to 0.3 mg l.^?1 and higher was evident during the acute toxicity bioassay and much of the post-treatment study. Post-treatment rate of growth for fish exposed to sublethal concentration of platinum for 96 h was less than that of the controls.All organs examined histopathologically were within normal ranges with the exception of the gills and olfactory organ. Lesions in fish exposed to concentration of 0.3 mg l.^?1 and higher were characterized by branchial epithelial hypertrophy and hyperplasia, and necrosis of olfactory epithelial cells.     

Acute toxicity of cadmium to Channa punctatus (BLOCH)

Effects of acute cadmium poisoning on survival, its residual values and histopathology in certain organs of a freshwater airbreathing fish, Channa punctatus (BLOCH ) were investigated. The threshold concentration, MATC and LC₅₀ values obtained from 96 h static bioassay, revealed that Channa is more susceptible to cadmium ions at higher temperature. The atomic absorption spectrophotometric determination of cadmium residues differed significantly in organs of specimens having survived and died after 96 h of exposure. The gill accumulate the highest amount of cadmium, the liver accumulated a slightly smaller amount than the gill, while the kidney accumulated the least. The histopathological lesions subjected to sublethal (5.2 mg/l Cd) and lethal (8.4 mg/l Cd) concentrations of cadmium were: detachment and rupture of lamellar epithelium, collapse of pillar cells and hypertrophy in mucus producing gland cells in the gill; vacuolization and coagulative necrosis in hepatic cells of the liver; and expansion, necrosis and accumulation of cellular debris in renal tubules of the kidney. The probable causes for death of fish due to cadmium ions have been discussed.     

Bioaccumulation of trace metals in the submarine hydrothermal vent crab Xenograpsus testudinatus off Kueishan Island,Taiwan

The concentrations of ten trace metals were determined in the gills, muscles, hepatopancreas, and exoskeleton tissues of Xenograpsus testudinatus that lives around shallow and acidic hydrothermal vents off Kueishan Island, northeastern Taiwan. The analytical results demonstrate that the metal concentrations vary significantly with the type of crab tissue. The concentrations of Al, Cd, Co, Cu, Fe, Ni, and Zn are highest in the gills, whereas the concentration of Mn is highest in the exoskeleton. Cr and Pb concentrations are similar across the different tissues. The enhanced accumulation of most metals in the gills suggests the metal accumulation via the respiration pathway rather than the uptake of food. The results also reveal that the distribution patterns of metals in tissues are similar in the hepatopancreas and muscles, but very different in both the gills and the exoskeleton, perhaps because of the different pathways of metal utilization in the different crab tissues.     

Protective effect of phospholipid hydroperoxide glutathione peroxidase (PHGPx) against lipid peroxidation in mussels Perna perna exposed to different metals

Levels of antioxidant defenses and lipid peroxidation were evaluated in mussels exposed to lead (200 mg/l), iron (500 microg/l), cadmium (200 microg/l) and copper (40 microg/l), for 12, 24, 72 and 120 h. Glutathione S-transferase (GST) activity was unchanged with all treatments. Catalase (CAT) increased after 120 h of exposure to all metals. Mussels exposed to Cd for 12 h, and to Cu and Fe for 120 h had increased lipid peroxidation, which might be associated to decreased levels of reduced glutathione (GSH) and glutathione peroxidase (GPx) activity. Pb exposure caused GSH depletion after 12 h and increased GPx activity after 120 h. Negative correlations were observed between the enzyme phospholipid hydroperoxide glutathione peroxidase (PHGPx) and malonaldehyde (MDA) levels after Fe and Cu exposure, indicating a protective role of PHGPx against lipid peroxidation, and suggesting the use of this enzyme as a new potential biomarker of toxicity associated with contaminant exposure in mussels.     

The Accumulation of Monocrotophos in Some Piscine Organs

Monocrotophos (0,0-dimethyl-0-(1-methyl-2-N-methyl-carbamoyl)-cis-vinylphosphate) as an active component of some insecticides is highly soluble in water at a low hydrolysis rate: half-time at 2 mg/l, pH = 7.0 and 38 °C 23 d. Individuals of the following fish species (individual weight in g) were kept during the static bioassay for 6… 72 h at 40 mg/l monocrotophos in the medium: Cyprinus carpio (70), Carassius carassius (45) and Leucaspius delineatus (1.5), and then the concentration of monocrotophos was gas-chromatographically determined in different tissues. Initially, till the 24th hour a high accumulation in the gills took place (in mg/kg fresh weight: Cyprinus 0.49, Carassius 3.88), then the concentration remained uniform or slightly falling here, whereas in the other organs (digestive organs, muscle, gonads) a further accumulation up to 1 mg/kg occurred. Leucaspius was investigated only as whole individuals, and after 72 h their fresh weight had increased to 6.96 mg/kg. The rates of accumulation decrease in the following order: Cyprinus: digestive organs, gills, muscle; Carassius: gills, digestive organs, gonads, muscle.     

Immunotoxicity of the xenoestrogen 4-nonylphenol to the cockle Cerastoderma glaucum

The in vivo effects of 4-nonylphenol (NP) on functional responses of haemocytes from the cockle Cerastoderma glaucum were investigated after 7 days exposure to sublethal NP concentrations (0, 0+acetone, 0.0125, 0.025, 0.05 and 0.1 mg/l NP). Haemocytes from both controls and exposed cockles were collected, and the effects of NP on total haemocyte count (THC) and volume of circulating cells, intracellular superoxide anion (O(2)(-)) levels, acid phosphatase and lysozyme-like activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. Exposure of cockles to 0.1mg/l NP significantly increased THC (p<0.05) with respect to controls. Analysis of haemocyte size frequency distribution showed that the haemocyte fraction of about 7-8 microm in diameter and 250 femtolitres in volume increased markedly in cockles exposed to the highest NP concentration tested. Apoptosis resulting in cell volume reduction in NP-exposed animals cannot be excluded. No statistically significant variation in intracellular O(2)(-) levels was observed. Conversely, significant increases (p<0.05) in acid phosphatase activity were observed in CFH from 0.05 and 0.1mg/l NP-exposed animals; no significant differences in enzyme activity were recorded in HL. Lysozyme-like activity also increased significantly in CFH from cockles exposed to 0.05 mg/l NP (p<0.05) and 0.1 mg/l NP (p<0.001). Instead, lysozyme-like activity decreased significantly (p<0.05) in the HL of animals exposed to 0.05 mg/lNP. Our results suggest that NP induces variations in the functional responses of haemocytes of C. glaucum, mainly by reducing cell membrane stability and promoting cell degranulation.     

Molecular cloning and β-naphthoflavone-induced expression of a cytochrome P450 1A (CYP1A) gene from an anadromous river pufferfish, Takifugu obscurus

In recent years, there has been a decline in the wild populations of river pufferfish, Takifugu obscurus. Besides overexploitation for commercial purposes, environmental pollution is believed to have contributed to its decline. However, almost no information exists about genes involved in metabolism of xenobiotics by this species. Nevertheless, there is interest in fugu fishes, since they possess the smallest genome among vertebrates. We cloned and characterized the full-length cDNA sequence of a cytochrome P450 1A (CYP1A) gene from T. obscurus. Phylogenic relationship of T. obscurus CYP1A was also compared to other fish species. The tissue distribution and time-dependant induction of CYP1A mRNA were studied by real-time PCR in T. obscurus exposed to an aryl hydrocarbon receptor (Ahr) agonist, β-naphthoflavone (BNF). The greatest basal expression in livers of control as well as BNF-treated individuals. However, brain, gill, gonad, intestine, and kidney also expressed CYP1A. Muscles expressed the least CYP1A. The results of the time-course study revealed induction in brain and gills after 6 h and at 12 h in most tissues. Except for gills, all other organs retained induced expression of CYP1A mRNA up to 96 h.     

Biomonitoring of the Contamination of Water by a Sublethal Concentration of Pesticides — A System Analysis Approach

Heteropneustes (Saccobranchus) fossilis (Pisces; Clariidae) is exposed to the biocides Chlordane, Aldrin, Metasystox, Dichlorvos, Carbofuran and Sevin for 30 d at a concentration of the 1/15 fraction of LC_50;96h with daily exchange of the medium. At intervals of 5 d the respiration rate was determined for 1 h. In addition, after the end of tests 13 haematological parameters as well as the activities of the adenosine triphosphatases and the acetylcholinesterase were determined in the liver, the kidney and the gills. During all exposures to biocides the respiration rate decreases significantly with the test time. At all exposures the blood concentrations of glucose, sodium- and chloride-ions show a significant increase, whereas the cholesterin content and the clotting time decrease significantly. The ATPase activity is significantly reduced only by Aldrin and Chlordane. A significant reduction of the acetylcholinesterase activity occurs in each of the three investigated tissues only for Metasystox and Dichlorvos and in the liver in addition also for Chlordane, Aldrin and Carbofuran. Therefore, the respiration rate and haematological criteria permit a general stress indication, enzymatic activities constitute a specific indication of individual groups of biocides, so that a systematic analysis of these three indication groups will probably provide a specific resultant statement in a biomonitoring system due to the specific inhibition pattern.     

Alteration in the Acid and Alkaline Phosphatase Activity of a Freshwater Prawn after in vitro Aldrin Exposure

In vitro effects of aldrin on acid and alkaline phosphatase activities in various tissues, viz. hepatopancreas, stomach, muscle, gill and brain, of a freshwater prawn Macrobrachium lamarrei have been studied. Aldrin exposure inhibited the activities of both the phosphatases in all the tissues and this inhibition in enzyme activity showed a dose-dependent response. The results were discussed in relation to in vivo findings and it is concluded that aldrin causes some enzymological changes and thereby creates widespread disturbance in the metabolism.     

Bioaccumulation of trace metals in the submarine hydrothermal vent crab Xenograpsus testudinatus off Kueishan Island, Taiwan

The concentrations of ten trace metals were determined in the gills, muscles, hepatopancreas, and exoskeleton tissues of Xenograpsus testudinatus that lives around shallow and acidic hydrothermal vents off Kueishan Island, northeastern Taiwan. The analytical results demonstrate that the metal concentrations vary significantly with the type of crab tissue. The concentrations of Al, Cd, Co, Cu, Fe, Ni, and Zn are highest in the gills, whereas the concentration of Mn is highest in the exoskeleton. Cr and Pb concentrations are similar across the different tissues. The enhanced accumulation of most metals in the gills suggests the metal accumulation via the respiration pathway rather than the uptake of food. The results also reveal that the distribution patterns of metals in tissues are similar in the hepatopancreas and muscles, but very different in both the gills and the exoskeleton, perhaps because of the different pathways of metal utilization in the different crab tissues.     

Histopathological Lesions in Heteropneustes fossilis Subjected to Acute Thiodan Toxicity

The common Indian freshwater catfish, Heteropneustes fossilis, was exposed to the 96 h-LC₅₀ (0.007-0.013 ppm) of the organochloride thiodan (endosulfan) for 4 days. The stressed fishes showed various histopathological changes. In the liver, extensive degeneration of cytoplasm, pycnosis of nuclei, and loss of glycogen was observed most commonly. Stasis of circulation and constriction of bile duct lumen was also seen frequently. In the kidney, shrinkage of glomeruli, cytoplasmic damage and epithelial desquamation in tubules, and extensive degeneration of hemopoietic stroma was commonly observed. In the intestine, the damage was comparatively negligible, the most common effect was an increased mucus cell activity leading to accumulation of mucoid exudates in the lumen.     

Molecular cloning and beta-naphthoflavone-induced expression of a cytochrome P450 1A (CYP1A) gene from an anadromous river pufferfish, Takifugu obscurus

In recent years, there has been a decline in the wild populations of river pufferfish, Takifugu obscurus. Besides overexploitation for commercial purposes, environmental pollution is believed to have contributed to its decline. However, almost no information exists about genes involved in metabolism of xenobiotics by this species. Nevertheless, there is interest in fugu fishes, since they possess the smallest genome among vertebrates. We cloned and characterized the full-length cDNA sequence of a cytochrome P450 1A (CYP1A) gene from T. obscurus. Phylogenic relationship of T. obscurus CYP1A was also compared to other fish species. The tissue distribution and time-dependant induction of CYP1A mRNA were studied by real-time PCR in T. obscurus exposed to an aryl hydrocarbon receptor (Ahr) agonist, beta-naphthoflavone (BNF). The greatest basal expression in livers of control as well as BNF-treated individuals. However, brain, gill, gonad, intestine, and kidney also expressed CYP1A. Muscles expressed the least CYP1A. The results of the time-course study revealed induction in brain and gills after 6h and at 12h in most tissues. Except for gills, all other organs retained induced expression of CYP1A mRNA up to 96h.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC₅₀ value was 0.3 mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days’ exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

Lethal and estrogenic effects of 4-nonylphenol in the cockle Cerastoderma glaucum

The lethal and sublethal effects of the xenoestrogen 4-nonylphenol (NP) were evaluated in the cockle Cerastoderma glaucum. In a 96-h lethality test, bivalves were exposed to 0, 0+ acetone, 0.19, 0.38, 0.75, 1.5 and 3.0 mg NP/l. The 96-h LC₅₀ value was 0.3 mg NP/l. No mortality was observed at 0.1 mg NP/l. The potential estrogenicity of NP was studied in both sexually undifferentiated (resting phase) and differentiated (pre-spawning phase) cockles, exposed for 7 and 14 days to 0, 0+ acetone, 0.0125, 0.025, 0.05, and 0.1 mg NP/l. Vitellogenin (Vg)-like protein levels were determined in both haemolymph and digestive gland by the alkali-labile phosphate (ALP) assay. In the resting phase, exposure for 7 days to 0.1 mg NP/l resulted in significant increases in ALP in both haemolymph and digestive gland, compared with controls. A significant increase was also observed in digestive gland of animals exposed to 0.0125 mg NP/l-exposed animals. After 14 days of exposure, haemolymph ALP levels were significantly increased in exposed animals at all NP concentrations tested, whereas no difference was recorded in digestive gland. In the pre-spawning phase, exposure for 7 days to NP significantly increased ALP levels in haemolymph from males exposed at all NP concentrations tested, whereas no significant variations were found in haemolymph from females. NP (0.05 and 0.1 mg/l) was also shown to increase ALP concentrations significantly in digestive gland of males, but not in those of females. Likewise, after 14 days’ exposure, ALP levels significantly increased in haemolymph from males only at 0.1 mg NP/l. Conversely, NP caused significant increases in ALP levels in digestive gland from both males (at all NP concentrations tested) and females (at 0.025 and 0.1 mg NP/l). These results demonstrate that NP induces Vg synthesis in C. glaucum. Interestingly, males were more responsive to NP than females.     

“Man and the Biosphere”-Studies on Sikkim Himalayas. Part 1: Acute Toxicity of Copper and Zinc to Common Carp Cyprinus carpio (Linn.) in Soft Water

In the daily exchange of the medium with only 7.1 mg/l Ca²⁺ and 0.7 mg/l Mg²⁺ during the batch experiment the LC_50,96h for Cyprinus carpio is 0.063 mg/l Cu²⁺ and 3.12 mg/l Zn²⁺ at water temperatures of 11 to 14°C. Significant differences in mortality as referred to the LC_50,12h occur for copper only after 72 h of exposure and for zinc after 48 h of exposure. The values of LC_50,96h are clearly lower for both metals in the investigations described than most data given in literature for different fish species.     

Accumulation and depletion of chromium in mystus vittatus (BLOCH)

Exposure against two ppm chromium from K₂Cr₂O₇ for 1 to 7 months produced increasing accumulation of chromium in the liver, kidneys, brain and bone of the freshwater fish Mystus vittatus. In gonads, the accumulation of chromium was progressive for the first 5 months (i.e., April to August), but not in the subsequent 2 months. After 4 months exposure, fish transferred to chromium–free freshwater had a significant depletion of chromium from all tissues at 1, 2 and 3 months.