Gas–liquid chromatography for fenvalerate residue analysis: alterations in the ionic concentration and associated adinosine triphosphatases in fish,Cirrhinus mrigala (hamilton)

In the present study, an attempt has been made to quantify the fenvalerate accumulated in different tissues (gill, muscle and liver) and observe changes involved in the levels of sodium, potassium and calcium ions and Na⁺–K⁺, Mg²⁺ and Ca²⁺ adenosine triphosphatase (ATPase) activities in the freshwater fish, Cirrhinus mrigala on short-term and long-term exposure to the median lethal and sublethal concentration of fenvalerate. Residue analysis using gas–liquid chromatography (GLC) technique revealed that fenvalerate accumulated in highest quantity in gill followed by liver and muscle under median lethal concentration (6?µg?L^?1). Whereas in sublethal concentration (0.6?µg?L^?1), muscle accumulated highest quantity followed by gill and liver, which might be due to the fact that fenvalerate is highly lyphophilic. The ion concentration and ATPase activity were found effected in fish exposed to lethal and sublethal concentrations of fenvalerate. Concentration of Na⁺, K⁺ and Ca²⁺ ions decreased in gill, muscle and liver on being exposed to median lethal concentration to a significant level. Whereas the changes were not highly pronounced at sub lethal level indicating low concentration of fenvalerate and its non-toxic effect at chronic exposure. Na⁺–K⁺, Mg²⁺ and Ca²⁺ ATPases activity were also found decreased in correspondence to the ionic change under median lethal and sub lethal concentrations in target tissues. This might have lead to behavioural changes and create wide-spread disturbance in the normal physiology, ultimately causing the death of the fish. The results suggest that in biomonitoring programmes, ions and associated ATPases can be a good diagnostic tool for fenvalerate toxicity.     

Effects of dietary copper exposure on accumulation,growth, and hematological parameters in Cyprinus carpio

The influence of dietary copper (Cu) exposure on accumulation, growth, and hematological parameters was investigated in Cyprinus carpio after sub-chronic ingestion of 0, 250, 500, 750 or 1000?mg?kg^?1 for 60 days. The profile of Cu accumulation among tissues in C. carpio was dependent on the exposure period and Cu concentration. Liver of C. carpio was the predominant storage tissue and the order of Cu accumulation in tissues was liver?>?intestine?> gill?>?kidney?>?muscle. Cu concentration at >125?mg?kg^?1 reduced growth rate, and was inversely related to growth. The RNA?:?DNA ratios were not affected by exposure and there was no correlation between growth rate and RNA?:?DNA ratio in liver and muscle. There were no significant effects of exposure on blood parameters except for magnesium. Cu exposure time and dose increased the serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT) activity levels.     

Study by evolution of nucleic acid content of prepuberal growth in the shrimp Crangon vulgaris

Growth of the shrimp Crangon vulgaris was studied by following the evolution of its nucleic acid concentration and total content. Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) analysis were effected on homogenates of whole shrimps. Cell multiplication was estimated from evolution of total DNA content, and cell size increment from evolution of fresh weight: DNA ratio. DNA puric bases ratio and RNA monoribonucleotides percentages were constant throughout the investigation period. A decrease in DNA concentration was observed from 50 to 20 M/g dry defatted weight (DDP), when fresh weight increases from 40 to 570 mg. This decrease was more marked in the first half of the period studied (prepuberal phase). The DNA content in the whole shrimp increases with fresh weight without slowing down at sexual maturity. However, at the beginning of the prepuberal phase the cell-multiplication rate seems to be graduated rather than continuous; this is still to be explained. Cell size, calculated from fresh weight: DNA ratio, increases until the shrimp weighs 300 mg; beyond this weight, cell increment proceeds much more slowly. The weight gain of shrimps throughout the growth period studied is mainly attributable to hyperplasia — which occurs constantly — while hypertrophy insures only 21% weight increment in the largest shrimps. As protein content, RNA content is a linear function of shrimp weight. RNA:DNA ratio evolution is similar to that of cell size and protein content. We suggest that cells have attained a physiological balance when shrimps reach a weight of 300 mg.     

汞和硒对剑尾鱼Na+/K+-ATPase活性的影响

应用浸浴法研究汞(Hg)对剑尾鱼(XiphophorushelleriHeckel)鳃和肝脏Na /K ATPase活性的影响以及硒(Se)对机体Na /K ATPase汞中毒的保护作用.结果表明,Hg和Se对剑尾鱼96hLC50分别为0.84mg/L和6.64mg/L.Hg对剑尾鱼鳃和肝脏Na /K ATPase活性的影响相似,与对照组相比,处理d1时低浓度Hg组鳃和肝脏ATPase活性没有明显变化(P>0.05),但高浓度Hg组ATPase活性变化显著(P<0.05),至d3和d5时,酶活性均极显著下降(P<0.01),鱼鳃和肝脏酶活性分别下降32%和60%,表明Hg处理对鳃和肝脏Na /K ATPase活性具有抑制作用.单独加Se组鱼鳃和肝脏Na /K ATPase活性在d3和d5显著提高(P<0.01).Hg Se组与Hg组相比,酶活性有明显差异(P<0.05或P<0.01),Se对剑尾鱼Hg中毒具有保护作用.剑尾鱼鳃和肝细胞Na /K ATPase活性可作为对水环境汞污染效应环境风险评价(ERA)的有效生物学标记.表2参16     

Impact of industrial effluents on the biochemical composition of fresh water fish Labeo rohita

In acute toxicity (96 hr) experiment the fingerlings of freshwater fish Labeo rohita was exposed to tannery, electroplating and textile mill effluents. The LC0 and LC50 concentrations were 15% and 20% for tannery effluents, 3% and 6% for electroplating effluents and 18% and 22% for textile mill effluents respectively. It was found that, electroplating effluent was more toxic than tannery and textile mill wastes. After acute toxicity experiments for different industrial effluents, various tissues viz. gill, liver, muscle and kidney were obtained separately from control, LC0 and LC50 groups. These tissues were used for biochemical estimations. The glycogen content in all the tissues decreased considerably upon acute toxicity of three industrial effluents except muscle in LC50 group of tannery effluent and kidney in LC50 group of textile mill effluent, when compared to control group. The total protein content decreased in all tissues in three effluents except gills in LC50 group of tannery effluent, kidney in LC50 group of electroplating effluent and kidney in LC0 group of textile mill effluent. In general total lipid content decreased in all tissues after acute exposure when compared to control group. The results obtained in the present study showed that, the industrial effluents from tannery, electroplating and textile mills caused marked depletion in biochemical composition in various tissues of the fish Labeo rohita after acute exposure.     

Effect of salinity adaptation on nitrogen metabolism in the freshwater fish Tilapia mossambica. I. Tissue protein and amino acid levels

Water, proteins and total free amino acids were estimated in different tissues of the euryhaline fish Tilapia mossambica after adaptation to various strenghts of sea water. The water content did not vary significantly in any tissue on salinity adaptation. The soluble and insoluble proteins displayed a general and considerable decrease in muscle, liver and heart; the decrease in the soluble fraction in the heart and the proteins of the muscle in 75% sea water (100% sea water=32.5 S) were significant. The gill proteins did not alter with salinity; the kidney proteins tended to increase slightly in 100% sea water (SW). The total free amino-acid content decreased insignificantly in all tissues on adaptation to 25% SW; in higher salinities, however, the content increased significantly. This increase was sudden and steep in 50% SW, and gradual and less steep in 75 and 100% SW. It is suggested that constancy in water content may contribute to the great adaptability of T. mossambica to heterosmotic media, and that the total free amino acids may be involved in isosmotic intracellular regulation. The possibility of amino acid increase as a result of protein breakdown is also indicated.     

不同性质天然水中铜在虹鳟(Oncorhynchus mykiss)鳃表的形态及对虹鳟的毒性

测定了在差异很大的4种水样(京密引水渠、北京地下水、长江南京段和汤旺河)中铜对虹鳟(Oncorhynchusmykiss)的半致死浓度,同步分析了各类水样参数和鳃铜蓄积量.用生物配位模型计算了铜在水相的形态分布和在鱼鳃表面的吸附量.研究了铜的水相形态、鳃表吸附量和鱼鳃蓄积量及毒性的关系.结果表明,铜的毒性与水相游离铜含量并无线性关系,但与鳃组织蓄积量成正比.图6参10     

Effect of mercury and methyl parathion on the ovaries of Labeo rohita (Ham)

Adult pre-spawning fish Labeo rohita were sublethally (1/5th 96h LC50) exposed to mercuric chloride and metacid-50 (methyl parathion). Accumulation of mercury and methyl parathion was studied and it was found that pre-spawning ovary appears as a potent organ for deposition of both the pollutants. RNA/DNA ratio of the control and treated fish were studied. It was found that the signifcant decrease in RNA/DNA ratio occurs after 9 and 30 days of exposure for mercury and 30 days for methyl parathion. Fluorescence microscopic studies by acridine orange staining method were also performed to show how much it is related to biochemical alterations. In some cases loss of metachromasia is correlated with the fall in RNA/DNA ratio. Some other abnormalities like fall in stage II: stage I oocyte ratio and necrosis was also observed.     

Relationship between growth and biochemical indices in laboratory-reared juvenile Japanese flounder (Paralichthys olivaceus), and its application to wild fish

In order to estimate growth rates based on biochemical indices of the liver of wild Japanese flounder (Paralichthys olivaceus), juveniles were reared at six ration levels (0, 0.5, 2, 4, 6 and 8% body weight day⁻¹) in the laboratory for 14 days, and the relationship between their growth rates and biochemical indices (RNA/DNA, protein/DNA, triglyceride/DNA, phospholipid/DNA and cathepsin D activities) were determined. Positive and approximately linear relationships were seen between growth rates and the indices of RNA/DNA, protein/DNA and phospholipid/DNA. The triglyceride/DNA ratio decreased with increasing growth rates up to approximately 1% body weight day⁻¹, then increased linearly with increasing growth rates. There was no significant correlation between growth rates and cathepsin D activity, and the highest values were obtained in the starved fish. Compared with laboratory-reared specimens, wild specimens of similar sizes were found to have significantly larger livers. The RNA/DNA, protein/DNA and phospholipid/DNA ratios of wild specimens fell in a broad range between ration groups of reared juveniles. The protein/DNA ratios of wild specimens were low and outside the range of the reared juveniles at six ration levels. In contrast, the levels of cathepsin D activity of wild fish were highest compared to the reared fish. Estimated growth rates of wild fish from the RNA/DNA, protein/DNA and phospholipid/DNA regressions obtained from the rearing experiment were 1.66, −1.74 and 0.10% day⁻¹, respectively. Based on our results, the RNA/DNA index may be regarded as the most valid and reliable growth estimator. It is noted that the larger liver size, the lower liver protein/DNA ratio and the unexpectedly high level of cathepsin D activities of wild specimens found in this study may reflect the different metabolic conditions of fish reared in the laboratory compared to those collected in the field. Received: 29 February 2000 / Accepted: 26 August 2000     

Essential fatty acid (docosahexaenoic acid, DHA) availability affects growth of larval herring in the field

Larval fish growth and survival depends not only on prey quantity, but also on prey quality. To investigate effects of prey fatty acid concentration on larval herring growth, we collected different prey organisms and larval herring (Clupea harengus L.) in the Kiel Canal during the spring season of 2009. Along with biotic background data, we analysed fatty acids both in prey organisms and in the larvae and used biochemically derived growth rates of the larvae as the response variable. Larval herring reached their highest RNA/DNA derived growth rates only at high docosahexaenoic acid (DHA) concentration. When the ratio of copepodids to lesser quality cirriped nauplii was low, larval growth and larval DHA concentration were both significantly negatively affected. This was true even as prey abundance was increasing. This finding indicates that even in mixed, natural feeding conditions, growth variations are associated with DHA availability in larval fish.     

Effects of environmental hypercapnia on hemato-immunological parameters,carbonic anhydrase,and Na+, K+-ATPase enzyme activities in rainbow trout (Oncorhynchus mykiss) tissues

In this study, the effects of environmental hypercapnia on hemato-immunological parameters and the activities of respiratory enzymes such as carbonic anhydrase (CA) and Na⁺, K⁺-ATPase were investigated in rainbow trout (Oncorhynchus mykiss) tissues (gill, liver and kidney). Batches of 12 fish were exposed to 4.5 mg L^?1 (control) and 14 mg L^?1 CO₂. No mortalities occurred during the 14 days of the experimental period. Red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels, and innate immune parameters such as nitro blue tetrazolium (NBT), lysozyme, and myeloperoxidase activities, and the melano-macrophage frequency were negatively affected by elevated CO₂ levels. Patterns of change in CA activity differed among the gill, liver, and kidney. Compared with the activities of CA in the control group, the CA enzyme was significantly stimulated at day 7 in the gill tissue, whereas it was stimulated at day 14 of the experiment in the liver tissue of fish exposed to 14 mg L^?1 CO₂ (P < 0.05). In contrast to the pattern of CA enzyme activities, the Na⁺, K⁺-ATPase enzymes were stimulated significantly in the liver after day 7 but inhibited in the kidney and gill (P < 0.05). These results suggest that a subchronic exposure to hypercapnia of rainbow trout tissues may lead to adaptive changes in the respiratory enzymes and negatively affects hemato-immunological parameters.     

Possible Use of Rna:Dna Ratio for Detecting Oil Induced Disturbance: A Field Report

The effects of oil contamination on the metabolic state of suspended participate and sedimented organic matter were investigated through the analysis of the RNA:DNA ratio on samples collected from January 1991 to January 1992 in the Ligurian Sea (northwestern Mediterranean). the ratio of RNA to DNA appeared to be highly sensitive to hydrocarbon stress since the oil disturbance resulted in an evident decline in the RNA:DNA ratio relative to the pre-pollution conditions. Such a pattern was unusual since it occurred in a period when we would expect a seasonal increase in bacterial activity. Differences are seen when comparing the temporal trend of DNA in suspended particulates and in sediments. However, the effects of oil on the RNA:DNA ratio were similar. Oiled samples showed a RNA:DNA ratio lower than 1 whereas RNA concentrations exceeded DNA content in undisturbed samples. Bacterial DNA accounted for a significant fraction (average 26%) of total DNA in sediments. It is likely that changes in RNA:DNA ratio in oiled sediments are due to the disturbance of bacterial communities. Consistent with the meiofaunal response to oil contamination, the hydrocarbons had only temporary and limited effects and RNA:DNA values recovered to pre-pollution conditions after a few weeks. the possible use of the RNA:DNA ratio in pollution monitoring is discussed.     

Toxicity of cypermethrin on total lipids and free fatty acids in liver,muscle, kidney and ovary of Clarias batrachus (L) and recovery response

Alterations in the levels of lipid metabolites, total lipids and free fatty acids, were studied in physiologically important tissues viz; liver, muscle, kidney and ovary of a freshwater edible fish Clarias batrachus. The fish was exposed to sublethal concentration (1/3 of LC50 value for 96?h) of a synthetic pyrethroid insecticide, cypermethrin for 1, 5 and 10 days. After 10 days of exposure, fish were released into fresh water and total lipids and free fatty acids were determined at intervals of 1, 5 and 10 days to assess fish recovery response after cessation of cypermethrin exposure. Fish showed an increased rate of lipogenesis in liver, muscle, kidney and ovary during the exposure period followed by a decrease after transfer of fish into fresh water. There was also an elevation in liver and kidney free fatty acids contents, while the muscle and ovary free fatty acid contents were reduced during exposure period. The alterations in free fatty acids were restored to near normal values after 10 days of cessation of exposure.     

Biochemical responses during starvation and subsequent recovery in postlarval Pacific white shrimp,Penaeus vannamei

Postlarval shrimp, Penaeus vannamei Boone, 1931, were held individually in cages and exposed to two feeding regimes. One group was starved for 12 d and then fed during the following 12 d. A second group was fed throughout the 24 d study. Four individuals were sampled from each of the two groups on Days 0, 1, 2, 4, 8, 12, 13, 14, 16, 20, and 24. Molting and growth among the starved-fed postlarvae stopped after 2 d starvation, while fed postlarvae increased significantly in size throughout the 24 d study. Among the starved-fed postlarvae, water content increased rapidly in response to starvation. DNA and sterol concentrations increased significantly during starvation due to selective catabolism of cellular components. After 12 d, RNA concentration was not significantly different between the fed and starved-fed postlarvae, but became significantly higher in the starved-fed postlarvae 48 h after feeding resumed. Triacylglycerol reserves were severely depleted during the first day of starvation, while protein concentrations began to decrease after the second day of starvation. RNA, protein, and the polyamines spermidine and spermine, when expressed as a ratio to DNA, decreased in response to starvation. Concentrations of all measured parameters in the starved-fed postlarvae returned to levels similar to those in the fed group 8 to 12 d after feeding resumed. Results of this study suggest that triacylglycerol provides energy during short periods of starvation, while protein is utilized during prolonged starvation. The ratios of RNA:DNA, protein:DNA, spermidine:DNA, spermine:DNA, two unidentified amine compounds, and percent water content are all useful indicators of prolonged nutritional stress in postlarval P. vannamei.     

Impact of malathion on the biochemical parameters of gobiid fish, Glossogobius giuris (Ham)

There is a dearth of information regarding the changes in heart muscle metabolites induced by pesticides. In the present study, the gobiid fish, Glossogobius giuris, was exposed to sub lethal concentrations of (0.05, 0.25 and 0.5 ppm) organophosphorus pesticide, malathion for short duration (24 to 96 hr). The cardiac muscles showed maximum depletion of glycogen and cholesterol content during 72 and 96 hr after treatment with 0.5 ppm malathion. Whereas a slight fluctuation of protein and glycogen content was observed in low concentration (0.05 ppm) of malathion. The levels of protein showed a significant decrease at high concentration (0.5 ppm) when treated for longer duration (96 hr). The present study reports metabolic dysfunction in response to malathion toxicity in the fish.     

RNA:DNA ratio during the critical period and early larval growth of the red drum Sciaenops ocellatus

Eggs from two separate spawning stocks of the red drum Sciaenops ocellatus (Linnaeus) were hatched, and the larvae were reared in the laboratory for 2 wk under closely controlled conditions. Total RNA, DNA, and soluble protein were measured in each population daily in triplicate pooled samples of larvae from each of three tanks. Growth rate in mm d^-1 was determined for each population at 2 d intervals. Growth rate explained 72 and 95% of the variation in the RNA:DNA ratios of the two populations individually, and 86% of the variation in the RNA:DNA ratio when data from the two populations were combined. The RNA:DNA ratio appeared to be most effective as an indicator of growth in rapidly growing larvae, and to lose some resolution when growth was intermittent. The rates of deposition of RNA, DNA, and protein into tissue were all highly correlated with growth rate and with each other. Mean population RNA:DNA ratios of red drum yolk-sac larvae decreased from Day 1 post-hatch until larvae initiated successful feeding behavior, and then increased steadily throghout the remainder of the experimental period. This pattern of change in the RNA:DNA ratios during the yolk-sac stage appears to be an intrinsic developmental pattern of red drum ontogeny. The lowest values for the RNA:DNA ratio were observed just prior to the initiation of feeding or during the critical period, indicating that red drum larvae experience a decrease in capacity for protein synthesis as they initiate feeding. Intrinsic variation in the RNA:DNA ratio during development suggests that caution be used when comparing the RNA:DNA ratios of yolk-sac larvae to a critical ratio calculated from Buckley's general model.     

Cytogenotoxicity and histopathological assessment of Lekki Lagoon and Ogun River in Synodontis clarias (Linnaeus, 1758)

The cytogenotoxicity and histopathological alterations induced by xenobiotics in Lekki Lagoon and Ogun River on Synodontis clarias were investigated. Fish from these water bodies and a fish farm (control) were examined for micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood. Also gill, liver, kidney, and ovary were processed for histopathology using hematoxylin-eosin staining. Concentrations of cadmium, zinc, lead and copper in the water were determined. There was significant (p < 0.05) increase in micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood of S. clarias from the lagoon and river compared to the reference site. Loss and disorganization of the primary and secondary lamellae, multifocal degeneration, hemorrhages, cellular infiltration, congestions, vacuolations, atresia, and necrosis were common lesions in the examined tissues of fish from the lagoon and river. Cd, Zn, Pb, and Cu in water samples from the lagoon and river were higher than the reference site. Xenobiotics in Lekki Lagoon and Ogun River, mostly metals, induced deoxyribonucleic acid (DNA) and pathological damage in S. clarias.     

苯并(a)芘对大弹涂鱼(Boleophthalmus pectinirostris)卵巢还原型谷胱甘肽含量的影响

分析了大弹涂鱼肌肉、鳃、肝脏、内脏 (不含肝脏 )以及卵巢等的GSH含量 ;并以GSH含量最高的卵巢为研究对象 ,在实验条件下 ,经 0 (CK) ,0 .0 5 ,0 .2和 0 .5mg/L等不同浓度的BaP暴露 7d ,分析其卵巢GSH含量变化的时间 -效应和剂量 -效应特征 ,结果显示 :0 .5mg/L浓度组的大弹涂鱼卵巢GSH含量迅速升高 ,暴露 12h即显著高于CK(P≤ 0 .0 5 ) ,但随暴露时间的延长 ,其GSH含量显著降低 ,7d的样品显著低于CK(P≤ 0 .0 5 ) ;在暴露 3d ,GSH水平与BaP浓度的剂量 -效应关系表现为正相关 ,而在 7d ,则呈负相关 ,这可能表明大弹涂鱼卵巢对BaP暴露由适应性反应到产生毒性效应的过程 .以上研究结果表明 ,卵巢GSH可对BaP暴露作出快速灵敏的反应 ,GSH水平有可能作为具氧化 -还原循环活性的污染物暴露的一种敏感的生物标记 .图 2表 2参 9     

Bioaccumulation of fenvalerate technical grade in different organs of the frog Haplobatrachus tigerinus (Daudin)

Bioaccumulation studies of fenvalerate were conducted on Indian bullfrog Haplobatrachus tigrinus (Daudin) after exposure to sublethal intraperitoneal dose of technical grade fenvalerate (1/3 LD50 i.e. 116.66 microg/kg body weight) at 3, 6, 12, 24, 48 and 72 hours schedule. The tissues viz., muscle, liver, kidney, testis, brain, and whole body accumulation was analysed for residue estimations. In all the tissues, analysed maximum residue was recovered in the initial stages of exposure (3 and 6 hours). However, in brain the residues remained up to 72 hours. In the whole body, analysis after 3 hours of exposure 78.65% residue was recovered and by the time 72 hours passed only, 9.4% residue was recovered. The decline in residue levels along with the period of exposure indicates the fast acting nature of fenvalerate and metabolites.     

Alterations in Carbohydrate Metabolism in Selected Tissues of Crab, Oziotelphusa Senex Senex (Fabricius) Under Fenvalerate-Induced Stress

Changes in carbohydrate metabolism in hepatopancreas and muscle of the crab, Oziotelphusa senex senex exposed to a sublethal concentration (0.2 ppm) of fenvalerate, a pyrethroid insecticide, were studied. the glycogen and total carbohydrate levels decreased significantly in the tissues of crab exposed to fenvalerate. an increase in phosphorylase 'a' and decrease in aldolase activity levels suggested increased glycogenolysis, and decreased glycolysis during fenvalerate toxicity. Krebs cycle enzymes such as NAD-isocitrate dehydrogenase, succinate dehydrogenase and malate dehydrogenase were decreased, suggesting reduced mitochondrial oxidative metabolism. Glucose-6-phosphate dehydrogenase activity was increased significantly, indicating enhanced oxidation of glucose through the hexose monophosphate shunt pathway. Lactate dehydrogenase activity was elevated indicating the development of anaerobic conditions at tissue level in the stressed crab. Cytochrome C oxidase and Mg2+ ATPase activity levels were also decreased, indicating the impaired energy synthesis and prevalence of energy crisis. These results suggest that fenvalerate has a profound effect on the glucose metabolism of crab.