Acute and chronic effects of endosulfan on the haemato-immunological and histopathological responses of a threatened freshwater fish, spotted murrel, Channa punctatus

Two experiments, one short-term and one long-term, were conducted to elucidate the acute and chronic effects, respectively, of endosulfan exposure on the haemato-immunological and histopathological responses of Channa puncatatus. In the short-term study, fish were exposed to sublethal endosulfan (8.1 μg l⁻¹) for 12, 24, 36, 48, 72 and 96 h. In the long-term study, fish were fed with normal diet and simultaneously either exposed to endosulfan (1.2 μg l⁻¹) for 90 days or not. Results showed that the ascorbic acid levels in both the liver and the muscle decreased significantly (P < 0.05) by acute and chronic endosulfan exposure. The haemoglobin (Hb) level reduced significantly (P < 0.05) by 15.5% within 12 h of acute endosulfan exposure, further decreased by 25.8% after 24 h of exposure, however, thereafter the values increased and at the end of 72 h returned to normal levels. Almost similar trend was observed for the erythrocyte (RBC) count. The WBC count and the nitroblue tetrazolium (NBT) value showed a general increasing trend with increase in the duration of acute endosulfan exposure. The chronic exposure of C. punctatus to endosulfan significantly (P < 0.05) lowered the Hb level, RBC and WBC counts, NBT reduction value and the plasma parameters such as plasma protein, albumin (A) and globulin (G) compared with that of the control (except for A/G ratio). Endosulfan exposure also severely altered the liver histological structure. Overall results indicated that both short-term acute and long-term chronic endosulfan exposure had a significant impact on the haemato-immunological parameters and tissue histopathology of C. punctatus.     

Dietary pyridoxine promotes growth and cellular metabolic plasticity of Chanos chanos fingerlings exposed to endosulfan induced stress

An attempt was made to assess the effect of dietary pyridoxine supplementation on growth performance and cellular metabolic enzymes of Chanos chanos fingerlings under endosulfan induced stress. Two‐hundred and twenty‐five fish were randomly distributed into five treatments, each with three replicates and fed four isocaloric and isonitrogenous diets with graded levels of pyridoxine for 45 days. Endosulfan concentration in the water was maintained at sublethal level of 1/40th of 96 h LC₅₀ (0.52 ppb). Dietary pyridoxine supplementation significantly enhanced growth performance viz. weight gain (%), specific growth rate, feed conversion ratio and protein efficiency ratio of fish compared to control diet fed group. The results of this experiment revealed that the activities of digestive and metabolic enzymes were significantly improved after feeding with dietary pyridoxine supplementation in C. chanos. Significantly increased muscle and brain ascorbic acid content was observed in fish fed diets containing pyridoxine. Histological studies on liver of fish fed pyridoxine‐supplemented diet showed the normal architecture of hepatocytes compared to non‐pyridoxine fed group. The results obtained in this study revealed that the inclusion of pyridoxine in diet has a protective effect against endosulfan induced stress in C. chanos fingerlings through metabolic enzyme activities and vitamin C content. This study has a potential in ameliorating the toxic effect of endosulfan induced stress using dietary supplementation of pyridoxine.     

Effects of bifenthrin on some haematological,biochemical and histopathological parameters of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.)

The aim of this study was to assess the effect of bifenthrin on common carp (Cyprinus carpio L.). The 96-h LC50 value of Talstar EC 10 (active substance 100 g l⁻¹ bifenthrin) was found to be 57.5 μg l⁻¹. Examination of haematological and biochemical profiles and histological tissue examination was performed on common carp after 96 h of exposure to Talstar EC 10 (57.5 μg l⁻¹). The experimental group showed significantly higher (P < 0.01) values of plasma glucose, ammonia, aspartate aminotransferase and creatine kinase as well as the relative and absolute monocyte count, compared with the control group. Histological examination revealed teleangioectasiae of secondary gill lamellae and degeneration of hepatocytes. The bifenthrin-based Talstar EC 10 pesticide preparation was classified as a substance strongly toxic for fish.     

Cypermethrin and λ-cyhalothrin induced alterations in nucleic acids and protein contents in a freshwater fish, <Emphasis Type="Italic"> Channa punctatus</Emphasis>

In this study, a freshwater fish Channa punctatus was exposed to subacute concentrations of synthetic pyrethroid insecticides (cypermethrin and λ-cyhalothrin) for 96 h to evaluate their impact on the levels of nucleic acids and protein in its different organs. Significant enhancement in the level of DNA was recorded in all tissues of the fish at high concentration of cypermethrin, whereas RNA and protein contents increased in tissues at all concentrations of cypermethrin tested. In contrast, λ-cyhalothrin treatment caused an increase in the level of DNA only in liver and brain, whereas increase of RNA and protein varied to different levels in different tissues. Cypermethrin treatment induced RNA/DNA ratio in all fish organs tested, whereas λ-cyhalothrin caused a sharp decrease in the ratio. Protein/DNA ratios were found to be tissue specific in treatments with both of the insecticides. The results clearly indicated that both of these pyrethroids exerted their effects in a similar manner in fish liver but differed in other tissues. These insecticides acted as potential biomodulators in C. punctatus, though following different routes. The results may be an indicator of aquatic pollution affecting freshwater fauna and flora and thus signaling the need for strict regulation on the indiscriminate input of pyrethroids from agricultural sites.     

Assessment of tissue-specific effect of cadmium on antioxidant defense system and lipid peroxidation in freshwater murrel, Channa punctatus

The effects of different concentrations of cadmium chloride on the extent of lipid peroxidation (LPO) and alterations in the antioxidant enzyme activities were studied in liver, kidney and gill tissues of freshwater murrel, Channa punctatus. The fish specimens were exposed to 6.7, 13.4 and 20.1 mg l⁻¹ sublethal concentrations of cadmium chloride and the oxidative stress was assessed after 24, 48, 72 and 96 h post-exposure. The biomarkers selected for the study were thiobarbituric acid reactive substances for assessing the extent of lipid peroxidation and antioxidant defense system such as reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GP_X), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) activities. In general, the cadmium exposure elevated the LPO in subject tissues of treated group and modulated the activities of GPx, GST, SOD, CAT, GR and level of GSH after given exposure as compared to the control. All enzymes activities, except CAT (in kidney and gills), and amount of LPO elevated significantly (P < 0.05) in treated group with respect to control in all tissues, while significant difference was not observed between the exposed concentrations and within exposure duration. The results indicated that increase in LPO level and the fluctuation in antioxidant defense system in fish could be due to cadmium-induced increase in the production of reactive oxygen species (ROS). The potential role of these parameters as biomarkers of heavy metal pollution in aquatic system is discussed.     

Energy balance of juvenile <Emphasis Type="Italic">Cyprinus carpio</Emphasis> after a short-term exposure to sublethal water-borne cadmium

Stress in fish can be assessed by means of a bioenergetic approach, based on the evaluation of changes in their physiological parameters. The objective of this study was to determine the impact of sublethal water-borne cadmium (Cd) on the energetic balance of juvenile Cyprinus carpio under laboratory conditions after a short-term exposure. Fish were exposed to a concentration of Cd (0.15 mg Cd l⁻¹) for 2 weeks. This concentration is environmentally realistic since it is usually found, even at higher values, in heavily polluted periurban water bodies of Argentina. No mortality was recorded among the animals used in the experiments. Food intake, food assimilation and assimilation efficiency, fecal production, liver glycogen content, oxygen consumption, oxygen extraction efficiency, specific metabolic rate, ammonia excretion and ammonia quotient (AQ), condition factor, and liver somatic index were determined. The overall balance was expressed as the scope for growth (SFG). The morphological indices and the liver glycogen content of Cd-exposed fish showed no significant differences when compared to those of controls. There was a significant decrease in the food intake, fecal production, and food assimilation rates as well as in AQ; the SFG exhibited a highly significant decrease. The remaining parameters (assimilation efficiency, oxygen consumption, oxygen extraction efficiency, specific metabolic rate, and ammonia excretion) increased after the exposure to Cd. We concluded that the sub-chronic exposure of Cyprinus carpio to a sublethal concentration of Cd causes important alterations in the energy-related homeostasis of fish. Most of the responses are indicative of physiological adaptations to compensate an increased energy requirement due to the impairments caused by the metal.     

Alterations in blood electrolytes of a freshwater catfish <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis> in response to treatment with a botanical pesticide, <Emphasis Type="Italic">Nerium indicum</Emphasis> leaf extract

The present study aimed at investigating the effects of Nerium indicum leaf extract on the blood electrolytes of Heteropneustes fossilis for short- and long term. Fish were subjected to Nerium indicum leaf extract for short term (11.27 mg/L i.e. 0.8 of 96 h LC₅₀) and long term (2.81 mg/L i.e. 0.2 of 96 h LC₅₀). Fish were killed on each time intervals from control and experimental (Nerium indicum) groups after 24, 48, 72, and 96 h in short-term exposure and after 7, 14, 21, and 28 days in long-term experiment. Blood samples were analyzed for calcium and inorganic phosphate levels. Acute exposure of Nerium indicum leaf extract caused a progressive decrease in the serum calcium levels after 48 h in fish H. fossilis, which persists till the close of the experiment (96 h). The serum inorganic phosphate levels remain unaffected till 48 h in the Nerium indicum leaf extract–exposed fish. After 72 and 96 h, the levels exhibit a decrease. Chronic Nerium indicum leaf extract treatment provoked a decrease in serum calcium levels at day 14. This decrease continues till 28 days. The serum phosphate level of the Nerium indicum leaf extract–treated fish decreases on day 14 and 21. However, on day 28, the levels become close to the normal values. We conclude that Nerium indicum leaf extract exposure alters the blood electrolytes of the fish, thus causing physiological disturbances which might affect seriously the normal vital functions, growth rate, reproduction, and their survival in nature.     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

Metallothionein biosynthesis as a detoxification mechanism in mercury exposure in fish,spotted scat (<Emphasis Type="Italic">Scatophagus argus</Emphasis>)

It is of crucial importance to study on the biomarkers types to assess the specification of the pollutants and health status of marine ecosystems in environmental evaluation projects. In this respect, total metallothionein biosynthesis and mercury bioaccumulation in the liver and gills under acute mercury exposure were investigated in fish, Scat (Scatophagus argus). Spotted scat was exposed to different mercury concentrations (0, 10, 20, 30) for 24, 48, 72 h. Total MT levels were determined by enzyme-linked immunosorbent assay (ELISA) method. Mercury contents were determined through cold vapor atomic absorption spectrometry (CVAAS). Induction of MT during exposure was tissue specific, displaying different response pattern in gills and liver. Mercury accumulated in liver much higher than in gills and the latter also showed lower MT level (P < 0.05). MT biosynthesis in liver showed a significant (P < 0.05) increase after exposure to different mercury concentration with increase in exposure time, whereas total MT content did not significantly (P > 0.05) change in gills except for 72 h exposure at 30 μg l⁻¹. Nonetheless, the relationship between MT biosynthesis and Mercury bioaccumulation in both tissues was significant (P < 0.05). The results suggest that this form of MT in S. argus was Hg inducible and could be extended as a biomarker of mercury pollution in marine ecosystems.     

Acute Toxicity and Sublethal Effects of Nitrite on Selected Hematological Parameters and Tissues in Dark-banded Rockfish, Sebastes inermis

Acute toxicity and sublethal effects of nitrite in dark‐banded rockfish, Sebastes inermis (83.3 ± 7.2 g), were studied under static conditions for a period of 96 h. The acute toxicity of nitrite evaluated for the 96‐h lethal concentration (LC₅₀) was 700 mg/L. The sublethal effects on selected hematological parameters of S. inermis, such as total erythrocyte count (TEC), hemoglobin, plasma glucose, and serum protein content, were measured after 0, 6, 12, 24, 48, 72, and 96 h of exposure to 0, 50, 100, 200, 400, and 700 mg/L of nitrite. Sublethal nitrite caused progressive reduction in the TEC, hemoglobin, and serum protein content in fish depending on the nitrite concentration and exposure period. The 96‐h exposure resulted in a 14–42% reduction in TEC and 25–33% reduction in hemoglobin content for 100–700 mg/L of nitrite compared to the control. A dose‐related reduction in plasma glucose (25.7–34.2%) was observed for concentrations of 200–700 mg/L of nitrite during 48 h of exposure, followed by an increase through 96 h. A significant reduction in serum protein (7.3–12.6%) was observed for 200–700 mg/L of nitrite after 96 h of exposure. Abnormal histological changes in skin, gill, liver, and kidney tissue were observed in fish exposed to 700 mg/L of nitrite after 96 h of exposure compared to the control. Although no mortality of S. inermis occurred at 500 mg/L of nitrite, all hematological parameters adversely responded to a nitrite dose of 200 mg/L for 96 h. These results showed that although acute toxicity concentration of nitrite in S. inermis is higher than 700 mg/L, sublethal concentrations of nitrite also negatively affect hematological parameters.     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

Effect of chronic exposure to ammonia on alterations of proteins and immunoglobulins in sea bass (Dicentrarchus labrax) serum

The relationship between serum protein, immunoglobulin concentrations and protein molecular weight profile (PMWP) alterations of sea bass (131.3 ± 4.3 g) reared in sea water with sublethal concentrations of ammonia was studied over two periods totalling 116 days. During the exposure period (62 days) the first group (group 1) lived in sea water with 0.204  mg.L^-1unionized ammonia nitrogen equivalent to 12 % of the lethal concentration for 50 % of a population exposed for 96 h to ammonia (96-h LC₅₀), whilst the second group (group 2) lived in sea water with 0.340  mg.L^-1UIA-N equivalent to 20 % of the 96-h LC₅₀ of ammonia. Then, the two groups were left for a recovery period (54 days) in the same water as the control group (group 0). The determination of the total immunoglobulin (Ig) concentration was carried out by enzyme-linked immunosorbant assay (ELISA). Gel filtration columns were used for the serum PMWP. Serum Ig concentration of the exposed fish (group 2: 1.76 ± 0.43 ; group 1: 1.19 ± 0.33  mg.mL^-1) was lower than the control group (3.39 ± 1.01  mg.mL^-1) after 21 days of exposure period but this difference was reduced at the end of this period (day 62) and treated group Ig concentrations switched to higher than the control group after the recovery period (day 116, group 2: 9.75 ± 1.84  mg.mL^-1; group 1: 7.50 ± 1.22  mg.mL^-1group 0: 6.38 ± 1.13  mg.mL^-1). In fact, at the end of the experiment, the cumulative Ig production difference between fish exposed to ammonia and the control was less than 10 %. Although a similar evolution of the Ig serum occurred with protein concentration, the serum protein concentration deficit of group 1 was restored at the end of the exposure period (group 2: 46.49 ± 2.34  mg.mL^-1; group 0: 46.74 ± 1.97  mg.mL^-1) and the cumulative production during the experiment was not significantly different between group 1 and group 0. However, this remained lower for group 2. During the exposure period, the PMWP of treated fish moved towards smaller molecular weight proteins. This alteration of the PMWP showed that the 0.2-kDa fraction increased, and another fraction appeared at the end of the exposure period. However, at the end of the recovery period, no difference was found between the PMWPs of control and treated fish. The alterations of the serum protein and Ig of fish reared, for a limited period (2 months), in water with sublethal concentrations of ammonia (20 % 96-h LC₅₀ or less) should disappear completely after a few months in normal rearing conditions.     

Effect of starving and feeding on some haematological and physiological responses of the Nile catfish, <Emphasis Type="Italic">Clarias gariepinus</Emphasis> exposed to copper at extreme seasons

The lethal concentration for 50% of fish for 96h (96h LC₅₀) of copper (Cu²⁺) was estimated for the Nile catfish (Clarias gariepinus) in extreme seasons, winter and summer, 4.31 and 4.79 mg/l, respectively. The Nile catfish was exposed to 96h LC₅₀ of copper for 7 days in extreme winter and summer. The body indices, haematological parameters as well as some plasma and liver enzyme activities and metabolite level were significantly differed in fish exposed to copper over than those of the control fish. Most of the tested parameters were not significantly different between the control fish of winter and summer (winter, water temperature 18 ± 2°C and summer, 27 ± 2°C). The effect of two ration sizes on copper toxicity in two different seasons on C. gariepinus was justified. It was found that the haematological parameters and the tested plasma activities of enzymes were significantly valid due to season differences. The blood parameters as well as plasma activities of enzymes were significantly differed in fishes fed elevated ration (3%) and exposed to copper challenge. On the other hand, the exploit of low feeding ration (0.5%) along with copper exposure during the examined seasons induced non-significant differences of the tested parameters, from those of the corresponding control. Therefore, the low feeding ration provides some tolerance against the possible water-borne copper exposure.     

Application of non-invasive techniques to differentiate sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>, L. 1758) quality cultured under different conditions

European sea bass, Dicentrarchus labrax, is one of the main species cultured in Europe. In this study, non-invasive techniques such as visible-near infrared spectroscopy and dynamometric analysis were used to discriminate concrete tank–cultured sea bass from sea cage–cultured sea bass. A total of 198 samples were collected at six time points from 3 Italian fish farms. A partial least squares analysis was conducted on individual animals at 48 and 96 h post-mortem to determine whether textural and spectral parameters can discriminate between concrete tank– and sea cage–reared sea bass. The results of this analysis demonstrated that spectral measurements better discriminate individual animals at 48 h post-mortem (87% in the independent test) with respect to 96 h post-mortem (66.7% in the independent test). Differences in texture were observed between the two groups of fish, but the texture changed in the time. The texture in sea cage–cultured fish showed higher values of measured force (N) with respect to fish reared in concrete tanks at 48 h post-mortem. Instead, opposite results were observed at 96 h post-mortem. Fillet colour were influenced by both farm and rearing condition.     

Dietary protein requirement for fingerling Channa punctatus (Bloch), based on growth, feed conversion, protein retention and biochemical composition

An 8-week feeding trial was conducted in a flow-through system (1–1.5 L min⁻¹) at 27°C to determine dietary protein requirement for Channa punctatus fingerlings (4.58 ± 0.29 g) by feeding six isocaloric diets (18.39 kJ g⁻¹, gross energy). Diets containing graded levels of protein (300, 350, 400, 450, 500 and 550 g kg⁻¹) were fed to triplicate groups of fish to apparent satiation at 09:00 and 16:00 h. Maximum absolute weight gain (AWG; 8.11 g fish⁻¹), specific growth rate (SGR; 1.82%) and best feed conversion ratio (FCR; 1.48) were recorded in fish fed diet containing 450 g kg⁻¹ protein, whereas protein efficiency ratio (PER; 1.52), protein retention efficiency (PRE; 25%), energy retention efficiency (ERE; 78%) and RNA/DNA ratio (3.01) were maximum for the group fed dietary protein at 400 g kg⁻¹. Second-degree polynomial regression analysis of AWG, SGR and FCR data against varying levels of dietary protein yielded optimum dietary protein requirement of fingerling between 462.24 and 476.72 g kg⁻¹, whereas the regression analysis of PER, PRE, ERE and RNA/DNA ratio data showed a lower protein requirement of 438.28–444.43 g kg⁻¹ of the diet. Considering the PER, PRE, ERE and RNA/DNA ratio as more reliable indicators, this protein requirement is recommended for developing quality protein commercial feeds for C. punctatus fingerlings.     

Difference Between Tetrodotoxin and Saxitoxins in Accumulation in Puffer Fish Takifugu rubripes Liver Tissue Slices

In vitro accumulation of tetrodotoxin (TTX) and paralytic shellfish toxin (PST) in tiger puffer fish Takifugu rubripes was investigated using liver tissue slices. When T. rubripes liver slices were incubated with Leibovitz’s L-15 medium containing 0.13 mM TTX at 20 °C in air with saturated humidity, they accumulated 21.5 ± 7.3 μg TTX g⁻¹ liver after the incubation for 12 h and increased to 55.3 ± 8.2 μg TTX g⁻¹ liver at 48 h. In the incubation of T. rubripes liver slices with 0.13 mM PST-containing medium, PST was detected 6.3 ± 0.9 μg g⁻¹ liver at 12 h and reached a plateau thereafter. These results reveal the difference between TTX and PST in accumulation in T. rubripes liver tissue slices. To examine the variation in PST accumulation among fish species, the liver tissue slices from tiger puffer fish T. rubripes, parrot-bass Oplegnathus fasciatus and green ling Hexagrammos otakii were incubated at a concentration of 0.027 mM PST. The toxin contents of 3.0 μg g⁻¹ liver were observed at 8 h regardless of fish species but were not increased subsequently, showing no variety among these three species as to accumulation patterns of PST. It is noted that the tiger puffer fish T. rubripes liver specifically accumulate TTX in preference to PST.     

Effects of the estrogen mimic genistein as a dietary component on sex differentiation and ethoxyresorufin-O-deethylase (EROD) activity in channel catfish (<Emphasis Type="Italic">Ictalurus punctatus</Emphasis>)

A number of aquaculture species, including channel catfish Ictalurus punctatus, are fed high proportions of soybean meal in their diet. We have investigated the potential for the most common phytoestrogen in soybean meal to alter phenotypic sex during sexual differentiation in channel catfish. Channel catfish were fed four dietary concentrations of the phytoestrogen genistein (0, 2, 4, and 8 mg g⁻¹) to determine its effect on gonadal sex differentiation. The four treatment diets were fed to sexually undifferentiated channel catfish between 5 and 140 days post hatch (dph) and between 60 and 150 dph. Phenotypic sex was determined by histological examination of the gonads. Ethoxyresorufin-O-deethylase activity was not significantly different among the treatment and control groups. Phenotypic sex was significantly dependant on dietary phytoestrogen concentration (P = 0.01). Additionally, logistic regression showed a significant relation between genistein concentration in the diet and gonadal sex (P = 0.02). Intersex individuals were present at all treatment concentrations, with increasing proportions of intersex fish as the genistein concentration increased for individuals fed treated diets between 5 and 140 dph. Increased proportions of phenotypically male individuals resulted from chronic dietary exposure to the estrogen mimic genistein. There were no significant differences in the proportions of males and females between feed treatment durations. These findings establish that dietary concentrations of genistein can alter sex ratios in cultured channel catfish populations and demonstrates the need to further understand the actions of this and other prominent phytoestrogens in aquaculture species.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.     

Study of muscle glycogen content in both sexes of an Indian teleost <Emphasis Type="Italic">Clarias batrachus</Emphasis> (Linn.) exposed to different concentrations of Arsenic

The present investigation has been conducted to study the effect of different sublethal concentrations of arsenic (5, 10 and 15 mg/L) on the muscle glycogen content in an Indian teleost, Clarias batrachus, during their post-spawning period (October to January).The species were exposed to various arsenic concentrations for six consecutive days i.e.,144 h. Significant differences in average muscle glycogen content were found in the treated male and female C. batrachus specimens. However, significantly higher (P < 0.01) muscle glycogen content was observed in male fish when compared to females in the untreated specimens. Arsenic caused muscle glycogenolysis in both sexes of C. batrachus. The depletions were not always dose (arsenic concentration) and time dependent, although higher concentrations were more glycogenolytic than the lower concentrations. After 96 h of treatment with arsenic, less depletion of muscle glycogen content was recorded in both sexes of fish.