Comparison of in vitro vitellogenin syntheses by three phenols in primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes

The in vitro effects of bisphenol-A (BPA), nonylphenol (NP) and 4-tert-octylphenol (OP) on vitellogenin (VTG) synthesis were examined using primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes. In vitro VTG syntheses were induced by exposure to 10^?5 M of BPA and 10^?4 M of both NP and OP, suggesting that BPA had the highest estrogenic potential among the three phenols.     

Structural and functional effects of early exposure to 4-nonylphenol on gonadal development of Nile tilapia (Oreochromis niloticus): b-histological alterations in testes

The present study investigates the effects of different doses (0, 40, 60, 100 μg/L) of 4-nonylphenol (NP) on testis histology and sperms motility of mature Oreochromis niloticus. A tendency for a dose-dependent reduction in the gonado-somatic index (GSI) in the NP-exposed groups was observed. Histological examination revealed that high doses of NP (60 and 100 μg/L) cause hyperplasia of interlobular connective tissue components coincide with decrease in the size of seminiferous lobules and amount of lumen spermatozoa in comparison with the control and low-dose (40 μg/L)-exposed groups. In addition, within the seminiferous lobules, rupture of spermatogenic cysts, vacuolation and necrosis of primary spermatocytes due to damage of sertoli cells were clearly observed and lead to decrease in advanced spermatogenic cells. The highest dose caused significant decrease in the GSI associated with appearance of testis–ova (intersex) state. The percentage of abnormal sperms increased with increasing the dose of NP reaching to 96 % for those subjected to 100 μg NP/L, while it was only 11 % for the control group specimens. The study indicates that NP has estrogenic potency induced marked alteration in the sexual development including inhibition in testicular growth and deformation in the sperms.     

Distinct expression of three estrogen receptors in response to bisphenol A and nonylphenol in male Nile tilapias (Oreochromis niloticus)

Environmental estrogens, such as bisphenol A (BisA) and nonylphenol (NP), have been shown to affect the estrogen receptor (ER) expression and induce male reproductive abnormalities. To elucidate molecular mechanisms of action of xenoestrogenic chemicals on the expression of estrogen receptors in the testes of Nile tilapia (Oreochromis niloticus), three full-length cDNAs respectively encoding ntERα, ntERβ1 and ntERβ2 were cloned from testes. The amino acid sequences of ntERα, ntERβ1 and ntERβ2 showed a high degree of similarity to the relevant fish species. Tissue-specific expression study showed that three receptors were highly expressed in pituitary, liver, testis, kidney and intestine tissues. The ntERα, ntERβ1 and ntERβ2 mRNA expressions were significantly higher at the sexual early recrudescing stage than at other recrudesced stages. After being exposed to xenoestrogens from weeks 2 to 4, the ntERα mRNA levels were increased significantly in testes after NP treatment at all sampling times or after 4 weeks of exposure to BPA. The ntERβ1 mRNA levels remained unchanged, while a significant decrease of the ntERβ2 mRNA level was observed in testes after exposure to NP and BPA. The present study demonstrates that the regulation of all three ntER subtypes in testes may act via different molecular mechanisms of exposure to NP and BPA.     

Effect of different avian egg yolk types on fertilization ability of cryopreserved common carp (Cyprinus carpio) spermatozoa

In spite of the fact that egg yolk from different avian species has successfully been used as an additive for the cryopreservation of sperm in mammalian species, its efficacy for cryopreserving fish sperm has not previously been tested comparatively. Therefore, the present study was carried out to determine the effect of egg yolks from different avian species, namely domestic chicken (Gallus gallus domesticus), turkey (Meleagris gallopavo) and quail (Coturnix coturnix), on post-thaw motility and fertilization ability of cryopreserved common carp spermatozoa. Egg yolks from chicken, turkey and quail were analysed for moisture, total fat, protein, cholesterol and phospholipid profile. Total fat and cholesterol contents of the turkey egg yolk were higher than chicken and quail egg yolks (p < 0.05). Semen was frozen according to conventional slow freezing procedure. The extender contained 350 mM glucose, 30 mM Tris and 5 % glycerol supplemented with different ratios of avian egg yolk (10, 15 and 20 %). Semen was equilibrated at 4 °C for 15 min and placed into 0.25-ml straws and frozen in liquid nitrogen vapour (for 10 min at ?120 °C) and finally stored in liquid nitrogen (?196 °C) tank. The frozen spermatozoa were thawed in a water bath at 35 °C for 30 s. Fertilization was conducted using a ratio of 1 × 10⁵ spermatozoa/egg. Cryopreservation experiments resulted in higher post-thaw motility and fertilization rates. Mean post-thaw motility of cryopreserved spermatozoa was between 45 and 80 %, and fertilization rates, expressed as the percentage of eyed embryos, ranged from 70 to 95 %. In conclusion, the present study showed that turkey and quail egg yolks are suitable alternatives to the chicken egg yolk for the cryopreservation of common carp spermatozoa.     

The role of Ca2+ and Na+ membrane transport in brook trout (Salvelinus fontinalis) spermatozoa motility

The role of environmental ion composition and osmolality in Ca²⁺ signaled activation was assessed in spermatozoa of brook trout Salvelinus fontinalis. Milt from ten mature males was obtained by abdominal massage. Spermatozoa motility was evaluated in 0, 100, and 300 mOsm/kg NaCl or sucrose solutions, buffered by 10 mM Tris–HCl pH 8.5. For investigation of spermatozoa reaction to external Ca²⁺ concentration, 2 mM ethylene glycol tetraacetic acid (EGTA) was added to the activation media as a calcium ions chelator. For investigation of the effect of external Na⁺ concentration in conditions of low external Ca²⁺, 100 µM amiloride was added to the EGTA-containing solutions as a Na⁺ transport blocker. Low motility was observed in sucrose (Na⁺ free) solutions containing 2 mM EGTA but not in Na⁺ solutions containing 2 mM EGTA. Addition of amiloride led to significantly increased motility (P < 0.05) compared with sucrose (Na⁺ free) solutions containing 2 mM EGTA. We conclude that Na⁺ transport in Ca²⁺-free solutions plays a regulatory role in brook trout spermatozoa activation. The influence of competitive Na⁺ and Ca²⁺ transport on the control of spermatozoa activation requires further study with respect to its application for improvement of artificial activation and storage media.     

Diel cyclic hypoxia alters plasma lipid dynamics and impairs reproduction in goldfish (<Emphasis Type="Italic">Carassius auratus</Emphasis>)

Diel cyclic hypoxia occurs with varying frequency and duration in freshwater habitats, yet little is known about its effects on reproduction of freshwater fishes. The present study shows that long-term exposure of goldfish (Carassius auratus) to cyclic hypoxia (0.8 ± 0.2 mg/l dissolved oxygen) for 9 h or more, per day, altered plasma lipid and sex steroid profiles, which in turn directly or indirectly suppressed ovarian growth and viable spermatozoa production. Hypoxia decreased total cholesterol and high density lipoprotein (HDL p < 0.05) and elevated triglycerides (TG; p < 0.05) in both sexes. Plasma steroid concentrations particularly of 17α-hydroxyprogesterone (17-HP), estradiol (E2), testosterone (T) in females, and T and 11-ketotestosterone (11-KT) in males were attenuated under diel hypoxic conditions. Intriguingly, both diel and continuous hypoxia elevated plasma E2 and vitellogenin levels in males. However, neither lipid nor steroid profiles recorded any variation in a dose-dependent manner in response to diel hypoxia. The reduced GSI, decreased number of tertiary oocytes, and motile spermatozoa in hypoxic fish clearly indicate suppression of gametogenesis. Thereby, prolonged diel cyclic hypoxia may affect valuable fishery resources and fish population structure by impairing reproductive performances and inducing estrogenic effects in males.     

Protection of common carp (Cyprinus carpio L.) spermatozoa motility under oxidative stress by antioxidants and seminal plasma

The protective influence of seminal plasma and the antioxidants catalase (CAT), superoxide dismutase (SOD), and glutathione (GTH) on quality parameters, oxidative stress indices, and antioxidant activity was studied in common carp (Cyprinus carpio) spermatozoa exposed to the xanthine–xanthine oxidase (X–XO) system. Fish spermatozoa were incubated for 5 and 20 min at 4 °C with X–XO concentrations of 1 mM X–0.1 U/mL, 0.6 mM X–0.05 U/mL, 0.3 mM X–0.025 U/mL, and 0.1 mM X–0.0125 U/mL. A dose-dependent reduction in spermatozoa motility and velocity was observed at concentrations of 0.1 mM X–0.0125 U/mL to 1 mM X–0.1 U/mL XO. Increase in spermatozoa motility parameters was recorded following treatment with antioxidants and seminal plasma. The level of the oxidative stress indices lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) was significantly reduced after addition of CAT, SOD, or GTH along with seminal plasma. Significant differences in SOD, glutathione reductase, and glutathione peroxidase activity were seen in spermatozoa incubated with, compared to that without, seminal plasma at all studied X–XO concentrations. The data demonstrated that CAT, SOD, or GTH in combination with SP can reduce reactive oxygen species stress in fish spermatozoa and improve spermatozoa quality.     

Changes of sperm morphology,volume, density,and motility parameters in northern pike during the spawning period

Sexually mature males (BW?=?1600?±?150 g and TL?=?235?±?30 mm) of northern pike (Esox lucius L.) were randomly selected from a pond to record changes in their sperm quality parameters (spermatozoa morphology, sperm volume, density, and motility parameters) during the spawning season. The morphological and motility parameters changed significantly during the reproductive season with following trends. Only, head width was not changed during the spawning season. The longest spermatozoa and its flagellar length were found at the middle of spawning period (TL?=?38.24?±?0.37 μm and 35.14?±?0.26 μm) and shortest at the beginning of spawning period (TL?=?34.81?±?0.29 μm and 32.53?±?0.18 μm). Other morphological characters were always the lowest at the beginning of spawning period. Sperm volume was changed from 0.33?±?0.3 ml in February, 0.43?±?0.2 ml in March to 0.24?±?0.1 ml in April, and density from 16.2?±?0.2?×?109 spermatozoa ml^?1 in February, 19.4?±?0.2?×?109 spermatozoa ml^?1 in March to 4.8?±?0.2?×?109 spermatozoa ml^?1 in April. Same sperm velocity was observed in all spawning terms at 10 and 20 s after activation. Higher velocity was found at 30 and 40 s after activation in sperm collected at the middle and the end of spawning period. Significantly, higher percentage of motile sperm was observed at 20, 30, and 40 s after activation in sperm sampled at the end of spawning period. This study supports the hypothesis that longer spermatozoa swim faster.     

Endocrine disruption,oxidative stress,and testicular damage induced by 4-nonylphenol in <Emphasis Type="Italic">Clarias gariepinus</Emphasis>: the protective role of <Emphasis Type="Italic">Cydonia oblonga</Emphasis>

Exposure to xenoestrogens like 4-nonylphenol (NP) is recognized by disrupting endocrine functions and causes reproductive dysfunction in male fish. The present study aimed at investigating the 4-nonylphenol propensity to induce oxidative stress and hormonal disturbances in male catfish and at studying the protective role of quince (Cydonia oblonga). To fulfill this aim, catfish Clarias gariepinus were exposed to pure 100 μg/L 4-NP and to quince the leaf extract added to 4-NP, both for 15 days. The 4-NP exposure induced a marked increase in 17ß-estradiol (E2), LH, and cortisol, while thyroid hormone (TSH, T3), testosterone (T), and FSH levels noticeably decreased; however, 4-NP had no effect on T4 level. Moreover, 4-NP exposure was accompanied by histological impairments in testes. Existence of 4-NP was associated with oxidative damage as evidenced by the significant increase (p < 0.05) of the enzymes, superoxidase dismutase (SOD), catalase (CAT), acetylcholinesterase (AchE), glutathione s-transferase, total antioxidant capacity (TAC), and malondialdehyde (MDA). Adding quince was effective to neutralize hormonal levels and to repair the testicular histological alterations. In response to quince remedy, the enzymes AchE and MDA reduced significantly (p < 0.05), while limited or no response was detected for other tested enzymes. Our results concluded that quince can antagonize 4-NP toxicity in catfish, confirming that quince leaf extract displayed antioxidant activities against the toxicity of hazardous chemicals.     

The performance of dry ice-transported cryopreserved spermatozoa in the artificial insemination of the hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

ABSTRACT

In this study, the cryopreserved spermatozoa of Epinephelus lanceolatus were transported using a novel method involving dry ice as the medium of preservation and a Styrofoam box. Five conditions were investigated for the cryopreserved sperm under different dry ice exposure times of (24, 48, and 72) h corresponding to treatment 1 (T1), 3 (T3), and 5 (T5), respectively. Meanwhile, the remaining treatments (T2 and T4) involved the same exposure to dry ice for (24 and 48) h followed by re-immersion into liquid nitrogen (LN). The performance of the cryopreserved spermatozoa of the hybrid grouper (E. fuscoguttatus ♀ × E. lanceolatus ♂) was evaluated through fertilization and hatching trials. The results showed no significant difference in fertilization for all five treatments. However, significantly poorer hatching rates than the fresh sperm were observed for spermatozoa exposed to dry ice after 48 h. This study recommends the use of the proposed method to successfully transport E. lanceolatus spermatozoa for the production of hybrid groupers via artificial insemination.     

Supplementation of microbial levan in the diet of Cyprinus carpio fry (Linnaeus, 1758) exposed to sublethal toxicity of fipronil: effect on growth and metabolic responses

A 60-day feeding trial was conducted to study the effect of dietary microbial levan on growth performance and metabolic responses of Cyprinus carpio fry exposed to sublethal dose (1/10th LC₅₀) of fipronil [(±)-5-amino-1-(2,6-dichloro-α,α,α-trifluoro-p-tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile]. Two hundred and twenty five fry were randomly distributed in five treatments in triplicates. Four purified diets were prepared with graded levels of microbial levan. Five different treatment groups were levan control L₀P₀ (basal feed + 0 % levan without exposure to pesticide); pesticide control L₀P₁ (basal feed + 0 % levan with exposure to pesticide); L_0.25P₁ (basal feed + 0.25 % levan with exposure to pesticide); L_0.50P₁ (basal feed + 0.50 % levan with exposure to pesticide); and L_0.75P₁ (basal feed + 0.75 % levan with exposure to pesticide). Weight gain% and specific growth rate were significantly higher (p < 0.05) in levan fed groups compared to their non-levan fed counterpart. Highest (p < 0.05) content of ascorbic acid in muscle, liver and brain tissues was observed with higher level of dietary levan. Glucose-6-phosphate dehydrogenase activity decreased with the increasing level of dietary levan in the liver and muscle. Aspartate aminotransferase activity exhibited a second order polynomial relationship with the dietary levan, both in liver (Y = ?1.001x ² + 5.366x + 5.812, r ² = 0.887) and muscle (Y = ?0.566x ² + 2.833x + 6.506, r ² = 0.858) while alanine aminotransferase activity showed third order polynomial relationship both in liver (Y = 1.195x ³ ? 12.30x ² + 35.23x + 9.874, r ² = 0.879) and muscle (Y = 0.527x ³ ? 8.429x ² + 31.80x + 8.718, r ² = 0.990). Highest (p < 0.05) superoxide dismutase activity in gill was observed in the group fed with 0.75 % levan supplemented diet. Overall results indicated that dietary microbial levan at 0.75 % in C. carpio fry ameliorated the negative effects of fipronil and augmented the growth.     

Effect of nitrite exposure on oxygen-carrying capacity and gene expression of <Emphasis Type="Italic">NF-κB</Emphasis>/<Emphasis Type="Italic">HIF-1α</Emphasis> pathway in gill of bighead carp (<Emphasis Type="Italic">Aristichthys nobilis</Emphasis>)

Nitrite (NO₂^?) contamination of water can severely impact the health of aquatic life and is a major concern for commercial aquaculture. In order to study the effect of nitrite on Aristichthys nobilis, we investigated the oxygen-carrying capacity, NF-κB/HIF-1α pathway, and the gill tissue structure under nitrite stress. In the current study, bighead carp (initial weight 180.05?±?0.092 g) were exposed to nitrite (48.634 mg/L) for 96 h and then for 96 h recovery test. After nitrite exposure for 6 h, hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) mRNA expression increased significantly in the gill of bighead carp (P?<?0.05). After nitrite exposure for 12 h, hemoglobin (Hb) and methemoglobin reductase (MHBR) content in blood decreased significantly (P?<?0.05); TLR4 mRNA expression increased significantly (P?<?0.05). After nitrite exposure for 24 h, methemoglobin (MetHb) content increased significantly (P?<?0.05). After recovery test, all the indicators except TLR4 mRNA expression level recovered to initial level. In conclusion, nitrite exposure can affect hemoglobin dynamics, as oxidization of nitrite by hemoglobin results in the reduction of Hb to MetHb leading to hypoxia and nitrite exposure can also result into gill tissue damage. In the face of nitrite exposure, NF-κB and HIF-1α mRNA expression level increased immediately to protect the body from oxidative damage and eased hypoxic condition caused by nitrite. It was also observed that nitrite damage is recoverable in Aristichthys nobilis, but it may be need more than 96 h.     

Immunotoxic responses of chronic exposure to cypermethrin in common carp

In the current study, laboratory evaluations were made to assess the immunomodulatory effect of cypermethrin on fingerlings of common carp (Cyprinus carpio L.). Results showed that 96-h LC50 of cypermethrin in common carp was estimated at 0.85 μg/L. Fish were exposed for 21 days to cypermethrin at three sub-lethal concentrations of 0.042, 0.085, and 0.17 μg/L that represented 5, 10, and 20%, respectively, of the 96-h LC50 of the pesticide for this fish species. Blood samples were taken after 7, 14, and 21 days of exposure. Immunological indices and resistance against bacterial infection were determined. Compared to the control group, the fish exposed to cypermethrin showed a significant increase in neutrophil ratio but exhibited a significant decrease in leukocyte number and lymphocyte ratio in treatments exposed to 0.17 and/or 0.085 μg/L after 21 days of exposure (p < 0.05). Serum protein level was significantly decreased in group exposed to 0.17 μg/L on day 14 and also in groups exposed to 0.085 and 0.17 μg/L on day 21 (p < 0.05). Immunoglobulin value was significantly reduced in groups exposed to 0.085 and 0.17 μg/L after 21 days of exposure (p < 0.05). Serum lysozyme activity and phagocytic activity were significantly decreased following exposure to 0.17 μg/L determined on days 14 and 21, post-exposure (p < 0.05). Mortality rate following the challenge with Aeromonas hydrophila significantly increased in fish exposed to 0.17 μg/L of cypermethrin. Overall, the present results indicate severe immunotoxicological effects of cypermethrin in common carp. Therefore, the use of cypermethrin in the proximities of common carp farms should be carefully considered.     

Effects of dietary selenium of organic form against lead toxicity on the antioxidant system in Cyprinus carpio

In this study was evaluated potential protective effect of organic selenium (Se) on heavy metal stress induced by lead (Pb) in Cyprinus carpio. For this reason, C. carpio was exposed to sublethal concentration of Pb (1.5 mg/L Pb(NO₃)₂) for 14 days. The fish were fed a basal (control; measured 0.55 mg/kg Se) diet or a basal diet supplemented with 2.50 mg/kg (measured 2.92 mg/kg Se) organic Se (Sel-Plex^®) during the experiment period. The variations in glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) activities, and levels of reduced glutathione (GSH) with malondialdehyde (MDA) in liver and brain tissues of C. carpio were investigated in experimental groups. GSH levels in liver and brain tissues were significantly decreased by exposure to Pb. GST activity was significantly increased (p < 0.05) in liver tissue, but decreased in brain of treated fish by exposure to Pb. Also, GSH-Px activity was significantly increased in liver tissue, but decreased in brain of Pb-treated fish. Levels of MDA were increased in liver and brain of Pb-treated fish. The organic Se treatment for Pb-intoxicated animals improved activities of GSH-Px, GST and levels of MDA within normal limits. Supplemented Se could be able to improve Pb-induced oxidative stress by decreasing lipid peroxidation and regulating antioxidant defense system in tissues.     

Thermal tolerance, oxygen consumption and haemato-biochemical variables of Tor putitora juveniles acclimated to five temperatures

A 30-day acclimation trial was conducted using Tor putitora to elucidate its thermal tolerance, oxygen consumption, haemato-biochemical variables and selected enzymatic activities at five acclimation temperatures (AT). Juveniles of T. putitora were randomly distributed among five treatment groups (20, 23, 26, 29 and 32 ± 0.5 °C). There was a significant curvilinear increase in critical thermal maxima (CT_max) (y = ?0.0693x ² + 1.7927x + 34.628, R ² = 0.996) and lethal thermal maxima (LT_max) (y = ?0.1493x ² + 2.3407x + 35.092, R ² = 0.991) with increasing AT. The oxygen consumption rate increased significantly with increasing AT. The Q ₁₀ values were 1.16 between 20 and 23 °C, 3.09 between 23 and 26 °C, 1.31 between 26 and 29 °^C and 1.76 between 29 and 32 °C of AT. The acclimation response ratios were ranged between 0.37 and 0.59. Catalase, superoxide dismutase and ATPase activities were increased linearly in liver, gill and kidney, while brain acetylcholine esterase activity decreased linearly with increasing AT. Blood glucose remained unchanged up to AT of 26 °C and increased significantly at AT of 29 and 32 °C. Haemoglobin content was increased linearly with increasing AT. The highest WBC count was observed at 20 °C, and no significant changes found till AT of 26 °C and significantly decreased at 32 °C. Total serum protein and globulin were significantly decreased with increasing AT. Highest values were observed at 20 °C and remained consistent till 26 °C, then decreased significantly. There was no significant change in A/G ratio through the AT 20–29 °C and increased significantly at 32 °C. The increase in CT_max, LT_max and oxygen consumption rate with increasing AT may suggest that the thermal tolerance of T. putitora is dependent on its prior thermal exposure history, and it could adapt to higher AT by altering its haemato-biochemical variables.     

Alteration in certain enzymological parameters of an Indian major carp, Cirrhinus mrigala exposed to short- and long-term exposure of clofibric acid and diclofenac

The extensive use of pharmaceuticals in human and veterinary medicine may enter the aquatic environment and pose a serious threat to non-target aquatic organisms like fish. In this study, Indian major carp Cirrhinus mrigala was exposed to different concentrations (1, 10 and 100 μg L^?l) of most commonly used pharmaceutical drugs clofibric acid (CA) and diclofenac (DCF) to evaluate its impacts on certain enzymological parameters during short- and long-term exposures. During short-term (96 h) exposure period, plasma glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and gill Na⁺/K⁺-ATPase activity were significantly altered at all concentrations of both the CA- and DCF-treated fish. In long-term exposure (35 days), gill Na⁺/K⁺-ATPase activity was found to be significantly increased at all concentration of CA and DCF exposures throughout the study period (except at the end of 7th day in 10 and 100 µg L-1) . However, a biphasic trend was observed in plasma GOT and GPT activity when compared to the control groups. In both short- and long-term exposure, a significant (P < 0.01 and P < 0.05) changes were observed in all enzymological parameters of fish C. mrigala exposed to different concentrations of CA and DCF. The alterations of these enzymological parameters can be effectively used as potential biomarkers in monitoring of pharmaceutical toxicity in aquatic environment and organisms.     

Mercuric chloride induced toxicity responses in the olfactory epithelium of Labeo rohita (HAMILTON): a light and electron microscopy study

Bioaccumulation of mercury and histomorphological changes in the olfactory epithelium of Labeo rohita were investigated after exposing the fish to two sublethal concentrations of HgCl₂ (66 and 132 μg/L) for 15 and 30 days. Mercury deposition increased in the tissue significantly (p < 0.05) with dose- and duration-dependent manner. Severe damage to the olfactory epithelium was evident. When fish exposed to 66 μg/L for 15 days, the histology of olfactory epithelium exhibited that mucous cell proliferation was upregulated and cell size was significantly increased from the control. Similar trends were found in 30 days exposure in both treated groups. Histology showed that mercury induced degeneration of columnar sensory cells, supporting cells and ciliated non-sensory cells and induced basal cell proliferation. Basal cell hyperplasia led to form intraepithelial proliferative lesion, thickening of epithelium, basal lamina disruption and cyst formation. Scanning electron microscopy revealed that mercury exposure at 66 μg/L caused clumping and loss of cilia, erosion in microridges on the supporting cells and proliferation of mucous cell opening. Complete degeneration of ciliated cells and cyst formation was observed in the fish when exposed to 132 μg/L HgCl₂. This result suggests that prolonged exposure to mercury might cause irreversible damage to the olfactory epithelium and impair the olfactory function of fish.     

The effect of yeast polysaccharide (YSP) on the immune function of Apostichopus japonicus Selenka under salinity stress

We evaluated the effects of yeast polysaccharide (YSP) on the immune function of Apostichopus japonicus exposed to a decrease in salinity (3, 5, or 9 ppt/day). Juvenile sea cucumbers were fed artificial feed containing 0.5 % YSP. We measured phagocytic activity and lysozyme, acid phosphatase, and alkaline phosphatase activity 24 h after each decrease in salinity. When salinity was decreased by 3 ppt/day, after 24 h, the immune index were significantly enhanced by supplementation of the diet with YAP (P < 0.05). There was no obvious change in the phagocytic index of animals fed a YSP supplemented diet and exposed to a decrease of 5 ppt/day, though the other immune indices were significantly elevated relative to the starting control (30 ppt). The immune index was significantly higher in animals fed a YSP supplemented diet and exposed to a decrease of 9 ppt/day than in animals held at 30 ppt. It is concluded that the addition of YSP to the basal diet improved the immunocompetence of A. japonicas during exposure to the salinity stressors.     

Structural and functional effects of early exposure to 4-nonylphenol on gonadal development of Nile tilapia (Oreochromis niloticus): a-histological alterations in ovaries

The present study investigated the gonads alterations of mature female Oreochromis niloticus caused by different concentrations (0, 40, 60, 100 μg/L) of 4-nonylphenol (NP) and also its effect on steroidogenesis. A tendency for a dose-dependent reduction in the gonadosomatic index, fecundity and oocytes diameter in the NP-exposed groups was observed. Histological examination revealed that NP impairs gonadal growth clearly shown in the oocyte development and differentiation. Also, the estrogenic activity of such NP was identified through 11-ketotestosterone, 17b-estradiol and vitellogenin which were affected significantly by the applied concentrations. The study indicates that NP has estrogenic potency-induced marked alteration in the sexual development including gonadal maturation, spawning time and egg production.     

Response of a freshwater air-breathing fish, Clarias batrachus to salinity stress: an experimental case for their farming in brackishwater areas in Andaman, India

The study was conducted to assess the effects of salinity on growth and biochemical composition of freshwater catfish, Clarias batrachus. A static nonrenewable acute toxicity bioassay test was conducted and LC₅₀ of salinity for 96-h exposure to the fingerling (14.5 cm) was 12.52 ‰. Based on these results, two sublethal salinity levels, viz. 4 and 8 ‰ were selected to study the long-term effects of salinity on C. batrachus for a period of 90 days. From the study, it was found that growth and survival rate were less in saline water (4 and 8 ‰). Maximum growth and survival were recorded in freshwater (0 ‰ salinity) and subsequently at 4 and 8 ‰. To assess the biochemical alteration, few important biomarkers were estimated. At the end of 90 days rearing period, glucose level in the brain and blood of C. batrachus was found to decrease with salinity. The level of liver and muscle glycogen in the fish reared at 4 ‰ was lower than that of control. Ascorbic acid in all organs under study was found to decrease with increasing salinity, which was attributed to stress mitigation effect of vitamin C. Acetylcholine esterase (AchE) activity recorded a gradual decrease with increasing salinity. Metabolic enzymes, alkaline phosphatase (ALP) activity and adenosine triphosphosphtase (ATPase) activity also reduced both in liver and muscle tissues with increasing salinity. From the present investigation, it can be concluded that exposure to higher salinity significantly (P < 0.01) affects the growth and physiological response of Clarias batrachus.