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.     

孔雀石绿及其主要代谢产物在欧洲鳗鲡体内的蓄积及消除规律

为了解孔雀石绿及其有毒代谢产物无色孔雀石绿在鱼体中的蓄积与消除规律,达到对孔雀石绿的禁用监控,本试验对初始体重为12.42±2.18 g的欧洲鳗鲡进行0.1 mg/L药浴24 h,再转移到清水中养殖120d,采用高效液相色谱法测定血液、肝脏、肾脏和肌肉组织中孔雀石绿(MG)及其代谢物无色孔雀石绿(LMG)的残留。结果表明:在药浴开始阶段,肝脏、肾脏和肌肉中的MG含量迅速上升,肝脏、肾脏和血液于浸浴6 h时即达到最高平均值,分别为859.8±127.0μg/kg、589.2±40.0μg/kg和88.6±51.3μg/kg,肌肉于浸浴12h时达最高值(720.5±192.6μg/kg),随后含量下降。鳗鲡各组织中LMG高峰出现时间都晚于MG,血液、肝脏和肾脏中的LMG都是在浸浴12 h时,达到最高平均值,分别为1 135.0±376.4μg/kg、1 730.9±538.5μg/kg和238.9±105.5μg/kg;肌肉组织LMG的高峰出现时间更晚,是清水养殖3 d(72 h)时,为960.1±251.0μg/kg。血液中的MG消除最快,于清水养殖的第2天(48 h)检测不到残留。肾脏于养殖10 d(240 h)、肝脏于养殖45 d(1 080 h)时检测不到残留MG,而肌肉中的MG在养殖90 d时才检测不到。LMG在鳗鲡血液和肌肉组织中消除时间与MG相比显著延长,血液中的LMG消除时间是养殖90 d(2 160 h),而肌肉中于养殖120 d时,仍能检测到一定含量的LMG。除了肾组织在整个试验阶段和肌肉组织在浸浴过程中,所含平均MG比LMG高以外,其余情况下都是LMG平均含量明显高于MG平均值。本试验表明,可以通过对鳗鲡肌肉中的无色孔雀石绿残留的检测达到对孔雀石绿禁用的监控。     

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.     

Effects of <Emphasis Type="Italic">Euphorbia hirta</Emphasis> plant leaf extract on growth performance,hematological and organosomatic indices of hybrid catfish, <Emphasis Type="Italic">Clarias macrocephalus </Emphasis>× <Emphasis Type="Italic">C. gariepinus</Emphasis>

This study was conducted to evaluate the effects of Euphorbia hirta leaf extract on the growth performance, hematological and organosomatic indices of hybrid catfish, Clarias macrocephalus?×?C. gariepinus. The fish were treated with 0 (control), 300, 500 and 800 mg/kg (ppm) for 90 days. The weight gain, average daily growth rate, and specific growth rate were at significantly higher levels in fish from all the treatment groups on days 75 and 90, while the feed efficiency and protein efficiency ratio were consistently higher in fish from all the treatment groups from day 60 up until day 90. The feed conversion rate significantly decreased from day 60 until day 90 in all treatment groups when compared with the control group, and the survival rate was significantly different from day 30 until day 90; a consistently higher rate was observed in fish fed 800 mg/kg. The highest viscerosomatic index and intraperitoneal fat were observed in the group fed 500 mg/kg (p?<?0.05). The hepatosomatic index was significantly increased alongside increasing levels of E. hirta extract. The total white blood cell count in the control group was significantly higher on day 30, but on day 90 all the treatment groups showed higher levels. Hematocrit percentage was significantly different on days 30 and 90. Lymphocyte, eosinophil and thrombocyte levels were shown to be significantly different (p?<?0.05) when different groups of fish were compared. In conclusion, 300 mg/kg of E. hirta leaf extract could improve growth performance, hematological and some organosomatic indices in hybrid catfish.     

Effects of calcium and copper exposure on lipogenic metabolism, metal element compositions and histology in Synechogobius hasta

The present study was conducted to evaluate the effects of calcium (Ca) and copper (Cu) exposure on growth performance, metal element composition, enzymatic activities and histology in Synechogobius hasta and tested the hypothesis that Ca could protect against Cu-induced toxicity in the fish species. Three hundred sixty S. hasta (initial mean weight 22.6 ± 0.2 g, mean ± SEM) were stocked in 18 circular fiberglass tanks (water volume: 300 l), 9 of which were pre-exposed to Ca at a rate of 600 mg Ca/l for 10 days and then exposed to concentrations of 0, 77 and 154 μg Cu/l for 30 days. Another 9 tanks were cultured in natural seawater (no extra Ca addition) for 10 days and then exposed to concentrations of 0, 77 and 154 μg Cu/l for 30 days. Both Ca exposure and then waterborne Cu exposure influenced the accumulation of metal elements (Cu, iron, zinc and manganese) in several tissues (muscle, intestine, gill, liver and spleen), changed hepatic lipogenic metabolism and induced histological alterations in liver, gill and spleen. In general, Ca exposure seemed to mitigate the severity of Cu-induced mortality and histopathological injuries, indicating that Ca exposure had the capacity to decrease Cu toxicity in S. hasta. To our knowledge, it is the first study involved in the effects of Ca and Cu exposure on hepatic lipogenic metabolism and metal element compositions in a wide range of tissues and organs in fish, which provides new insight into the protective effect of Ca on toxic metal elements in fish.     

Aflatoxin B<Subscript>1</Subscript> (AFB<Subscript>1</Subscript>) reduces growth performance,physiological response,and disease resistance in Tra catfish (<Emphasis Type="Italic">Pangasius hypophthalmus</Emphasis>)

Triplicate groups of one hundred Tra catfish (8 g?±?0.2) were fed seven test diets containing increasing levels of AFB₁ (0, 50, 100, 250, 500, and 1000 μg AFB₁ kg^?1). Additionally Mycofix® Secure was added at 1.5% to one diet containing 500 μg AFB₁ kg^?1. Results showed that Tra catfish are sensitive to AFB₁. Reduction in weight gain (P?<?0.05) was observed for fish fed 50 μg AFB₁ kg^?1 and declined further with increasing levels of AFB₁ in the diets. Fish fed diets contaminated with 500 and 1000 μg AFB₁ kg^?1 showed increased (P?>?0.05) hepatosomatic index (HIS), while an increase in adipose somatic index (ASI) was observed in fish fed 50 μg AFB₁ kg^?1 and above when compared to the control and Mycofix® diets. After 12 weeks, blood serum analysis revealed higher alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in fish fed the 50, 100, and 250 μg AFB₁ kg^?1 suggesting occurrence of liver damage. Disease resistance of fish exposed to Edwardsiella ictaluri was also compromised by the presence of AFB₁ in the feed and was directly related to the contamination level. Seven days after Edwardsiella ictaluri exposure, survival rates were 50, 41.7, 31.7, and 8.3% for fish fed control, 50, 100, and 250 μg AFB₁ kg^?1, respectively. This trial shows that AFB₁ at a level of 50 μg AFB₁ kg^?1 and above can affect fish performance and disease resistance. Application of an effective mycotoxin management in the feed seems to be useful to prevent the negative effects of AFB₁.     

Modulatory effect of lycopene against carbofuran toxicity in African catfish, <Emphasis Type="Italic">Clarias gariepinus</Emphasis>

In the current study, African catfish, Clarias gariepinus, was exposed to a sublethal concentration of carbofuran (CF) to investigate its negative effects on biochemical and oxidative stress biomarkers. Also, the putative role of lycopene (LYC) administration in alleviating these negative effects was evaluated. Fish were divided into six groups in triplicates as follows: group I was without treatment, group II was orally administered corn oil, group III was orally administered 18 mg LYC/kg body weight, group IV was exposed to 0.121 mg CF/L, group V was orally administered 9 mg LYC/kg body weight and exposed to 0.121 mg CF/L, and group VI was orally administered 18 mg LYC/kg body weight and exposed to 0.121 mg CF/L for 4 weeks. At the end of this period, blood and tissue (liver and kidney) samples were collected and biochemical and oxidative stress biomarkers were analysed. Also, histopathological changes were determined. Carbofuran caused significant increments of glucose, cortisol, aspartic amino transferase, alanine amino transferase, cholesterol, urea, and creatinine; meanwhile, serum acetylcholinesterase, total protein, albumin, and total lipids were significantly reduced. Significant increments in hepatic and renal malondialdehyde (MDA) and superoxide dismutase (SOD) levels and marked reduction in hepatic and renal catalase (CAT), glutathione (GSH), and total antioxidant capacity (TAC) levels were observed in CF-exposed fish comparing to the control group. Treatment with LYC attenuated the CF-induced oxidative stress, and this improvement was more pronounced in fish received the high LYC dose (18 mg/kg body weight). Further, congestion of the central vein with infiltration of mononuclear inflammatory cells, vacuolar necrosis, and haemorrhage was observed in the livers of CF-exposed fish. Oral administration of LYC reduced behavioural changes and histopathological alterations. All the altered biochemical parameters and antioxidant biomarkers were also restored to be near the normal levels. The obtained results evoked that LYC administration alleviated the destructive effects of carbofuran and reduced its toxicity effect on African catfish.     

Effects of fish size on the response of antioxidant systems of Oreochromis niloticus following metal exposures

The size of a fish is an important factor in its physiology, and metal uptake is affected by animal physiology. In this study, small and large tilapias (Oreochromis niloticus) differing approximately twofold in length and fivefold in weight were compared for their antioxidant response. Both groups were exposed to Cu or Cr (1.0 μg/mL) in a freshwater (?80 mg CaCO₃/L, conductivity 1.77 mS/cm) using 2 exposure protocols (20 μM for 48 h and 10 μM for 6 days). Following the exposures, the antioxidant enzyme activities (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; glutathione reductase, GR and glutathione S-transferase, GST) and glutathione (GSH) levels were measured in the liver of fish. Results showed that small fish was affected from exposure conditions much more than large ones as their antioxidant parameters significantly decreased even in controls. Metal exposures of small fish caused significant increases in SOD and CAT activity in acute Cu or Cr exposures. Subchronic Cr exposure of small fish also caused significant increases in CAT, GPx and GST activities, while there was no significant change in Cu-exposed ones. Large fish, however, showed different antioxidant responses as their levels mostly decreased. This study demonstrated that the response of antioxidant system in the liver of tilapia varied in relation to fish sizes and emphasized using different size groups in environmental monitoring and also in evaluation of fish biomarkers.     

Mercury distribution in target organs and biochemical responses after subchronic and trophic exposure to Neotropical fish Hoplias malabaricus

In the present study, we investigated the mercury distribution, mercury bioaccumulation, and oxidative parameters in the Neotropical fish Hoplias malabaricus after trophic exposure. Forty-three individuals were distributed into three groups (two exposed and one control) and trophically exposed to fourteen doses of methylmercury each 5 days, totalizing the doses of 1.05 μg g^?1 (M1.05) and 10.5 μg g^?1 (M10.5 group). Autometallography technique revealed the presence of mercury in the intestinal epithelia, hepatocytes, and renal tubule cells. Mercury distribution was dose-dependent in the three organs: intestine, liver, and kidney. Reduced glutathione concentration, glutathione peroxidase, catalase, and glutathione S-transferase significantly decreased in the liver of M1.05, but glutathione reductase increased and lipid peroxidation levels were not altered. In the M10.5, most biomarkers were not altered; only catalase activity decreased. Hepatic and muscle mercury bioaccumulation was dose-dependent, but was not influenced by fish sex. The mercury localization and bioaccumulation corroborates some histopathological findings in this fish species (previously verified by Mela et al. in Ecotoxicol Environ Saf 68:426–435, 2007). However, the results of redox biomarkers did not explain histopathological findings previously reported in M10.5. Thus, fish accommodation to the stressor may reestablish antioxidant status at the highest dose, but not avoid cell injury.     

Effect of taurine supplementation on hepatic metabolism and alleviation of cadmium toxicity and bioaccumulation in a marine teleost,red sea bream, <Emphasis Type="Italic">Pagrus major</Emphasis>

This study was performed to unravel the mechanism of the beneficial action of taurine on marine teleost fish, red sea bream (Pagrus major), by analyzing the hepatic metabolism. Moreover, the ameliorative effects of the nutrient against cadmium toxicity and bioaccumulation were further evaluated. The fish were fed a diet containing 0 % (TAU0 %), 0.5 % (TAU0.5 %), or 5.0 % (TAU5.0 %) taurine for 40–55 days (d) and subjected to cadmium acute toxicity and bioaccumulation tests. Taurine deficiency in feed severely affected growth and the hepatic metabolic profiles of the fish, including a remarkable increase in myo-inositol, aspartate, and ß-alanine in the TAU0 % group, which indicates a complementary physiological response to taurine deficiency. For the acute toxicity test, fish were fed the test diets for 55 d and were then exposed to different dose of cadmium ranging from 0 to 5.6 mg/L for 96 h. Fish fed taurine had a higher tolerance to cadmium than those not fed taurine. For the bioaccumulation test, fish were fed the test diets for 40 d and then were chronically exposed to 0.2 mg/L of cadmium for 28 d followed by depuration for 21 d. Cadmium concentrations in the liver and muscle of fish fed TAU5.0 % were significantly lower than those of fish fed TAU0 % for the first 7 d of exposure and the first 7 d of elimination. Our findings suggest a possible mechanism for the beneficial role played by taurine and that the inclusion of taurine in fish aquaculture feed may reduce cadmium contamination of fish intended for human consumption.     

Effects of dietary exposure to brominated flame retardant BDE-47 on thyroid condition, gonadal development and growth of zebrafish

Little is known about the effects of brominated flame retardants in teleosts and some of the information currently available is inconsistent. This study examined effects of dietary exposure to 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) on thyroid condition, body mass and size, and gonadal development of zebrafish. Pubertal, 49-day-old (posthatch) fish were fed diets without BDE-47 (control) or with 1, 5 or 25 μg/g BDE-47/diet. Treatments were conducted in triplicate 30-L tanks each containing 50 zebrafish, and 15 fish per treatment (5 per tank) were sampled at days 40, 80 and 120 of exposure. Measurements were taken of body mass, standard length, head depth and head length. Sex (at 40–120 days of exposure), germ cell stage (at 40 days) and thyroid condition (at 120 days; follicular cell height, colloid depletion, angiogenesis) were histologically determined. Whole-body BDE-47 levels at study completion were within the high end of levels reported in environmentally exposed (wild) fishes. Analysis of variance was used to determine differences among treatments at each sampling time. No effects were observed on thyroid condition or germ cell stage in either sex. Reduced head length was observed in females exposed to BDE-47 at 80 days but not at 40 or 120 days. In males, no apparent effects of BDE-47 were observed at 40 and 80 days, but fish exposed to 25 μg/g had lower body mass at 120 days compared to control fish. These observations suggest that BDE-47 at environmentally relevant whole-body concentrations does not affect thyroid condition or pubertal development of zebrafish but does affect growth during the juvenile-to-adult transition, especially in males.     

Effect of enrofloxacin and emodin on heat-shock proteins’ expression in hepatic cells of grass carp (Ctenopharyngodon idellus)

The aim of the present study was to compare the expression pattern of heat-shock protein 60 (HSP60), 70 (HSP70), and 90 (HSP90) mRNA in hepatic cells of grass carp exposed to enrofloxacin and emodin concentrations. The expression pattern of different genes encoding heat-shock proteins in hepatic cells of grass carp was exposed to graded levels of enrofloxacin (12.5, 25, 50, 100, and 200 μg/ml) or emodin (0.04, 0.2, 1, 5, and 25 μg/ml) for 24 h and were investigated by real-time quantitative PCR. When cells were exposed to up 50 μg/ml enrofloxacin, both HSP60 and HSP70 mRNA levels firstly were increased and thereafter significantly dropped at high concentrations (P < 0.05), while HSP90 decreased with increasing enrofloxacin concentration. Besides, HSP60 and HSP70 expressions were significantly inhibited in the high-concentration groups. In addition, the HSP90 mRNA levels in the treatment exposed to 100 and 200 μg/ml enrofloxacin were significantly lower than those of the control group (P < 0.05). Furthermore, when cells were exposed to graded levels of emodin, HSPs (HSP60, 70, and 90) mRNA levels significantly increased in the groups exposed to 5 and 25 μg/ml of emodin. The different expression pattern of HSPs implied that enrofloxacin could inhibit the expression levels of HSPs, while optimal level of emodin could trigger higher expression levels of HSPs in hepatic cell of grass carp to protect against further damage.     

Effect of vitamin C dietary supplementation in reducing the alterations induced by fenitrothion in Oreochromis niloticus

The objective of this study was to investigate the dietary effect of vitamin C in amelioration some of studied alterations induced by fenitrothion in Nile tilapia (Oreochromis niloticus). Nile tilapia was exposed to sub-lethal concentration of fenitrothion 0.04 mg/l (96 h LC₅₀ value was 0.8 mg/l), and basal diet was supplemented with two different dose of vitamin C (500 and 1200 mg/kg B wt/day) for 30 days. Vitamin C supplemented groups showed significant decrease in plasma cortisol and glucose level, enzymes activity of liver and gills (catalase, glutathione-S-transferase and superoxide dismutase) and % of tail DNA damage compared to exposed group. Moreover, fish revealed significant increase in total plasma protein, albumin, globulin and A/G ratio. High dose of vitamin C dietary supplementation (1200 mg/kg B wt/day) returns these parameters to its normal levels with no significant difference compared to non exposed control group. These results indicated that incorporation of high dose of vitamin C (1200 mg) in aqua feed for 30 days could be potentially less expensive and effective in reducing the alterations induced by fenitrothion in Nile tilapia.     

Hepatoprotective action of celery (Apium graveolens) leaves in acetaminophen-fed freshwater fish (Pangasius sutchi)

Acetaminophen (APAP)-induced liver damage is one of the most common problems among the population. Therefore, the study was aimed to investigate the hepatoprotective effect of celery leaves on APAP-induced toxicity in a freshwater fish, Pangasius sutchi. Fish were divided into four experimental groups of 6 fish each. Group 1 served as control. Group 2 fish were exposed to APAP (500 mg/kg) for 24 h. Groups 3 and 4 fish were exposed to APAP + celery leaf powder (CE) (500 mg/kg) and CE for 24 h, respectively. The severity of liver damage, hepatic lipid, glycogen, ions status and histological alterations was examined. The characterization of CE extract was also performed. APAP-exposed fish showed elevated levels of both circulating and tissue hepatotoxic markers (AST, ALT and ALP), reduced hepatic glycogen and lipid contents (TG and cholesterol), increased tissue lipid peroxidation markers (TBARS, LHP and PCO), altered tissue levels of enzymatic (SOD, CAT, GPx and GST) and non-enzymatic (GSH) antioxidants and cellular thiol levels (T-SH, P-SH and NP-SH), and reduced hepatic ions (Na⁺, K⁺ and Ca²⁺) and abnormal liver histology. The abnormalities associated with APAP exposure were reversed on treatment with CE. The TLC separation and HPLC quantification of petroleum ether/acetone extract of CE showed the peaks for highly efficient flavonoids such as rutein, quercetin and luteolin. The observed hepatoprotective effect of CE might be due to its rich flavonoids.     

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.     

Effect of hormone injection frequency on the lipid content and fatty acid compositions in gonad,muscle and liver of Anguilla japonica during artificial maturation

Two groups of Anguilla japonica were treated with hormone, one on a weekly basis, the other on a biweekly basis. Intramuscular injection was applied at a dose of 0.75 mg/kg BW carp pituitary extract (CPE) plus 150 IU/kg BW human chorionic gonadotropin (hCG) for female fish, while males received half of this dose. The average total lipid content of the gonads from the weekly treated group, i.e. 20.6 ± 1.2 % for females and 18.0 ± 2.3 % for males, was significantly higher than the biweekly treated group, 16.8 ± 0.7 and 15.5 ± 1.3 %, respectively (p < 0.05). For both muscle and liver, the readings were not significantly different. The gonads from the weekly treated fish had more fatty acids, particularly saturated fatty acids, polyunsaturated fatty acids, highly unsaturated fatty acids, eicosapentaenoic acid (20:5n-3, EPA), docosahexaenoic acid (22:6n-3, DHA) and arachidonic acid (20:4n-6, ARA). Histological examination showed that the ovaries of both the weekly and the biweekly treated fish were mainly at stage IV. However, the weekly treated females had bigger oocyte diameter (722.0 ± 60.9 μm) than the biweekly females (611.6 ± 22.6 μm). These results suggest that CPE and hCG promoted the maturation process for both scheduled induction and that the frequency of hormone injection influenced the biochemical composition of gonads, especially their lipids. Our study describes for the first time the effect of hormone injection frequency on the lipid content and fatty acid composition in the gonads of A. japonica during artificial maturation.     

养殖用水中孔雀石绿和结晶紫及其代谢物的测定

建立了用高效液相色谱紫外、荧光检测器同时检测养殖用水中的孔雀石绿、结晶紫及其代谢物隐色孔雀石绿和隐色结晶紫的分析方法。孔雀石绿和结晶紫用紫外检测器588 nm波长检测;隐色孔雀石绿和隐色结晶紫用荧光检测器检测,激发波长:265 nm,发射波长:360 nm。孔雀石绿和结晶紫的线性范围5.0~2500μg/L;隐色孔雀石绿和隐色结晶紫的线性范围1.0~1 000μg/L,四种物质的检出限均为0.5μg/L,回收率均在70%以上,RSD15%,此方法也可用于地下饮用水中孔雀石绿、结晶紫、隐色孔雀石绿和隐色结晶紫的检测。     

Influence of leucine-enkephalin on pituitary-ovary axis of the cichlid fish <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis>

The present investigation was conducted to elucidate the influence of an opioid peptide, leucine-enkephalin (L-ENK), on the reproductive axis of the tilapia Oreochromis mossambicus. In the first experiment, administration (i.p.) of 25, 100, and 300 μg L-ENK to the stripped female tilapia, for a period of 22 days, resulted in a significantly higher number of stage I follicles compared to those of initial controls and experimental controls, whereas the mean number of stage II and III follicles and serum levels of E₂ did not significantly differ among different experimental groups. A significant increase in the number of stage V (fully ripened) follicles was concomitant with significant reduction in the follicular diameter in 25 or 100 μg L-ENK-treated fish compared to those of experimental controls. However, significant reduction in the mean number and diameter of these follicles was observed in 300 μg L-ENK-treated fish compared to those of experimental controls and 25 or 100 μg L-ENK-treated fish. In the second experiment, the stimulatory effect of 25 μg L-ENK on the ovary was abolished in combination with gonadotropin-releasing hormone antagonist (GnRH-A). In conclusion, these results suggest that L-ENK exerts stimulatory as well as inhibitory effects on the ovary in a dose-dependent manner, and that these effects are possibly mediated through the GnRH, for the first time in fish.