Melatonin in the regulation of annual testicular events in carp Catla catla: evidence from the studies on the effects of exogenous melatonin, continuous light, and continuous darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February-March), pre-spawning (April-May), spawning (July-August), and post-spawning (September-October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid-day and mid-night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin-injected fish during the preparatory phase and in LL carps during the pre-spawning phase. In contrast, testicular functions in both the melatonin-treated and DD fish were inhibited during the pre-spawning and spawning phases, while the testes did not respond to any treatment during the post-spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub-tropical surface-dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Development and characterization of a new cell line CF from caudal fin of knifefish, Chitala chitala (Hamilton-Buchanan)

A new cell line was successfully obtained from caudal fin tissue of the economically important freshwater fish Chitala chitala. The cell line was optimally maintained at 28°C in Leibovitz’s L-15 supplemented with 20% fetal bovine serum (FBS). The effects of temperature and concentration of FBS on the growth of CF cells were examined. The CF cell line consisted predominantly of fibroblastic-like cells. Moderately low plating efficiencies 8%, 11%, and 17% were observed, with CF cell line in L-15 Medium with 20% FBS. Chromosomal analysis of the cell line revealed a diploid number of 42 chromosomes in C. chitala. Molecular characterization of mitochondrial 16S rRNA and cytochrome oxidase subunit I confirmed the origin of the cell line. The cells were successfully cryopreserved in liquid nitrogen (?196°C) for 6 mo, and more than 85–90% of CF cells were revived.     

秸秆隔层对盐碱土水盐运移及食葵光合特性的影响

在内蒙古河套灌区通过微区试验,研究了秸秆深埋(S)、上盖秸秆下埋秸秆(简称上秸下秸,S+S)、上盖地膜下埋秸秆(简称上膜下秸,P+S)和地膜覆盖(对照,CK)4种耕作措施对0-40 cm土层水盐运移及食葵光合特性的影响.结果表明:(1)不同措施对土壤水盐调控的效果与作用时期差异较大.P+S在整个生育期内土壤盐分含量和盐溶质浓度较低,控盐效果显著;S+S仅在苗期能保墒控盐,但控盐效果比P+S差,后期出现水减盐增现象,保墒控盐效果也不佳;S在整个生育期内土壤盐分含量和盐溶质浓度最高,控盐效果最差;CK在整个生育期内土壤水分含量变化不大,而盐分含量较高,控盐效果也不明显.(2)不同措施对土壤水盐运移调控程度的差异,导致食葵光合特性也有明显变化.与CK、S、S+S相比,P+S由于其较低的盐溶质浓度环境,明显改善了其光合特性,在苗期、蕾期和花期能提高净光合速率(Pn)和蒸腾速率(Tr),增加气孔导度(Gs),降低胞间CO2浓度(Ci),从而使其作物长势和干物质积累明显高于其它措施.综合试验结果,P+S是内蒙古河套灌区盐碱地改良中优选的控抑盐耕作措施.     

Inhibition of spawning and associated suppression of sex steroid levels during confinement in the substrate-spawning Tilapia zillii

Substrate-spawning Tilapia zillii failed to spawn in crowded holding tanks but exhibited a marked tendency to spawn soon after transfer to individually partitioned aquaria. Confined conditions suppressed the levels of serum 17 β -oestradiol (E₂) and testosterone (T) in females, which remained low throughout the period of confinement. Levels of both steroids rose significantly following transfer of fish to individual aquaria and were maintained at consistently higher levels than those of fish which remained confined. Proportions of stage 3 (late perinucleolar) oocytes were significantly lower ( P <0.001) in individual fish 21–30 days after transfer, coincident with significantly higher ( P <0.01) proportions of stage 6 or 7 oocytes (late vitellogenic or maturing oocytes). However, no significant differences ( P ≥0.05) were detected between individual or confined groups of fish in the relative proportions of stage 2 (early perinucleolar), stage 4 (cortical alveolar) or stage 5 (early vitellogenic) oocytes. Atresia increased as the period of confinement increased. Following return of individual fish to confined conditions, blood steroids fell rapidly to levels seen in fish that had remained confined throughout. It is suggested that the reduced sex steroid levels detected during confinement were insufficient to allow developing oocytes to complete oocyte growth and maturation. The detection of significantly lower proportions of stage 3 and significantly higher proportions of stage 6/7 oocytes soon after transfer suggest that fish were preparing one batch of oocytes for spawning (stage 6/7 oocytes) whilst recruiting from a pool of previtellogenic oocytes (stage 3) for the ensuing spawning cycle.     

Influence of acclimation temperature and season on acute temperature preference of adult mountain whitefish,Prosopium williamsoni

Synopsis Temperatures preferred by four groups of adult mountain whitefish, Prosopium williamsoni, collected in October before spawning, in December after spawning, in late winter, and in spring were determined in the laboratory in a horizontal gradient. Acute temperature preference based on fish tested soon after capture and final preferendum estimates were 12.8 and 17.7°C, respectively (pre-spawning), 9.6 and 11.9°C (post-spawning), 10.8 and 9.9°C (winter), and 16.4 and 16.3°C (spring). Seasonal influence on temperature preference was evident on the basis of differences in final preferenda, covariance analysis of responses of laboratory-acclimated fish, and temperature preference of fish held at ambient river temperatures. Fish of the post-spawning and winter groups preferred lower temperatures than did those of pre-spawning and spring groups. Temperatures preferred by pre-spawning fish were too high for embryo survival. Caution is necessary in predicting thermal preference on the basis of a sample collected at one time of year or stage of sexual development.     

Short- and long-term effects of a dietary yeast beta-glucan (Macrogard) and alginic acid (Ergosan) preparation on immune response in sea bass (Dicentrarchus labrax)

The present study investigated the immunomodulatory activity of Ergosan, an algal extract containing alginic acid, and Macrogard, a yeast extract containing beta-glucans, on innate and specific immunity in sea bass (Dicentrarchus labrax). Four cycles of experimental feeding using normal fish feed formulation (control group) supplemented with Ergosan (0.5%) or Macrogard (0.1%) were performed at 60-day intervals (15 days of treatment+45 days of suspension). Serum complement, lysozyme, total proteins and heat shock protein (HSP) concentrations were measured at 15, 30 and 45 days from the end of the first 15-day feeding cycle (short term) and 45 days after the end of each feeding cycle over a 35-week period (long term). The percentage of B- and T-lymphocytes in peripheral blood leucocytes and gut were measured over long-term trial. Significant elevation (P < 0.05) in serum complement activity occurred in sea bass fed with alginic acid and glucans, at 15 days from the end of first cycle of treatment. Significant elevation (P < 0.05) in serum lysozyme, gill and liver HSP concentration were observed in the same experimental groups at 30 days from the end of treatment, whereas a significant increase (P < 0.05) of complement activity was only observed in fish that received an Ergosan diet. At 45 days from the end of treatment, complement, lysozyme and HSP concentration did not differ among groups. Over the long-term period, no significant differences were observed in innate and specific immune parameters, survival, growth performances and conversion index in treated and control fish. A dramatic decrease of both innate and acquired immune parameters was observed during the winter season in all groups, followed by a partial recovery when water temperature increased. Reduction in complement and lysozyme activities was significatively correlated (p < 0.01) to water temperature variation. The results suggested the potential of alginic acid and beta-glucans to activate some innate immune responses in sea bass, and particularly under conditions of immunodepression related to environmental stress.     

闽江河口湿地植物枯落物立枯和倒伏分解主要元素动态

采用分解袋法,对闽江河口湿地2种挺水植物——芦苇(Phragmites australis)和互花米草(Spartina alterniflora)花和叶枯落物的立枯和倒伏分解过程及C、N、P元素动态进行研究。结果表明:(1)立枯分解是2种湿地盐沼植物重要的分解阶段,干物质损失率在13.26%—31.89%之间。多项式模型能较好描述2种植物花和叶的枯落物分解残留率动态。(2)立枯分解阶段,芦苇花和叶的C含量主要为波动下降,互花米草较为稳定;倒伏阶段后期,2种植物都以升高为主。立枯分解阶段2种植物枯落物N含量略有下降,而倒伏阶段逐渐上升。分解过程中枯落物P含量的波动较大。(3)2种植物花和叶C、N的NAI值在分解过程中<100%。芦苇的花和叶中P的NAI值在立枯和倒伏分解阶段都经历了明显下降和升高的过程,而互花米草在立枯阶段变化不大,倒伏阶段下降较为明显。(4)与芦苇相比,互花米草的花和叶枯落物C库较高,N库较低,P库差异不大。     

Toxicity of cypermethrin in Labeo rohita fingerlings: biochemical,enzymatic and haematological consequences

The effect of exposure to sub-lethal concentrations of cypermethrin, a synthetic pyrethroid pesticide, on biochemical parameters of muscle, blood and enzyme activities in brain, liver and kidney of the Indian major carp, Labeo rohita was studied. The sub-lethal exposure studies were done for up to 45 days at 1/10 and 1/50 of 96 h LC(50) of cypermethrin. The 96 h LC(50) was found to be 0.139 ppm. RNA levels decreased while DNA levels were elevated. Acid phosphatase was unchanged while alkaline phosphatase was depleted. Brain acetylcholinesterase activity was decreased significantly (P<0.05) over a period of 45 days at both cypermethrin concentrations. Lactate dehydrogenase activity in brain and liver was elevated, but inhibited in kidney. Succinate dehydrogenase and ATPase activities were depleted in brain, kidney and liver. There was a decrease in serum protein level over control at both concentrations of the pyrethroid. Blood glucose level and total leucocytes were elevated compared with controls at either concentration from day 15 to day 45. Haemoglobin percentage and total erythrocytes decreased in both sub-lethal concentrations. Extracts of the herb Datura stramonium were effective in countering the toxicity of this pesticide. Our data suggest that sub-lethal exposure of cypermethrin alters the biochemical, haematological parameters and enzymes of organs tissue and exert stress on the fish. Plant extracts may be useful in counteracting some of these effects.     

In vivo effects of the anesthetic,benzocaine, on liver microsomal cytochrome P450 and mixed-function oxidase activities of gilthead seabream (Sparus aurata)

Gilthead seabreams were exposed to benzocaine, 4-aminobenzoic acid ethyl ester, 57 mg/l in sea water for 3 min, daily, for 2 or 3 consecutive days. The fish were killed 20 hr after the last treatment. Benzocaine treatment for 2 or 3 days resulted in 57% and 67% inhibition of liver microsomal aniline 4-hydroxylase and ethylmorphine N-demethylase activities,respectively. The total cytochrome P450 content of fish liver microsomes was unaltered following the 2-day benzocaine treatment. However, additional 3 min benzocaine treatment on day 3 reduced cytochrome P450 level by 50%. Benzocaine produced type II difference spectra with rabbit liver microsomes. Difference spectra of fish liver microsomes elicited by benzocaine were complex. The position of peak and intensity were greatly influenced by the concentration of benzocaine.     

The effect of sublethal endrin exposure on rainbow trout, Salmo gairdneri Richardson. II. The effect of altering serum cortisol concentrations on the immune response

The effect of altering serum cortisol concentrations on the immune response was elucidated in endrin- and non-endrin-exposed rainbow trout, Salmo gairdneri. Fish were immunized with 10 μg of Yersinia ruckeri O-antigen following 30 days of treatment. The migration inhibition factor assay (MIF), plaque-forming cell assay (PFC) and serum agglutination titres (SAG) were performed 2, 14 and 30 days post-antigen inoculation. Endrin exposure was continued subsequent to antigen inoculation. Control fish were fed 20 and 35 mg kg⁻¹ body weight day⁻¹ of cortisol and metyrapone, respectively. Endrin-exposed fish received 35 mg kg⁻¹ body weight day⁻¹ of metyrapone in their diet. Control fish receiving cortisol had significantly reduced MIF, PFC and SAG responses. The MIF response was completely restored in endrin-exposed fish receiving dietary metyrapone. The PFC response and SAG titres were partially restored, 61 and 69% respectively, in endrin-exposed fish receiving metyrapone. The results indicate that elevated serum cortisol concentration obtained in endrin-exposed fish has a central role in repression of the immune response.     

Plasmid-mediated mineralization of carbofuran by Sphingomonas sp. strain CF06.

A bacterial strain (CF06) that mineralized both the carbonyl group and the aromatic ring of the insecticide carbofuran and that is capable of using carbofuran as a sole source of carbon and nitrogen was isolated from a soil in Washington state. Phospholipid fatty acid and 16S rRNA sequencing analysis indicate that CF06 is a Sphingomonas sp. CF06 contains five plasmids, at least some of which are required for metabolism of carbofuran. Loss of the plasmids induced by growth at 42 degrees C resulted in the inability of the cured strain to grow on carbofuran as a sole source of carbon. Introduction of the plasmids confers on Pseudomonas fluorescens M480R the ability to use carbofuran as a sole source of carbon for growth and energy. Of the five plasmids, four are rich in insertion sequence elements and contain large regions of overlap. Rearrangements, deletions, and loss of individual plasmids that resulted in the loss of the carbofuran-degrading phenotype were observed following introduction of Tn5.     

Characterization of progesterone profile,physiological responses,milk composition and blood biochemical and hematological indices at the early stage of lactation in goats

The aim of this research was to describe the progesterone profile, physiological response, milk composition, and blood chemistry at the early stage (DIM 30-90 days) of lactation (E₁:30-50; E₂:50-70; E₃:70-90 days) in Baladi goats under Egyptian conditions. The serum progesterone level was significantly reduced with the progress of the early lactation period to reach the least concentration (0.24 ng/ml) at the E₃ stage of lactation (P=0.001). The daily milk yield was significantly increased with the progress of the lactation to reach the peak at the E₂ stage of lactation (P=0.015). The percentage of milk fat was significantly decreased with the progress of the early lactation period (P=0.032). However, the percentages of total solids and SNF were significantly increased in E₂ and E₃ stages when compared with the E₁ stage (P=0.001 and 0.018, respectively). The serum glucose and total antioxidant capacity were considerably reduced with the progress of the lactation period (P=0.047 and 0.039, respectively). In conclusion, Baladi goats showed a decreased serum progesterone level with the progress of the early lactation period and produce milk with a relatively stable protein and lactose contents. Furthermore, Baladi does able to maintain the stability of most blood biochemical parameters during such period.     

Acute toxicity and sub-lethal effects of a pyrethroid (cypermethrin) on survival,development and fitness of Helicoverpa armigera

The chickpea pod borer, Helicoverpa armigera (Hübner) is one of the most destructive pests of India and most of the countries. In this experiment, sub-lethal effects of cypermethrin were evaluated against sixth instar larvae of this pest that had ingested insecticide-treated chickpea pods. The LC₅₀, LC₃₀ and LC₁₀ values of cypermethrin were 80.38, 26.65 and 5.41?mg a.i.?L^?1, respectively, against sixth instar. The sub-lethal effects on fitness, eggs, larvae, pupae, adult longevity and reproduction were observed in H. armigera population that survived exposure to the sub-lethal doses of LC₅₀, LC₃₀ and LC₁₀ of cypermethrin. Survivorship was reduced to 36?days if H. armigera exposed to the sub-lethal dose of LC_50, whereas it was 42?days in unexposed groups. Larvae treated with LC₅₀ dose exhibit lowest intrinsic rate of increase (r_m) (0.0413 females/female/day) and highest (0.0517) with the unexposed group. Time taken by H. armigera to complete one generation (T_c) was 40.09?days, while this time significantly lowered to 32.85?days when exposed to the sub-lethal dose of LC₃₀. Hatching eggs were greatly reduced in the larvae exposed to cypermethrin at all the sub-lethal doses. About 430 eggs/female/generation were recorded after treatment with 80.38?mg/L as compared with untreated females (1390). Larvae that were exposed to sub-lethal concentrations of cypermethrin exhibited lower pupal weight and prolonged pupal developmental times compared control larvae. The overall fitness performance was better in the unexposed population as compared to the population exposed to sub-lethal doses.     

Influence of water temperature and net tending frequency on the condition of fish bycatch in a small-scale inland commercial fyke net fishery

To date, most studies of commercial fisheries bycatch have focused on mortality at time of capture as an endpoint. However, sub-lethal indicators of organismal condition have the potential to reveal mechanisms associated with mortality (both at time of capture and post-release) and opportunities for improving fish welfare. In this study, we simulated commercial fishing efforts in inland lakes with fyke nets during a typical fishing season (early April to late June) in southeastern Ontario, Canada, where bycatch of non-target fish species had previously been documented. Using non-target gamefish (i.e., largemouth bass [Micropterus salmoides, Lacépède], northern pike [Esox Lucius, L.]), as well as a target species (i.e., bluegill [Lepomis macrochirus, Rafinesque]), we examined the sub-lethal consequences of capture (e.g., blood physiology, reflex impairment, and injury) and compared the effects of being retained in the net for two different durations (i.e., two or six days) over a range of water temperatures (i.e., 3–28 °C). Sub-lethal physiological disturbances (i.e., blood glucose and lactate) in largemouth bass and bluegill tended to be greater at higher water temperatures. However, fish retained for six days generally did not exhibit greater stress than those retained for two days, with the exception of plasma glucose in largemouth bass. Reflex impairment was similar among temperature and retention periods. Fish retained in nets experienced a range of injuries (including fin frays, scale loss, and mouth damage) that had the potential to facilitate the development of opportunistic pathogenic infections. Greater incidences of injury on fish bycatch tended to be associated with higher temperatures and longer retention. To reduce physiological disturbances and injury that could lead to delayed mortality, we suggest that regulations for inland commercial fishers require them to check their nets more frequently as water temperatures increase. We suggest that future studies of bycatch incorporate sub-lethal endpoints given that they serve as an objective measure of fish welfare and can provide quantitative mechanistic information to support management actions.     

Granular Nematicides for Control of the Yam Nematode,Scutellonema bradys, and Relevant Residues in Raw Tubers

Four granular nentaticides were evaluated for control of the yam nematode, Scutellonema bradys (Steiner &LeHew) Andrassy, on Guinea yam, Dioscorea rotundata Poir, under field conditions prevelant in the tropics. A single application of nematicides (sidedressing) at the rate of 2 kg ai/ha as postplanting treatment at the onset of the rainy season depressed numbers of S. bradys attacking yams during the growing season and significantly increased tuber yields over untreated. The efficacy, based on the regression coefficient values of evaluated nematicides, was in the order of miral, carbofuran, aldicarb, and oxamyl (b = -75.9, -75.5, -72.1, and -65.9, respectively). Yam tuber yields increased by 136.9, 90.6, 87.9, and 85.3% over untreated (P = 0.05) in aldicarb, carbofuran, oxamyl, and miral treated plots, respectively. Residues in raw tubers pretreated with aldicarb, carbofuran, or miral were negligible (front less than 0.02 to 0.3 ppm) and far below the established tolerance levels (l.0 and 1.3 ppm for aldicarb and carbofuran, respectively) of a related crop in the United States. This is the first report on residues of systemic pesticides in yams.     

Attenuation of cellular antioxidant defense mechanisms in kidney of rats intoxicated with carbofuran

Carbofuran, an anticholinestrase carbamate, is commonly used as an insecticide. Its toxic effect on kidney is less established. The present study was designed to investigate the effect of carbofuran on kidneys and to understand the mechanism involved in its nephrotoxicity. Male Wistar rats were divided into two groups of eight animals each; control animals received sunflower oil (vehicle) and carbofuran exposed animals were treated with carbofuran (1 mg/kg body weight) orally for 28 days. At the end of the treatment, significant increase was observed in urea and creatinine levels in serum along with the inhibition of acetylcholinesterase, suggesting nephrotoxicity. The antioxidant defense system of animals treated with carbofuran was altered in terms of increased lipid peroxidation, reduced glutathione, and total thiols and decreased activity of antioxidant enzymes (superoxide dismutase and catalase). The results indicate that carbofuran is nephrotoxic and increased oxidative stress appears to be involved in its nephrotoxic effects. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:393–398, 2012; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21433     

两种温度下大黄素对团头鲂生长、血液指标及肝脏 HSP70 mRNA表达的影响

实验采用2×2双因子设计, 研究两种温度下在饲料中添加大黄素对团头鲂(Megalobrama amblycephala)生长、血液生理反应及肝脏HSP70 mRNA表达的影响。选取300尾健康的团头鲂初均重(7.29±0.01) g, 两种温度下(25℃、30℃)分别投喂不添加和添加25 mg/kg的大黄素配合饲料, 养殖时间为77d。结果显示: 在长期30℃高温下, 鱼体增重率(WGR)和特定生长率(SGR)、红细胞积压(HCT)、乳酸脱氢酶(LDH)显著升高, 饵料系数(FCR)和脏体比(VSI)、谷草转氨酶(AST)、总胆汁酸(TBA)显著下降(P0.05); 添加25 mg/kg大黄素, SGR和肝体比(HSI)、红细胞积压(HCT)、乳酸脱氢酶(LDH)显著升高, 饵料系数(FCR)和摄食率(FR)显著下降(P0.05)。温度和大黄素对谷丙转氨酶(ALT)和血糖(GLU)无显著影响(P0.05), 对尿素(UREA)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、补体3 (C3)、补体4 (C4)和肝脏HSP70 mRNA的相对含量存在显著交互作用(P0.05)。与常温对照组相比, 高温对照组UREA、TG、TC显著下降; 在高温下, 添加大黄素组UREA、TG、TC有升高趋势(P0.05), C3、C4和肝脏HSP70 mRNA的相对含量显著升高(P0.05)。以上结果表明, 在饲料中添加25 mg/kg大黄素有助于缓解长期高温对团头鲂血液生理的影响, 提高补体水平和肝脏HSP70 mRNA的相对含量, 促进鱼体的生长。       

Angling-Induced Barotrauma in Snapper Chrysophrys auratus: Are There Consequences for Reproduction?

In response to concerns regarding the potential for sub-lethal impacts of barotrauma on reproductively active Chrysophrys auratus during catch and release, 90 males and 90 females representing five reproductive stages (immature or resting-28%, developing-8%, developed-7%, ripe or spawning-23% and spent-34%) were angled from 8–70 m and macroscopically assessed (on-board and then in a laboratory). Irrespective of sex, all fish exhibited various clinical signs of barotrauma, including a prolapsed cloaca (60% of fish); gastric herniation (46%); ruptured swim bladder (73%); organ displacement (48%); and kidney (3%), liver (73%) and coloemic-cavity haemorrhaging (33%);with the frequency of nearly all positively associated with capture depth. Reproductive stage was also an important barotrauma predictor (reflecting related morphological changes) with a general trend towards spent fish least likely to incur the various clinical signs—especially for a prolapsed cloaca (also common among immature or resting fish and significantly affected by food in the digestive tract) and a ruptured swim bladder (common among ripe or spawning fish). The only macroscopically visible gonad damage was haemorrhaging, which was least common among immature or resting and spent fish and, irrespective of reproductive stage, temporally reduced in frequency, and more quickly among males than females. While further research is required to accurately describe the effects of angling at each stage of the reproductive cycle and the physiological consequences of barotrauma on the gonads of C. auratus, given the observed influences of reproductive stage and depth on barotrauma found in this study, any adverse effects might be partially managed by regulating either temporal or spatial fishing effort.