菊黄东方魨河口区海水全人工繁育技术研究

利用杭州湾河口区的天然海水,对人工繁养的菊黄东方鲀亲鱼进行强化培育,使其达到性成熟,挑选其中性腺发育良好的进行催产,并采用半干法人工授精.在水温21～22℃下,受精卵经过100～120 h的静水充气孵化后仔鱼出膜;在水温24～25℃下,经过约25 d的培育,乌仔的全长达15 mm.试验结果表明2004年分4批共计催产18组,催产率为94.4%,获得受精卵67.5万粒、初孵仔鱼58.1万尾、乌仔29.3万尾,受精率为70.2%,孵化率86.1%,育苗成活率50.4%;2005年催产12组,催产率83.3%,获得受精卵110.3万粒、初孵仔鱼90.8万尾、乌仔49.6万尾,受精率为77.5%,孵化率为82.3%,育苗成活率54.6%.试验结果还表明两个不同盐度(15,25)对菊黄东方鲀受精卵孵化和苗种培育的影响不明显.     

牙鲆人工育苗试验

2000年12月20日至2001年2月15日在日本静冈栽培中心进行牙鲆人工育苗试验。4.3万粒受精卵经50多h后，共计孵出仔鱼2.9万尾。在水温17-18℃，pH8.0-8.2，海水比重1.021的条件下，经55d培育，共育出平均全长25mm的鱼苗1.6万尾，育苗成活率的55%。     

双棘黄姑鱼鱼苗培育技术

采用牡蛎受精卵-轮虫-丰年虫幼体-桡足类-鱼肉糜育苗饵料系列,双棘黄姑鱼的育苗水温为23.5℃以上,2003年出苗全长4.11cm鱼苗12万尾;2004年出苗全长2.7 cm鱼苗8万尾;2005年出苗全长3.41 cm鱼苗33.7万尾,2005年育苗单位水体出苗率为3 510尾/m3,育苗成活率为41.6%.双棘黄姑鱼苗的特点为前期生长很慢而15日龄后生长极快,全长1.0 cm开始出现严重的自相残杀现象.     

美洲鲥室内人工育苗技术初探

为探究美洲鲥室内人工育苗的技术方法,对其受精卵孵化及苗种培育进行了初步的研究。在美洲鲥苗种培育过程中,用空气能制冷制热机组将受精卵孵化温度控制在17～19℃,苗种培育温度控制在19.0～20.5℃,并以池塘培养的生物饵料(轮虫、枝角类和桡足类等)作为仔鱼的开口饵料。试验结果:2017—2019年连续3年,每年4—8月收集美洲鲥卵,数量分别为277.96万、471.48万、2 226.32万粒,获得的受精卵分别为18.58万、74.47万、195.85万粒,受精率分别为6.68%、15.80%、8.80%;受精卵孵化后,仔鱼经30～50 d的室内水泥池培育,最终分别获得体长30～50 mm并已成功驯化摄食配合饲料的幼鱼9 392、40 083、78 600尾,育苗成活率分别为11.01%、15.00%、15.72%。试验获得了1套适合上海地区美洲鲥室内人工育苗的模式,可以为美洲鲥后续的人工育苗研究和规模化苗种培育生产提供技术参考。     

花鲈人工育苗及淡化试验

１９９６年１２月１７日收集花鲈受精卵１５６万粒于室内育苗池孵化，出仔鱼１２２３万尾，经７～１０ｄ室内培育后移入土池育苗，９０ｄ后共收获体长５～７ｃｍ大规格淡化鱼种１３６６万尾。     

南美白对虾的性腺发育、交配、产卵和受精

对池养南美白对虾 (Penaeusvannamei)亲虾进行人工促熟 ,并使之交尾、产卵、受精。实验在上海金山区杭州湾河口低盐地区一套自行设计的水循环系统中进行 ,育苗用水通过兑配使盐度为 2 7～ 2 8。育苗期间基本不换水和移动亲虾。亲虾 2 0 5尾 ,雌雄比约 1∶1。雌虾切除眼柄后 3～ 4d ,性腺即开始发育 ,6～ 7d产卵 ,但未受精。手术后约 30d ,亲虾出现追尾现象、随之发生交配并出现受精卵。实验进行了约 6 0d ,共收集虾卵 4 5 6 .7万粒 ,其中受精卵出现后所产卵为 137万粒 ,通过设置在水循环系统内的集卵网袋 ,可收集到 95 %以上的卵 ,共获受精卵 6 1万粒 ,平均日受精率为 4 4 .5 % ,无节幼体 2 5 .1万尾 ,受精卵的平均孵化率为 4 1.2 % ,最高达 87.5 %。文中还重点讨论了雌雄亲虾性腺发育的特点 ,产卵交配规律 ,以及亲虾规格大小、饵料种类、水质状况对南美白对虾交配和受精的影响     

棕点石斑鱼池塘人工育苗试验

利用对虾养殖高位池进行棕点石斑鱼(Epinephelus fuscoguttatus)仔、稚鱼和鱼苗中间培育试验。共进行5批育苗试验,其中每批购受精卵量均为1 kg,平均孵化率为91.5%,5批共孵出仔鱼593.5万尾。共培育出体长2～3 cm的鱼苗82.3万尾,仔鱼平均育苗成活率为13.9%。5批鱼苗进行池塘网箱培育试验,共培育出体长8～10 cm的大规格鱼种70.9万尾,中间培育平均成活率为85.4%。     

菊黄东方鲀河口区海水全人工繁育技术研究

利用杭州湾河口区的天然海水,对人工繁养的菊黄东方鲀亲鱼进行强化培育,使其达到性成熟,挑选其中性腺发育良好的进行催产,并采用半干法人工授精。在水温21~22℃下,受精卵经过100~120h的静水充气孵化后仔鱼出膜;在水温24~25℃下,经过约25d的培育,乌仔的全长达15mm。试验结果表明:2004年分4批共计催产18组,催产率为94.4%,获得受精卵67.5万粒、初孵仔鱼58.1万尾、乌仔29.3万尾,受精率为70.2%,孵化率86.1%,育苗成活率50.4%;2005年催产12组,催产率83.3%,获得受精卵110.3万粒、初孵仔鱼90.8万尾、乌仔49.6万尾,受精率为77.5%,孵化率为82.3%,育苗成活率54.6%。试验结果还表明:两个不同盐度(15,25)对菊黄东方鲀受精卵孵化和苗种培育的影响不明显。     

乐清湾斑鱼祭鰶人工繁殖和育苗技术研究

为了进一步了解乐清湾斑鰶的繁殖习性,实现斑鰶苗种规模化人工繁育,于2018年在浙南乐清湾斑鰶繁殖季节捕捞产卵亲鱼,采用湿法人工授精获得受精卵,在室内水泥池进行了育苗试验。人工繁育时海水水温16.3～20.1℃,比重1.017～1.019。育苗饵料系列为轮虫—丰年虫幼体—配合饲料。海上人工授精共获得发育出现胚体的斑鰶受精卵1 998 g,每千克受精卵约53.7万粒。人工授精单日的次日浮卵率最高为13.4%,单日的每千克鱼次日浮卵量最高为15.2 g。2018年共育出平均全长为4.97 cm的斑鰶苗28.16万尾,育苗成活率约为31%。     

美洲鲥室内人工育苗技术初探

为探究美洲鲥室内人工育苗的技术方法,对其受精卵孵化及苗种培育进行了初步的研究。在美洲鲥苗种培育过程中,用空气能制冷制热机组将受精卵孵化温度控制在17~19℃,苗种培育温度控制在19.0~20.5℃,并以池塘培养的生物饵料(轮虫、枝角类和桡足类等)作为仔鱼的开口饵料。试验结果:2017-2019年连续3年,每年4-8月收集美洲鲥卵,数量分别为277.96万、471.48万、2 226.32万粒,获得的受精卵分别为18.58万、74.47万、195.85万粒,受精率分别为6.68%、15.80%、8.80%;受精卵孵化后,仔鱼经30~50 d的室内水泥池培育,最终分别获得体长30~50 mm并已成功驯化摄食配合饲料的幼鱼9 392、40 083、78 600尾,育苗成活率分别为11.01%、15.00%、15.72%。试验获得了1套适合上海地区美洲鲥室内人工育苗的模式,可以为美洲鲥后续的人工育苗研究和规模化苗种培育生产提供技术参考。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.