乌苏里拟鳞的染色体组型研究

采用体内注射PHA和秋水仙素,体内肾细胞短期培养,常规空气干燥法制备乌苏里拟鳞的染色体玻片标本。对100个中期分裂相记数统计,确定乌苏里拟鳞的染色体数为2n=52。由测得的核型参数和按Levan等提出的染色体划分标准得出：有12对中部着丝点染色体（m）,9对亚中部着丝点染色体（sm）,5对亚端部着丝点染色体（st）。染色体臂数（NF）为94。乌苏里拟鳞组型公式为2n=24m＋18sm＋10st。     

蓝色鳞（Cyprinus carpio blue vat）的细胞遗传学分析

采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备蓝色鳞鲤染色体,对100个中期分裂相记数统计,确定蓝色鳞鲤（Cyprinus carpio blue var）的染色体数为2n=100。测得核型参数按Levan等的染色体划分标准得出：蓝色鳞鲤有15对中部着丝点染色体（m）;13对亚中部着丝点染色体（sin）;22对端部和亚端部着丝点染色体（st,t）,其染色体总臂数（NF）为156,蓝色鳞鲤核型公式为2n=30m＋26sm＋44st,t。采用流式细胞分析仪测定了蓝色鳞鲤的DNA含量,与鸡血细胞标准对照相比为1．73±0．10,以鸡红细胞DNA含量2．3Pg．N^-1计,则蓝色鳞鲤的体细胞DNA含量为3．99Pg．N^-1。     

乌苏里拟鲿的染色体组型研究

采用体内注射PHA和秋水仙素,体内肾细胞短期培养,常规空气干燥法制备乌苏里拟鳞的染色体玻片标本.对100个中期分裂相记数统计,确定乌苏里拟鲿的染色体数为2n=52.由测得的核型参数和按Levan等提出的染色体划分标准得出:有12对中部着丝点染色体(m),9对亚中部着丝点染色体(sm),5对亚端部着丝点染色体(st).染色体臂数(NF)为94.乌苏里拟鳞组型公式为2 n=24m+18 s m+10st.     

赤点石斑鱼(Epinephelus akaara ♀)与云纹石斑鱼(E.moara ♂)杂交子一代染色体核型分析

以赤点石斑鱼(Epinephelus akaara)为母本、云纹石斑鱼(E.moara)为父本进行杂交,通过人工授精技术可获得正常发育的杂交子一代,俗称红云石斑鱼(红云斑)。本文通过活体注射植物血球凝集素(PHA)及秋水仙素溶液,采用头肾细胞直接制片法,研究红云石斑鱼染色体核型。结果表明,红云石斑鱼染色体数目为48(即2n=48),其中中部着丝点染色体(m)和亚中部着丝点染色体(sm)各1对,2对为亚端部着丝点染色体(st)和20对为端部着丝点染色体(t),其染色体臂数(NF)为56,染色体核型公式为2n=48=2m+2sm+4st+40t, NF=56。本研究为红云石斑鱼的亲缘关系分析、遗传变异,以及杂交后代种质鉴定、性状选育和种质改良提供了重要的科学根据。     

淡水自鲳（Colossoma brachypomtim）染色体组型分析

淡水白鲳的染色体数目2n=54，全都为中部着丝粒和亚中部着丝粒染色体。核型公式20m 7sm,染色体臂数(NF)=108．     

淡水白鲳(Colossoma brachypomum)染色体组型分析

淡水白鲳的染色体数目2n=54,全部为中部着丝粒和亚中部着丝粒染色体。核型公式20m+7sm,染色体臂数(NF)=108。     

阿德利尔虹鳟染色体组型分析

选用蒸汽固定法对阿德利尔虹鳟的染色体核型进行了研究。结果表明:阿德利尔虹鳟具有染色体60条,中部着丝点染色体(m)20对,亚中部着丝点染色体(sm)2对,端部着丝点染色体(t)8对,核型公式为:2n=4sm+40m+16t,NF=104。未发现异形染色体。     

贵州虹彩光唇鱼的核型研究

以肾脏细胞制作染色体标本,研究了贵州虹彩光唇鱼的核型,结果表明:虹彩光唇鱼中部着丝点染色体(m)9对;亚中部着丝点染色体(sm)3对;亚端部着丝点染色体(st)3对;端部着丝点染色体(t)10对。染色体臂数(NF)80,核型公式2n=50,14m 16 sm 10 st 10 t,NF=80。     

黄颡染色体组型的初步分析

采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备黄颡鱼的染色体玻片,对100个中期分裂相记数统计,确定黄颡的2倍染色体数为2n=52。由测得的核型参数和按Levan等提出的染色体划分标准得出：有14对中部着丝点染色体(m);5对亚中部着丝点染色体(sm);4对亚端部着丝点染色体(st);3对端部着丝点染色体(t)。其染色体总臂数(NF)为90,黄颡核型公式为2n=28m 10sm 8st 6t。     

条斑星鲽染色体核型分析

采用体内注射植物血细胞凝集素（PHA）和秋水仙素法制备条斑星鲽染色体。实验鱼头肾细胞经体内短期培养,取出体外经低渗处理、卡诺氏固定液固定后,常规空气干燥法制作染色体标本。Giemsa染液染色后获得中期分裂相进行核型分析。结果表明,条斑星鲽有46条染色体,其核型为2n=46=2sm＋44t,即有1对亚中部着丝点染色体和22对端部着丝点染色体,染色体臂数为NF=48,尚未发现有多倍体的现象,也未发现异型性染色体和随体染色体。     

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.