两种孵化温度对中华鳖性别分化的影响

为分析不同孵化温度对中华鳖性别比例的影响,本实验设置两个温度(27、32℃)孵化同一批中华鳖受精卵。结果表明,当孵化温度为27℃时孵化率较低,约为91.25%,雌性个体多于雄性个体,雄性个体只占36.17%;孵化温度为32℃时,孵化率高,达95.3%,雄性个体比例明显提高,达到49.06%。结果表明,孵化温度可能影响中华鳖的性别比例,在一定温度范围内,孵化温度越高,雄性所占比例越大。     

温度对中华鳖孵育率及性别决定影响的研究

温度对中华鳖孵育率及性别决定明显影响,在一定范围内孵育率随温率升高而升高,最适温度下孵育率最高可达１００％,性别分化出率也随温度变化而变化,在２４℃～３２℃范围内,随温度升高,雌性孵化率降低,雄性孵育率升高,在２４℃时雌性孵育率高达９５％以上,而在３２℃雌性孵育率高达８５％。     

中华鳖高雄性物理控温法人工孵化技术

<正>现有研究表明,中华鳖性别决定由遗传因素(GSD)和环境因素(TSD)双重决定。采用物理控温法孵化中华鳖受精卵,在适宜的范围内提高孵化温度将会缩短中华鳖的孵化周期以及提高孵化率,孵化温度不仅能决定中华鳖的孵化周期和孵化率还能影响稚鳖的雌雄比例。可根据生产需要,设置相应性别比例的孵化温度。     

半滑舌鳎的性腺分化和温度对性别决定的影响

半滑舌鳎(Cynoglossus semilaevisGünther)是近年来新开发的一种理想的增养殖对象,雌性个体生长速度比雄性快2～3倍。由于温度可影响多种鱼类的性腺分化,因而探索温度对该鱼的性别决定和性腺分化的影响,获得性逆转雄性亲本,用于培育全雌后代,具有重要的应用前景和经济效益。通过石蜡组织切片,对半滑舌鳎早期性腺分化进行组织学观察。发现在24℃饲养条件下,孵化后30 d半滑舌鳎性腺开始分化,其中具有裂隙的性腺原基未来发育为卵巢,而不具裂隙的原基未来发育为精巢。在孵化后25～100 d对半滑舌鳎进行不同温度(16℃、20℃、24℃、28℃、32℃)处理,在9月龄时利用石蜡组织切片鉴定表型性别,观察温度对性腺分化的影响,结果表明,28℃和32℃高温能显著提高群体中的雄性比例,使其分别达到69.2%和66.7%;而低温16℃和20℃对性腺分化影响不大,群体中雄性比例分别为56.5%和57.1%;24℃处理群体中雌雄个体比例接近1∶1。半滑舌鳎雌性特异的遗传性别鉴定技术也检测到高温和低温处理组出现了由基因型雌性向表型雄性逆转的雄性个体。这些结果表明温度影响了半滑舌鳎性腺分化方向。     

全自动恒温恒湿孵化中华鳖的技术措施

全自动恒温恒湿孵化中华鳖的技术措施潘东甫（湖北省荆沙市水产学校）冯盛义（湖北省沙市电子科技实验厂）鳖卵的孵化在自然状况下受温度、湿度、敌害等因素的影响．孵化率低，一般为３０—４０％。在人工控制条件下，集中孵化．采用增温．喷水保湿等措施，孵化率一般在８...     

中华鳖SOX9基因在胚胎期性腺中的表达模式及在性逆转中的表达变化

SOX9基因在脊椎动物性别决定和性腺分化中扮演重要的调控作用。从mRNA和蛋白水平分析中华鳖SOX9基因在不同组织中的差异性表达、在胚胎性腺和成年睾丸中的细胞定位以及在性逆转中的表达变化,研究SOX9基因在中华鳖性别分化中的调控作用。Real-time PCR结果显示,SOX9基因在中华鳖雄性性腺中特异性表达。免疫荧光染色分析显示,SOX9蛋白在雄性18期胚胎性腺中开始表达,随着性腺的发育,SOX9蛋白定位于性腺Sertoli前体细胞细胞核中;而在雌性胚胎性腺并未见其表达。此外,在雌激素诱导的雄性向雌性性逆转胚胎中,SOX9基因显著下调,而在芳香化酶抑制剂诱导的雌性向雄性性逆转胚胎中,SOX9基因表达则显著上升。研究表明,SOX9基因为中华鳖雄性特异性基因,参与雄性性腺的发育过程,可能在中华鳖早期性别分化过程中起调控作用。     

孵化温度对中华鳖幼体早期生长的影响

在温度分别为 2 6、30、32、34℃ ,湿度为 - 12KPa的条件下孵化中华鳖 (Trionyxsinensis)卵 ,孵化期分别 6 6 1、4 5 2、4 2 0和 39 4天将各温级中孵出幼体在实验室恒温 (2 9± 1℃ )饲养 ,测定其早期生长。结果表明 :孵化温度明显影响中华鳖幼体的早期生长 ,其中 2 6℃组生长最快 ,30、32℃组居中 ,34℃组生长最慢     

温度对红鳍东方鲀早期生长和性别分化的影响

研究了全同胞家系的红鳍东方鲀（Takifugu rubripes）早期幼苗在4个不同温度（15℃、17℃、19℃和正常养殖温度21～25℃）条件下,水温对其生长性状和雌雄性别组合的影响。结果表明,培育温度越低,鱼苗生长速度越慢。对照组（正常养殖温度21～25℃）的幼苗生长最快,其次依次为19℃、17℃和15℃。30～90日龄,不同温度处理组绝对增长率和生长指标具有相似变化趋势。70日龄前,正常养殖温度组的绝对增长率和生长指标最高;80～90日龄,19℃组最高。统计180日龄各温度组雄性和雌性个体比例,利用组织学切片法辨别雄雌。15℃组雄性和雌性比例为86.67%和13.33%,17℃组为70.00%和30.00%,19℃组为56.67%和43.33%,正常养殖温度组为53.33%和46.67%。低温能够有效提高红鳍东方鲀家系雄性个体比例,15℃组为最佳诱导温度处理组。     

恒温恒湿孵化中华鳖技术

恒温恒湿孵化中华鳖技术潘东甫（湖北省荆沙市水产学校）潘泳冯盛义（湖北农学院水产系）（湖北省荆沙市电子科技实验厂）鳖卵的孵化在自然状况下受温度、湿度、敌害等因素的影响，孵化率低，一般为３０─４０％。在人工控制条件下集中孵化，采用增温、喷水保温等措施，孵...     

不同性别鳖的生长差异及孵化温度控制鳖性别的初步研究

同样的饲养条件,雄、雌性鳖在一定体重范围内,生长速度有差异。在同一养殖群体中,个体重超过平均体重的鳖85%以上为雄性。雄鳖相对于雌鳖有生长优势.孵化温度对鳖性别有一定影响.     

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.