药物杀除极大螺旋藻培养液中的轮虫的初步研究

通过向极大螺旋藻（Spirulina maxima）培养液中添加“轮虫克星Ⅰ号”以杀除褶皱臂尾轮虫（Brachionus plicatilis）的试验，表明该药物能有效杀灭藻液中的褶皱臂尾轮虫；后者的死亡时间、死亡率以及杀灭轮虫后螺旋藻的恢复情况与药物的浓度有关；在试验范围内既能杀灭轮虫、对螺旋藻生长的影响又较小的最佳药物浓度为1．25％。     

聚维酮碘对褶皱臂尾轮虫消毒的浓度

为研究聚维酮碘在褶皱臂尾轮虫培育中消毒时的最适添加量,采用不同浓度聚维酮碘对褶皱臂尾轮虫Brachionus plicatilis进行消毒试验,研究了聚维酮碘对于轮虫生长和繁殖的影响。结果表明:在聚维酮碘(药品浓度)1 mg/L的情况下,对褶皱臂尾轮虫的存活、种群增长、繁殖、带卵率影响不大,即与对照组无显著性差异(P0.05)。     

褶皱臂尾轮虫室内培养过程中聚缩虫的防治

通过不同浓度甲醛溶液对褶皱臂尾轮虫的种群密度和聚缩虫成活的影响的研究结果表明:(12.5~15)×10-6的甲醛溶液能在2~4d内有效杀灭聚缩虫的成体;使轮虫种群增殖速度保持持续增长。     

基于ND4和ND5基因序列研究八种臂尾轮虫的系统关系和分类地位

通过对尾突臂尾轮虫、裂足臂尾轮虫、剪形臂尾轮虫、方形臂尾轮虫、红臂尾轮虫、镰形臂尾轮虫、矩形臂尾轮虫和十指臂尾轮虫8种臂尾轮虫的ND4和ND5基因序列进行扩增、测序,结合Gen Bank数据库中的褶皱臂尾轮虫、萼花臂尾轮虫、转轮虫和旋轮虫的ND4和ND5基因序列,使用MEGA6.0软件构建这12种轮虫系统发生树(NJ树、ME树、UPGMA树),探讨了8种臂尾轮虫之间的系统关系。结果显示,本研究所涉及的轮虫ND4和ND5基因序列差异百分比均值为40.3%,可作为分子标记应用于轮虫属内种间系统关系研究;系统树所显示亲缘关系与形态学研究基本一致,均支持将十指臂尾轮虫、裂足臂尾轮虫归入臂尾轮属。     

光照、pH及盐度对褶皱臂尾轮虫培养效果的影响

研究了在不同光照周期,pH和盐度突变条件下褶皱臂尾轮虫的存活和增殖率.试验结果表明,光照对褶皱臂尾轮虫种群增殖的促进作用明显,持续光照条件下轮虫种群增殖率为持续黑暗状态的3.5倍;pH存活上限9.5,下限5.5;种群增殖最适pH为7.5～8.5;种群繁殖的最适盐度为30～35,存活的下限为15,上限为40.研究结果显示,光照对于褶皱臂尾轮虫的增殖是必要的;pH的不同对褶皱臂尾轮虫带卵量、孵化率和孵化时间均有影响.盐度突变盐差越大,褶皱臂尾轮虫的适应能力(适应所需时间和存活率)越弱.     

提高室内精养褶皱臂尾轮虫增殖率的技术要点

从褶皱臂尾轮虫(Brachionus plicatilis)的基础环境因子和控制因子等各方面阐述如何提高轮虫增殖率。褶皱臂尾轮虫的适宜繁殖条件:最适pH值7~8,最适温度35℃,最适盐度18‰,比重1.016,光照4 400~10 000lx,溶氧1.5mg/L以上。轮虫接种密度以1个/mL为宜,池中轮虫繁殖密度以100个/mL为宜。提倡用光合细菌+扁藻投喂轮虫,慎用酵母培养褶皱臂尾轮虫。在适宜条件下,经10d的培养,褶皱臂尾轮虫可由原来的1个/mL增殖到1 500~2 000个/mL。     

褶皱臂尾轮虫对提高暗纹东方鲀鱼苗成活率的作用

利用褶皱臂尾轮虫和暗纹东方鲀鱼苗在5‰的咸淡水中都能够存活的特点,用面包酵母培育褶皱臂尾轮虫经海水小球藻强化培育后在5‰的盐度条件下投喂暗纹东方鲀鱼苗,结果表明,褶皱臂尾轮虫较之淡水轮虫可使暗纹东方鲀鱼成活率显著提高。     

褶皱臂尾轮虫规模化培养新模式

褶皱臂尾轮虫(Brachionus plicatilis)(下文简称轮虫)隶属于轮虫纲、单巢目、臂尾轮虫科;是广盐性种类,广泛分布于温带到热带半咸水和海水水域。它是目前唯一能集约化培养的一种海水轮虫。作为海产苗种良好的开口饵料,随着近年来海产鱼类养殖和人工育苗工作的深入开展,其规模化培养技术愈发显示出重要作用。     

几种轮虫需精卵休眠时间的初步研究↑（*）

本文报道了角突臂尾轮虫，萼花臂尾轮虫，褶皱臂尾轮虫部分新产需精卵的自然休眠期分别为９天，６－７天，大于１２天，晶囊轮虫部分需精卵的休眠显８－１１ｈ。光照是轮虫新产需精卵萌发的必要条件。     

褶皱臂尾轮虫(Brachionus Plicatilis)室内高密度培养技术

<正> 褶皱臂尾轮虫(以下简称轮虫)属轮虫纲(Rotatoria)、单巢目(Monogononta)、游泳亚目(Ploima)、臂尾轮虫科(Brachionidae)。它营养丰富,大小适口(100～300μm),是鱼类、虾蟹人工育苗中优良的动物性饵料,在育苗生产中需求量很大。由于浙南地区的轮虫天然资源不足,不能满足生产需要,特别是在河蟹人工育苗期间,外界气温低,在室外大面积培养轮虫繁殖速度慢、密度低,需     

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.