虾夷扇贝与风向标扇贝种间杂交的初步研究

针对虾夷扇贝在养殖过程中出现的问题,本研究以风向标扇贝和虾夷扇贝为亲本进行了种间杂交实验,培育出了虾夷扇贝（♀）×风向标扇贝（♂）（PY♀×PC♂）及风向标扇贝（♀）×虾夷扇贝（♂）（PC♀×PY♂）两种杂交一代,并对其早期发育及幼虫期和稚贝期生长进行了比较。结果显示,PY♀×PC♂杂交一代的受精率、孵化率和幼虫期的生长和存活率介于双亲之间,而壳高和壳长的生长均高于双亲,表现出显著的杂种优势;PC♀×PY♂杂交一代的受精率、孵化率、幼虫期生长和存活率均低于双亲,表现为杂种劣势。在养殖第1年,PY♀×PC♂杂交一代的壳高、壳长、壳宽和体质量增长率均高于双亲,杂种优势显著。研究表明,卵子来源对后代的表现具有极其显著的影响,以雌性虾夷扇贝与雄性风向标扇贝进行种间杂交从而改良虾夷扇贝种质是可行的。     

壳金长牡蛎自交和杂交家系生长与存活比较

选择具有生长优势的长牡蛎(Crassostrea gigas)G2(A)家系及具有明显生长和存活优势的G19(B)和G28(C)家系进行完全双列杂交,得到包括3个自交组(AA、BB、CC)和6个杂交组(AB、AC、BA、BC、CA、CB)共9个壳金长牡蛎实验组,分析各实验组在幼虫期及稚贝期不同日龄的各生长指标和存活性能,并评估了杂交组的杂种优势。结果表明,在整个培育阶段,大多数杂交组在不同生长时期均表现出较高的生长和存活性能,其中,浮游幼虫期5日龄,所有杂交组的壳高和壳长均显著大于自交组(P0.05);10日龄后,杂交组CB、BC和AC的壳高显著大于相应的自交组(P0.05),杂种优势明显;稚贝期85和130日龄,杂交组CB的壳高性能大于其他实验组。浮游幼虫期10日龄,CB组存活率显著大于AA、CA组(P0.05);稚贝期85日龄后,杂交组AC、CA和CB的存活率杂种优势逐渐增大;190日龄,杂交组AC、BC与自交组AA、CC之间存活率差异显著(P0.05)。研究结果为壳金长牡蛎新品种的培育及杂种优势的充分利用奠定了重要基础。     

不同pH对海湾扇贝胚胎发育及幼虫生长发育的影响

为探究pH对海湾扇贝(Argopecten irradia)胚胎发育及幼虫生长发育的影响,本研究以pH 8.0~8.2为对照组,分析不同pH (分别为7.6~7.7、7.3~7.4、7.0~7.1和6.7~6.8)对海湾扇贝胚胎及幼虫的影响。结果显示,随着pH的降低,各实验组孵化率降低,畸形率增高,孵化时间延长,且幼虫个体明显小于对照组(P<0.05)。D形幼虫期与壳顶幼虫期幼虫存活率与生长速率均显著低于对照组(P<0.05)。幼虫发育至第8天时,pH 7.0~7.1与pH 6.7~6.8实验组的存活率分别为6.1%和5.6%,而对照组为75.2%,除pH 7.6~7.7组外,各实验组生长速率均在3.4 μm/d及以下,基本呈现生长停滞状态。眼点幼虫附着率与对照组差异显著(P<0.05),第7天时,pH 6.7~6.8实验组附着率仅为31.7%。研究表明,海水pH变化对海湾扇贝早期胚胎及幼虫生长发育具有显著影响,该研究为评估pH变化对海湾扇贝等经济贝类资源和养殖产业的影响提供了参考资料。     

墨西哥湾扇贝选育系早期发育与生长

在水温28~29℃、盐度24~25条件下,研究墨西哥湾扇贝(Argopecten irradians concentricus)选育系的早期发育及幼虫和稚贝的生长。受精卵经50 min发育至卵裂,经22~24 h发育至D形幼虫,经11 d发育至眼点幼虫并出现足,进入附着变态阶段。选育系的胚胎发育时间和浮游幼虫前期壳长、壳高与对照系比较差异不显著(P0.05);浮游幼虫后期及至附着变态阶段,选育系与对照系的壳长和壳高差异显著(P0.05)。选育系的附着变态率和存活率与对照系比较均差异显著(P0.05),且优势明显。本研究表明,墨西哥湾扇贝浮游幼虫发育前期缓慢,幼虫后期、稚贝期生长迅速;壳长和壳高的生长速度不同步,幼虫前期壳长生长快于壳高,后期壳高生长快于壳长,并于第28~30天时壳高超过壳长。墨西哥湾扇贝选育系在幼虫和稚贝阶段生长速度明显快于对照系,具有明显的生长优势及生长潜力。     

海湾扇贝自交家系的建立和自交效应

以1992年12月从加拿大引种到中国、以群体交配的方式在中国繁养了7代的海湾扇贝(Argopecten irradians)为材料，随机选取15个性腺发育成熟的个体，分为A、B、C3组，分别通过自体受精建立15个自交家系，每组设各自混群对照。结果表明，海湾扇贝自体交配家系的受精率较高，并与对照组相近；但孵化率变化较大，15个自交系的孵化率变化范围为1．35％～68．23％，平均为31．44％。自交家系F。面盘幼虫的生长速率明显低于对照组，附着变态时间也迟于对照。在15个自交家系中有14个获得成体子代，共1311个个体，但各家系间差异很大，从13个到186个不等。自交家系子代个体可以发育到性成熟并可繁殖F2，证明通过自交培育海湾扇贝家系在技术上可行。方差分析表明，除A组自交家系体重与其对照差异不显著(P≥0．05)外，其余绝大多数自交家系和对照组相比，在壳长、壳高和体重方面的差异显著(P≤0．05)或极显著(P≤0．01)。本研究还表明，海湾扇贝的自交可导致自交衰退，表现为发育迟缓和个体小型化。在面盘幼虫阶段自交衰退率为25．9％，成体阶段平均为12．7％。     

青蛤家系的建立及早期生长发育比较

选取5个同一批苗种培育的亲贝(3个母本记为A、B、C,2个父本记为1、2)建立6个家系,并对各家系早期生长发育进行了比较.结果表明, 2个父本的壳长差异显著(P＜0.05),3个母本差异不显著(P＞0.05);不同母本产卵量不同,差异显著(P＜0.05);3个母本的卵径不同,但差异不显著(P＞0.05),各家系的受精率、孵化率与母本密切相关,以母本建立的家系之间差异极显著(P＜0.01).幼虫期(0-8日龄),D形幼虫的大小不同,家系A1(G♀1×G♂1)、A2(G♀1×G♂2)、B1(G♀2×G♂1) 与 B2(G♀2×G♂2)、C1(G♀3×G♂1) 、C2(G♀3×G♂2)差异显著(P＜0.05),8日龄时,以A1家系幼虫壳长最小,与其他家系无显著差异(P＞0.05),不同母本建立的家系的存活率差异极显著(P＜0.01).变态期(8-12日龄),幼虫生长缓慢,变态率不同,除C1 、C2无显著差异外,其他家系差异极显著(P＜0.01),变态持续2.67 d.稚贝期(12-30日龄),12-15日龄,A2、B1、B2家系稚贝略大于A1、C1、C2,差异不显著(P＞0.05);20日龄时,A2、C1家系稚贝大于其他家系,差异显著(P＜0.05);25日龄时,B1家系稚贝最小,与其他家系差异显著(P＜0.05);30日龄时,各家系稚贝生长速度、存活率依次为:C2 ＞C1 ＞A2＞A1 ＞B2 ＞B1,其中,B1 家系稚贝生长慢、存活率低,与A2、B2 家系差异显著(P＜0.05),与其他家系差异极显著(P＜0.01).各家系的面盘幼虫、变态规格、单水管稚贝规格几乎相同,无显著性差异(P＞0.05).各家系子代的早期生长发育,受亲本及相互作用的共同影响,以母本影响为主.     

不同壳型菲律宾蛤仔杂交家系的建立及早期生长发育比较

利用3种壳型的菲律宾蛤仔,即壳宽型（H）、中间型（M）和壳扁型（P）,采取双列杂交方式,成功建立3种杂交组合（PH、HM和MP）,每个杂交组合由4个杂交家系组成,共12个杂交家系。结果表明,不同壳型亲本形态差异显著（P〈0.05,n=4）;各杂交组合的D形幼虫、附着规格、变态规格和单水管稚贝的大小彼此间差异不显著（P〉0.05,n=120）。幼虫浮游期间,3种杂交组合表现出不同程度的生长、存活优势。PH杂交组合表现出明显的生长优势,与HM、MP组幼虫大小差异显著（P〈0.05,n=120）,生长速度分别为（10.21±0.42）、（9.96±0.52）和（9.29±0.52）μm/d;从存活率上看,9日龄时,PH杂交组合存活率最高,与HM、MP组合差异极显著（P〈0.05,n=12）。变态期间,幼虫生长缓慢,PH、HM和MP杂交组合的生长速度分别为（1.72±0.48）、（1.93±0.53）和（2.08±0.39）μm/d,差异显著（P〈0.05,n=120）;变态率分别为（83.20±8.47）%、（6.45±3.06）%和（10.75±3.70）%,差异极显著（P〈0.01,n=12）。稚贝培育期间,3组稚贝的大小差异极显著（P〈0.01,n=120）,生长速度分别为（16.74±3.06）、（13.08±2.24）和（15.20±2.55）μm/d;本阶段3组存活率均较高,分别为（93.25±2.99）%、（90.75±2.22）%和（87.25±4.86）%,彼此间差异显著（P〈0.05,n=12）。     

虾夷扇贝獐子岛红品系家系建立及生长发育的比较

采用不平衡巢式设计(♂∶♀=1∶3),建立了12个金贝广场挑选的獐子岛红虾夷扇贝父系半同胞(A、B、C……L)和36个母系全同胞(A1、A2、A3,B1、B2、B3,C1、C2、C3……L1、L2、L3)家系,比较各家系间浮游幼虫期和稚贝期的生长及存活指标,以评估虾夷扇贝"獐子岛红"品系的生长性状。结果表明,K3家系5日龄幼虫生长速度最快,比生长最慢的B3平均壳长大9.40%;I1家系在15日龄时生长速度最快,壳长为164.83μm,C3、D3家系25日龄壳长最大,较平均壳长高17.71%;D2家系120日龄稚贝壳高生长最快,较生长最慢的C1和D1家系平均壳高大40.41%;D2家系180日龄壳高依然最高,较H1平均壳高(18.3mm)高38.96%。I1家系幼虫存活率最高,较D1家系平均存活率高17.67%;F2家系稚贝存活率最高,显著高于其他试验组(P0.05,n=108)。综合36个同胞家系的各指标,D2、I1家系生长、抗逆性状最优,可作为优良品种选育的育种材料。     

祖国各地

”中科红”海湾扇贝是由中国科学院海洋研究所张国范课题组培育成功的，先在山东、河北、辽宁等地推广并获得成功。今年11月24日和27日，又有两批移植湛江市中海水产养殖有限公司养殖基地试养，目前长势良好，标志着”中科红”海湾扇贝南移试验取得了初步成功。     

温度和盐度对华贵栉孔扇贝幼虫存活联合效应研究

利用中心复合实验设计法研究了温度和盐度两个环境因子对华贵栉孔扇贝幼虫存活的影响。采用响应曲面分析法确立了存活的二次回归方程,同时进行优化处理得到温度、盐度的最佳组合。结果显示在华贵栉孔扇贝幼虫存活中温度与盐度在一定的范围内的互作效应不显著,其中温度效应较为明显。温度对存活率影响的一次项效应和二次项效应均达到显著水平(P0.0001)。盐度对存活率影响的一次项效应不显著(P0.05),然而,盐度对存活率影响的二次项效应达到显著水平(P0.0001)。响应曲面法对华贵栉孔扇贝幼虫存活率不同日龄同时进行优化,其结果显示最佳温度、盐度组合为24.68℃和28.03,此条件下20日龄存活率为48.25%,其满意度函数值达到93.04%。     

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).     

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.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.