菲律宾蛤仔家系的建立及早期生长发育

采用不平衡巢式设计建立33个菲律宾蛤仔家系(11个父系半同胞家系和33个全同胞家系),并对各家系蛤仔的卵径、受精率、孵化率及生长、存活和变态的相关指标进行了分析。结果表明:各家系蛤仔的卵径、受精率无显著差异(P>0.05,n=90),但孵化率有明显差异(P<0.05,n=90)。在不同时期各家系蛤仔的生长情况不同。幼虫期间,9日龄C2生长最快比生长最慢的F2平均壳长大28.24%,且差异显著(P<0.05,n=90)。D3绝对生长最大比平均值高37.24%。稚贝期间,40日龄I1生长最快比生长最慢的B3平均壳长大78.29%,且差异显著(P<0.05,n=90)。I1绝对生长也最大比平均值高87.61%,其中400～500μm个体占30%,500μm以上个体占53.33%,家系内个体趋于大型化,而B1、B3家系内个体生长性状出现衰退现象,趋于小型化,300μm以下个体分别占整个家系的83.33%、90%。在相同时期各家系蛤仔的存活率不同。幼虫期间,9日龄I2存活率最高比平均存活率高94.14%,E3存活率最低比平均存活率低72.65%。稚贝期间,40日龄时各家系稚贝的存活率较高,都在85%以上。变态期间,同胞...     

菲律宾蛤仔獐子岛群体两种壳型表型性状比较

以大连獐子岛野生菲律宾蛤仔为材料,对"壳宽型"、"壳扁型"蛤仔两个壳型的壳长、壳宽、壳高、活体质量、软体质量、产卵量及其子代的生长和存活进行比较。试验结果表明,相同壳长的两种壳型蛤仔,"壳宽型"蛤仔的活体质量及软体质量均显著高于"壳扁型"蛤仔(P0.05);两种壳型蛤仔的壳宽均是影响活体质量的主要因素,壳长均是影响软体质量的主要因素;两种壳型蛤仔初孵D形幼虫、3日龄、6日龄幼虫壳长生长差异不显著(P0.05);15日龄"壳扁型"蛤仔幼虫壳长生长显著快于"壳宽型"蛤仔(P0.05);稚贝期,除30日龄外,"壳宽型"蛤仔壳长生长显著快于"壳扁型"蛤仔(P0.05);"壳宽型"蛤仔幼虫及稚贝期存活率均显著高于"壳扁型"品系(P0.05)。     

胶州湾菲律宾蛤仔个体生长模型的构建与验证

为更好地掌握胶州湾菲律宾蛤仔(Ruditapes philippinarum)的动态生长情况,本研究基于动态能量收支(Dynamic energy budget, DEB)理论,利用python软件构建了菲律宾蛤仔的个体生长模型,以2018年4月24日~2019年1月9日观测的胶州湾海域叶绿素a和水温为强制函数,通过现场实验和已有文献报道获取模型参数,模拟了菲律宾蛤仔软体组织质量和壳长的生长情况,并根据胶州湾海域菲律宾蛤仔生长的实测数据对模型进行了验证。结果表明,构建的个体生长模型能够很好地模拟胶州湾海域菲律宾蛤仔软体组织干重和壳长的生长,软体组织干重和壳长的模拟值与实测值呈显著线性相关关系(P<0.01),R2分别为0.9374和0.9168。敏感度指数最高的是阿伦纽斯温度TA和参考温度T1,如果TA和T1分别改变10%,菲律宾蛤仔软体组织干重增加高达8.86%。研究结果为后续开展基于生态系统动力学模型的养殖容量动态评估提供了基础模块和数据支撑。     

胶州湾移植底播菲律宾蛤仔的生长和死亡特性

2004年5月至2005年4月对胶州湾菲律宾蛤仔（Rudimpes philippinarum）底播增殖区进行了逐月定点采样，将样品带回放于实验室内水族箱暂养，每站随机取样，共对3269个个体进行了基本生物学特性测定，研究了移植底播菲律宾蛤仔的生长、死亡等渔业生物学特性。结果表明，底播增殖菲律宾蛤仔苗种的平均个体质量0．26g、平均壳长11．1mm；底播蛤仔与野生蛤仔在生长规律上基本一致，春末至秋初（4～9月）是其主要生长期；1～3龄期间个体生长速度较快，1～2龄、2～3龄，个体质量分别增长3．88g和4．02g；1～2龄生物量增长最快，2～3龄由于死亡率增大，其生物量增长缓慢；1～3龄蛤仔软体部的生长速度快于贝壳，5～6月是菲律宾蛤仔的繁殖肥育期。根据个体生长特性，3龄为最佳采捕年龄；根据目前的养殖状况，2龄蛤仔已达到商品规格，从生物量上分析，采捕2龄蛤仔收益最高；6月是最佳的捕获时期。蛤仔生长具有明显的季节变化，水温是影响菲律宾蛤仔生长的主要环境因子。[中国水产科学，2006，13（4）：642—649]     

不同近交系数蛤仔橙色品系的生长和成活研究

以不同遗传背景的蛤仔（Ruditapes philippinarum）橙色品系为材料,采用建立半同胞家系子一代H1（F=0.125）、全同胞家系子一代F1（F=0.250）、全同胞家系子四代F4（F=0.594）及自然对照组C1（F=0）的方法,进行了不同近交系数蛤仔橙色品系生长和成活的研究.结果表明,在整个养殖周期中F1组的壳长生长最慢,在稚贝期和养成期均显著小于F4和C1组（P＜0.05）,H1、F4和C1组之间的壳长生长差异不显著（P＞0.05）.C1组的成活率在整个养殖周期中为最小,除幼虫期3日龄外,其他各日龄均显著小于其他试验组（P＜0.05）,而H1、F1和F4组之间的成活率则差异不显著（P＞0.05）.试验组F1壳长生长的衰退率为2.92％ ～ 18.33％,平均值为（12.05±6.54）％,试验组H1在稚贝期、养成期显现出一定的衰退现象,而试验组F4在整个养殖周期中均未表现出近交衰退现象;各试验组的成活性状在各日龄均未出现近交衰退.该研究为橙色品系的选育奠定一定的理论基础.     

不同地理群体菲律宾蛤仔的选择反应及现实遗传力

对不同地理群体的3龄菲律宾蛤仔(Pp莆田群体、Dp大连群体、Tp东京群体)进行了混合选择。测量了各实验组的壳长,计算了不同地理群体菲律宾蛤仔的选择反应和现实遗传力。结果表明,3个地群体菲律宾蛤仔子代的上选组壳长显著大于对照组(P<0.05)。在不同生长发育阶段,菲律宾蛤仔的选择反应(R)和现实遗传力(hR2)随着日龄的增大而减小,即R幼虫培育期0.804±0.084>稚贝期0.705±0.039>养成期0.671±0.024;hR2幼虫期0.458±0.051>稚贝期0.402±0.025>养成期0.382±0.013。从总体水平上分析,菲律宾蛤仔R为0.726±0.1074,莆田群体、大连群体、东京群体的R分别为0.758±0.101、0.690±0.049、0.732±0.059;hR2为0.414±0.044,莆田群体、大连群体、东京群体的hR2分别为0.432±0.058、0.393±0.028、0.417±0.033。地理群体间的R和hR2次序为莆田群体>东京群体>大连群体,且彼此间无显著差异。     

桑沟湾楮岛大叶藻床区域菲律宾蛤仔的生态贡献

桑沟湾楮岛大叶藻(Zostera marina L.)床周边存在大量的底栖菲律宾蛤仔(Ruditapes philippinarum),为摸清菲律宾蛤仔的生理活动与大叶藻的相互作用,2016年5~7月,在菲律宾蛤仔和大叶藻集中分布区,评估了菲律宾蛤仔种群资源量,现场流水法测定了菲律宾蛤仔个体水平的摄食、代谢生理,围隔实验法探讨了种群水平蛤仔与大叶藻的相互作用。结果显示,桑沟湾楮岛大叶藻床海区菲律宾蛤仔的平均生物量为(572.00±20.23) ind./m2,大(壳长为3.50~4.10 cm)、中(壳长为3.00~3.50 cm)、小(壳长为2.00~3.00 cm)规格各占9.01%、43.60%和47.38%。菲律宾蛤仔的排氨率、耗氧率、滤水率、摄食率分别为(0.44±0.15)~(1.40±0.35) μmol/(ind.·h)、(0.21±0.02)~(0.33± 0.08) mg/(ind.·h)、(0.69±0.38)~(0.83±0.66) L/(ind.·h)和(2.57±0.41)~(3.41±0.68) mg/(ind.·h),且都随体重的增加而增大。围隔实验设有3个实验组(蛤仔组、大叶藻组和大叶藻+蛤仔组),1个空白组,每组3个平行(大叶藻30茎枝左右、蛤仔15个左右),实验进行4 h。研究表明,蛤仔组、大叶藻+蛤仔组和大叶藻组间的溶氧浓度存在显著差异(P<0.05);蛤仔组与其他3组的氨氮浓度存在显著差异(P<0.05);蛤仔组、大叶藻+蛤仔组与空白组的水体颗粒物浓度存在显著差异(P<0.05),大叶藻组与空白组差异不显著(P>0.05)。桑沟湾楮岛海区菲律宾蛤仔养殖面积约为0.5 km2,蛤仔每天可以过滤46 t海水中的悬浮颗粒物,并为大叶藻提供0.4 t的氨氮。本研究为深入揭示大叶藻海区菲律宾蛤仔的生态作用提供了基础数据。     

基于叶绿素a时空分布的胶州湾菲律宾蛤仔养殖容量评估

2017年7月~2019年4月期间,本研究采用大面观测、现场模拟实验与生长情况跟踪相结合的手段,基于Dame指标和Herman模型估算了胶州湾菲律宾蛤仔(Ruditapes philippinarum)的养殖容量。结果显示,调查期间,胶州湾水体的叶绿素a浓度为2.09~4.28 mg/m3,均值为3.07 mg/m3;不同规格(壳长2.29~3.59 cm)的菲律宾蛤仔单位个体的平均滤水率为0.45 L/(h?ind.),单位重量菲律宾蛤仔的平均滤水率为2.52 L/(g?h);菲律宾蛤仔1龄、2龄和3龄的平均干重分别为0.18、0.30和0.42 g;胶州湾的水团停留时间为52 d,初级生产时间为1.58 d,贝类滤水时间为2.09 d;1龄、2龄和3龄蛤仔的养殖容量分别为637、378和272 ind./m2。目前,菲律宾蛤仔养殖量已超过养殖容量,建议若以2龄蛤为采捕对象,适宜的播苗密度为582 ind./m2;若以3龄蛤为采捕对象,适宜的播苗密度为789 ind./m2。本研究结果可为保障胶州湾菲律宾蛤仔养殖产业的绿色高质量发展提供理论依据和数据支撑。     

波纹巴非蛤早期发育的扫描电镜观察

本实验使用扫描电镜显微技术观察了波纹巴非蛤胚胎发育和幼虫发育变态的早期发育过程,并获得了清晰的电镜照片,描述了波纹巴非蛤配子、胚胎和早期幼虫发育的形态变化特征。观察结果还表明,波纹巴非蛤卵子卵径为58.49μm,D形幼虫壳长87.38μm,附着幼虫壳长为200~220μm,以上形态规格均小于菲律宾蛤仔、缢蛏和硬壳蛤等双壳贝类。     

不同地理群体菲律宾蛤仔表型性状的相关性与通径分析

随机选取繁殖时期的南方群体北方养殖菲律宾蛤仔(SN)、北方土著品种(NN)和南方群体南方养殖蛤仔(SS),测量其壳长(SL)、壳高(SH)、壳宽(SW)、活体质量(mL)、软体质量(mM)和烘干后软体部干质量(mF),采用相关与通径分析方法分析不同地理群体的菲律宾蛤仔表型性状对软体质量的作用效果,为菲律宾蛤仔的选择育种提供理论依据和测度指标。试验结果表明,不同地理群体菲律宾蛤仔的形态学指标无显著差异(P>0.05);不同地理群体各数量性状之间的相关关系均达到极显著水平(P<0.01);决定系数分析表明,不同地理群体菲律宾蛤仔影响软体质量的主要表型性状并不相同,南方群体北方养殖蛤仔群体的壳宽直接作用最大,壳宽通过壳高的间接作用是次要因素;北方土著品种群体壳长是主要因素,壳长通过壳高的间接作用是次要因素;南方群体南方养殖蛤仔群体壳高的直接作用是主要因素,壳长通过壳高的间接作用是次要因素。对通径系数检验不显著的自变量进行删除,利用逐步回归的方法,建立不同地理群体菲律宾蛤仔软体质量的最优回归方程:SN群体,mM1=-4.276+0.211SH+0.116SW;NN群体,mM2=-2.806+0.088SL+0.087SH;SS群体,mM3=-3.101+0.290SH。回归方程分析表明回归关系均达到极显著水平(P<0.01)。     

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.