中华绒螯蟹卵巢快速发育期内脂类积累以及对抱卵的影响

本文报道中华绒螯蟹卵巢快速发育期内（９月初至１２月初）脂类积累及对抱卵的影响。结果表明：在这一时期,卵巢重量和脂肪含量都迅速增加,增加的脂肪中,中性脂（ＮＬ）主要是甘油三酯和胆固醇磷脂（ＰＬ）主要是卵磷脂和脑膦脂,且ＮＬ或ＰＬ在总脂的比值比较稳定;无论是ＮＬ还是ＰＬ,ω３系列的多不饱和脂肪（主要是Ｃ２０：５和Ｃ２２：６）的百分组成熟显著增加,而ω６系列的（主要是Ｃ２０：４）百分组成则显著降低;不能     

克氏原螯虾大颚器对卵巢发育的影响

采用埋植和离体方法进行克氏原螯虾的大颚器（ＭＯ）对卵巢发育作用的研究,埋植大颚器７次能显著提高成熟系数和促进卵径增大,处于不同卵巢发育时期的克氏原螯虾大颚器提取物（ＭＯＥ）的离体研究发现,卵黄发生期的ＭＯＥ对初级和次级卵黄发生期卵径增大均有极显著的作用,而对卵黄发生前期的卵径增大无作用,处于卵巢发育早期的ＭＯＥ对卵黄发４生期卵巢小块总ＲＮＡ含量升高无显著作用,处于卵黄发生前期,卵黄发生期和恢复期卵     

中华绒螯蟹多倍体的诱导研究I.细胞松驰素B诱导中华绒螯蟹三倍体和四倍体胚胎

采用细胞松驰素Ｂ（Ｃ．Ｂ）处理法，研究了诱导中华绒螯蟹（Ｅｒｉｏｃｈｅｉｒｓｉｎｅｎｓｉｓ）三倍体、四倍体的可能性和处理条件。实验所用Ｃ．Ｂ浓度为０．５～２．０ｍｇ／Ｌ，处理的起始时间分别在受精后１０～２５ｍｉｎ（诱导三倍体）和８．５～９．８ｈ（诱导四倍体），处理的持续时间为１０～２５ｍｉｎ。三倍体的适宜诱导条件为卵子在受精后１５ｍｉｎ，浓度为１．５ｍｇ／Ｌ的Ｃ．Ｂ溶液处理１８ｍｉｎ。三倍体诱导率最高可达５８．１８％。根据诱导三倍体的适宜Ｃ．Ｂ处理浓度和处理时间，用１．５ｍｇ／ＬＣ．Ｂ溶液处理卵子１８ｍｉｎ，获得了２９．４６％～５７．８９％的四倍体胚胎。另外，还对中华绒螯蟹三倍体、四倍体胚胎的诱导技术、诱导参数对胚胎发育的影响等有关问题进行了分析讨论     

中华绒螯蟹卵黄发生期卵母细胞和卵泡细胞超微结构观

通过透射电镜技术观察了中华绒螯蟹第二次卵巢发育过程中卵巢的超微结构变化。结果表明：（1）中华绒螯蟹第二次卵巢发育过程中卵黄发生期可分为初期和后期；（2）卵黄发生初期（雌蟹第一次排卵后的16 d内），卵黄生成以卵母细胞内源性合成为主，此时卵母细胞胞质中存在大量内质网囊泡、高尔基体和线粒体，这些细胞器参与胞内卵黄物质的合成。内源性合成后期，卵母细胞膜形态多样，呈现触手状、波浪状和断裂状，为外源合成期做准备。此期卵泡细胞还未向卵母细胞靠近，两类细胞间存在着由淋巴细胞吐出的絮状物；（3）卵黄发生后期，首先为卵泡细胞与卵母细胞的结合阶段（排卵后16~21 d），此后，卵泡细胞胞质中含有大量内质网囊泡、卵黄颗粒和脂滴，卵母细胞与卵泡细胞膜变为链珠状便于物质交换，卵母细胞的卵黄合成能力减少，转由卵泡细胞进行外源性物质吸收和卵黄物质合成（21~36 d）；（4）卵黄发生结束后，双层卵膜形成，卵黄体和脂肪滴均匀分布在卵母细胞胞质中。     

中华绒螯蟹对蛋白质和脂肪消化率的初步研究

在室内控温条件下，以鱼肉粉，小麦粉等为主要原料配成精制饵料，测定了中华绒螯蟹对蛋白质和脂肪的消化率。结果表明，中华绒螯蟹对饵料的蛋白质表观消化率和实际消化率随饵料中小麦粉含量增加而下降，在２４％－５４％饵料蛋白范围内，蛋白质表观消化率（ＡＰＤ）与蛋白质含量（Ｐ）呈正比关系：ＡＰＤ＝７８．１３０３＋０．２４９８Ｐ（ｒ＝０．８５０８，ｄｆ＝４，Ｐ＜０．０２５）；脂肪消化率（ＡＬＤ）则随饵料含脂量（Ｌ）     

3种不同栖息环境下蟹鳃的超微结构、脂类组成及含量的比较

为了探讨三疣梭子蟹、无齿相手蟹和中华绒螯蟹的鳃与其所处生境的关系,分别采用电镜和生化手段观察比较了3种蟹鳃丝的超微结构,并分析了其总脂和脂肪酸组成及含量。结果表明,3种蟹鳃丝均具有相似的超微结构,而其鳃中线粒体个数和微绒毛结构差异显著。中华绒螯蟹前鳃的线粒体个数显著高于其他两者(P＜0.05),后鳃中三疣梭子蟹的线粒体个数显著低于其它两者(P＜0.05);三疣梭子蟹鳃的微绒毛较其余两者稀疏。3种蟹总脂及脂肪酸含量存在一定差异性,其中三疣梭子蟹中甘油三酯(TG)含量最高,前鳃中TG含量高出其余两种蟹3倍以上(P＜0.05);无齿相手蟹的胆固醇(Cho)含量显著高于其余两者(P＜0.05);磷脂(PL)在3种蟹鳃中的含量均无显著性差异(P＞0.05)。花生四烯酸(ARA)和高不饱和脂肪酸(HUFA)在无齿相手蟹鳃中的含量显著高于其他两者(P＜0.05)。实验结果表明,3种蟹鳃丝超微结构和脂类含量的差异与其生活的环境密切相关,这更利于其生理功能的正常发挥和对环境的适应。     

中华绒螯蟹线粒体DNA 12S rRNA基因片段的序列研究

首次进行了中华绒螯蟹（Ｅｒｉｏｃｈｅｉｒ ｓｉｎｅｎｓｉｓ）线粒体ＤＮＡ１２ＳｒＲＮＡ的序列分析。参考果蝇、蚤状序列对中华绒螯蟹线粒体ＤＮＡ１２ＳｒＲＮＡ基因片段做了引物设计、ＰＣＲ扩增及序列测定,结果得到４５７ｂｐ的碱基序列,其Ａ、Ｔ、Ｇ、Ｃ含量分别为１５９ｂｐ（３４．７９％）、１７７ｂｐ（３８．７３％）、５１ｂｐ（１１．１６％）、７０ｂｐ（１５．３２％）、与果蝇、蚤状的相同基因片段的序列含量基     

早熟期间中华绒螯蟹肝胰腺指数、肝脂含量及脂肪酸组成的变化

实验采用称量法和气相色谱法分别测定了性早熟前后中华绒螯蟹的肝胰腺指数(HI)、肝脂含量(LC)和肝胰腺脂肪酸组成。雌雄蟹的HI和LC在早熟前后的变化趋势一致,都是早熟个体低于未成熟个体。在性早熟前,雌雄蟹的肝胰腺脂肪酸组成基本一致,只是雌蟹中C20∶5n-3较高(P<0.05)。然而,对早熟前后的个体作比较时,发现雌雄蟹变化情况有很大差异:(1)早熟雌蟹中C16∶1n-7、C16∶0和C18∶0高于未成熟雌蟹(P<0.01),而C20∶5n-3、C20∶4n-6和C22∶6n-3则低于后者(P<0.01);(2)早熟雄蟹中C18∶1高于未成熟雄蟹(P<0.05),而C14∶0(P<0.05)、C16∶1n-7(P<0.05)和C20∶5n-3(P<0.01)皆低于后者。以上结果表明:雌雄蟹在性早熟前后存在肝胰腺脂肪酸组成变化的差异,且C20∶5n-3可能主要参与成熟蜕壳和膜的构建,而C22∶6n-3可能主要参与卵巢发育和卵黄的发生。     

中华绒螯蟹线粒体DNA l2S rRNA基因片段的序列研究

首次进行了中华绒螯蟹（ Ｅｒｉｏｃｈｅｉｒ ｓｉｎｅｎｓｉｓ）线粒体 ＤＮＡ １２ＳｒＲＮＡ的序列分析。参考果蝇、蚤状溞序列对中华绒螯蟹线粒体ＤＮＡ １２ＳｒＲＮＡ基因片段做了引物设计、ＰＣＲ扩增及序列测定,结果得到 ４５７ ｂｐ的碱基序列,其Ａ、Ｔ、Ｇ、Ｃ含量分别为 １５９ｂｐ（３４．７９％）、１７７ｂｐ（３８．７３％）、５１ｂｐ（１１．１６％）、７０ｂｐ（ １５． ３２％）,与果蝇、蚤状溞的相同基因片段的序列含量基本相似。     

鲤亚急性铜中毒的血液生化指标研究

以水中铜离子（Ｃｕ２＋）浓度分别为０．０３、０．０５、０．１０、０．１８、０．３２ｍｇ／Ｌ对鲤（Ｃｙｐｒｉｎｕｓｃａｒｐｉｏ）进行５周的染毒试验，研究血液生化指标的变化。结果表明，随着Ｃｕ２＋浓度的增加和试验时间的延长，中毒鲤血液红细胞数和血红蛋白数量逐渐减少，血浆超氧化物歧化酶、铜兰蛋白、碱性磷酸酶活性逐渐降低，谷丙转氨酶活性逐渐升高。试验组的各项指标与对照组存在显著差异（Ｐ＜０．０５）或极显著差异（Ｐ＜０．０１）。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

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 performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

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).     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg^?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg^?1 protein) than essential amino acids (425.82 ± 9.15 g kg^?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.     

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.     

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.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.