泥鳅的性腺分化及温度对性腺分化的影响

通过组织学方法观察泥鳅(Misgurnus anguillicaudatus)原始生殖细胞(PGCs)的迁移、生殖嵴的生成和性腺分化,并且研究了不同温度下性腺分化的差异。结果表明:1日龄仔鱼PGCs位于消化管上方的脏壁中胚层中;5日龄时PGCs迁移到背部的腹膜上皮上;10日龄时生殖嵴生成;20日龄时PGCs迁入生殖嵴,形成原始性腺;25日龄时原始性腺开始向卵巢分化;30日龄时原始性腺向精巢分化。46日龄时,双侧卵巢已经合并成1个卵巢。仔鱼孵化后的25~30 d为性腺分化的关键时期。在自然水温(23~25)℃和较低水温(22±1.0)℃中培育的幼鱼,雄性率分别为(51.1±1.5)%和(48.6±1.8)%,而培育于较高水温[(27±1.0)℃,(32±1.0)℃]中的泥鳅,雄性率分别达到了(82.0±3.4)%和(90.0±2.5)%,有些个体出现了雌雄同体的现象。研究表明,温度影响泥鳅的性腺分化,高温会诱导产生雄性。[中国水产科学,2007,14(1):74-82]     

黑脊倒刺鲃性腺早期分化的研究

对黑脊倒刺鲃(Spinibarbus caldwelli)早期性腺分化的过程进行了显微和亚显微研究。结果显示:在5日龄仔鱼中,观察到生殖嵴出现;在28日龄稚鱼中,观察到精巢的早期分化;在32日幼鱼中,观察到明显分化;而直到42日龄幼鱼中,才观察到早期卵巢的早期分化;在90日龄幼鱼中,观察到明显分化。结果表明,黑脊倒刺鲃的精巢分化要明显早于卵巢的分化。     

施氏鲟的性腺分化及养殖水温对其性腺分化的影响

利用组织学方法观察了养殖施氏鲟(Acipenser schrenckii)原始生殖细胞(PGCs)的起源、迁移和原始生殖腺的形成及分化。结果表明,施氏鲟PGC最早出现于1.5 dph(days post-hatching)的仔鱼中肾管(又称吴尔夫氏管)原基一侧的中胚层中。3~7 dph,PGCs数目增多,并聚集成细胞索,17~31 dph,PGCs沿肠系膜向肾管区下方迁移,并与生殖嵴细胞一起形成原始性腺;40～60 dph时成群的小细胞团由肾区向原始性腺迁移,原始性腺上皮细胞柱状;90~120 dph,原始性腺中出现生殖上皮和性细胞,脂肪细胞和血管增多。180 dph以后,施氏鲟幼鱼性腺在组织学水平上出现光滑型(精巢)和褶皱型(卵巢)2种性腺组织结构,标志施氏鲟幼鱼性腺在解剖学水平上已经分化。本实验结果表明,孵化后31～180 d为其性腺分化的关键期。将孵化后56 d的仔鱼分别在不同的水温(27℃±0.5℃、24℃±0.5℃、21℃±0.5℃、18℃±0.5℃和15℃±0.5℃)中培育至210 d,组织学观察其性腺的发育和分化,结果发现,温度可影响施氏鲟幼鱼性腺分化的时间,但不影响其性腺分化的方向。高温(24℃和27℃)可使施氏鲟幼鱼提前性腺分化,其中27℃组较对照组(21℃)提前50 d,24℃组提前30 d;低温(15℃组)则延长幼鱼性腺分化的时间。施氏鲟幼鱼的性别分化不属于温度依赖型。     

西伯利亚鲟性腺早期发生、分化、发育的组织学观察

利用组织学方法研究人工繁育、养殖的西伯利亚鲟(Acipenser baerii)仔、幼、成鱼性腺发育过程中原始生殖细胞(Primordial germ cells,PGC)的迁移,生殖褶的生成,性腺雌雄分化和发育过程。结果表明,西伯利亚鲟于3dph(Days post-hatch)时,PGC以单细胞的形式存在于仔鱼卵黄囊上方两中肾管之间,10dph时其迁移到背侧的腹膜上皮上,13dph时其沿腹膜上皮朝肾管区下方迁移,22dph时生殖嵴形成,31dph时原始性腺形成,40～60dph时原始性腺发育形成裂腔,PGCs沿腔壁排列,裂腔经过进一步发育并从背离腹膜上皮的底端慢慢愈合成实体,90dph时原始性腺中出现生殖上皮和性细胞,150dph时脂肪细胞和血管增多,210dph时性腺出现2种不同的表面结构,性腺开始分化。本研究利用组织学方法借鉴鱼类性腺分期标准,将西伯利亚鲟性腺分化划分为0期和Ⅰ～Ⅵ期,并描述了各期精巢、卵巢的结构变化以及细胞形态学特征。     

石鲽仔、幼鱼性腺发育的组织学观察

利用组织学方法对人工培育的石鲽(Kareius bicoloratusBasiewsky)仔、幼鱼性腺发育进行研究。结果表明,性腺的发育与体长密切相关。刚孵化石鲽的原生殖细胞数目为2个,孵化3 d数目增至8个,之后经过迁移,至孵化9 d到达生殖嵴。在全长为7.2～8.5 mm(孵化9～11 d)的仔鱼中,性腺原基中的体细胞迅速增殖并包围原生殖细胞,后者在全长10～15 mm(孵化后10～35 d)的仔幼鱼中增殖成为生殖干细胞。原始性腺在全长15～30 mm(孵化40～60 d)的幼鱼中逐渐发育完善,呈细线状,位于腹腔后部中肾管下方紧贴体壁。雌性性腺最早在全长32.5 mm(孵化66 d)的个体中出现分化特征,至全长89～102 mm时雌性性腺特征完全分化。雄性性腺的分化较雌性性腺晚,最早在全长为91 mm的幼鱼中开始,至全长为114～118 mm时雄性性腺分化特征已经十分明显。     

17α-甲基睾丸酮诱导白斑狗鱼性逆转初步研究

实验以白斑狗鱼(Esox lucius)同一家系人工授精孵化的仔鱼为研究对象,用经浓度为20 mg/L 17α-甲基睾丸酮(17α-MT)浸泡的饵料分别在仔鱼生长的不同时期(出膜10、20、30、40 d)投喂,持续投喂30 d,待仔鱼培育至7月龄将其解剖取出性腺,进行组织学观察。结果显示:投喂17α-MT对其有显著致死效应;在10d喂药组出现较高的雄性率(40.8%),其它3个试验组雄性率相对较低,而对照组雄性率仅为26.3%,由此推断白斑狗鱼性腺分化转雄的时间节点在出膜10~20 d。在试验过程中发现白斑狗鱼性腺分化在稚鱼阶段存在精巢组织与卵巢组织共存现象,根据其性腺的组织学特征推断白斑狗鱼性腺分化方式可能是未分化型雌雄异体型。     

黑尾近红鲌胚胎发育和原始生殖细胞的迁移研究

用胚胎固定脱膜方法对黑尾近红鲌胚胎发育进行连续观察,用整胚原位杂交技术对原始生殖细胞的起源和迁移进行研究.试验结果表明,黑尾近红鲌受精卵为粘性,淡黄色,圆形,直径(0.94±0.05) mm.在水温24.5 ℃的孵育条件下,受精卵经25 min开始胚胎发育,45 min进入2细胞期,5 h 15 min进入囊胚期,7 h 25 min进入原肠期,10 h 15 min进入神经胚期;50 h,50%以上的仔鱼孵出.刚出膜的仔鱼长约(4.10±0.02) mm.黑尾近红鲌原生殖细胞最早发现于原肠期;在原肠中期和神经胚期,其位于胚盾靠近卵黄囊的内胚层;在胚层分化期,原生殖细胞迁移到脏壁中胚层;在孵化期,原生殖细胞迁移到体两侧的生殖嵴中,与生殖嵴共同组成胚胎的未分化性腺.     

中华锯齿米虾内外性征分化时间与特征

通过对中华锯齿米虾(Neocaridina denticulate sinensis)早期性腺发育的组织学观察和第一腹肢的形态学观察,研究了其雌雄性腺分化的时间、分化过程中生殖细胞及第一腹肢变化的特征。实验结果表明,卵原细胞出现在孵化后发育12～18 d的幼体,此时期所有幼体的第一腹肢内肢均具有刚毛。孵化后发育18～22 d幼体的性腺出现了两种类型:类型1和类型2,类型1具有典型的卵巢特征,之后沿着卵巢发育方向分化;类型2兼备精巢和卵巢的一些特征,之后沿着精巢发育方向分化。精巢的分化要经过一个类卵巢的阶段,在该时期还出现了雄性生殖肢,其第一腹肢内肢刚毛数量减少且为卵圆形,而雌性第一腹肢的内肢在形状与刚毛数量上无明显变化,雄性生殖肢的出现与精巢的分化时间是一致的。     

半滑舌鳎性腺分化的组织学观察

运用组织学方法,对半滑舌鳎性腺的发育和分化过程进行了观察。半滑舌鳎的性腺发育为雌雄异体的分化类型,可以分为3个阶段:性腺的原始阶段、性腺分化前期阶段和性腺分化完成阶段。在50日龄以前,原始性腺形成但未分化。62日龄,个体达到40.5mm时,一种性腺形成了成簇发育的原始细胞群,性腺迅速增大变宽,后来发育成为卵巢;另一种仍处于休止阶段,相对较小,后来发育为精巢。雌鱼在120日龄形成明显的卵巢腔,开始了细胞学的分化;精巢在100日龄精原细胞开始快速增殖,体细胞有丝分裂,形成输精管原基,在150日龄左右开始细胞学的分化,并形成精小叶,190日龄左右出现初级精母细胞,性腺分化基本完成。雌鱼性腺分化时期早于雄鱼。     

菊黄东方鲀性腺分化的组织学观察

运用组织切片法,对菊黄东方鲀性腺分化过程进行了观察。在40日龄前,原始性腺未分化。40日龄(21.60～24.71mm),约半数个体开始形成卵巢腔,分化为卵巢;其他约半数个体的性腺仍未分化。雌鱼在60日龄(35.51～43.60mm),形成封闭的卵巢腔;70日龄(46.30～54.75mm),卵原细胞形成卵母细胞;120日龄(94.83～107.29mm),产卵板充分发育,卵巢结构已近完善。雄鱼在60日龄(35.51～43.60mm),形成精小囊及精原细胞,开始分化;90日龄(71.59～80.54mm),形成精小管和精小叶;120日龄(94.83～107.29mm),初级精母细胞形成,性腺分化基本完成。雌鱼性腺分化时期早于雄鱼。     

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.     

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).     

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.     

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.     

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.