四指马鲅卵巢发育和卵子发生的组织学观察

为了解四指马鲅（Eleutheronema tetradactylum）的卵巢发育和卵子发生过程，本研究运用组织切片HE染色方法对其卵巢发育的组织结构变化和卵子发生过程中各时相卵母细胞的结构进行观察。结果显示，四指马鲅卵巢为被膜型卵巢，紧贴中肾腹面，两支卵巢前端分离于后端融合，呈"Y"字形。卵子发生过程分可分为5个时相，在II时相中期卵母细胞周围开始出现滤泡细胞，II时相晚期形成单层滤泡细胞，直至III时相中期滤泡细胞层外形成鞘膜细胞层；卵黄核在II时相中期开始出现至III时相早期消失；卵黄泡在III时相早期的细胞核附近及胞质边缘出现；卵黄颗粒在III时相晚期开始出现于卵黄泡之间，并在IV时相早期填充卵黄泡，至IV时相晚期形成卵黄小板。卵巢发育过程分为6个时期，每个时期都存在不同时相的卵母细胞；V期卵巢的卵径呈双峰分布，分别在50.00~100.00 μm和300.00~350.00 μm区间出现峰值。四指马鲅卵巢发育模式为非同步发育-分批产卵类型。     

长鳍吻卵巢发育组织学及周年变化的初步研究

采用常规解剖和组织学切片的方法,对长江上游宜宾江段长鳍吻卵巢发育、卵子发生以及成熟系数的周年变化进行了研究。研究结果表明,长鳍吻卵巢发育分为Ⅵ期,卵母细胞发育分为6个时相,第2时相中期出现卵黄核,2时相晚期迁移至皮质层破碎、消失;第3时相早期卵黄核消失位置开始出现卵黄泡,第3时相中后期放射带出现后卵黄颗粒开始积累;第5时相卵母细胞体积最大,胞径(1805±60.99)μm。卵巢成熟系数周年变化范围为0.78%~18.22%,一年内仅在4月份出现一个峰值,4—5月大部分卵巢处于Ⅳ、Ⅴ期,综合成熟系数测定和性腺组织学观察结果,初步推测长鳍吻繁殖期为3—5月,产卵盛期为4月上旬至5月上旬。     

黄海高眼鲽卵巢发育特征及卵径分布

本研究通过组织学观察,描述了黄海高眼鲽(Cleisthenes herzensteini)卵母细胞发育特征及其退化过程。高眼鲽卵母细胞发育分为5个时相:第1时相为卵原细胞,细胞体积小,细胞质少,细胞核明显;第2时相卵母细胞细胞核附近出现卵黄核;第3时相由胞质外缘向内层逐渐产生液泡并生成卵黄颗粒,出现双层滤泡膜;第4时相卵母细胞内充满卵黄,细胞核向动物极移动,放射膜增厚;第5时相细胞核溶解,卵母细胞从滤泡膜中释放出来并发生水合作用;产卵期过后,卵巢发生退化,卵黄颗粒逐渐被吞噬,放射膜溶解断裂。通过比较卵巢中各时相卵母细胞组成比例,表明卵母细胞发育具有非同步性。Ⅳ、Ⅴ、Ⅵ-Ⅳ'期卵巢内卵径(长径)呈单峰分布,优势粒径组分别为0.45~0.55 mm、0.60~0.65 mm和0.40~0.50 mm;Ⅴ'期卵巢,卵径分布呈双峰型,峰值分别为0.50~0.55 mm和0.90~0.95 mm,水合卵母细胞与卵径较小的小生长期卵母细胞比例增大,呈现出明显的双峰分批产卵型特征。     

广东鲂性腺发育组织学研究

利用组织学方法,对珠江下游野生广东鲂(Megalobrama terminalis)性腺进行了显微观察和研究,描述了其卵巢、精巢的结构特征以及各类型生殖细胞形态。结果显示,广东鲂属于一次性产卵类型,卵巢发育类型为部分同步型。其卵巢为细线或条状,卵巢发育经历卵原细胞期、单层滤泡细胞期、卵黄泡期、卵黄充满期、成熟期和退化期。卵细胞发生经历3个阶段:卵原细胞、初级卵母细胞和次级卵母细胞;卵巢初级卵母细胞中核仁外排现象出现在第2时相末期至第3时相早期;精巢呈线状或直棒状,精巢为叶状型结构。精细胞发生经历5个阶段:精原细胞、初级精母细胞、次级精母细胞、精子细胞及精子;繁殖季节精子充满于精小叶内,精小囊消失。     

细鳞鱼卵巢滤泡细胞的发育及功能

通过光镜和透射电镜对细鳞鱼（Brachymystax lenok）的卵巢滤泡细胞进行了研究。结果表明,滤泡细胞起源于非生殖细胞,并且其整个发育过程可分为4个时期：零散细胞期、单层扁平细胞期、多层扁平细胞期和颗粒细胞分泌期。滤泡细胞在初级卵母细胞早期并不具备任何功能,到卵母细胞卵黄积累初期,滤泡细胞上皮协同其合成部分卵膜;卵黄积累旺盛时期,滤泡细胞在此过程中起到信号介导、物质转运和储存的作用;在卵巢退化阶段,滤泡细胞与未发育的卵母细胞形成闭锁卵泡,对卵细胞残体和碎片还具有吞噬、消化等作用。     

养殖金乌贼卵子发生和卵巢发育的组织学观察

为了深入了解金乌贼(Sepia esculenta Hoyle)的生殖生理特性,本研究采用解剖学方法及组织切片技术,观察和描述了金乌贼卵巢形态和卵子各个时相结构特征及其变化规律。研究结果表明,金乌贼卵子发生过程的主要特征表现为,卵子成熟之前滤泡细胞一直存在且伴随着其结构变化,滤泡细胞解体与卵黄物质出现同步发生,卵黄物质从无到有再到大量积累直至充满整个卵母细胞,细胞核则随着滤泡细胞内折逐渐发生偏移直至最后消失。研究还发现金乌贼卵巢内卵子发生不同步且形态差异显著,未成熟的卵子粘连在卵巢基质上,依次环布于卵巢基质周围,越靠近中心,卵母细胞发育程度越低;成熟后的卵子游离在卵巢腔中,经过输卵管排出体外。根据卵巢内卵子的大小和形态、滤泡细胞形态及卵黄形成情况,将金乌贼卵子发生分为增殖期(卵原细胞期)、小生长期(无滤泡期,单层滤泡期,双层滤泡期)、大生长期(滤泡内折期,卵黄形成早期,卵黄形成晚期)、成熟期、退化吸收期共5个时期9个时相。卵巢发育分期则是以卵子时相为基础,根据卵巢的外观形态、性腺系数以及切面上优势卵子的时相,把卵巢分为Ⅰ~Ⅵ共6个时期。金乌贼卵巢发育和卵子发生的动态观察与描述,对丰富头足类繁殖生物学理论及提升人工繁殖技术都具有重要意义。     

金钱鱼性腺发育及其组织结构观察

采用常规石蜡切片,H.E染色研究了金钱鱼性腺发育及组织结构特征.结果显示,2+龄的金钱鱼性腺发育成熟.金钱鱼精巢为管型,可分为:精原细胞增殖期、精母细胞生长期、精母细胞成熟期、精子细胞变态期、精子成熟期等5个时期.从Ⅱ期精巢起,金钱鱼精巢的成熟系数(GSI)为0.2％～1.5％,精巢成熟系数在发育到精子细胞成熟期(Ⅴ期)达到峰值,肝重指数(HSI)在精子细胞变态期(Ⅳ期)达到峰值.金钱鱼卵巢的卵母细胞发育过程可分为5个时相,相对应的卵巢发育亦分为5个时期.从Ⅱ期卵巢起,金钱鱼卵巢的成熟系数为1.2％～14.9％,在发育到Ⅴ期时达到峰值,HSI在Ⅳ期达到峰值.Ⅱ时相卵母细胞出现卵黄核和滤泡膜.Ⅲ时相卵母细胞中开始出现油滴,卵黄颗粒.Ⅳ时相卵母细胞中卵黄颗粒与油滴的数量迅速增多.Ⅴ时相卵母细胞中卵黄颗粒融合成片,在卵母细胞中卵黄颗粒与油球之间在数量上没有明显的差异.根据切片观察,Ⅴ期卵巢中,除了Ⅴ时相卵母细胞外,还有一定数量的Ⅱ、Ⅲ和Ⅳ时相卵母细胞,卵母细胞发育呈现非同步型.并且发现在多数产完卵后的卵巢中,除空的滤泡外亦存在一定数量的Ⅱ、Ⅲ和Ⅳ期卵母细胞,因此推测,金钱鱼的产卵类型为分批非同步产卵类型.     

嘉陵江瓦氏黄颡鱼卵巢组织学的周年变化

采用形态学和连续组织切片技术对瓦氏黄颡鱼(Pelteobagrus vachelli)卵巢的组织形态学进行了研究。结果显示:瓦氏黄颡鱼卵巢发育分为6个时期,卵母细胞发育分为6个时相。从10月至次年的2月份,卵巢处于第Ⅱ期;3—4月份卵巢处于第Ⅲ期;4—5月初卵巢处于第Ⅳ卵期;5—7月卵巢处于第Ⅴ期;8—9月卵巢处于第Ⅵ期。在第2时相的卵母细胞中出现卵黄核和滤泡细胞;第3时相的卵母细胞出现液泡,数目和层数随着卵巢的发育逐渐增加;第4时相的卵母细胞充满卵黄颗粒,并出现了受精孔和精孔细胞。瓦氏黄颡鱼是一次产卵类型。     

孔雀鱼雌性生殖器官组织结构及卵子发生的研究

应用光镜技术研究了孔雀鱼（Poecilia reticulata）雌性生殖器官组织结构和卵子发生的特点,以期为孔雀鱼的良种繁育工作提供基础资料。结果表明,孔雀鱼雌性生殖器官由卵巢、卵子输送管和泄殖孔构成。卵子发生过程经历了6个时相：第Ⅰ时相主要为卵原细胞;第Ⅱ时相是处于小生长期的初级卵母细胞,形成生长环、卵黄核和单层扁平滤泡细胞膜;第Ⅲ时相是进入大生长期的初级卵母细胞,卵黄开始积累,形成2层滤泡细胞膜和2层卵包膜;第Ⅳ时相为发育晚期的初级卵母细胞,细胞发生了极化,卵包膜发育完善;第Ⅴ时相发育成熟,卵黄颗粒凝结呈块状,卵黄泡主要分布于细胞膜内缘,滤泡细胞膜萎缩;第Ⅵ时相是闭锁卵泡,被滤泡细胞消化和吸收转变为卵巢内结缔组织。     

池养鲻的卵巢发育和卵子发生过程

通过卵巢切片的组织学分析揭示，幼鲻在土池养殖3个月后可见线状卵巢，大约5个月后卵原细胞进入第一次成熟分裂前期的双线期转变为早期初级卵母细胞。接着卵母细胞生发泡(核)和胞质体积增加，核质比从3．5：1减少至2：1。此后卵巢中卵母细胞停滞发育持续至养殖的第3年。在第3年卵巢切片看出卵母细胞进入脂肪泡时相，在第3年秋季进入卵黄发生时相。但在人工养殖条件下卵母细胞仅能发育到卵黄发生后期，即卵母细胞胞质充满卵黄颗粒，生发泡居中而不移位。这些结果对于用人工养殖鲻为亲鱼开展人工繁殖提供重要的科学依据。并讨论了卵子发生6个时相的生物学特点及其重要的细胞器在卵黄发生中可能的生理作用。     

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.