Gonadotropins, gonadotropin receptors and their expressions during sexual maturation in yellowtail, a carangid fish

To study the physiological roles of gonadotropins (GtHs) in the yellowtail, the cDNAs encoding each GtH subunit (GPHα, FSHβ and LHβ) and their receptors (FSHR and LHR) were isolated from the pituitary gland and gonads using the polymerase chain reaction (PCR). In addition, thyrotropin (TSH) and its receptor (TSHR) cDNAs, were isolated from the pituitary gland, ovary and testis. The changes in the mRNA levels of each subunit were determined at different stages of maturation. The isolated cDNAs of GPHα, FSHβ, LHβ and TSHβ were 662, 545, 595 and 879 bp long, respectively. The amino acid sequence identity of the yellowtail GPHα, FSHβ, LHβ and TSHβ subunits was 85–63, 68–33, 93–65 and 74–46%, respectively, as compared with other fish species. Northern blot analysis showed that GPHα and FSHβ were strongly expressed in pituitary at the early vitellogenic stage and during spermatogenesis, whereas LHβ was expressed significantly in the late vitellogenic stage, and in both spermatogenesis and spermiation. Full-length cDNAs encoding FSHR, LHR, and TSHR were obtained from the testes and ovaries. The FSHR, LHR and TSHR cDNA encoded a protein of 680, 702 and 778 amino acids, and showed the highest identity with tilapia FSHR (76%), tilapia LHR (84%) and striped bass TSHR (94%), respectively. Northern blot analyses indicated that all of these receptors are expressed differently at different stages in the ovaries and testes.     

东莞生态园湿地鱼类群落结构

为了解东莞生态园湿地水域群落结构特征,探讨2种类型刺网(沉网和浮网)及不同网目网片渔获物组成的差异性,实验利用多网目(1.0、1.25、1.6、2.0、2.5、3.1、3.9、4.8、5.8、7.0、8.6和11.0 cm)单层刺网对该湿地的鱼类进行了调查。研究期间共采集到13种鱼类,相对重要性指数显示,海南似和莫桑比克罗非鱼为优势种,前者密度最高(占84.8%),后者生物量居多(40.2%)。渔获物物种组成沉浮网间差异显著,但单位努力捕捞数量(NPUE)、单位努力捕获重量(BPUE)及物种数与沉浮网间均无显著差异;海南似的NPUE在2种类型刺网间无显著差异,但莫桑比克罗非鱼和尼罗罗非鱼的NPUE浮网显著低于沉网。12个网目网片的渔获物物种聚成2类,1.0～2.0 cm网目网片渔获物聚为1类,海南似为指示种;2.5～11.0 cm片的聚为另1类,莫桑比克罗非鱼和尼罗罗非鱼为指示种。渔获物大小结构聚为2类,1.0～3.9 cm网目网片渔获物聚为1类,4.8～11.0 cm网目网片的聚为另1类。随着网目大小增加,渔获物平均体长线性递增,NPUE呈幂函数式下降。研究表明,为对内陆水域鱼类群落...     

鱼类加工副产物中胶原提取技术与性质分析

鱼类加工产业发展的同时,不可避免地产生大量鱼皮、鱼鳞、鱼骨、鱼鳍、鱼鳔等不同鱼类加工副产物,如何对这些副产物进行高值加工利用引起广泛关注。目前,鱼类加工副产物的利用途径主要包括制作休闲食品、饲料鱼粉、胶原等。其中,因胶原产品的附加值空间大,以鱼类加工副产物为原料提取胶原成为当前研究的热点之一。在介绍不同鱼类加工副产物组成特点的基础上,阐述了近年来国内外以不同鱼类加工副产物为原料提取胶原的研究现状,总结分析了从不同鱼类加工副产物中提取胶原的提取率、产物纯度及性质,展望了以鱼类加工副产物为原料提取胶原的发展前景和方向,分析认为酸-酶结合法更适用于鱼胶原的提取,并具有广阔前景。     

海州湾及其邻近海域鱼类群落结构冗余度

过度捕捞、环境污染等人类活动以及气候变化等影响着海洋鱼类群落动态及其稳定性。为深入研究海州湾海域鱼类群落结构及其变化,根据2011年、2013—2016年春、秋两季在该海域进行的渔业资源与环境调查数据,应用多元统计分析和结构冗余度分析方法,研究了海州湾海域春、秋季鱼类群落结构冗余度。结果显示,海州湾及邻近海域春、秋季鱼类均可划分为南北两个群落。春季北部群落的典型种包括方氏云鳚、细纹狮子鱼、黄鮟鱇和大泷六线鱼;南部群落的典型种为小黄鱼、短吻红舌鳎、方氏云鳚和星康吉鳗。秋季北部群落的典型种为棘绿鳍鱼、长蛇鲻和星康吉鳗;南部群落典型种包括短吻红舌鳎、棘头梅童鱼、鮸、矛尾虾虎鱼、小黄鱼和皮氏叫姑鱼。春、秋季鱼类群落响应单元数均为1,群落结构冗余度水平低;响应单元中种类与南、北鱼类群落典型种组成相似,表明海州湾春、秋季鱼类群落空间结构特征主要受方氏云鳚、细纹狮子鱼、小黄鱼、棘绿鳍鱼和长蛇鲻等少数种类决定。决定海州湾及邻近海域鱼类群落结构特征的可替代种类较少,当主要种类受损时,群落结构会发生较大变化,鱼类群落结构稳定性较低。     

剑尾鱼 2 种卵黄蛋白原全长 cDNA 的克隆及序列分析

卵黄蛋白原是环境雌激素效应研究的重要生物标志物,广泛应用于水环境内分泌干扰物污染的评价。本研究利用RT-PCR和cDNA末端快速扩增法(RACE)克隆获得了剑尾鱼(Xiphophorus helleri)的2种卵黄蛋白原基因—VgA和VgB,并对其基因结构和系统进化进行分析。VgA基因cDNA序列全长5159bp,开放阅读框(ORF)5058bp,编码1685个氨基酸。VgB基因cDNA序列全长5349bp,开放阅读框(ORF)5067bp,编码1688个氨基酸。2种cDNA序列均具有与已发现的卵黄蛋白原分子相似的主要特征和功能域,包括Vitellogenin结构域、VWFD结构域和多聚丝氨酸区域,且与已知其他鱼类卵黄蛋白原基因有较高的同源性。     

Phenotypic and genotypic characterization of Flavobacterium psychrophilum from Finnish fish farms

Finnish isolates ( n =37) of Flavobacterium psychrophilum isolated from farmed salmonids were studied using phenotypic and genotypic characteristics. The characteristics of isolates were compared with the characteristics of Swedish and Estonian F. psychrophilum isolates and the type strain, F. psychrophilum NCIMB 1947^T. Multiple isolates from eight disease outbreaks were examined to determine differences between isolates from a single outbreak. The F. psychrophilum isolates represented a biochemically homogeneous group. However, some minor differences in biochemical and physiological characteristics were observed. Seven different antigenic patterns among Finnish isolates were detected and the results suggest a new serotype of F. psychrophilum . Using Cla Ι, Hae ΙΙΙ and Pvu ΙΙ restriction enzymes in ribotyping analyses 13 different genotypes were demonstrated and a possible relationship between serotype Fd and genotype F1 was determined. There were no significant differences between the isolates from Finland, Estonia, Sweden and the type strain of F. psychrophilum .     

Benthic fish behavior characterization with a mechanically scanned imaging sonar

A point stationary, acoustic-based surveying system was proposed to fulfill the necessities of benthic fish behavior assessment. The surveying system can be split into two major and inter-dependent techniques, i.e., acoustic image acquisition and image post-processing. The first part comprises a high-frequency, mechanically scanned imaging sonar (MSIS) with bottom-fixed, side-looking working configuration. Major modules of the proposed image processing procedure include: stationary objects subtraction, region and textural feature extraction, unsupervised classification, fish target identification and quantification. For the specific case study conducted in a deep water fishpond, it is evident that both individual and school fish could be discriminated by image frames collected at a randomly selected point with scanning range setting at 5 m and frame rate at 75 s. Based on spatial and temporal analyses on position and area of the discriminated fish targets, it was concluded that fish movement pattern in the scanned area followed two distinctive corridors with significant different passage rate, i.e., a discrete, periodic and high passage rate mode in Corridor#1 and a continuous, steady and low passage rate mode in Corridor#2. Environmental features, such as bank slope, substrate mound and float raft represented specific meeting points for dynamic aggregations and schooling. Fish tended to converge into prominent schools subsequent to interactions with these features. The proposed system represented a practical and cost effective tool in acquiring image frames with sufficient spatial and temporal resolution for the characterization of fish behavior.     

鱼源嗜水气单胞菌多重耐药菌株整合子的分子特征

为研究嗜水气单胞菌多重耐药菌株整合子-基因盒分布及分子特征, 首先采用K-B纸片扩散法检测28株鱼源嗜水气单胞菌对18种抗生素的耐药性, 然后利用PCR方法检测菌株中Ⅰ、Ⅱ、Ⅲ型整合酶基因并对其携带基因盒序列进行分析。结果表明, 分离到的鱼源嗜水气单胞菌呈多重耐药性, 对b-内酰胺类、大环内酯类、氯霉素类和四环素类药物的耐药率超过60%, 而对氟喹诺酮类药物较敏感, 且菌株间耐药谱差异较大。53.57%的菌株Ⅰ类整合子阳性, 21.43%的菌株Ⅱ类整合子阳性, 整合子阳性菌株对多种药物的耐药率均高于阴性株, 且Ⅰ类整合子阳性株多重耐药率明显高于Ⅱ类整合子阳性株, 表明整合子系统在嗜水气单胞菌多重耐药性中发挥重要作用。Ⅰ类整合子基因盒以aadA、dfrA、catB家族为主, 分别介导氨基糖苷类、甲氧磺胺嘧啶类和氯霉素类药物耐药; 基因盒的排列以aadA2+dfrA12类型为主。此外, Ⅰ类整合子阳性的嗜水气单胞菌多重耐药性在不同个体间也存在较大差异, 提示多重耐药菌株的耐药表型与基因盒的类型无直接相关性。

     

Molecular characterization of Portuguese strains of Yersinia ruckeri isolated from fish culture systems

A total of 23 Portuguese strains of Yersinia ruckeri , the causative agent of enteric redmouth disease (ERM), were comparatively studied by means of lipopolysaccharide (LPS) and outer membrane protein (OMP) analysis, plasmid profiling and ribotyping in order to investigate the heterogeneity among isolates and the usefulness of these methods as epidemiological markers. Only two LPS profiles were observed among the isolates studied, corresponding with the serotypes O1 and O3 of Y. ruckeri . A higher heterogeneity was detected analysing the OMP, seven different patterns being observed. Although some isolates carried different small plasmids, all the serotype O1 isolates showed a plasmid band of 50 MDa and all the serotype O3 strains shared in common a extrachromosomic DNA band of 30 MDa. The analysis of the ribopatterns obtained using three different enzymes, separated the strains into 10 ribotypes, indicating genetic heterogeneity among the isolates. The heterogeneity was greater within serotype O1 isolates, six ribotypes being detected, than within serotype O3 in which only three ribotypes were found. Although OMP analysis and ribotyping can be useful for the differentiation of strains on the basis of farm and/or season of isolation, we consider that ribotyping is the best candidate for epidemiological studies because it is easier to perform and offers a slightly better discriminative power.     

Phenotypical and genotypical characterization of Shewanella putrefaciens strains isolated from diseased freshwater fish

Between 2007 and 2012, a variety of disease outbreaks most often characterized by skin disorders were observed among different species of freshwater fish in Poland. In most cases, the clinical signs included focally necrotized gills, necrotic skin lesions or ulcers. Internally, haemorrhages, oedematous kidney and abnormal spleen enlargement were generally noted. The disorders were accompanied by increased mortality. Most of the problems concerned cultured common carp Cyprinus carpio L. and rainbow trout Oncorhynchus mykiss (Walbaum). Fish have been examined from a number of these farms, and additionally, the wild and ornamental fish with similar clinical signs of diseases were also tested. Bacteria were isolated consistently from lesions and internal organs. They had characteristic orange-pigmented colonies which grew in pure culture or constituted 55–95% of total bacterial flora. One hundred and eighteen isolates were collected and biochemically identified as Shewanella putrefaciens group, and this was confirmed by sequencing. Challenge tests confirmed the pathogenicity of these bacteria. This is the first report characterizing and describing S. putrefaciens as a pathogen of different species of freshwater fish in Europe.     

2株海水鱼源潜在益生菌的分离、鉴定及特性分析

为筛选用于微生态制剂研发的海水鱼源益生菌,本研究对海捕野生许氏平鲉(Sebastes schlegelii)和大泷六线鱼(Hexagrammos otakii)的消化道内壁黏膜样品进行细菌分离纯化,获得80株可培养细菌。使用选择性培养基对菌株的产酶能力进行测定,选取海水鱼常见病原菌为指示菌测定分离菌株产物的抑菌活性,筛选出2株潜在益生菌TS2和TH8,并进行菌株的生理生化检测、16S rDNA序列分析、生长特性及其对宿主安全性的研究。结果显示,TS2产蛋白酶(protease)、淀粉酶(amylase)和脂肪酶(lipase),其无菌培养产物可显著抑制鳗弧菌(Vibrio anguillarum)、副溶血弧菌(Vibrio parahaemolyticus)、哈维氏弧菌(Vibrio harvey)和假交替单胞菌(Pseudoalteromonas nigrifaciens)的生长。TH8产蛋白酶和脂肪酶,其无菌培养产物可显著抑制鳗弧菌、溶藻弧菌(Vibrio alginolyticus)、副溶血弧菌、假交替单胞菌、嗜水气单胞菌(Aeromonas hydrophila)、金黄色葡萄球...     

Identification and characterization of outer membrane vesicles from the fish pathogen Vibrio ordalii

Vibriosis outbreaks due to Vibrio ordalii occur globally, but Chilean salmon aquaculture, in particular, has suffered significant monetary losses in the last 15 years. Little is known about the virulence mechanisms employed by V. ordalii. However, most Vibrio pathogens (e.g., Vibrio anguillarum, a very close taxonomic species) present outer membrane vesicles (OMVs) that are released extracellularly and implicated in the delivery of virulence factors to host cells. This study provides the first reported evidence of the fish pathogen V. ordalii producing and releasing OMVs under normal growth conditions. Analyses were conducted with the V. ordalii strain Vo-LM-18 and the type strain ATCC 33509^T. For comparative purposes, the reference strain V. anguillarum ATCC 43307 was employed. The average size for the three Vibrio strains was 0.215 ± 0.6 µm (via scanning electron microscopy) or between 0.19 and 1.8 µm (via dynamic light scattering), with each bacterium presenting a wide range. SDS-PAGE revealed similarities in OMV patterns, but neither total nor external proteins were identical. Comparing V. ordalii ATCC 33509^T and Vo-LM-18, bands were most evident in the total proteins, and the greatest degree of similarity in OMV profiles was between 37 and 50 kDa. The purified OMVs demonstrated haemolytic enzyme activity, which could play a role during V. ordalii infection. These data represent an initial step towards gaining new insights into this virulence factor, of which a lot is known in other pathogenic microorganisms.     

Isolation and characterization of a rhabdovirus from co-infection of two viruses in mandarin fish

Co-infection of two viruses has been observed in mandarin fish (Siniperca chuatsi), but the two viruses have not been characterized. In this study, a rhabdovirus has been isolated from the co-infected two viruses extracted from the diseased mandarin fish, and its morphological structure and partial biochemical and biophysical characteristics have been observed and analyzed. The isolated rhabdovirus has a typical bullet shape, and is therefore called S. chuatsi rhabdovirus (SCRV). And, the isolated rhabdovirus produced a higher titer (10^8.5 TCID₅₀ ml^− 1) than did the co-infecting viruses (10^6.5 TCID₅₀ ml^− 1). Subsequently, the viral genome RNA was extracted, and used as template to clone the complete nucleoprotein (N) gene by RT-PCR amplification. Cloning and sequencing of the SCRV N protein revealed 42%-31% amino acid identities to that of trout rhabdovirus 903/87 and the rhabdoviruses in genus Vesiculovirus. SDS-PAGE separation of the isolated SCRV and other two rhabdoviruses also revealed obvious polypeptide profile difference. Moreover, the anti-SCRV N protein antibody was prepared, and the anti-SCRV N protein antibody only could recognize the SCRV N protein, whereas no antigenicity was detected in other two rhabdoviruses. The data suggested that the SCRV should be a rhabdovirus member related to the genus Vesiculovirus in the Rhabdoviridae.     

Diversity and distribution of early life stages of carangid fishes in the northwestern Mediterranean: responses to environmental drivers

The diversity and distribution of the early stages of carangid fishes were investigated in relation to environmental conditions off the Catalan Coast (NW Mediterranean). Data were obtained during four oceanographic cruises conducted in spring–summer 2003 and 2004. A total of 4743 larvae from seven species and five genera were identified. Most were thermophilic species very abundant in the warm waters of the south and eastern Mediterranean. The presence of larvae and juveniles of Caranx crysos, Caranx rhonchus, Seriola dumerili and Trachinotus ovatus in the northwestern Mediterranean is reported for the first time. Except for Trachurus trachurus, which spawns in winter–spring, all species reproduce in spring–summer with a temporal succession in their spawning peaks. On average, the spatial patterns of different species (except T. trachurus) showed common features: a clear preference for spawning in coastal areas, high abundance of larvae in the south warmer zone in relation to the surface productive waters of Ebro river runoff and relatively high larval concentrations south of the thermal front and its associated anticyclonic eddy. The presence of larvae of thermophilic species of carangids in the northwestern Mediterranean indicates successful reproduction and establishment of these species in the cold part of the basin.     

Development,characterization, conservation and storage of fish cell lines: a review

Cell lines provide an important biological tool for carrying out investigations into physiology, virology, toxicology, carcinogenesis and transgenics. Teleost fish cell lines have been developed from a broad range of tissues such as ovary, fin, swim bladder, heart, spleen, liver, eye muscle, vertebrae, brain, skin. One hundred and twenty-four new fish cell lines from different fish species ranging from grouper to eel have been reported since the last review by Fryer and Lannan (J Tissue Culture Methods 16: 87–94, 1994). Among the cell lines listed, more than 60% were established from species from Asia, which contributes more than 80% of total fish production. This includes 59 cell lines from 19 freshwater, 54 from 22 marine and 11 from 3 brackish water fishes. Presently, about 283 cell lines have been established from finfish around the world. In addition to the listing and a scientific update on new cell lines, the importance of authentication, applications, cross-contamination and implications of overpassaged cell lines has also been discussed in this comprehensive review. The authors feel that the review will serve an updated database for beginners and established researchers in the field of fish cell line research and development.     

Improvement of the extraction procedure for hyaluronan from fish eyeball and the molecular characterization

The object of this study was to improve the isolation procedure of hyaluronan and to compare characteristics of hyaluronan from the eyeball of bigeye tuna Thunnus obesus with other sources. General sources of hyaluronan are from Streptococcus zooepidemicus and rooster comb. Hyaluronan can be also obtained from the vitreous of fish eyes. Pure hyaluronan of higher molecular weight was obtained by the following improved extraction procedure: the frozen vitreous of a tuna eyeball was used to avoid contamination with blood, muscle tissue, and other factors; extracting was carried out over a long time period under cold conditions; cetylpyridinium chloride was used in order to separate mucopolysaccharides containing hyaluronan in the initial procedure without the process of removing fat and protein by reagents. The hyaluronan obtained was characterized by gel permeation chromatography, dynamic light scattering measurements, and viscometry. The characteristics of hyaluronan from tuna eyeballs were similar to those from other sources. However, the viscosity was lower. The possible reason could be ascribed to the wide distribution of molecular size in the vitreous humor of fish eye.     

Occurrence and molecular characterization of betanodaviruses in fish and invertebrates of the Greek territorial waters

Betanodaviruses are small ssRNA viruses that cause viral encephalopathy and retinopathy, a severe neuropathological infectious disease in marine fish species worldwide. In the present study, the occurrence of betanodaviruses was investigated in wild and cultured populations of fishes and invertebrates of the Greek territorial waters. Betanodaviruses were detected in 35 species belonging to 21 families and 12 orders. To our knowledge, 23 of those are reported for the first time in Greek waters, while 11 of them are reported for the first time globally. The positive samples were subjected to sequencing and phylogenetic analysis of partial segments of RNA1 and RNA2 genes. Almost all the viruses circulating in Greece fell within RGNNV genotype, while reassortant viruses were detected in three samples, namely two inter‐RGNNV and one RGNNV/SJNNV. A novel unclassified Betanodavirus sequence was also identified. Most of the Greek sequence types have a restricted geographic distribution except for two RNA1 and one RNA2 sequence types that are widespread throughout the Mediterranean basin. The results of this study indicate the range of reservoirs/hosts of betanodaviruses and also their wide spread in the Greek territorial waters and reinforce the hypothesis that wild fish species transmit the virus to cultured ones and vice versa.