杂交鳢(斑鳢♀×乌鳢♂)内脏类结节病病原菌的分离、鉴定与特性分析

从广东佛山、广州两地养殖场患内脏类结节病杂交鳢(Channa maculata♀×C.argus♂)内脏器官分离到2株细菌,纯化培养后获得2个分离株,编号为WL-1和WL-2,对分离菌株进行了细菌鉴定、致病性分析及药敏实验。应用常规生理生化鉴定和ATB系统细菌自动鉴定仪对分离菌株进行细胞形态学、理化特性分析,初步判定所分离菌为舒伯特气单胞菌。采用16S rRNA基因、DNA促旋酶的B亚单位蛋白(gyrB)基因对分离菌株进行DNA分子鉴定,结果显示,两个菌株间的16S rRNA基因序列相同,gyrB基因序列同源性为99.7%;分离菌株与GenBank上登录的舒伯特气单胞菌16S rRNA基因序列和gyrB基因序列同源性均最高,达99%以上;分离菌株在系统进化树上与舒伯特气单胞菌聚为一族,进一步确认分离株为舒伯特气单胞菌。人工感染健康鱼后出现与自然发病相似的内脏类结节病症状,从发病鱼内脏组织再分离的细菌特性与原感染菌相同。综合理化特性分析、基因鉴定和人工感染实验确认舒伯特气单胞菌是杂交鳢内脏类结节病的致病菌。药敏实验发现分离菌株对头孢唑啉、庆大霉素等14种药物敏感;对青霉素G、苯唑西林2种药物耐受。     

Identification of Mycobacterium spp. isolated from snakehead, Channa striata (Fowler), and Siamese fighting fish, Betta splendens (Regan), using polymerase chain reaction–reverse cross blot hybridization (PCR–RCBH)

A variety of methods have been used to identify Mycobacterium spp. isolated from snakehead and Siamese fighting fish, including biochemistry, mycolic acid profiles and antibody-based methods. However, these methods are unable to differentiate between different species of Mycobacterium . Polymerase chain reaction (PCR) followed by reverse cross blot hybridization (RCBH) was adapted in this study to speciate aquatic mycobacteria. The method was highly specific for Mycobacterium spp. and identified the bacteria to species level with a detection limit of 100 fg DNA, equivalent to 20 mycobacteria. Twenty-nine isolates previously collected and cultured from Siamese fighting fish (10 isolates) and snakehead (19 isolates) during outbreaks of mycobacteriosis were analysed using PCR–RCBH. Six of the Siamese fighting fish isolates and nine of the snakehead isolates were identified as Mycobacterium fortuitum , while the remainder were classified as M. marinum . Notably, two isolates recovered from snakehead and Siamese fighting fish, previously identified as M. poriferae and M. piscicida , respectively, were confirmed to be M. fortuitum .     

Isolation and characterization of Aeromonas schubertii from diseased snakehead, Channa maculata (Lacepède)

Pure bacterial cultures were isolated from diseased snakeheads, Channa maculata (Lacepède), suffering high mortality in a farm in Zhongshan, southern China. Three isolates, namely ZS20100725, ZS20100725-1 and ZS20100725-2, were identified as Aeromonas schubertii. All the isolates showed high 16S rRNA sequence similarities with A. schubertii. The isolates exhibited strong virulence to snakeheads in experimental challenges with LD(50) ranging between 1.4 × 10(4) and 6.4 × 10(6) CFU g(-1). Two of the isolates were positive for haemolysin, elastase, lipase and lecithinase by phenotypic determination, which was further confirmed by PCR amplification of the haemolysin and elastase genes. In sterile liquid medium, the best growth conditions of strain ZS20100725 were 30 °C, pH 7 and 0.5% salinity (w/v). Antibiotic susceptibility tests showed that strain ZS20100725 was susceptible to cefoxitin, cefoperazone and chloramphenicol. Furthermore, histopathology of diseased snakeheads infected with A. schubertii showed necrosis and congestion in liver, kidney and spleen and also damage to the cardiac muscle, intestine and gills.     

Aeromonas shuberti as a cause of multi‐organ necrosis in internal organs of Nile tilapia,Oreochromis niloticus

A disease with white spots in internal organs of Nile tilapia occurred in Zhanjiang, southern China. Multiple, white nodules, 0.8–2.2 mm in diameter, were scattered throughout the liver, spleen and kidney of diseased fish. Signs of nodules reproduced after artificial infection with the isolated strain. Isolated bacteria were Gram‐negative, facultative anaerobic, motile, short rod‐shaped, with a length of 1.2–2.2 μm. Morphological and biochemical tests, as well as phylogenetic analysis, all strongly indicated that the isolate from tilapia is identical to Aeromonas schubertii (A. schubertii) which temporary named LF1708 strain. Antibiotic sensitivity assays showed the LF1708 is sensitive to 24 of 27 tested antibiotics. Pathogenicity test revealed that the isolate at the dose of 3.75 × 10⁶ CFU/g killed 100% of experimental tilapia within 2 days and the dose of 1 × 10⁷ CFU/g killed 100% of experimental zebrafish within 1 day. Histopathology of diseased tilapia infected with A. schubertii showed numerous necrotic lesions widely distributed in spleen, liver and kidney, and infiltration with a large number of bacteria. To our knowledge, this was the first report that associated A. schubertii with mortality in tilapia.     

池塘网箱养鱼的可行性及技术研究

１９９５、１９９６年在精养成鱼塘设置网箱养殖商品湘鲫与乌鳢鱼种，采取灌排水形成微流、启动增氧机、投放耐低氧吃食性鱼类、投喂颗粒馈料和动物肉、勤刷钢箱养措施，结果：湘鲫养殖１３６天，从１５０ｇ长到６７０ｇ，平均日增重３．８２ｇ，２５ｍ＾２的网箱产成鱼３３４．３３ｋｇ。平均１３．３ｋｇ／ｍ＾２，成活率９９．８％，乌鳢种培育４２天，由体长１ｃｍ到１０ｃｍ，平均日增长２．１４ｍｍ，２０ｍ＾２的网箱，共育鱼     

Study on the pathogenicity of Nocardia asteroides to the Formosa snakehead, Channa maculata (Lacepède), and largemouth bass, Micropterus salmoides (Lacepède)

Abstract. The Formosa snakehead, Channa maculata (Lacepède), and the largemouth bass, Micropterus salmoides (Lacepède), were given an intraperitoneal (i.p.) and/or intramuscular (i.m.) inoculation of Nocardia asteroides isolated from cultured Formosa snakehead, Typical granulomatous lesions induced by the experimental fish were similar to the naturally infected cases. The Formosa snakehead showed 100% and 93% mortality at 14 and 35 days after i.m. inoculation with 8 mg and 0·8 mg of bacterial suspension, respectively. The largemouth bass was slightly resistant to N. asteroides but still had 95% mortality at 35 days after inoculation (i.m.) with 8 mg bacterial suspension. The bacteria were easily reisolated from spleen and kidney of the experimental fish. Many typical multifocal granulomata were found at the sites of injection and the internal organs such as liver, spleen, kidneys, pancreas, intestine, gills, eyes and brain. The result indicated that the N. asteroides isolated from the field ease was pathogenic to Formosa snakehead and largemouth bass.     

Ulcerative disease rhabdovirus: cell-line susceptibility and serological comparison with other fish rhabdoviruses

Abstract. Five isolates of a rhabdovirus associated with a severe ulcerative disease of striped snakehead, Ophicephalus striatus, and other freshwater fish in S.E. Asia were examined for growth in a range of cold and warm water fish cell lines and for serological relationships to other pathogenic fish rhabdoviruses. All five isolates grew in AS, BF-2, snakehead, O. striatus and O. micropeltis, and climbing perch, Anabas testudineus, cell lines. CHSE-214, RTG-2, EPC, Nile tilapia, Oreochromis niloticus, and grass carp Ctenopharyngodon idella, cell lines were all refractory to infection. Cross neutralization tests showed the ulcerative disease rhabdovirus (UDRV) to be serologically distinct from VHSV, IHNV, EVA, EVX, SVCV, PFRV and perch rhabdovirus. UDRV appears to be a new distinct pathogen of freshwater fish in S.E. Asia.     

Effect of body weight on the chemical composition and collagen content of snakehead fish <Emphasis Type="Italic">Channa striata</Emphasis> skin

Snakehead fish (Channa striata) skin is a fishery by-product that has the potential for further processing because it contains a high amount of organic matter. This study investigates the effect of body weight on the chemical composition and collagen content of snakehead fish skin. This study used fresh snakehead fish of either gender. Their body weights were divided into three groups: small, medium, and large size. The characteristics of snakehead fish skin included proximate composition, amino acid content, collagen content, microstructure, and minerals. Snakehead fish skin from fish of different body weights indicated that the moisture content and ash tended to decrease, the protein content was relatively stable, and the lipid content tended to increase with increasing body weight. Glycine and proline comprised the highest percentages of amino acids at all levels of body weight, and the presence of hydroxyproline showed that snakehead fish skin was the source of collagen. There was no significant difference in body weight observed on collagen protein content of the skin. This study on the microstructure and mineral content of snakehead fish skin can be used as supporting information to promote the potential utility and economic value of the skin.     

Pathogenicity and culture studies comparing the Aphanomyces involved in epizootic ulcerative syndrome (EUS) with other similar fungi

Pathogenicity and cultural experiments described here provide futher evidence that a distinct species of Aphanomyces is responsible for much of the characteristic pathology of epizootic ulceration syndrome (EUS). Zoospores from 58 fungal isolates were injected intramuscularly in snakehead fish, Channa striata (Bloch). These fungi comprised: Aphanomyces strains isolated from EUS-affected fish; saprophytic Aphanomyces , Achlya and Saprolegnia spp. from infected waters; and further saprolegniaceous fungi involved in other diseases of aquatic animals. Only the Aphanomyces strains isolated from fish affected by EUS, Australian red spot disease (already considered synonymous with EUS) or mycotic granulomatosis described from Japan were able to grow invasively through the fish muscle and produce the distinctive EUS lesions. In contrast to Aphanomyces astaci Schikora, the EUS- Aphanomyces was shown to be unable to infect noble crayfish, Astacus astacus L. The snakehead-pathogenic strains were further distinguished from all the other fungi under comparison by their characteristic temperature-growth profile and inability to grow on certain selective fungal media.     

桁杆虾拖网选择性装置的试验分析

设计并研制了一种内、外圆环形刚性框架和柔性六角形网片组合而成的桁杆虾拖网选择性装置。所研制选择性装置在东海区近海桁杆虾拖网中,采用对比试验法开展了海上选择性试验,并针对试验期间主要渔获品种哈氏仿对虾,以Logistic曲线为选择性模型,对六角形网目水平缩结长度分别为D1=31.00 mm,D2=27.50 mm,D3=21.25 mm三种规格选择性装置的选择性效果进行分析。结果表明,3种规格选择性装置均具有选择性效果,其中安装六角形网目水平缩结长度为31.00 mm和27.50 mm的选择性装置的目标网囊内哈氏仿对虾的优势体长均为75~95 mm,相应非目标网囊内哈氏仿对虾的优势体长分别为25~45 mm和35~55 mm,具有较显著的差异性;而安装六角形网目水平缩结长度为21.25 mm的选择性装置的目标网囊内哈氏仿对虾的优势体长35~55 mm,相应非目标网囊内哈氏仿对虾的优势体长同为25~45 mm。六角形网目水平缩结长度31.00 mm的选择性装置对哈氏仿对虾的50%选择体长L50最大,为48.89 mm,而选择性范围SR最小,为16.75 mm,在试验规格范围内的选择性效果最显著。     

乌鳢群体遗传多样性和遗传结构分析

为了解乌鳢群体遗传变异规律,本研究对8个群体共212个个体的mtDNA控制区全序列进行群体遗传多样性、遗传结构和群体历史动态分析。结果显示,乌鳢控制区全序列长度为907 bp,乌鳢群体单倍型多样性水平变化较大,中国黄河及以北的乌鳢群体单倍型多样性水平比淮河和长江等南方群体相对较低,所有群体表现出较低的核苷酸多样性水平(h0.5%)。基于单倍型构建的系统发育树和群体聚类树结果均未显示出与地理位置相对应的谱系结构。单倍型网络图显示存在多个主单倍型。遗传结构分析显示,不同水系间存在显著的遗传差异,相同水系间遗传差异较小。群体历史动态分析显示,中国乌鳢所有群体有效种群数量在中更新世晚期到晚更新世0.222—0.050百万年出现了一次较明显的快速增长,之后在晚更新世末次冰期0.050—0.010百万年出现了有效种群下降,伴随着全新世到来,在0.010百万年之后,乌鳢群体又发生了一次较小的有效种群增长。洞庭湖群体则发生一次有效种群的快速增长,增长时间大约在0.160百万年。研究表明,青藏高原隆起后,东亚季风在中国南北方气候的差异和秦岭山脉屏障对季风的阻断作用加大了这一差异,可能对乌鳢群体的遗传多样度差异造成一定影响。乌鳢群体不存在显著的谱系结构可能与乌鳢遗传分化时间较短有关,但是地理隔离等因素导致了不同水系间确切的遗传差异。第四纪更新世气候的波动,尤其是中更新世间冰期气候的转暖、末次盛冰期的降温和冰后期全新世的到来可能对乌鳢群体的数量和栖息地的扩缩起着重要影响。     

阿克苏地区三种食用鱼肉质特性的比较分析

以新疆阿克苏地区的三种食用鱼黄颡鱼(Pelteobagrus fulvidraco)、鲶(Silurus spp)、黑鱼(Channa argus)为材料,对它们成体鱼肉质特性,包括pH值、滴水损失、熟肉率、肌纤维直径、肌纤维密度、肌纤维面积和肌内肌纤维与结缔组织体积比进行了测定。结果表明,黄颡鱼除滴水损失逊于鲶和黑鱼外,其它肉质特性指标均优于鲶和黑鱼,说明其肉质细嫩,食用品质相对较好。     

东海蓝圆鲹的摄食习性

以2008年5月至2009年2月东海灯光围网采集到的叉长范围为52～276 mm的444尾蓝圆鲹为对象,对其胃含物进行分析,应用K-W非参数检验、卡方检验、聚类分析等方法,对不同季节和发育阶段条件下蓝圆鲹的食性进行了研究。结果表明,蓝圆鲹的摄食种类有80余种,浮游甲壳类(主要包括浮游甲壳类幼体、磷虾和桡足类等)和小型鱼类为其主要食物类群。若按相对重要性指数百分比(IRI%)排序,优势饵料种类最高的是七星底灯鱼(IRI%为40.44%),其次是口虾蛄Alima幼体(IRI%为14.24%)、太平洋磷虾(IRI%为11.26%)等。统计分析检验表明,蓝圆鲹的空胃率和平均胃饱满系数有显著的季节变化和叉长变化(P<0.01),摄食强度春季最高,秋季和夏季相对较高,而冬季最低;叉长50～99 mm和200～299mm的蓝圆鲹的摄食强度较高,其余叉长的强度相对较低。食物组成也随季节和发育阶段的不同而有所变化,在春季主要以七星底灯鱼和口虾蛄Alima幼体为食,夏季主要以甲壳类幼体为食,秋季主要以七星底灯鱼和太平洋磷虾为主食,冬季食物匮乏,小型桡足类丽隆剑水蚤和糠虾类占其食物组成的很大比例。聚类分析的结果表明,蓝圆鲹叉长200 mm是其摄食取向的拐点,小于这个叉长的蓝圆鲹以浮游甲壳类为主,大于这个叉长的以捕食小型鱼类为主,随着叉长的增大,蓝圆鲹更多的以游泳动物为食。根据蓝圆鲹摄食饵料的类群变化,东海蓝圆鲹的营养级为3.55,属于浮游动物和游泳动物混合食性的肉食性鱼类。     

Isolation and characterization of Edwardsiella ictaluri strains as pathogens from diseased yellow catfish Pelteobagrus fulvidraco (Richardson) cultured in China

Yellow catfish Pelteobagrus fulvidraco (Richardson) is a commercially important fish generally distributed in Southeast Asian countries. The well‐known aetiological agent of enteric septicaemia of catfish, Edwardsiella ictaluri, was isolated from diseased yellow catfish P. fulvidraco (Richardson) reared at two commercial fisheries in China. The economic losses due to the high mortalities (about 50%) caused by this bacterium have been increasing annually. The affected fish presented two different, typical symptoms: pale gills, slight exophthalmia and a ‘hole in the head’, and haemorrhage on the opercula, in the skin under the jaw, creating a ‘hole under the jaw’. These diseases were found frequently in cultured yellow catfish throughout China. The isolates from both outbreaks were all Gram negative, facultatively anaerobic and short rod. Morphological and biochemical tests and phylogenetic analysis based on the 16S rDNA sequences all strongly indicated that these yellow catfish isolates were highly identical to the known E. ictaluri. In addition, the isolates possessed the typical plasmid profile of E. ictaluri. Experimental infection assays were conducted and pathogenicity (by an intraperitoneal injection) was demonstrated in yellow catfish and channel catfish Ictalurus punctatus. The results showed that yellow catfish isolates were quite conservative phenotypically and genetically, and were able to cause two different, typical symptoms in this fish under unknown conditions and mechanism.     

泥鳅胚胎和幼鱼发育的研究

本文研究了用人工授精、孵化方法获得的泥鳅胚胎、幼鱼的形态特征。此项研究于1983年3—5月份在珠海市进行。卵圆球形，具弱粘性，卵径0．72-0．84毫米。在水温19．5—23℃情况下，多数受精卵经26小时40分后孵化出苗。初孵仔鱼全长1．95—2．4毫米，体节26-32对。在水温19—28℃情况下，孵化2天后，幼鱼具长条状外鳃。5天后，幼鱼卵黄囊大部分被吸收。15天后，外鳃消失，内鳃形成。45天后，幼鱼外部形态发育完全，形似成体。     

四种常见鱼卵对丝氨酸蛋白酶抑制活性的比较

选择胰蛋白酶、弹性蛋白酶、枯草杆菌蛋白酶、胰凝乳蛋白酶及其特异底物,建立比色法检测,对沙塘鳢、鲫鱼、草鱼、乌鳢鱼卵的丝氨酸蛋白酶抑制活性进行了比较。试验结果表明,胰蛋白酶抑制活性依次为乌鳢草鱼沙塘鳢鲫鱼;弹性蛋白酶抑制活性依次为沙塘鳢鲫鱼草鱼乌鳢;枯草杆菌蛋白酶抑制活性依次为沙塘鳢鲫鱼草鱼乌鳢,胰凝乳蛋白酶活性只受乌鳢抑制。研究表明这可能与抑制剂P1位点的残基种类有关。此外还对鱼卵中的抑制剂成分进行热稳定性与酸碱稳定性的检测,结果表明其具有很强的热稳定与酸碱稳定性。     

龙溪河张氏■生长与繁殖初步研究

基于2011年7-12月和2012年3-6月采集的样本,对长江上游支流龙溪河张氏■(Hemiculter tchangi)种群生长和繁殖特征进行研究。研究共采集张氏■样本227尾,采用鳞片鉴定年龄,测量鳞片半径,进行体长退算,选取成熟鱼卵描绘卵径分布,估算繁殖力,研究其年龄结构、生长特征和繁殖习性。结果显示:龙溪河张氏■体长101~187 mm,平均体长为(141±15) mm,体重12.5~82.8 g,平均体重为(39.3±13.2) g。群体雌雄性比为1.38∶1。群体年龄组成为1~5龄,其中3龄最多,占总样本的48.5%。群体体长与鳞径呈线性关系,雌雄无显著差异,其关系表达式为L=32.08 R+43.08;体长与体重的关系呈幂函数关系,其关系表达式为W=10-5×0.13.035。张氏■种群生长式型属于等速生长,生长方程表达式为:Lt=242.69[1-e-0.164(t+2.71)];Wt=170.66[1-e-0.164(t+2.71)]3.035。种群繁殖时间为4-8月,雌雄群体1龄达性成熟,其中雌性生物学最小型体长为110 mm,体重为28.7 g;雄性生物学最小型体长为105 mm,体重为14.6 g。龙溪河张氏■种群卵径为单峰型,平均卵径为(0.72±0.05) mm;种群绝对繁殖力平均为(32 688±11 426)粒;体长相对繁殖力FL平均值为(219±76)粒/mm;体重相对繁殖力FW平均值为(680±215)粒/g。     

网箱养殖大黄鱼诺卡氏菌病的初步研究

首次报道了大黄鱼诺卡氏菌病的发生情况。病鱼以体表和心、脾、肾等内脏出现白色结节为主要症状，平均死亡率15％。对病鱼进行细菌分离培养和光镜、电镜检查，均发现长或短的丝状分枝状杆菌。用分离菌株作回归感染，证实为该大黄鱼结节病的病原菌。对病原菌的基本生物学特性进行了研究，确定病原为诺卡氏菌。     

Weaning methods using formulated feeds for snakehead (Channa striata and Channa micropeltes) larvae

The culture of snakehead fish (Channa striata and Channa micropeltes) in Vietnam is limited, and snakehead culture has been banned in Cambodia, because traditional practices include capture of fingerlings from the wild as seed, as well as capture of small‐size (also known as trash fish or low‐value) fish. As hatchery breeding technology has improved, we investigated the optimal weaning practices for these two species. Both laboratory experiments and farm trials were conducted. For C. striata, the optimal weaning procedure is to begin at 17 days after hatch (dah) and wean the fish at 10% replacement of live feed with formulated feed per day. However, for C. micropeltes, the optimal procedure is to wait until 40 dah to begin weaning and then to wean the fish with a 10% replacement of live feed with formulated feed every 3 days. These results should enable farmers to domesticate snakehead culture in Vietnam and Cambodia and eliminate reliance on fish captured from the wild as both seed and feed.     

Diet of non‐native northern snakehead (Channa argus) compared to three co‐occurring predators in the lower Potomac River,USA

Abstract – Introductions of large, non‐native, carnivorous fishes continue to occur worldwide and represent a substantial management concern to global biodiversity. One of the most recent non‐native fishes to successfully establish in North America is the northern snakehead (Channa argus), found in the lower Potomac River catchment. Dispersal of the northern snakehead throughout this system has been well documented since its original discovery in May 2004; however, little is known about the foraging habits of this species and its interactions with co‐occurring predators. Here, we quantify northern snakehead diet in comparison with the diets of naturalised largemouth bass (Micropterus salmoides), and native American eel (Anguilla rostrata) and yellow perch (Perca flavescens) collected from tidal freshwaters bordering Virginia and Maryland near Fort Belvoir, Virginia. Over 97% of northern snakehead gut contents were fishes, with fundulid and centrarchid species consumed most frequently. Dietary overlap was biologically significant only between northern snakehead and largemouth bass. Aquatic invertebrates were >10 times more common in native predator diets, reducing dietary overlap with northern snakehead. Ontogenic shifts in adult northern snakehead diet were also detected, which may be explained by optimal foraging rather than true prey specificity. Northern snakehead may be occupying a novel niche based on a piscivorous diet, therefore limiting competition with resident predators in the lower Potomac River. Further research into interactions between largemouth bass and northern snakehead is needed to inform management decisions and understand the ecological impacts of this non‐native species.