鳗弧菌、溶藻胶弧菌外膜蛋白的分离及特性

分别用SDS、Sarkosyl及PMSF法分离制备鳗弧菌（Vibrio anguillarum）、溶藻胶弧菌（Vibrio alginolyticus）主要外膜蛋白（MOMP），通过SDS-PAGE分析比较2种弧菌主要外膜蛋白的组成结构。结果表明，SDS、Sarkosyl和PMSF法分离提取的鳗弧菌和溶藻胶弧菌外膜蛋白中，SDS-PAGE电泳图谱不同，鳗弧菌外膜蛋白分子量为27-138kD；溶藻胶弧菌外膜蛋白分子量为27-104kD，其中，45kD、30kD和27kD外膜蛋白为鳗弧菌和溶藻胶弧菌所共有。经比较，Sarkosyl法对2种弧菌外膜蛋白的提取效果较好。对3种方法提取的2种弧菌的主要外膜蛋白进行SDS PAGE后，分别与吸附后的兔抗鳗弧菌、抗溶藻胶弧菌血清的Western-blotting印迹显示，兔抗鳗弧菌血清与其菌株的外膜蛋白主要有3条免疫反应带，其分子量分别为97kD、51kD和30kD，说明其可能与鳗弧菌抗原的特异性有关；兔抗溶藻胶弧菌血清与其菌株的外膜蛋白主要有2条免疫反应带，其分子量分别为51kD和27kD，说明可能与溶藻胶弧菌抗原的特异性有关，而51kD外膜蛋白可能是2种弧菌共有的特异性抗原。     

温度对平鲷胚胎发育的影响

<正> 前言有关鱼类胚胎对于温度的敏感性,国外已有不少报道。国内学者在淡水鱼类方面作了许多研究,然而,对海水经济鱼类这方面的专门研究尚不多见。平鲷Rhabdosargus sarba(Forskal)     

平鲷消化系统形态学、组织学及组织化学研究

对平鲷(Rhabdosargus sarba)消化系统结构进行观察,采用HE和AB-PAS染色方法分别从组织学和组织化学方面进行研究.结果表明,平鲷口咽腔宽大,具不同类型发达齿;食道粗短,内表面分布纵行粘膜褶;胃分为贲门、胃旨囊和幽门,贲门胃小凹和胃腺最多,胃盲囊厚度最大;肠道具丰富粘膜褶;幽门与前肠交界处有幽门盲囊8～10个.肝小叶分界不明显,管状结构丰富.胰腺腺泡和胰岛分布于肝脏内大静脉周围,腺泡细胞染色深蓝色.根据AB-PAS染色结果将消化道粘液细胞分为4型,Ⅰ型为红色,Ⅱ型为蓝色,Ⅲ型为紫红色,Ⅳ型为蓝紫色.胃部町见极少量Ⅱ型及Ⅲ型粘液细胞,其余部位均以Ⅱ型粘液细胞为主.结果表明,粘液细胞消化系统不同部位的分布密度与对应部位消化功能相关.     

养殖牙鲆弧菌病病原菌初步研究

1998年 7月自荣成市寻山水产集团总公司养鱼场养殖牙鲆病死鱼体内分离到 1株细菌 ,经人工感染试验证明是牙鲆弧菌病病原菌。该菌主要特征为 :革兰氏阴性 ,弧状 ,以极生单鞭毛运动。氧化酶阳性 ,发酵葡萄糖产酸不产气 ,不产生 H2 S,不产生色素。生长温度范围 10～ 4 2℃ ,p H范围 6～10 ,盐度范围 0 .5～ 6。经 Biolog Microstation System鉴定为河川弧菌 - 。该菌株对环丙沙星、氟哌酸、利福平、呋喃唑酮、呋喃妥因、红霉素、链霉素等药物敏感。     

平鲷胚胎发育及仔,稚,幼鱼的形态观察

本文介绍以人工繁殖所获得的平鲷受精卵进行孵化,培育的结果。连续观察并描述该种鱼的胚胎发育过程及仔、稚、幼鱼的形态。     

平鲷的人工繁育技术

平鲷（Rhabdorsargus sarba）属于鲈形目、鲷科、平鲷属鱼类。广东俗称圆头[鱼立]、丝[鱼立],金丝[鱼立]等。     

养殖大黄鱼溶藻酸弧菌病病原菌传染途径的研究

从养殖大黄鱼病鱼的肝脏和脾脏内分离、纯化到溶藻酸弧菌，经回归感染试验证实它是该病的致病菌。使用法国生物一梅里埃公司生产的自动细菌鉴定仪(ATB)进行细菌鉴定，该菌株的生理生化特性符合溶藻酸弧菌的特性。研究结果表明，养殖大黄鱼溶藻酸弧菌病病原菌传染途径，主要是从水经伤口传染到体内，在试验条件下传染率为60％。病原菌在30℃时生长很快，20℃时生长慢，10℃时基本不生长，对氟苯尼考、四环素和乙酰螺旋霉素等7种药物敏感。     

若干环境因子及抗菌素对平鲷受精卵孵化率的影响研究

平鲷受精卵孵化时的最适水温为22℃,最适盐度为27.6‰,最适pH值为8.00。青霉素或氯霉素的浓度为1.0—1.5mg/时,可提高平鲷受精卵孵化率10％—20％。平鲷受精卵的孵化密度为10—20×104粒/m~3时孵化率较高。     

溶藻弧菌鞭毛蛋白flaC基因DNA疫苗对红笛鲷的免疫保护研究

为研究溶藻弧菌鞭毛蛋白flaC基因DNA疫苗对红笛鲷的免疫保护作用,实验构建了重组真核表达质粒pcDNA-flaC并将该质粒肌肉注射红笛鲷,采用PCR、RT-PCR、ELISA和攻毒试验等方法检测了该真核表达质粒在红笛鲷组织内的分布、表达和对红笛鲷的免疫保护.PCR结果显示,免疫接种7和28 d,注射点周围肌肉、鳃、肾脏、肝脏和脾脏都存在质粒分布;RT-PCR结果显示,免疫接种后第7天、14天和28天,红笛鲷不同组织内均有目的基因表达.ELISA结果表明,鱼血清内产生了抗FlaC蛋白的抗体,表明DNA疫苗免疫后鱼体表达了目的蛋白,并诱导产生了相应抗体.攻毒实验表明,免疫后的红笛鲷能较好地抵抗致病性溶藻弧菌的感染.结果表明,质粒pcDNA-flaC可能是抵抗溶藻弧菌感染的有效的疫苗候选物.     

中国对虾病原菌(鳗弧菌)的研究

1987年7～10月福建省平谭县对虾养殖场发生了一起严重的流行病。病虾活动力减弱,食欲下降,个体消瘦,头胸甲心区附近呈白色或浅桔红色,血淋巴液稀薄、混浊、不能凝固。因病虾步足、游泳肢及尾扇呈鲜红色,被称为“红腿病”。死亡率可高达90％以上。从五只垂死病虾的心脏血淋巴液中分离到细菌作纯焙养,经肌肉注射和浸泡的人工感染方法,得到了与虾池中患病虾相同的症状。菌原菌是革兰氏阴性短杆鼠。以极生单鞭毛运动。发酵葡萄糖,产酸不产气。氧化酶、过氧化氢酶阳性。能还原硝酸盐成亚硝酸盐。精氨酸脱羧;鸟氨酸、赖氨酸不脱羧。V.P.阳性.在42℃中不能生长。不能在无盐和含有8％NaCl 的胨水中生长,但在含有3％和6％NaCl的胨水中生长良好。对弧菌抑制剂0/129敏感。经鉴定为鳗弧菌(Vibrio anguillarum Bergman)。     

中国对虾糠虾幼体病原菌（非01群霍乱弧菌）的研究

本文对中国对虾糠虾幼体的一种病原菌———非０１群霍乱弧菌作了研究报道。这种病的症状是病虾运动能力差、趋光性弱、镜检发现肠道肿胀。从垂死病虾中分离到５株细菌，经感染健康糠虾幼体得到与病虾相同症状。５株细菌的４７项形态及生理生化特性与霍乱弧菌相同，但不被０１群霍乱弧菌多价血清凝集。血清学分型结果均为不同的ＶＢＯ血清型ＶＢＯ５，１４，２６，４７，５６。用ＤＮＡ霍乱ＣＴ基因探针菌落原位杂交及小鼠肠结扎法测肠毒素，表明菌株有强毒力。分离菌株对小鼠的ＬＤ５０为１５８×１０８个／只。     

香港河魨的毒性研究

产于我国的河魨鱼类（即科）大部分具有毒性［伍汉霖等１９７８］。１９０９年日本的田原良纯首先从河卵巢中提取了粗品毒素并命名为河毒素（ｔｅｔｒｏｄｏｔｏｘｉｎ）［Ｙｏｋｏｏ１９５０］。后来有更多学者在其它生物中发现河毒素,例如在美国加州蝾螈［Ｍｏ...     

对虾发光病病原菌的致病性及病理学观察

本文报道了对虾发光病病原菌的致病性、发病症状和病理变化。结果表明:哈维氏弧菌对虾类有极强的致病性,并引起组织和器官不同程度的病变。病理变化表现为:肌纤维颗粒变性,纵裂,坏死:肝胰腺空泡变性,坏死;淋巴器官坏死等。注射感染的发病虾,肝胰腺出现血细胞浸润,说明对虾对细菌感染有一定的防御能力。     

THE ARTIFICIAL PROPAGATION AND CULTURE OF BEAR SHRIMP,Penaeus semisulcatus de HAAN,IN HONG KONG

Penaeus semisulcatus (bear shrimp) was used for biological and culture studies. A peak of reproductive periodicity was shown in the months of May, June and July. Increase in body weight and in gonad weight coincided with the increase in sea water temperature. This suggested that sexual maturation might be a direct response to increased sea water temperature. Fecundity of mature female shrimp was estimated as 415,000 to 479,000 ova. In most cases, half-spent spawnings led to the production of poor eggs, characterized by irregular cytoplasmic formation and final autolysis. The embryonic and larval development of P. semisulcatus proceeded satisfactorily in a slightly alkaline medium (pH range from 7.5–8.5), and in water of salinity ranging from 28 to 35 ppt for egg and nauplius stages, and thereafter 25–35 ppt for zoea and mysis stages. The shrimps exhibited faster increase in body length than body weight in early growth. Later, after the shrimps had reached a body length of about 7.0 cm, the growth rate of body weight increased more markedly than body length. The rate of daily increase in weight was 1.01% in 7.0 cm shrimp. The feed efficiency of the formulated shrimp pellets was found to be 31.4%. During the nutritional study of P. semisulcatus, it was found that the combination of high dietary protein (about 40%) and low dietary lipid produced best growth and survival of bear shrimp. High increase of biomass of the shrimp fed with clam meat and high quality fish meal demonstrated the favorable response of shrimp to these diets. The use of cages for culturing bear shrimp was found to be practicable, but not efficient.     

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot,Scophthalmus maximus,and Flounder,Paralichthys olivaceus

Swollen abdomen disease (SAD) seriously threatens the aquaculture of turbots and flounders. Two dominant bacterial strains, FS1 and FS2, were isolated from the livers and kidneys of fish with diagnosed SAD. Applications of biochemical analyses, sequence analyses of 16S ribosomal RNA gene and heat shock protein 60 gene revealed two distinct pathogenic bacterial species, Edwardsiella tarda and Vibrio alginolyticus. These two hypothesized SAD‐causing pathogens were validated by challenge trials on flounder, Paralichthys olivaceus. Challenges with E. tarda and V. alginolyticus demonstrated lethal dose 50 (LD₅₀) values at 1.51 × 10⁵ colony‐forming units (CFU) and 1.05 × 10⁵ CFU, respectively. To develop a rapid SAD diagnosis method in flounders and turbots, a multiplex polymerase chain reaction (PCR) assay method was developed to simultaneously detect E. tarda and V. alginolyticus. Our multiplex PCR assay successfully detected as low as 10⁵ CFU/mL E. tarda and V. alginolyticus in flounders and turbots. No other common fish pathogens were detected with the multiplex PCR, suggesting a high specificity of this assay. The multiplex PCR assay developed in this study showed a great reliability in detecting SAD‐causing bacterial pathogens. Further optimization of this assay may contribute to the development of a novel SAD diagnosis tool in aquaculture.     

Putative apolipoprotein A‐I,natural killer cell enhancement factor and lysozyme g are involved in the early immune response of brown‐marbled grouper,Epinephelus fuscoguttatus,Forskal, to Vibrio alginolyticus

The gram‐negative bacterium, Vibrio alginolyticus, has frequently been identified as the pathogen responsible for the infectious disease called vibriosis. This disease is one of the major challenges facing brown‐marbled grouper aquaculture, causing fish farmers globally to suffer substantial economic losses. The objective of this study was to investigate the proteins involved in the immune response of brown‐marbled grouper fingerlings during their initial encounter with pathogenic organisms. To achieve this objective, a challenge experiment was performed, in which healthy brown‐marbled grouper fingerlings were divided into two groups. Fish in the treated group were subjected to intraperitoneal injection with an infectious dose of V. alginolyticus suspended in phosphate‐buffered saline (PBS), and those in the control group were injected with an equal volume of PBS. Blood samples were collected from a replicate number of fish from both groups at 4 h post‐challenge and analysed for immune response‐related serum proteins via two‐dimensional gel electrophoresis. The results showed that 14 protein spots were altered between the treated and control groups; these protein spots were further analysed to determine the identity of each protein via MALDI‐TOF/TOF. Among the altered proteins, three were clearly overexpressed in the treated group compared with the control; these were identified as putative apolipoprotein A‐I, natural killer cell enhancement factor and lysozyme g. Based on these results, these three highly expressed proteins participate in immune response‐related reactions during the initial exposure (4 h) of brown‐marbled grouper fingerling to V. alginolyticus infection.     

鳗鲡气单胞菌病一新种

鳗鲡气单胞菌病一新种ＡＮＥＷＳＰＥＣＩＥＳＯＦＡＥＲＯＭＯＮＡＳＡＳＰＡＴＨＯＧＥＮＩＮＥＥＬＳ王广和王坚（海安县卫生防疫站,江苏省２２６６００）钱晓明朱永祥王建明陆仁海（南通龙洋水产有限公司,江苏省２２６６３４）ＷＡＮＧＧｕａｎｇＨｅ,ＷＡＮＧＪ...     

养殖大黄鱼溃疡病的病原菌及其防治药物研究

从症状典型的网箱养殖患病大黄鱼体内分离到1株细菌824-1,经人工感染试验证实为是引起该病的病原菌。经鉴定,菌株824-1为溶藻弧菌(Vibrio alginolyticus)。利用杯碟法研究了其胞外产物的性质,结果表明:其胞外产物具有淀粉酶、明胶酶、酪蛋白酶、卵磷脂酶、脂酶和几丁质酶活性以及溶血活性,其中以淀粉酶、明胶酶和酪蛋白酶的活性最强,但没有脲酶活性。研究了20种化学药物和15种中草药对病原菌的抑菌作用,结果表明:复方新诺明、磺胺 TMP和庆大霉素等3种化学药物和石榴皮、地榆、五味子、大黄等4种中草药的抑菌能力最强,可作为防治该病的有效药物。     

中国对虾（Penaeus chinensis）养成期虾池水体和底质及虾体异养菌和弧菌含量的变化

本文报道了辽宁省瓦房店市邓屯乡高家养虾场对虾养成期虾池水体和底质及虾体异养菌和弧菌含量的变化。结果表明外海水中异养菌数量为１０￣４～１０￣５个／毫升，弧菌数量为１０￣２～１０￣３个／毫升，虾池水中异养菌数量为１０￣４～１０￣５个／毫升，弧菌数量为１０￣２～１０￣４个／毫升，底质中异养菌数量为１０￣５～１０￣７个／克，弧菌数量为１０￣３～１０￣４个／克。虾体肌肉中异养菌数量为１０￣３～１０￣５个／克，弧菌数量为１０￣３～１０￣５个／克；肝胰脏中异养菌数量为１０￣３～１０￣７个／克，弧菌数量为１０￣２～１０￣７个／克；肠道中异养菌数量为１０￣６～１０￣７个／克，弧菌数量为１０￣６～１０￣７个／克。