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1.
网箱养殖青石斑鱼的溃疡病病原   总被引:31,自引:3,他引:28  
覃映雪 《水产学报》2004,28(3):297-302
2002年夏季厦门同安湾一带气候炎热,降雨量少,海水盐度偏高,同安湾刘五店的网箱养殖石斑鱼大面积暴发溃疡病.本研究调查了石斑鱼溃疡病暴发的特征和症状,主要表现为肌肉溃疡坏死、眼球脱落、鱼骨暴露等.从病鱼体表及内脏分离出优势菌群命名为TS-628,经回归感染证实TS-628就是引发本次石斑鱼溃疡病的病原菌.对病原菌进行鉴定发现,该菌革兰氏染色呈阴性,电镜下观察菌体呈短杆状,极端单鞭毛,综合研究该菌在形态、生理生化、16S rDNA同源性及药物敏感性等方面的特性,基本确认分离到的病原菌为哈维氏弧菌(Vibrio harveyi),该菌对氯霉素、壮观霉素等多种抗生素敏感,对万古霉素、青霉素G等抗生素不敏感.哈维氏弧菌是水产养殖病害常见病原菌,但作为养殖石斑鱼的病原菌在国内属首次报道.  相似文献   

2.
为研究乌鳢诺卡氏菌的致病机理,本实验通过病原分离鉴定、组织病理学和基因表达水平分析对病原菌的致病性、药物敏感性及乌鳢的免疫抗性进行了研究。结果显示,患病乌鳢主要感染了命名为SDAT 0011病原菌,通过菌落形态观察、革兰氏染色鉴定、生理生化鉴定、16S rRNA鉴定及诺卡氏菌特异序列扩增鉴定,结果均显示该病原菌为诺卡氏菌。将分离的SDAT 0011感染健康乌鳢后,1×105~1×108 CFU/mL注射组的死亡率均为100%,感染乌鳢出现明显的诺卡氏菌病症状,如内脏出血,肝脏、脾脏和肾脏中有大量大小不等的结节,组织病理切片进一步检测发现,结节分界清晰,结节中有大量的淋巴细胞、受损或死亡的组织细胞。药物敏感性实验发现,诺卡氏菌对利福平等抗生素较为敏感,对青霉素等具有较强的抗性。基因表达分析显示,在感染初期(48 h)及中后期,Toll样受体2基因(TLR2)和Toll样受体13基因(TLR13)在脾脏和头肾的表达水平显著上调,而趋化因子受体9基因(CCR9)在脾脏和头肾中显著下调,这表明乌鳢Toll样受体和趋化因子受体信号通路可能在其抵抗诺卡氏菌感染中起重要作用。本实验为乌鳢诺卡氏菌病的...  相似文献   

3.
采用YEPD培养基诱导发酵生产重组抗菌肽。经 72h培养 ,由重组酵母表达并分泌到培养基中的抗菌肽活性可高达 5 0 0 0U/ml。用纸片扩散法和试管稀释法测定重组抗菌肽对水产养殖常见病原菌的抑菌效果。结果表明 ,抗菌肽对水产养殖常见病原菌嗜水气单胞菌 (Aeromonashydrophila)、梅氏弧菌 (Vibriometschnikovi)、迟钝爱德华氏菌 (Edwardsiellatarda)及温和气单胞菌 (A .sobria)有较好的抑菌效果 ,最小抑菌质量浓度为 4 μg/ml、杀菌质量浓度为 8μg/ml。对抗菌肽抑菌过程的超微结构观察表明 ,抗菌肽的抑菌方式是通过作用于细菌的细胞膜而导致菌体裂解死亡。本研究目的在于探讨基因工程抗菌肽在水产养殖中的应用。  相似文献   

4.
一株分离自浙江台州、舟山海水网箱养殖患结节病大黄鱼(Larimichthys crocea)的病原菌,经生理生化试验及形态结构观察显示,该菌株革兰氏阳性,好氧,具有弱抗酸性,菌体呈长或短杆状,或细长分枝状,过氧化氢酶阳性、氧化酶阴性,还原硝酸盐,不水解酪素、黄嘌呤、酪氨酸、淀粉和明胶,能以柠檬酸盐为唯一碳源生长.对其16S rRNA基因进行PCR扩增、测序和系统发育分析,结果表明,该菌株与诺卡氏菌属的菌株亲缘关系最近,与(鱼师)鱼诺卡氏菌(Nocardia seriolea JCM 3360^T)的16S rRNA基因序列相似性达99.9%.据此鉴定结果认为,该菌株为(鱼师)鱼诺卡氏菌(Nocardia seriolea).[中国水产科学,2006,13(3):410-414]  相似文献   

5.
耶尔森氏菌对水产养殖的危害及防治   总被引:1,自引:0,他引:1  
耶尔森氏菌是肠杆菌科、耶尔森氏菌属的总称,属兼性厌氧革兰氏阴性杆菌群。该属包含鼠疫耶尔森氏菌、假结核耶尔森氏菌、小肠结肠炎耶尔森氏菌等共11个种。其中鲁氏耶尔森氏菌是引起冷水性鲑鳟负类红嘴病的主要病原菌,是淡水养殖鱼类暴发性败血症的常见致病菌(气单胞菌、河弧菌、鲁氏耶尔森氏菌)之一。  相似文献   

6.
半滑舌鳎皮肤溃疡病病原研究   总被引:5,自引:0,他引:5  
为了对半滑舌鳎皮肤溃疡病的病原进行研究, 从患有严重皮肤溃疡病的半滑舌鳎病灶中分离病原菌, 并经人工感染确认病原。对引起此次疾病的病原菌的形态特征、理化特性、胞外产物酶活性与溶血活性及其对抗菌类药物的敏感性等生物学特性进行了研究和分析, 还测定了16 S rRNA、gyrB基因序列, 分析了相应序列的同源性并构建了系统发育树。结果从病灶中分离得到4株优势菌, 经人工注射感染证实菌株A3为引起养殖半滑舌鳎皮肤溃疡病的病原菌, 其半数致死量LD50为1.5×104.2 CFU/mL。其中, 16 S rRNA、gyrB在GenBank中的登录号分别是JN391271、JN168881。从基于16 S rRNA与gyrB基因序列构建的系统发育树看, 分离筛选出来的病原菌与哈维氏弧菌同源性最高, 结合生理生化特征和16 S rRNA、gyrB基因序列分析结果, 认为该病原菌为哈维氏弧菌。胞外产物酶活性及溶血活性检测结果表明,该病原菌具有淀粉酶、尿素酶、脂肪酶、蛋白酶和卵磷脂酶活性而不具有明胶酶活性, 在4%羊血TSA平板上呈β溶血活性。病原菌的药物敏感性试验结果显示, 该菌对四环素、强力霉素、诺氟沙星、磺胺异恶唑等敏感, 而对其他用于试验的抗生素敏感度低或具有一定的抗性。  相似文献   

7.
牛蛙爱德结氏菌病病原菌的鉴定和致病因素的研究   总被引:5,自引:0,他引:5       下载免费PDF全文
肖克宇  黄志坚 《水产学报》1997,21(3):316-321
从患爱德华氏菌病的牛蛙肌肉、肝、肾、血液和腹水中分离到8株细菌,根据其形态和生理生化特性鉴定为野生型迟钝爱德华氏菌。人工感染实验均为该病的病原菌,毒素检测试验表明,致病因素主要是内毒素而不是外毒素。分离菌 株的主要特性为杆状、革兰氏阴性、周生鞭毛、兼性厌氧。接触酶、甲基红试验和硝酸盐还原均为阳性。在三糖铁琼脂上产H2S。氧化酶、丙二酸盐利用、V.P试验、明胶液化、尿素酶。苯丙氨酸脱氨酶为阴性。分解  相似文献   

8.
网箱养殖大黄鱼诺卡氏菌病的初步研究   总被引:6,自引:1,他引:5  
王国良 《水产学报》2006,30(1):103-107
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。  相似文献   

9.
随着我国水产养殖业的发展,爱德华氏菌属细菌对水产养殖鱼类特别是鲇形目鱼类危害日益凸显。被认为是鱼类的一类重要病原。其中迟缓爱德华氏菌还是一种重要的人畜共患病的病原菌,对人类健康也造成威胁。因此,积极开展关于鱼类爱德华氏菌病的病原生物学、流行病学和诊断治疗技术研究具有重要意义。目前,国内外已对爱德华氏菌开展了大量研究,但其治病机理仍没有彻底理清,  相似文献   

10.
从大菱鲆(Scophthalmus maximus)肠道内分离出8株乳酸菌,经16S r DNA鉴定为Lactobacillus sp.,以对水产病原菌的抑制和对抗生素的敏感性为指标,研究乳酸菌的特性。其中,菌株N5对鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(V. harveyi)、副溶血弧菌(V. parahaemolyticus)和金黄色葡萄球菌(Staphylococcus aureus)均具有较强的抑制作用,对氨苄青霉素、庆大霉素和罗红霉素敏感。菌株N5发酵上清液经加热、排酸和过氧化氢酶处理后,仍有抑菌作用;经蛋白酶处理后失去抑菌能力,推测菌株N5分泌的抑菌物质为耐热的乳酸菌素,其可以抑制病原菌生物膜的形成。菌株N5在凡纳对虾(Litopenaeus vannamei)肠道定植5 d后,加入鳗弧菌和哈维氏弧菌,继续养殖7 d,死亡率显著降低。本研究结果可为菌株N5制备水产养殖益生菌剂提供实验依据。  相似文献   

11.
采用试管二倍稀释法测定了16种中草药单方及5种复方制剂对鲁氏耶尔森菌(Yersinia ruckeri)的最小抑菌浓度(MIC)。结果表明,黄芩(Radix Scutellariae)和诃子(Fructus Chebulae)对鲁氏耶尔森菌的抑菌作用最强,MIC均为0.028g·mL^-1,可作为防治水产养殖动物耶尔森菌病害的主要备选中草药;大黄(Radix et Rhizoma Rhei)的抑菌作用较强,其MIC值为0.227g·mL^-1,也可作为候选药物;青蒿(Herba Artemisiae)、金樱子(Fructus Rosae Laevigatae)、连翘(Fructus Forsythiae)、茵陈(Herba Artemisiae Scopariae)、五倍子(Galla Chinensis)及金银花(Flos Lonicerae)的抑菌作用较弱,其MIC值均为0.455g·mL^-1。5种复方制剂中,大黄、黄柏(Cortex Phel-lodendri)和黄芩3种中草药以5:3:2的比例配伍对鲁氏耶尔森菌具有最佳抑菌作用,其MIC值为0.057g·mL^-1,抑菌效果弱于黄芩,但远强于大黄单独使用,可为该细菌性疾病的防治提供复方配伍参考。  相似文献   

12.
采用通用引物PCR配合SSCP和RFLP技术检测鱼病病原菌   总被引:8,自引:0,他引:8  
彭宣宪 《水产学报》2000,24(4):345-348
采用通用引物PCR(UPPCR)、PCR-RFLP、PCR-SSCP技术,研究快速鉴别鱼病病原菌的分子生物学诊断技术。结果发现,采用细菌16S rRNA基因保守区特异性引物,以嗜水气单胞菌、鲁克氏耶尔森菌、鳗弧菌、柱状曲挠杆菌、乙型链球菌、荧光假单胞菌等部分常见鱼病病原菌为对象,可以建立一种UPPCR技术。该技术能在保证实验条件不变的基础上,检出上述所有细菌,并还可检出大肠杆菌和双歧杆菌等非鱼病病原菌。并且认为,该法与SSCP配合即采用UPPCR-SSCP技术能较好地鉴别被检菌而用于鱼病病原菌的快速诊断。  相似文献   

13.
Molecular cloning of Yersinia ruckeri aroA gene: a useful taxonomic tool   总被引:1,自引:0,他引:1  
The aroA gene of Yersinia ruckeri , which encodes 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase was cloned by complementation of the aroA mutation in Escherichia coli AB2829 by using pUC18 plasmid as a vector. Nucleotide sequence of the aroA gene revealed an open reading frame of 427 amino acids showing a high degree of homology to other bacterial AroA proteins. A pair of primers with 23 and 20 nucleotides were selected from the 5' and 3' termini, respectively, and formed the basis of a specific polymerase chain reaction (PCR) assay. A 1165-bp deoxyribonucleic acid (DNA) fragment was amplified from all lysed Y. ruckeri strains. An identical size fragment was also amplified from lysed Y. pseudotuberculosis , Y. aldovae , Salmonella enteritidis and E. coli , but not from other enterobacteria. Alu I restriction fragment length polymorphism (RFLP) of the PCR amplified products allowed for differentiation between Y. ruckeri and the other bacteria. Specificity and sensitivity make this PCR assay a useful method for rapid identification and diagnosis of Y. ruckeri infections.  相似文献   

14.
中国对虾杆状病毒垂直传播途径的初步探讨   总被引:9,自引:0,他引:9       下载免费PDF全文
汝少国 《水产学报》1998,22(1):49-55
中国对虾杆状病毒垂直传播途径的研究对切断病毒的传播途径,培育健康的对虾种质资源,具有重要的理论和实践意义。本文利用电镜技术,对中国对虾亲虾的卵巢,卵细胞和无节幼体,溲状幼体,糠虾,仔虾,幼成虾进行病毒检测,结果发现卵巢和卵细胞中有似病毒粒子,无节幼体,糠虾,仔虾,幼成虾有杆状病毒感染,并初步探讨了中国对虾杆状病毒垂直传播的可能性。  相似文献   

15.
Inhibition of bacterial fish pathogens by Tetraselmis suecica   总被引:3,自引:0,他引:3  
Abstract. Supernatants and extracts derived from a commercial heterotrophically grown spray-dried preparation of Tetraselmis suecica were observed to inhibit Aeromonas hydrophila, A. salmonicida, Lactobacillus sp., Serratia liquefaciens, Staphylococcus epidermidis, Vibrio anguillariim, V. salmonicida and Yersinia ruckeri type I, but not Planococcus sp., Pseudomonas fluorescens, Streptococcus sp., V. ordalii or Y. ruckeri type II by in vitro methods. When used as a food supplement, the algal cells inhibited laboratory-induced infections in Atlantic salmon. Moreover, there was a reduction in bacterial numbers in tank water. When used therapcutically, algal cells and their extracts reduced mortalities caused by A, salmonicida, Ser. liquefaciens, V. anguillariim, V. salmonicida and Y. ruckeri type I.  相似文献   

16.
Abstract. Seventeen strains of Yersinia ruckeri were examined for plasmid DNA by means of agarose gel electrophoresis of partially cleared lysates. All 10 serotype I strains examined contained a 70 Megadalton (Mdal) plasmid, whereas only one of the 7 serotype II strains examined contained a plasmid of about 2–3 Mdal. Yersinia ruckeri exhibited distinctive colonial morphology and serological reactivities which suggested a possible correlation with plasmid content. The luminol-enhanced Chemiluminescent (CL) responses of striped bass, Morone saxatilis (Walbaum), phagocytes to different strains of Y. ruckeri were also studied. Some variability was observed; however, the greatest phagocyte CL responses were elicited by serotype II strains without the plasmid while serotype I strains harbouring the plasmid consistently elicited weak CL responses from phagocytes.  相似文献   

17.
Abstract. Rainbow trout, Salmo gairdneri Richardson, (average weight 100g) were vaccinated intraperitoneally (i.p.) with Yersinia ruckeri bacterin in saline or in oily adjuvant. Agglutinating antibody kinetics were followed during 445 days before challenge (1.2×107 bacteria i.p.). Fourteen days after challenge 88.5% of the control fish had died while few mortalities were observed with vaccinated fish regardless of their agglutinating titre. Protection against Y. ruckeri does not seem to be due to antibodies.  相似文献   

18.
Bacteriocin‐producing bacteria in aquaculture may prevent spreading of potentially pathogenic microorganisms, and could be used as an alternative to the empirical use of antimicrobial drugs, especially for prophylaxis. Bacteriocinogenic bacteria inoculated as probiotics should not carry undesirable traits, such as antibiotic resistance. This study aimed to find potentially bacteriocinogenic bacteria in an aquaculture system and evaluate their antimicrobial susceptibility patterns. Selective cultures for enterobacteria, non‐fermenting Gram‐negative rods and Gram‐positive cocci were obtained from water samples before bacterial isolation and biochemical identification. Overall, 160 representative strains were recovered and for 57 antagonism was observed against selected strains such as Staphylococcus aureus, with antagonism being expressed better on Brain Heart Infusion medium. After exclusion of interfering factors, bacteriocin or bacteriocin‐like substances were suggested to be related to the antagonism observed. Higher drug‐resistant rates were observed among potentially bacteriocinogenic bacteria for different antimicrobials of clinical relevance. Although antibiotic resistance is a global health problem and bacteriocins are attractive alternatives to classical antibiotic even to multiresistant bacteria, the data obtained suggest that bacteriocin‐producing bacteria may harbour resistance genes available for transference in different environments. From the ecological and biotechnological perspective, antimicrobial susceptibility tests must be always performed when prospecting potentially bacteriocinogenic bacteria as probiotic candidates in the environment.  相似文献   

19.
闵洁  汪开毓  刘韬  贺扬  胡伟  罗梦笛 《水产学报》2017,41(12):1858-1866
鲁氏耶尔森菌是一种具有广泛致病性的条件致病肠杆菌。三型分泌系统(T3SS)是该菌的重要毒力系统,其中invF基因是T3SS功能表达的重要调控因子。为探讨invF和T3SS对Y.ruckeri致病作用的影响,本研究构建了Y.ruckeri SC09株invF基因的无痕缺失株,并对其生物学特性进行研究。通过融合PCR方法,将invF基因的上、下游片段A、C融合,构建同源臂AC;将获得的同源臂AC连接入自杀质粒pLP12,构建pLP12-invF同源重组载体;pLP12-invF电转化进入供体菌株大肠杆菌β2163,并利用接合转移方法转入受体菌株Y.ruckeri SC09,利用抗生素正向筛选和vmt反向筛选分别对插入突变株和缺失突变株进行筛选,并利用PCR技术和序列测定对Y.ruckeri invF缺失株进行鉴定;对突变株和野生株进行菌体菌落形态观察、生化特性鉴定和生长曲线测定。结果显示,融合PCR、AC片段经氯霉素抗性正向筛选和vmt反向筛选,PCR鉴定和测序鉴定后,成功获得了Y.ruckeri SC09 invF基因的无痕缺失突变株,突变株和野生株菌体菌落形态和生化特性基本一致,突变株菌落较野生株小,各个时期突变株的生长浓度较野生株低。研究表明,采用自杀质粒pLP12和大肠杆菌β2163接合转移系统,利用抗生素正向筛选和vmt反向筛选技术,在对其基本生物学特性无显著影响的情况下,可简捷高效地获得invF基因的无痕缺失突变株。  相似文献   

20.
The aroA gene of Yersinia ruckeri, which encodes 5-enolpyruvylshikimate 3-phosphate synthase, was insertionally inactivated with a DNA fragment containing a kanamycin resistance determinant and reintroduced by allelic exchange into the chromosome of Y. ruckeri 21102 O1 by means of the suicide vector pIVET8. The Y. ruckeri aroA::Kan(r) mutant was highly attenuated when inoculated intraperitoneally into rainbow trout, with a 50% lethal dose of >5 x 10(7) CFU. The mutants were not recoverable from the internal organs 48 h post-inoculation or later. The vaccination of rainbow trout with the AroA mutant as a live vaccine conferred significant protection (relative percentage survival = 90%) against the pathogenic wild-type strain of Y. ruckeri.  相似文献   

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