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1.
Biolog GN法对不同地区养殖对虾弧菌区系的比较研究   总被引:2,自引:2,他引:0  
采用Biolog细菌鉴定技术,分析来自5个国家的4个对虾养殖品种苗期及部分养成期虾体上的185株弧菌(其中24株来自成虾).结果表明:来源、种类不同的养殖对虾苗期的主要弧菌的区系组成相似,溶藻弧菌(Vibrio alginolyticus)和鲨鱼弧菌(V. carchariae)(即哈维氏弧菌V. harveyi)是普遍存在的种类,同一种对虾在不同地区养殖,其区系组成略有差异;哈维氏弧菌多为对虾苗期致病菌,副溶血弧菌(V.parahaemolyticus)主要为成虾致病菌;在健康虾苗和发病虾苗体内都可分离到溶藻弧菌.  相似文献   

2.
张红见 《畜禽业》2010,(4):40-42
应用常规细菌分离法对西宁市某几个农贸市场采集的70份仔虾样品进行了副溶血性弧菌的检测,经细菌形态观察、嗜盐性试验、细胞色素氧化酶试验和生化特性测定,鉴定为副溶血性弧菌。结果检出阳性菌株4株,阳性率为5.7%(4/70),同时通过致病性试验证明4株分离菌对小白鼠有致死效应。  相似文献   

3.
中国对虾病原菌(鳗弧菌)的研究   总被引:30,自引:3,他引:30  
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)。  相似文献   

4.
高杨 《河北渔业》2019,(2):18-20
由丹东东港市某养殖场患病的南美白对虾(Penaeus vannamei)幼虾组织体内分离出一株优势菌,通过全自动细菌分析仪及16srDNA序列分析对该菌株进行鉴定,命名为DS180707。并进行人工感染试验和药敏实验。结果显示,本次患病幼虾致病菌为溶藻弧菌,且具有较强的致病性。该菌对氟苯尼考、氯霉素、恩诺杀星、诺氟沙星、氧氟沙星、左氧氟沙星等6种抗生素敏感。  相似文献   

5.
斜带石斑鱼3种致病性弧菌的分子生物学鉴定   总被引:17,自引:3,他引:14  
李宁求 《水产学报》2005,29(3):356-361
从患病斜带石斑鱼分离到3株病原菌EcGS020802、EcGS021001、EcGS020801,经常规生理生化鉴定均属于弧菌属的种类。为了进一步确定其分类地位,测定了3株病原菌的16SrRNA和HSP60(heat shock protein,HsP60)基因部分序列。16SrRNA基因系统进化分析表明,3株病原菌与哈维氏弧菌、溶藻弧菌、副溶血弧菌亲缘关系较近,相互之间同源性均大于98.6%,差异不明显。HSP60基因序列分析表明,菌株EcGS020802、EcGS021001、EcGS020801 HSP60基因序列分别与哈维氏弧菌(AF230934)、溶藻弧菌(Ab230931)、副溶血弧菌(AF230951)HSP60基因的同源性最高,依次为95.7%、99.8%和99.8%,而与其它弧菌HSP60基因的同源性均低于90.6%,3株病原菌相互之间的同源性低于91.0%。HSP60基因构建的系统进化树表明,EcGS020802、EcGS021001、EcGS020801分别与Vibrio harveyi、Vibrio alginolyticus、Vibrio parahaemolyticus聚类。综合上述结果,菌株EcGS020802、EcGS021001、EcGS020801可分别鉴定为哈维氏弧菌、溶藻弧菌、副溶血弧菌。结果表明,HSP60基因比16SrRNA基因更适合用于海水鱼致病性弧菌种问的分类研究。  相似文献   

6.
采用K-B纸片扩散法和微量稀释法对从天津地区养殖的南美白对虾(Penaeus vannamei)中分离的弧菌进行耐药性调查,结果显示2020年在养殖季节从天津市患病的南美白对虾中共分离出弧菌33株,其中副溶血弧菌15株、溶藻弧菌8株、创伤弧菌7株、霍乱弧菌3株。在天津市南美白对虾以副溶血弧菌污染最为严重,约占总数的45%左右。药敏结果显示分离的33株弧菌对2种及2种以上药物耐药的有27株,约占总数的81.8%,说明天津地区弧菌已经出现了多重耐药的现象;通过对33株弧菌对药物的MIC值测定,33株弧菌对恩诺沙星、氟甲喹和盐酸多西环素比较敏感,对磺胺类药物比较耐药。结果提示应加强对天津地区南美白对虾弧菌病主要病原菌耐药性监控。  相似文献   

7.
从对虾养殖池中分离出1株编号为2013082515(简称菌株15)的菌株,以鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(Vibrio harveyi)和副溶血弧菌(Vibrio parahaemolyticus)为指示菌,分析了菌株15对指示菌的抑菌效果及最低抑菌浓度,同时,分析了菌株15对其他7株弧菌的抑菌效果.结果显示,菌株15对3株指示菌均具有抑菌效果,对鳗弧菌、哈维氏弧菌及副溶血弧菌的最低抑菌浓度分别为2.50×104、2.50×105、2.50×105 CFU/ml;对其他弧菌也具有一定的抑菌效果.采用注射及浸泡感染方法分析了菌株15对对虾的生物安全性,结果显示,在高浓度时,菌株15对对虾具有潜在毒性.分别用细菌全细胞脂肪酸气相色谱法和16S rDNA序列分析比对法对该菌进行分类鉴定,表明菌株15为一株假交替单胞菌(Pseudoalteromonas sp.).  相似文献   

8.
副溶血弧菌对南美白对虾致病力及黄酮抗菌作用初步研究   总被引:1,自引:0,他引:1  
研究副溶血弧菌3种不同的感染方式对南美白对虾致病力影响,发现注射感染虾只在高密度菌液中有死亡,24 h的LD50为1.2×107/mL,95%的置信限为5.61×106~2.04×107/mL;而创伤感染虾在不同密度菌液中都有死亡,浸泡感染虾则都没有死亡。表明创伤程度及其导致的健康状况恶化是弧菌致病的关键。同时发现肌肉创伤处是弧菌致病的主要部位。初步探讨了黄酮作为饲料添加剂对副溶血弧菌的抗菌作用,为养虾生产的防病治病提供新的途径。  相似文献   

9.
虾夷扇贝脓胞病病原的分离、鉴定与致病性   总被引:3,自引:1,他引:2  
开展对大连市长海县附近养殖海域近几年夏季出现的大规模虾夷扇贝死亡现象的研究,能为虾夷扇贝养殖过程中的疾病防治提供基础理论支持。以患病虾夷扇贝为研究对象,通过对病变组织进行细菌分离、提纯培养,并采用人工感染试验确定虾夷扇贝脓胞病的病原性质;通过生理生化特性、16SrRNA基因部分序列和药物敏感性测定,对虾夷扇贝脓胞病进行了鉴定。结果表明,从患病组织中分离出的其中一株细菌经人工感染试验证实为虾夷扇贝脓胞病的病原菌,在水温19℃、注射浓度为1.09×105CFU/mL的条件下,该菌有较强的致病性。通过对该病原菌的生理生化特性测定和16SrRNA基因部分序列分析,确定虾夷扇贝脓胞病的病原为查氏弧菌。  相似文献   

10.
健康和患病凡纳滨对虾幼虾消化道菌群结构的比较   总被引:2,自引:1,他引:1  
凡纳滨对虾养殖过程中,早期阶段是病害易感阶段,而消化道菌群结构与对虾健康关系密切。因此,探讨幼虾的消化道菌群尤其是弧菌类细菌与对虾发病的关系对病害有效防治具有重要意义。本实验采用Illumina测序研究了凡纳滨对虾健康和患病幼虾的消化道细菌群落结构,并基于纯培养和16S r DNA,rec A和pyr H基因序列比对,分析了幼虾消化道中弧菌的主要种类组成。结果发现,健康幼虾消化道中α-变形菌纲和厚壁菌门丰度较高,而患病幼虾中γ-变形菌纲、拟杆菌门、放线菌门和β-变形菌纲较高,其中放线菌门丰度差异显著。基于科水平的响应比分析,发现患病幼虾消化道中动性球菌科和噬菌弧菌科的丰度显著降低,而弧菌科的丰度显著升高。相似度分析发现,驱动群落变异的OTU主要来源于弧菌属、海洋杆状菌属、冷杆菌属、假交替单胞菌属以及未分类至属的红杆菌科和微杆菌科。健康幼虾消化道弧菌组成以锡那罗亚州弧菌为主,而患病幼虾消化道弧菌组成以坎氏弧菌为主。尽管健康和患病幼虾消化道内细菌群落结构整体差异不显著,但一些重要类群丰度变化显著,其特征与已知的生态功能一致。  相似文献   

11.
In this study, the phenotypic and pathogenic properties in freshwater host were characterized in 14 strains of halophilic Vibrio harveyi isolated from infected marine black tiger shrimp, Penaeus monodon. The lysogenic phenotype was assayed via prophage excision and mitomycin C induction. The bacteria were grouped into two types, corresponding to lysogenic and non‐lysogenic strains. The pathogenicity was determined via direct injection of bacterial cultures into post‐larval juvenile giant freshwater prawns, Macrobrachium rosenbergii De Man. All of the infected prawns showed similar symptoms and inflamed hepatopancreas. The V. harveyi isolates derived from the first‐injected infected prawns were re‐isolated and re‐injected into healthy giant freshwater prawns, in which they retained similar infectivity. Both lysogenic and non‐lysogenic Vibrio spp. showed identical virulence associated with 100% mortality within one day post‐injection. TEM micrographs showed hepatopancreatic nuclear deformation and lipid breakdown caused by lysogenic γ‐hemolytic VL19 and non‐lysogenic β‐hemolytic V33. However, the V33 strain was associated with severely disrupted mitochondria. None of the V. harveyi strains was able to produce a biofilm. Together, our findings indicate that the lysogenic and non‐lysogenic halophilic V. harveyi isolated from marine shrimps may use different virulence factors that are responsible for their pathogenicity in freshwater prawns.  相似文献   

12.
郝婧薇  华昕彤  傅松哲  周灿  刘鹰  胡惠秩 《水产学报》2023,47(3):039416-039416
为了阐明引起急性肝胰腺坏死病(acute hepatopancreatic necrosis disease, AHPND)副溶血性弧菌的接合型质粒在对虾养殖环境中的遗传多样性,实验从中国5个沿海省份的虾场收集了100个底泥样品,以质粒上编码接合转移蛋白的保守基因为目标,利用PCR法检测相关质粒的存在情况,并对质粒进行测序。结果显示,100个样品中有39个样品含有质粒的接合转移蛋白片段。从100个底泥样品中分离出15株副溶血性弧菌,其中13株含有1~2个质粒。质粒序列测序结果显示,这些质粒可分为8种类型/谱型,其中7种不携带pirAB,但均含有编码接合转移的基因簇。根据分离副溶血性弧菌携带质粒的8种谱型,分别选择8株副溶血性弧菌进行凡纳滨对虾攻毒实验,发现这些菌株对凡纳滨对虾的毒性有显著差异,实验虾死亡率为15%~100%。只有pirAB阳性菌株会对实验虾产生AHPND症状,死亡率为100%。对质粒组成进行分析表明,质粒之间遗传物质交换频繁,大部分质粒的遗传组成都来自一个183 kb的超大质粒pVP2HP。综上,本实验通过探究对虾养殖场底泥中结合性质粒的多样性,增强了人们对副溶血性弧菌...  相似文献   

13.
致病性哈维氏弧菌溶血素基因克隆及其检测   总被引:1,自引:1,他引:1       下载免费PDF全文
从山东沿海的发病鲈鱼(Lateolabrax japonicus)分离到1株致病性哈维氏弧菌(Vibrio harveyi),从该菌的染色体DNA扩增出一条长约1.4kb的特异性片段。DNA序列分析表明,该克隆片段含有完整的1254bp溶血素基因,该溶血素基因与哈维氏弧菌VIB645的溶血素基因VhhA和VhhB的相似性分别为99.0%和98.5%;与副溶血弧菌(V.parahaemolyticus)热稳定性溶血素基因(TDH)的相似性为74.5%;与拟态弧菌(V.mimicus)、创伤弧菌(V.vulnificus)、霍乱弧菌(V.chderae)磷脂酶基因的序列相似性分别为57.4%、59.2%、53.0%;与最小弧菌、创伤弧菌、霍乱弧菌、霍氏弧菌(V.hollisae)、河流弧菌(V.tguvialis)、鳗弧菌(V.anguillarum)的溶血素基因的相似性仅为19.9%~24.8%。根据溶血素基因的保守区段,设计了1对特异引物。分析表明,该引物能特异检测哈维氏弧菌。其可检测的DNA最小量为0.001ng。用该方法对55株不同来源的患病鱼分离的疑似病原菌进行检测,结果检出8株哈维氏弧菌,检出率为14.55%,从健康动物中检出数占所检弧菌数的6.25%,海洋环境为10.53%,海水养殖环境为26.53%,表明哈维氏弧菌在不同的海水环境及健康海洋动物中普遍存在,在养殖水体中的数量高于其他海水环境。  相似文献   

14.
为了解流水式养殖场养殖动物中弧菌和气单胞菌的感染状况,分别针对霍乱孤菌肠毒素A亚单位(ctxA)基因、0139和01群群特异基因、毒力协同调节菌毛A亚单位基因(tcpA)、中心调节蛋白基因(toxR)、副溶血弧菌不耐热溶血素基因(tlh)、创伤弧菌Vibrio vulnificus溶细胞素基因(vvhA)和嗜水气单胞菌的气溶素基因(aerA)设计引物,建立了聚合酶链式反应(PCR)检测技术,PCR产物经电泳,根据扩增条带的大小和数目,检测和区分霍乱孤菌、嗜水气单胞菌和副溶血孤菌.对南昌沿岸霍乱流行水域及附近养殖场的养殖动物进行了18个月的连续监测,对1 440份水产品的增菌液进行PCR检测,17份检出副溶血弧菌,25份检出霍乱弧菌,创伤弧菌和嗜水气单胞菌均未检出.扩增结果与传统培养检测法的结果一致.监测结果表明,南昌附近地区霍乱弧菌和副溶血弧菌检出率分别为1.74%和1.18%,总体感染状况处于较低水平,但个别养殖品种弧菌检出率较高.  相似文献   

15.
Juvenile western king prawn P. latisulcatus were fed 105 colony-forming units (CFU)/mL of two probiotics Pseudomonas synxantha and P. aeruginosa for 28 days. P. latisulcatus were then challenged with V. harveyi at 0 (control), 103, 105, and 107 CFU/mL. During the seven days of challenge, disease resistance of the probiotic-fed prawns was compared with that of prawns not fed probiotics. The immunological responses of the prawns did not improve during the challenge period in terms of total haemocyte count, hyalinocyte, semi-ganulocyte, granulocyte, clotting time, bacteraemia, and intestinal bacterial load. Overall, when prawns were challenged with V. harveyi, the LT50 values got shorter as V. harveyi concentration increased. LT50 values of prawns fed probiotics were significantly longer (P < 0.05) than those not fed probiotics. At a V. harveyi concentration of 103 CFU/mL, the 100% survival of the prawns fed probiotics was three times more likely than those of the prawns not fed probiotics.  相似文献   

16.
采用试管二倍稀释法测定二氟沙星对鳗弧菌W-1、副溶血弧菌1614和溶藻弧菌1833的最小抑菌浓度(MIC);采用菌落计数法测定二氟沙星对鳗弧菌、副溶血弧菌和溶藻弧菌的抗菌后效应(PAE)。结果显示,二氟沙星在1×MIC、2×MIC和4×MIC浓度时,对鳗弧菌W-1、副溶血弧菌1614和溶藻弧菌1833的PAE分别为:0·64、1·09和2·16h;0·74、1·73和2·64;0·54、1·08和2·05h。二氟沙星对这3种弧菌具有明显的抗菌后效应,并且随着二氟沙星浓度的增加,抗菌后效应的时间也明显延长。  相似文献   

17.
2020年6—10月,从辽宁地区患病大菱鲆(Scophthalmus maximus)体内共分离到394株菌株.基于16S rRNA序列鉴定分离株,采用微量稀释法分析随机挑选的18株大菱鲆弧菌(Vibrio scophthalmi)对8种抗生素的敏感性,检测其耐药基因携带情况,并基于ERIC-PCR进行分型研究.结果显...  相似文献   

18.
Immune cells were identified and their interaction towards Vibrio alginolyticus, V. parahaemolyticus and V. anguillarum was studied in vitro in the penaeid shrimp, Penaeus indicus. Haemocytes were divided into agranulocytes, semi-dense granulocytes and dense granulocytes according to their morphology. Agranulocytes (100%) and 0.3–0.7% of granulocytes were actively involved in coagulation. Granulocytes were involved in in vitro phagocytosis and encapsulation of foreign materials. Phagocytosis was enhanced by prior opsonization of bacteria with cell-free shrimp haemolymph. Semi-dense granulocytes were phagocytic towards V. alginolyticus with and without opsonization at the rate of 91.1% and 83.1%, respectively ( P < 0.05 ). Granulocyte death observed after 2 h with opsonized haemolymph was 26.1%. About 64.5% of dense granulocytes and 23.2% of semi-dense granulocytes were actively involved in encapsulation, forming capsules. A spectrophotometric nitroblue tetrazolium (NBT) reduction assay was used to demonstrate the production of superoxide anions (O2) by shrimp haemocytes. All the Vibrio spp. were able to induce superoxide anions (O2) during phagocytosis. Live Vibrio sp. induced O2 production in haemocytes in a dose-dependent manner. Significant activity was detected with a 40:1 bacteria to haemocyte ratio ( P < 0.05 ). NBT reduction assay for measuring the post-phagocytic killing mechanism in shrimp haemocytes might be a valuable tool for monitoring shrimp health and immunological studies.  相似文献   

19.
ABSTRACT

Fishing is among the main economic activities of the people of Zanzibar. Few fish dealers are transforming this sector into mariculture. Among the farmed fish is milkfish. Diseases are among the limiting factors in the development of the mariculture industry. Among other zoonotic diseases, vibriosis is caused by bacteria from the genus Vibrio. This study aimed to establish the occurrence of Vibrio cholerae and Vibrio parahaemolyticus among milkfish farms in Zanzibar. A total of 380 milkfish were sampled. Swabs were collected from gills, intestine, and kidney of each sampled milkfish. Preliminary identification of V. cholerae and V. parahaemolyticus was done by biochemical tests. PCR was run on 16S rRNA, outer membrane protein W, and collagenase genes to confirm Vibrio species, V. cholerae, and V. parahaemolyticus respectively. Almost one-third (32.1%) of all sampled milkfish were found to contain targeted Vibrio; 18% and 29.5% of the sampled milkfish were positive for V. cholerae and V. parahaemolyticus respectively.  相似文献   

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