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1.
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
2.
Antibacterial abilities of intestinal bacteria from larval and juvenile Japanese flounder against fish pathogens 总被引：5，自引：0，他引：5
HARUO SUGITA RYUJI OKANO YUKIKO SUZUKI DAISAKU IWAI MASANOBU MIZUKAMI NOBUHIKO AKIYAMA SEIJU MATSUURA 《Fisheries Science》2002,68(5):1004-1011
The present study was undertaken to examine the antibacterial abilities of intestinal bacteria isolated from juveniles and larvae of Japanese flounder Paralichthys olivaceus . Newly hatched larvae of flounder were held in a 100 L plastic circular tank and fed rotifiers, Artemia nauplii and commercial feeds, depending on the developmental stage of the fish. Genera Aeromonas , Moraxella and Vibrio were predominantly isolated from the intestinal tracts of Japanese flounder at larval and juvenile stages, whereas Aeromonas , Bacillus , coryneforms, Moraxella , Pseudomonas and Vibrio were detected at high densities in live diets and artificial feeds. Antibacterial bacteria accounted for 1.7–24.3% of the intestinal isolates against Lactococcus garvieae , Pasteurella piscicida , Vibrio anguillarum and V. vulnificus . In particular, as much as 53.3% of Vibrio spp. other than Vibrio -swarmer isolated from 197-day-old juveniles inhibited the growth of P. piscicida . These results suggest that intestinal bacteria having antibacterial activity play a role in the prevention of infectious diseases. 相似文献
3.
Characterization of Vibrio spp. Associated with Diseased Shrimp from Culture Ponds of Andhra Pradesh (India) 总被引：1，自引：0，他引：1
Abstract.— Surveys undertaken on diseases caused by Vibrio spp. in Penaeus monodon from culture ponds of coastal Andhra Pradesh recorded the occurrence of five types of diseases: tail necrosis, shell disease, red disease, loose shell syndrome (LSS), and white gut disease (WGD). Among these, LSS, WGD, and red disease caused mass mortalities in shrimp culture ponds. Six species of Vibrio—V. harveyi , V. parahaemolyticus, V. alginolyticus, V. anguillarum , V. vulnificus , and V. splendidus —are associated with the diseased shrimp. The number of Vibrio spp. associated with each disease ranged from two to five. Additionally, shrimp with red disease had concurrent infections with white spot syndrome virus. Vibrio harveyi in the case of LSS and WGD, V. parahaemolyticus for red disease, and V. alginolyticus for shell disease are the major etiologcal agents. Differences occur in the degree of virulence of different species of Vibrio and also different isolates of the same species. Vibrio harveyi isolated from LSS shrimp is the most virulent. In general, all the Vibrio isolates from LSS shrimp tend to be more virulent as compared to their counterparts from other diseased shrimp. It is apparent that the degree of virulence of various Vibrio isolates depends on its source and the pond environmental conditions. Most of the Vibrio isolates showed susceptibility to oxytetracycline, norfloxacin, and ciprofloxacin. The luminous V. harveyi exhibited resistance to many antibiotics and susceptibility to only three drugs. Considering the emergence of antimicrobial resistant strains of Vibrio , the need for using probiotics in place of antibiotics for disease control is stressed. 相似文献
4.
斜带石斑鱼3种致病性弧菌的分子生物学鉴定 总被引：14，自引：3，他引：14
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
5.
溶藻弧菌相关分离株的分子及VITEK鉴定 总被引：1，自引：0，他引：1
哈维群弧菌是弧菌属的核心菌群,包括溶藻弧菌在内的6个种在表型和遗传型上均十分相似,要准确鉴定各种有一定难度。看家基因的研究及生化鉴定系统的出现为弧菌鉴定提供了多种方法,本文比较了16S rRNA基因、toxR基因和pyrH基因以及VITEK 2 COMPACT GN鉴定卡对溶藻弧菌相关分离株的分辨力。哈维群弧菌基因组内16S rRNA基因是多拷贝的,且拷贝间序列差异大于种间差异,不适用于种的鉴定。单拷贝基因toxR和pyrH序列种内差异均小于种间差异。基于toxR基因相似性比较可清楚地将18个疑似溶藻弧菌分离株和5个参比株归并到4个种;toxR基因系统发育学分析也显示,哈维群弧菌各种独立地聚类分支,且溶藻弧菌种内存在两个明显的聚类分支,说明两个分支的溶藻弧菌具有独立的进化方向。pyrH基因的相似性比较和发育学分析也得到类似的结果,但pyrH基因具有较强的保守性,因而分辨力稍低于toxR基因。VITEK 2 COMPACT GN鉴定卡在鉴定溶藻弧菌时也有一定的错误率,且鉴定谱较窄。因此,建议在实际应用中采用toxR基因比对作为溶藻弧菌快速鉴定的主要手段,可再选用pyrH基因对鉴定结果进行验证。 相似文献
6.
Bacteria associated with early life stages of halibut, Hippoglossus hippoglossus L., inhibit growth of a pathogenic Vibrio sp. 总被引：2，自引：0，他引：2
Ø. BERGH 《Journal of fish diseases》1995,18(1):31-40
Abstract Bacteria capable of inhibiting growth of a pathogenic Vibrio sp. were isolated from the gastrointestinal tract of cultured halibut larvae during the first feeding and weaning stages. No such bacteria were found among isolates from the surface of eggs or the gastrointestinal tract of yolk sac larvae. The fraction of pathogen-inhibitors among the total number of isolates ranged between 0–100% (first feeding) and 0–66% (weaning). All pathogen-inhibitors were Gram-negative rods, and 95% were oxidase and catalase positive fermentative isolates, capable of producing acid aerobically from a varying range of carbohydrates. These isolates possessed the characteristics of the Vibrio/Aeromonas -group, but only 19% were sensitive to the vibriostatic agent 0/129. Isolates from eggs and yolk sac larvae were dominated by bacteria belonging to the Cytophaga/Flexibacter/Flavobacterium - group. The high fraction of isolates with the ability to inhibit growth of the pathogenic Vibrio sp. among the total number of isolates indicates that pathogen inhibition may be an important mutualistic role of the intestinal flora of early life stages of halibut. 相似文献
7.
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１５８×１０８个／只。 相似文献
8.
In situ adherence of Vibrio spp. to cryosections of Atlantic salmon, Salmo salar L., tissue. 总被引：1，自引：1，他引：0
Pathogenic and presumed non-pathogenic bacteria isolated from fish were tested for their adhesion to cryosections from different mucosal surfaces of Atlantic salmon, Salmo salar L. Adhered bacteria were detected by immunohistochemistry. Mucus was stained and fixed with Alcian blue after incubation of bacteria. The majority of the bacteria tested, i.e. Vibrio anguillarum serotype O1 , Vibrio salmonicida , Vibrio viscosus, Flexibacter maritimus and 'gut vibrios', i.e. Vibrio iliopiscarius and intestinal isolates of V. salmonicida , all adhered to mucus on all salmon epithelial surfaces tested, including sections from the foregut, hindgut, pyloric caeca, gills and skin. In contrast, V. anguillarum serotype O2, including both serotypes O2a and O2b, did not adhere to mucus, but did adhere to all other components of the tissues. As a positive control for adhesion of bacteria on cryosections, Escherichia coli was bound to piglet ileal mucosal lining, and as a negative control for adhesion, Staphylococcus aureus was found not to bind to any of the tissues tested. The present study shows that adhesion to mucus was not restricted to pathogenic bacteria, and furthermore, that not all pathogenic bacteria studied adhered to mucus. Hence, on the basis of these findings, the present authors suggest that V. anguillarum O2 may have an invasion strategy which does not involve adhesion to mucus, and thus, differs from the other pathogenic bacteria in the present study, which all bound to salmon mucus. 相似文献
9.
This study examined the diversity of siderophore‐producing bacteria in the intestinal tracts of coastal fish in Japan and screened candidate bacterial strains for probiotic use in aquaculture. Of the 2637 bacteria isolated from the 27 fish specimens (13 species) and six environmental samples collected in this study, 266 isolates exhibited the ability to produce siderophores. Siderophore producers were detected in the intestines of 18 of the fish specimens caught (68%) at densities of 2.3 × 104–2.3 × 108 CFU g?1, in all three seawater samples at 2.0 × 102–1.3 × 103 CFU mL?1 and in all three sand samples at 2.6 × 101–2.8 × 104 CFU g?1. These findings suggest that siderophore‐producing bacteria are widely distributed in the intestinal tracts of coastal fish and their environments. Analysis of 16S rRNA gene sequences revealed that the siderophore producers belonged to 38 species, of which Vibrio splendidus, Vibrio ichthyoenteri and Vibrio crassostreae accounted for 32.7%, 19.5% and 11.3% of the 266 isolates, respectively, suggesting that these bacteria are indigenous to the intestinal tract of coastal fish. Six bacterial species, Enterovibri norvegicus, Photobacterium leiognathi, Photobacterium phosphoreum, Photobacterium rosenbergii, V. crassostrea and Vibrio scophthalmi were identified as possible candidates for use as probionts in fish aquaculture. 相似文献
10.
Evaluation of the AQUARAPID-Va, AQUAEIA-Va and dot-blot assays for the detection of Vibrio anguillarum in fish tissues 总被引：1，自引：0，他引：1
The comparative accuracy of the serological assays AQUARAPID-Va, AQUAEIA-Va (BIONOR AS), and dot-blot for a rapid diagnosis of vibriosis in fish was evaluated. Twenty-one Vibrio anguillarum strains, representative of pathogenic and environmental serotypes, and 13 strains of other fish pathogenic bacteria were used to assess the sensitivity and specificity of the detection methods. The serological assays tested detected all the strains of V. anguillarum serotypes O1 and O2. The dot-blot assay was the most specific and sensitive method, detecting almost all isolates from serotypes O1, O2 and O3, with an average sensitivity of 1 x 10(6) bacteria g(-1) of fish tissue. The AQUARAPID-Va and the AQUAEIA-Va systems were able to detect 5 x 10(6) and 5 x 10(7) bacteria g(-1) of fish tissue, respectively. The simplicity, effectiveness and speed of the AQUARAPID-Va system confirmed this method as the most suitable serological test for the detection of V. anguillarum in field analysis and small-scale laboratory studies. 相似文献
11.
《水生生物资源》1999,12(5):343-350
Haemocytes of Mytilus edulis, allowed to attach to the plastic surface of a tissue culture plate in the presence of haemolymph, were observed by time-lapse video recording. When bacteria were added at concentrations of 10 or 50 bacteria per haemocyte, certain bacterial strains caused rounding of the cells within 2–3 h. Haemolymph was necessary for the rounding to occur; if bacteria were added in sterile seawater there was no significant difference in the number of rounded cells between control and bacteria-treated cultures for up to 4 h. The haemolymph factor required for this activity was active at 1/64 dilution in seawater, was sensitive to trypsin treatment, and activity was halved on heating at 56 or 100 °C for 30 min. For the most toxic bacteria tested, Vibrio alginolyticus NCMB 1339 and Vibrio anguillarum A7, haemocyte cell rounding appeared to be induced by a very small number of bacterial cells. Bacteria-free culture supernatant of V. anguillarum 2981 induced rounding of haemocytes in a dose-dependent manner, with 50 % of cells being rounded at a dilution of approximately 1/500 of the culture supernatant. In a survey of 226 bacterial isolates, those isolated from incidents of disease in a bivalve hatchery were significantly more toxic towards haemocytes than bacteria isolated from hatcheries without disease or from turbot hatcheries. 相似文献
12.
对青岛近海度假区、港口区、养殖区、工业区4类排污口邻近海域异养细菌的组成与分布进行了调查研究,采用海洋异养细菌平板涂布培养法分离细菌,基于16S rDNA的扩增性DNA限制性酶切片段分析(ARDRA)进行聚类筛选并测序鉴定。结果显示,青岛排污口邻近海域可异养培养的菌株分布于变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)和拟杆菌门(Bacteroidetes)4个细菌类群,变形菌门细菌数量占总测序数量的68%,其中γ-变形菌纲的数量在变形菌门中占88%;不同海域中细菌组成与分布有所不同,放线菌门和拟杆菌门的细菌种类较少且各海域分布有所不同,但γ-变形菌纲的弧菌属细菌(Vibrio sp.)在4类海域中均被发现,占总检测量的18.54%,其中溶藻胶弧菌(Vibrio alginolyticus)占弧菌总量的25%。不同海域也检测到V.parahaemolyticus、V.harveyi、V.campbellii、V.parahaemolyticus等水产养殖业的潜在致病菌。根据16S rDNA系统进化关系和细菌门类聚类分析得到的结论推断,工业区与养殖区海域的细菌组成和分布相似度较高,推断各海域的细菌组成与排污口类型有一定相关性。 相似文献
13.
用溶藻弧菌诱导后,收集耳鲍血淋巴,利用微量测定法进行抗菌活性分析.结果发现,耳鲍血淋巴具有抗革兰氏阴性菌-溶藻弧菌、副溶血弧菌、河流弧菌、嗜水气单胞菌、大肠杆菌的活性,对革兰氏阳性菌的抑制活性较低.利用胰蛋白酶对耳鲍血淋巴水解后,其抗菌活性消失,表明其中抗菌成分为蛋白质.将耳鲍血淋巴在不同温度(37～95 ℃)和不同pH(2.5～5.0)下保温,发现其抗菌活性成分对热及酸耐受性较强.耳鲍血淋巴在低浓度无溶血活性.该抗菌组分在耳鲍的免疫防御中具有一定的作用. 相似文献
14.
Abstract. A disease of cultured juvenile European oyster, Ostrea edulis , and American oysters, Crassostrea virginica , was characterized by infection of conchiolinous ligament and periostracum by surface-coating Vibrio spp. bacteria. A similar bacterium was isolated from diseased cultured hard clams, Mercenaria mercenaria , held in a facility about 300 km distance from the oyster hatcheries. The oysters demonstrated fragile, undercalcified peripheral shell and apparently excessive chalky deposits. Histological and scanning electron microscopical examination revealed that the oysters' eroded ligaments and perforated peripheral periostracum were infected with uniform rod-shaped bacteria. The infection appeared to interfere with normal proteinaceous and calcareous shell deposition and possibly with normal ligament function. Histological examination of the digestive system organs suggested that normal digestive processes were also interrupted. Two predominant bacteria were isolated from oyster samples and one predominant bacterium was isolated from clam samples. One of the isolates from oyster culture facilities was found almost exclusively associated with oyster shell surfaces and culture tank surfaces, this isolate was similar to the one taken from clam shell surfaces. The other predominant bacterium was found in water column samples in oyster culture facilities at levels from about 105 to 107 per ml. This isolate had a DNA base ratio (moles % G + C) of 43.6; the two surface-associated isolates had base ratios of 43.6 and 44.5%. These Gram-negative, oxidase positive, fermentive rods have been identified as a Vibrio spp. group. Specific antibody was produced which differentiated these bacteria from each other and from other similar isolates. Practical procedures for controlling the bacteria in shellfish culture systems are described. 相似文献
15.
16.
Phyu Phyu THAN Carmelo S . DEL CASTILLO Takeshi YOSHIKAWA Taizo SAKATA 《Fisheries Science》2004,70(4):659-666
ABSTRACT: Fourteen bacterial strains isolated from marine environments exhibited antagonistic action against a wide range of bacteria including Vibrio spp. A double layer agar method was used for preliminary screening to determine the relative degree of growth inhibition or bacteriolysis exhibited by the isolates. Most of the antagonistic isolates were found to be Gram-negative, motile rods and were oxidase positive, and oxidative in the oxidation and fermentation test, suggesting that they are belong to the genera Pseudomonas . The antagonistic isolates lyzed the dead cells of marine Gram-negative bacteria in both plate and liquid methods. Bacteriolytic and casein hydrolytic activities were observed in the culture supernatant of the isolates. Anion exchange column chromatography (Toyopearl DEAE-650 M) was used to purify the extracellular protease produced by an antagonistic strain A1-J25a. The active fractions of protease collected from the eluted solution also exhibited bacteriolytic activity. 相似文献
17.
Abstract Healthy lemon sharks, Negaprion brevirostris (Poey), were inoculated with Vibrio carchariae and V. damsela , two vibrios known to cause disease in other shark species. Vibrio damsela was not recovered from the inoculated sharks; V. carchariae infected the healthy lemon sharks, but did not produce clinically observable disease. However, histological evaluation revealed active splenic and hepatic disease with Gram-negative bacteria found in association with mononuclear cells. When the experiment was repeated with a physiologically compromised lemon shark, V. carchariae was lethal for the test animal, but V. damsela was again cleared. Vibrio carchariae was also isolated from skin lesions of a lemon shark infested with monogeneans ( Dermophthirius nigrellii ). Histological evidence indicated that this shark died of protozoan infection. Thus, lemon sharks are susceptible to infection with V. carchariae which can cause subclinical disease in otherwise healthy specimens. We suggest that D. nigrellii may act as a vector of V. carchariae and that stress plays a role in the clinical manifestation of this disease. 相似文献
18.
马氏珠母贝鳃组织抗菌活性的研究 总被引：2，自引：0，他引：2
用纸片琼脂扩散法和微量测定法对制备的马氏珠母贝鳃组织匀浆物进行抗菌活性分析.结果表明,马氏珠母贝鳃组织匀浆物具有抗革兰氏阴性菌-副溶血弧菌、河流弧菌、嗜水气单胞菌、大肠杆菌、绿脓杆菌活性;对革兰氏阳性菌-金黄葡萄球菌具一定抑制活性,但对枯草芽孢杆菌无抑制活性.利用胰蛋白酶对马氏珠母贝鳃组织匀浆物水解后,其抗菌活性消失,表明其中抗菌成分为蛋白质.将马氏珠母贝鳃组织匀浆物在不同温度(37～95 ℃)和不同pH(2.5～5)保温,发现其抗菌活性成分对热及酸耐受性较强.马氏珠母贝鳃组织匀浆物在低浓度无溶血活性.该抗菌组分在马氏珠母贝的免疫防御中具有一定的作用. 相似文献
19.
Simultaneous occurrence of Vibrio salmonicida and antibiotic-resistant bacteria in sediments at abandoned aquaculture sites 总被引：2，自引：0，他引：2
B. HUSEVÅG B. T. LUNESTAD P. J. JOHANNESSEN Ø. ENGER O. B. SAMUELSEN 《Journal of fish diseases》1991,14(6):631-640
Abstract. The present authors have examined the survival of the fish pathogenic bacterium Vibrio salmonidda and the occurrence of antibiotic resistant bacteria in sediments from abandoned aquaculture sites. Survival of V. salmonidda was determined by fluorescent monoclonal antibody labelling combined with 4',6-diamidino-2-phenylindole (DAPI) staining. Using this method, the authors were able to detect V. salmonidda in sediment samples from six out of 12 abandoned fish farm locations. One out of the six positive stations was abandoned in 1982. Vibrio salmonidda was not detected at the three stations not affected by aquaculture (control locations) or in sediment samples taken 200–250m away from aqua-culture sites. When applying high-performance liquid chromatography, oxytctracyclinc or furazolidonc residues were not detected in sediment samples from control or abandoned aquaculture sites. Nevertheless, the percentage of bacteria resistant to oxytctracycline was higher in aquaculture sediments, as compared to control sediments. The simultaneous occurrence of fish pathogens and high levels of resistant bacteria in sediments increases the risk of transfer of resistance to the pathogens. 相似文献