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1.
基于嗜水气单胞菌气溶素(aerolysin)基因的序列,设计了1套引物,通过条件优化,成功建立了针对致病性嗜水气单胞菌的环媒恒温基因扩增(Lamp)检测法。采用Lamp法对病原菌进行了扩增,并对病鱼血样进行了检测。结果表明,该法只检出致病性嗜水气单胞菌。对不同浓度的细菌悬液扩增结果表明,Lamp检测嗜水气单胞菌的最低菌液浓度为1.4~14/μL。  相似文献   

2.
为研究在饲料中添加天然植物复合提取物对罗非鱼感染嗜水气单胞菌后成活率及头肾组织酶活力的影响,试验设计了5个天然植物复合提取物浓度梯度的等氮等能饲料。饲料中天然植物复合提取物的添加量分别为0、0.05%、0.10%、0.15%和0.20%,饲喂8周后,每组取15尾鱼注射嗜水气单胞菌液。试验结果表明,饲料中添加天然植物复合提取物有助于提高感染嗜水气单胞菌后罗非鱼的成活率,并能显著提高头肾溶菌酶、超氧化物歧化酶和酸性磷酸酶的活力。以感染嗜水气单胞菌后的成活率和头肾组织酶活力为指标,罗非鱼饲料中天然植物复合提取物的适宜添加量为0.10%~0.15%。  相似文献   

3.
嗜水气单胞菌作为水产养殖业中的主要病原菌,严重危害该产业的发展。为探究单宁酸对嗜水气单胞菌的抑菌作用及其转录组的影响,本实验将单宁酸2倍倍比稀释(8μg/mL~8 192μg/mL)后,测定单宁酸对嗜水气单胞菌ATCC 7966株的最小抑菌浓度(MIC);将嗜水气单胞菌分别与不同浓度的单宁酸共培养,根据所测细菌OD600nm值(共测24 h),绘制细菌的生长曲线。结果显示,单宁酸对嗜水气单胞菌的MIC为2 048μg/mL;浓度低于64μg/mL (临界值)单宁酸处理后不影响嗜水气单胞菌的生长。因此本实验采用64μg/mL的单宁酸与嗜水气单胞菌ATCC 7966株共培养,每组重复4次,12 h后提取各组嗜水气单胞菌总RNA,反转录为cDNA后构建cDNA文库,再利用随机引物,经PCR扩增、定量后,采用Illumina HiSeq测序平台进行转录组测序(RNA-Seq)。通过计算每个样品中r RNA的占比(rRNA%)及碱基的错误率对测序数据质控;采用Bowtie2将获得的各组嗜水气单胞菌测序数据与GenBank中嗜水气单胞菌ATCC 7966株参考基因组比对,分析它们之间的相似性。结果...  相似文献   

4.
为了了解野生麋鹿死亡原因及生活地区存在的病原菌,找出安全高效的药物,提高麋鹿存活率,对其生存地区疫病进行合理防控,试验采用细菌分离培养、显微镜检、PCR扩增、16S rRNA基因测序鉴定,并通过细菌耐药性试验对2016年8月份湖北石首市野生麋鹿自然保护区随机采集的10份麋鹿粪便样本进行了研究。结果表明:从该批粪便样本中分离得到6株大肠杆菌、1株嗜水气单胞菌和1株变形假单胞菌。6株大肠杆菌和1株嗜水气单胞菌对庆大霉素、链霉素、环丙沙星、新霉素、恩诺沙星、阿奇霉素、头孢拉定、头孢曲松、丁胺卡那、氟苯尼考、诺氟沙星均敏感;嗜水气单胞菌对氨苄西林耐药;6株大肠杆菌对氨苄西林的敏感率为0,中介率为50%,耐药率为50%。大肠杆菌和嗜水气单胞菌都有一定程度的致病性。  相似文献   

5.
致病性嗜水气单胞菌耐喹诺酮类药物菌株的分离鉴定   总被引:4,自引:0,他引:4  
对吉林省内12个水域采集和送检的66尾病鱼进行了细菌分离鉴定,共检出46株嗜水气单胞菌疑似菌,通过生化试验结合PCR扩增气溶素基因Aer,确定其中22株为致病性嗜水气单胞菌。进一步采用纸片扩散法和双倍稀释法测定嗜水气单胞菌分离株对多种抗生素的耐药性,其中有4株对喹诺酮类药物耐药,并且为交叉耐药,耐药菌检出率为18.2%,纸片扩散法检测其对喹诺酮类药物的抑菌圈均小于19mm。将病料中分离的8株嗜水气单胞菌负染后电镜观察,耐药菌表面有许多大小不同的球形结构,而敏感菌表面则为细丝状菌毛结构。  相似文献   

6.
疑似草鱼嗜水气单胞菌的分离鉴定及药敏试验   总被引:2,自引:1,他引:1  
试验对疑似嗜水气单胞菌草鱼的肝脏、脾脏、肾脏组织进行嗜水气单胞菌的分离鉴定。通过普通营养琼脂、胰蛋白大豆琼脂及生化试剂等的分离培养与生化鉴定后,获得3株细菌;16SrRNA通用引物检测3株细菌的基因序列比对确定菌株后,对确定的草鱼嗜水气单胞菌进行回归试验确认其致病性,然后采用临床上常用的阿奇霉素、阿米卡星、链霉素等10种抗生素进行药敏试验。结果表明,经过分离培养、生化鉴定及16SrRNA通用引物鉴定后确定3种菌株分别为嗜水气单胞菌、嗜麦芽窄食单胞菌和腐败斯瓦尼菌,确定的嗜水气单胞菌回归试验证明其具有致病性;药敏试验显示阿奇霉素对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感,其抑菌直径分别为105、95mm,阿米卡星对嗜麦芽窄食单胞菌、嗜水气单胞菌和腐败斯瓦尼菌这3种菌都较敏感,抑菌直径分别为105、95和93mm。结果表明,疑似为嗜水气单胞菌病的草鱼有3种细菌感染,其中鉴定的嗜水气单胞菌是致病菌,常用抗生素阿米卡星对3种菌都有抑制作用,而仅对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感的是阿奇霉素。  相似文献   

7.
嗜水气单胞菌广泛存在于环境中,可致人、家畜和水生动物的多种疾病,已成为重要的人兽共患病的病原菌。从甲鱼的红脖子、烂甲板病中分离到一株嗜水气单胞菌,试验旨在探究这株细菌的血凝特性以及用血凝和血凝抑制试验来检测这类细菌的可能性。  相似文献   

8.
罗非鱼出血病病原鉴定及疫苗   总被引:2,自引:0,他引:2  
从惠出血病罗非鱼血液、肝脏、脾脏中分离到一株细菌,经理化鉴定,该菌为嗜水气单胞菌,动物回归试验阳性,认为该罗非鱼出血病是嗜水气单胞菌引起。用该分离菌制成的饵料及附型疫苗田间免疫罗非鱼成活率达87.1%,未免疫组成活率仅59.9%。  相似文献   

9.
嗜水气单胞菌临床分离株胞外毒力因子的检测   总被引:9,自引:0,他引:9  
嗜水气单胞菌在自然界,尤其是水环境中广泛存在,其毒力菌株能够引致人和多种动物传染性疾病,在公共卫生上占有一定的地位。自20世纪90年代以来引起的我国淡水养殖鱼类的暴发性败血症,造成重大的经济损失。嗜水气单胞菌有致病性与非致病性菌株之分。嗜水气单胞菌在生长繁殖过程中,不断地向外界环境中释放代谢产物,这对于细菌消化吸收营养物质、吸附、侵袭宿主细胞,在宿主体内定居、繁殖以及抵抗宿主免疫系统都有极其重要的意义。  相似文献   

10.
嗜水气单胞菌和维氏气单胞菌是鱼类常见条件致病菌,采用传统的形态分类和生理生化反应进行鉴别费时费力,有时还会错误判断,为了提高鉴别效率和准确率,有必要建立一种快速鉴别两种细菌的分子生物学方法。选取12株鱼源致病气单胞菌使用细菌16SrDNA通用引物扩增细菌目的片段,将所测得序列校正后构建系统发育树以明确菌种,并依据种间序列差异筛选合适的限制性核酸内切酶,对嗜水气单胞菌和维氏气单胞菌进行PCR-RFLP酶切图谱的鉴别分析。筛选出嗜水气单胞菌16SrDNA的特异性限制性核酸内切酶SnaBⅠ和可以鉴别维氏气单胞菌的限制性核酸内切酶BseNⅠ,建立了一种快速鉴别嗜水气单胞菌和维氏气单胞菌的PCR-RFLP方法。研究结果对于淡水养殖鱼类嗜水气单胞菌和维氏气单胞菌细菌性病害的病原快速鉴别具有参考和应用价值。  相似文献   

11.
嗜水气单胞菌选择培养基鉴别效果的比较   总被引:4,自引:0,他引:4  
本试验比较了嗜水气单胞菌(Aeromonashydrophila,Ah)三种选择培养基AHM、APM和RS培养基的鉴别效果,并用于121株细菌检测,Ah的阳性检出率分别是54.76%、81.33%和83.54%。经过交叉比较分析,确定AHM和RS可作为适用于生产实践的Ah的选择培养基。  相似文献   

12.
13.
企鹅源嗜水气单胞菌的分离鉴定   总被引:2,自引:0,他引:2  
从上海某水族馆因血痢、厌食、消瘦而死亡的秘鲁帝企鹅的心脏、肝脏中分离到2株细菌SH051237和SH060104,其培养性状及生化特性基本一致,经生化试验、VITEK试验等鉴定为嗜水气单胞菌.动物试验表明,分离株对小鼠均具有较强的致病性,且从死亡鼠回收到相应细菌.结果表明,分离到的菌株为致病性嗜水气单胞菌.  相似文献   

14.
本试验根据GenBank已登录的致病性嗜水气单胞菌保守序列16S rDNA和Aero,设计2对引物,以嗜水气单胞菌纯培养物为起始材料,建立PCR检测方法。从12株分离物中均扩增到16S rDNA片段,从3株分离物中均扩增到Aero片段,经序列测定和分析,所扩增的片段均为嗜水气单胞菌的核苷酸序列。结果表明,建立的PCR方法可用于检测致病性嗜水气单胞菌。  相似文献   

15.
In this study a total of 140 broiler carcasses and carcass parts purchased at different supermarkets in Ankara including 50 whole carcass, 30 wing, 30 leg and 30 breast samples were analysed for the presence of motile Aeromonas species. According to analysis findings, motile Aeromonas spp. were isolated from 116 (82.9%) of total 140 samples. The distribution of the isolates were 94%, 86.6%, 80%, 63.3% in broiler carcass, wing, leg and breast samples, respectively. Aeromonas hydrophila was isolated the most prevalent species with 56% the range followed by Aeromonas sobria with 29.3% and Aeromonas caviae with 14.7% from all of the carcass and carcass part samples, respectively. Consequently, it was supposed that, examined broiler carcass and carcass parts have been contaminated to important level with motile Aeromonas species and it has been risk for public health.  相似文献   

16.
Abstract

To determine the scope of antibacterial resistance among Aeromonas spp., bacterial cultures were taken from a variety of tropical fish species imported from Singapore. The samples were plated on Rimler-Shotts medium for bacterial isolation and identified with both the API20E and Nonfermenter Test Strip systems. Aeromonas hydrophila, A. sobria, and A. caviae were identified in monomicrobic and concomitant infections. Following identification of bacterial isolates, 11 antimicrobials routinely used in the tropical fish industry and 1 new fluoroquinolone were tested for their effectiveness against Aeromonas spp. with the Kirby-Bauer disk-diffusion technique. Aeromonas sobria proved to be the most resistant, often showing susceptibility to only 3 of 12 test drugs. Aeromonas hydrophila was consistently the least resistant. Based on these results, antimicrobial resistance appears to be a rapidly emerging problem in the pet fish industry.  相似文献   

17.
The presence of potentially human pathogenic strains of Aeromonas was investigated in 84 samples of seafood which were purchased from retail traders in Berlin, Germany in spring 2000. A total of 134 Aeromonas strains were isolated on selective [GSP agar and Aeromonas (Ryan) agar] and unselective (standard count agar and enterohaemolysin agar) media from 27 (32.1%) of the samples and were classified as Aeromonas hydrophila (67.9%), A. caviae (26.1%) and A. sobria (6.0%) by biotyping. Thirteen (48.1%) of the 27 positive samples contained more than one species of Aeromonas. Production of haemolysins on enterohaemolysin agar was found with 132 (98.5%) of the strains at 28 degrees C and with 130 strains (97.0%) at 37 degrees C growth temperature. Vero cytotoxins were produced by 99 (73.9%) of the strains when grown at 28 degrees C but only by 24 of the strains (17.9%) at 37 degrees C. The latter strains were identified as A. hydrophila (n = 22) and A. sobria (n = 2) which came from 17 (20.2%) samples of raw seafood and from ready-to-eat salted herring 'Matjes' products. Cytotoxin-encoding genes for aerolysin (aer) and haemolysin A (hlyA) were investigated by PCR. Aer and hlyA genes were detected in both, strains which produced toxins only at 28 degrees C and strains which produced toxins at 37 degrees C. Our data indicate that raw seafood and ready-to-eat fish products can harbour potential human pathogenic, cytotoxin producing Aeromonas strains.  相似文献   

18.
为了研究嗜水气单胞菌重组弹性蛋白酶的酶学性质,试验根据GenBank中的嗜水气单胞菌弹性蛋白酶基因ahyB设计1对含酶切位点的特异引物,以嗜水气单胞菌J-1(AhJ-1)株为模板,经PCR扩增得到不含信号肽的成熟弹性蛋白酶基因片段(787 bp),并与pMD18-T载体连接、测序,再用DNAStar软件分析。结果表明:该基因片段与豚鼠气单胞菌胞外蛋白酶同源性高达95%,与嗜水气单胞菌AG2株弹性蛋白酶ahyB基因同源性为92%,与铜绿假单胞菌LasB基因同源性为82%;将PCR产物连入表达载体pET-32a,转化至大肠杆菌BL21菌株中进行诱导表达,出现50 ku的融合表达蛋白,该表达产物纯化复性后表现出酶的活性。  相似文献   

19.
To ensure the cause of suspected bacteria disease for channel catfish, the bacteria from clinical cases were isolated and cultured, then the biochemical and molecular biology assay, and animal regression test were used to identify the species of the isolated bacteria, bacteria medicine sensitive test was used to detect the bacterial drug resistance. The results showed that the bacteria were gram negative bacilli. The colony morphology and biochemical characteristics of the isolated bacteria were same as Aeromonas hydrophila. The sequencing test of the 16S rDNA also confirmed the isolated bacteria were Aeromonas hydrophila. The isolated bacteria could infect the healthy channel catfish and cause the same symptoms as the clinical case. The isolated bacteria were sensitive to floroquinolones and resistant to penicillin and cephalosporin drugs. The results showed that the pathogen of case catfish bowel disease was Aeromonas hydrophila, and the floroquinolones could use to prevent and treat the disease.  相似文献   

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