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1.
旨在建立一种快速检测迟钝爱德华氏菌的斑点免疫金渗滤诊断方法.试验选用硝酸纤维素膜作固相栽体,以胶体金标记羊抗兔IgG,确定迟钝爱德华氏茵免疫血清和金标抗体最佳工作浓度,以出现明显清晰斑点者判定为阳性.结果表明,用该方法检测时,迟钝爱德华氏茵呈阳性,温和气单胞菌、嗜水气单胞菌、河流弧菌、溶藻弧菌、鳗弧菌、腐败希瓦氏菌、产碱普罗威斯登菌、阪崎肠杆菌和大肠杆菌均呈阴性.试验表明,DIGFA方法简便、特异、快速、结果直观,便于在基层推广使用.  相似文献   

2.
鱼类迟钝爱德华菌病诊断与防治研究进展   总被引:1,自引:0,他引:1  
迟钝爱德华菌(Edwardsiella tarda)是目前水产养殖中危害极大的病原菌,它可引起鱼类产生迟钝爱德华菌病,使鱼类腹部积水肿胀、体表出血、肠内出现黏液,造成鱼类的大量死亡,严重危害了鱼类的养殖,带来了巨大的经济损失.迟钝爱德华菌能够侵染鱼类宿主细胞,抵抗宿主免疫机制,并能分泌毒素使正常细胞发生病变.防治鱼类爱德华菌病的方法主要为化学治疗法、疫苗防治法和微生态制剂防治法.论文对国内外有关迟钝爱德华菌病的发病情况、致病性研究、诊断方法及防治等诸方面的研究概况进行了系统的综述.  相似文献   

3.
为探讨牙鲆致病性迟钝爱德华菌毒力基因携带与分布,以毒力基因为分子靶标研究其致病机理及建立快速检测方法,根据 GenBank 上的基因序列,设计2对引物扩增致病性迟钝爱德华菌毒力基因 fimA和小肠结肠炎耶尔森菌毒力基因 irp2,结果在7个供试牙鲆病例的130株致病性迟钝爱德华菌中,irp2毒力基因的阳性率为46.15%(60/130),fimA 毒力基因的阳性率为100%,且均与已发表的参考菌株相应序列高度同源,同源性分别为97.32%和97.59%。可见耶尔森菌强毒力岛(HPI)在牙鲆致病性迟钝爱德华菌中广泛分布,但不同病例分离菌株间存在差异;fimA 毒力基因存在于所有的被检致病性迟钝爱德华菌中,可以作为快速检测致病性迟钝爱德华菌的标志物。  相似文献   

4.
试验旨在研究蒲公英内生菌对由嗜水气单胞菌和迟缓爱德华氏菌引起鱼类疾病的作用效果,分为2部分:体外抑菌试验和攻毒试验.试验所用的嗜水气单胞菌菌液质量浓度为890万CFU/mL,迟缓爱德华氏菌菌液质量浓度为780万CFU/mL.体外抑菌试验结果显示,蒲公英内生菌对体外培养的2种细菌均有显著抑制效果,嗜水气单胞菌的抑菌圈直径为24.50 mm,对迟缓爱德华氏菌的抑菌圈直径为30.53 mm.攻毒试验以黄河鲤鱼为试验鱼,注射量为0.2 mL/尾.攻毒试验设5个处理组,即对照组、注射嗜水气单胞菌攻毒组、嗜水气单胞菌攻毒后投喂蒲公英内生菌饲料组、迟缓爱德华氏菌攻毒组和迟缓爱德华氏菌攻毒后投喂蒲公英内生菌饲料组.攻毒试验显示,迟缓爱德华氏菌攻毒后投喂含有蒲公英内生菌饲料组累积病死率显著高于未注射致病菌的对照组,低于注射致病菌投喂普通饲料的试验组.嗜水气单胞菌攻毒组与嗜水气单胞菌攻毒后投喂蒲公英内生菌饲料组试验鱼累计病死率无显著性差异.  相似文献   

5.
为了确定黄颡鱼发病的原因,试验从患病黄颡鱼体内分离得到1株病原菌,命名为GDYM20160809,通过形态学观察、生理生化特性鉴定、16S rDNA分析及系统进化树构建等方法对分离菌进行鉴定,采用滤纸片扩散法对分离菌进行药敏试验。结果显示,该分离菌为革兰氏阴性短杆菌,PCR扩增其16S rDNA片段,经测序及BLAST比对,显示其与迟钝爱德华菌(Edwardsiella tarda)的同源性达100%,结合生理生化鉴定结果确定分离菌为迟钝爱德华菌。药敏试验结果显示,分离菌对磺胺类药物、头孢类药物、青霉素、复方新诺明等药物敏感,对磺胺间甲氧嘧啶钠、新生霉素、多黏菌素B、阿米卡星、卡那霉素和万古霉素6种药物耐药;人工感染试验结果证实,菌株对罗非鱼、禾花鲤、草鱼和黄颡鱼都有很强的致病性。本试验结果为有效防控黄颡鱼细菌性疾病提供了理论依据,并可指导养殖户合理用药。  相似文献   

6.
对蛙三毛滴虫(Trichomonas bactrachorumPerty)与爱德华氏菌(Edwarsiellatarda)混合感染引发牛蛙疾病的临床症状、病原特征等作描述,对该病的诊断和防治作初步探讨。结果表明:用甲硝唑(用量1.6g/kg饲料)+氟苯尼考(用量0.5-1g/kg饲料)拌饵投喂,连续5d,并结合水体消毒处理,对发病早、中期的养殖牛蛙有很好的治疗效果。  相似文献   

7.
为查明广东省肇庆市某牛蛙养殖场患病牛蛙的致病原及有效的治疗药品,试验取病蛙的肝脏、肾脏、腹水划线培养,分离得到菌株N1和N2,在纯化后通过细菌生理生化和细菌16S rRNA序列分析对其进行鉴定,并进行人工感染试验及药敏试验.结果 表明:菌株N1为迟缓爱德华氏菌(Ed-wardsiella tarda),菌株N2为弗氏柠...  相似文献   

8.
2021年上海市一家鳜鱼养殖场暴发出血病,初始典型症状为下颌出血。本研究通过细菌分离、分子鉴定、生化反应以及健康鳜鱼感染试验等,确认鳜鱼发病死亡由维氏气单胞菌(Aeromonas veronii)和迟缓爱德华氏菌(Edwardsiella tarda)共同感染所致。分离的维氏气单胞菌鸟氨酸脱羧酶(ODC)和赖氨酸脱羧酶(LDC)反应均为阴性,表明其既不属于维氏气单胞菌维氏生物群(A. veronii biogroup veronii),也不属于维氏气单胞菌温和生物群(A. veronii biogroup sobria),但在遗传上与温和生物群更近;迟缓爱德华氏菌生化反应不产生H2S。分离菌腹腔注射感染健康鳜鱼幼鱼后,被感染幼鱼均出现了死亡现象,且相同数量级病原回感鳜鱼,维氏气单胞菌感染后,鳜鱼发病更快,死亡率更高,感染量为1×107 cfu/尾时即可导致鳜鱼幼鱼在24 h内100%死亡。两种病菌都对硫酸新霉素和红霉素耐药,而对氟苯尼考都高度敏感。本研究为养殖鳜鱼出血病的治疗和控制提供了参考。  相似文献   

9.
从河北某养鸡场80日龄左右鸡群所发生的禽霍乱病死鸡中,分离到了相应病原菌多杀巴斯德氏菌(pasteurella multocida)。对分离获得的16株菌(HPs-1至HPs-16)进行了形态特征、培养特性、理化特性等方面的鉴定,同时选取代表菌株(HPs-1)进行了16SrRNA基因的分子鉴定,测定了16SrRNA序列,构建了系统发育树。结果表明分离鉴定的16株细菌均为多杀巴斯德氏菌败血亚种(P.multocida subsp.septica),所测代表菌株的16SrRNA基因长度142bp,在GenBank登录号为AY999017,该菌株与检索出的22株巴斯德氏菌属(Pasteurella Trevisan,1887)细菌16SrRNA基因序列同源性均在98%~100%。另外,药敏试验结果显示分离株对供试的青霉素G等33种抗菌药物高敏、对克林霉素敏感、对苯唑青霉素等3种耐药。  相似文献   

10.
为探明仔猪细菌性腹泻肠道致病性大肠埃希氏菌和沙门氏菌流行的血清型、耐药表型及耐药基因型,本试验采集了贵州省5个地(州)市7个规模化养猪场的128份腹泻仔猪肠道样本,并对采集的样本进行了大肠埃希氏菌和沙门氏菌分离与鉴定,通过动物试验鉴定菌株的致病性,利用血清学方法鉴定其血清型,并通过药敏纸片法对主要致病菌进行耐药性研究,采用PCR技术检测各致病菌株耐药相关基因,分析细菌耐药表型和耐药基因型相关性。结果显示,本研究共分离鉴定到78株致病性大肠埃希氏菌与21株沙门氏菌,致病性大肠埃希氏菌血清型以O138、O87为主,沙门氏菌血清型以鼠伤寒沙门氏菌、肠炎沙门氏菌居多;药敏试验结果表明,本试验分离到的78株致病性大肠埃希氏菌对β-内酰胺类药物耐药率达80%以上,对其他种类的抗菌药耐药率均超过40%,分离鉴定的21株沙门氏菌对氨基糖苷类药物耐药率达50%以上,对其他种类的抗菌药耐药率均达20%以上;本试验分离鉴定的致病性大肠埃希氏菌共检出12种耐药相关基因,沙门氏菌共检出10种耐药相关基因,两种细菌耐药基因型与耐药表型符合率均达60%以上,且均为多重耐药。本研究为仔猪腹泻的综合防控提供了理论依据。  相似文献   

11.
A genetic typing method utilizing PCR for the identification of Actinobacillus pleuropneumoniae serotype 2 isolates has been developed based on the in vitro amplification of a 1.4 kb DNA segment of the serotype 2 capsular polysaccharide genes cps2AB. The assay was tested with all serotype reference strains and a collection of 92 different A. pleuropneumoniae strains of all 15 serotypes of both biovars I and II, originating from 18 different countries worldwide. The cps2 based PCR identified the serotype 2 reference strain and all 12 serotype 2 collection strains contained in this set. DNA was not amplified from the remaining A. pleuropneumoniae reference and collection strains, indicating the PCR assay was highly specific. Furthermore, the PCR method detected all 31 A. pleuropneumoniae serotype 2 field isolates from diseased pigs that were identified in parallel as serotype 2 by agar gel diffusion. The serotype 2 PCR assay proved to be highly specific and reliable for the identification of serotype 2 isolates of A. pleuropneumoniae.  相似文献   

12.
Between April 2001 and 2007, 18 Yersinia pseudotuberculosis outbreaks occurred in breeding monkeys at 12 zoological gardens in Japan, and 28 monkeys of 8 species died. A total of 18 Y. pseudotuberculosis strains from the dead monkeys, comprising one strain per outbreak, were examined for serotype and the presence of the virulence genes virF, inv, ypm (ypmA, ypmB and ypmC) and irp2. Of the 18 Y. pseudotuberculosis strains, 7 (38.9%) were serotype 4b, 7 (38.9%) were serotype 1b, and there was one each of serotypes 2b, 3, 6 and 7. All the 18 strains examined harbored virF and inv. Sixteen (88.9%) strains, including the strain of serotype 7, harbored ypmA. However, no strain harbored ypmB, ypmC and irp2.

This study demonstrated that among other pathogenic factors, almost all the Y. pseudotuberculosis isolated from the outbreaks had the ypm gene encoding the superantigenic toxin, YPM. As most of the monkeys who died in those outbreaks originated from South America and other regions, where the presence of the ypm gene have not been reported, YPM might be the cause, or at least the most important factor for, the high mortality of the breeding monkeys infected by Y. pseudotuberculosis in Japan. This is also the first report of a fatal case due to Y. pseudotuberculosis serotype 7 infection in the world.  相似文献   


13.
The pathogenic potential of five strains of serotype 2 infectious bursal disease virus (IBDV) for specific-pathogen-free chickens was examined. There were no gross or microscopic lesions in the inoculated chickens. Bursa-to-body-weight ratios of IBDV-infected chickens were not significantly different from those of uninfected controls. Virus-neutralizing antibodies to IBDV of serotype 2, but not serotype 1, were detected in infected chickens. This study indicated that the serotype 2 viruses examined were infectious but not pathogenic in chickens.  相似文献   

14.
160 ovine isolates of Pasteurella haemolytica, representing each of the 16 serotypes and also untypable strains, were examined for plasmid content. Plasmid DNA was identified in, and prepared from, strains of serotypes A2, T3, A14 and A16 and also from an untypable strain. The relationship between the plasmids present in the different strains was examined both by restriction fragment profile analysis and by DNA/DNA hybridisation. Both methods gave broadly similar results and showed that each serotype tended to contain either a single plasmid species, or a limited range of species, and that structural similarities could traverse serotype boundaries. None of the plasmid-bearing strains showed any significant level of resistance to a range of antibiotics.  相似文献   

15.
对猪链球菌2型、7型、9型的标准菌株和7株猪链球菌2型分离菌的谷氨酸脱氢酶(GDH)基因进行PCR扩增,经回收纯化后克隆到PMD18-T载体,筛选阳性克隆菌后测序并对其进行序列分析。结果,PCR扩增出1300bp左右的片段,包括GDH基因的整个开放阅读框,而测序结果表明,其序列与GenBank中的猪链球菌GDH基因序列一致,进一步序列分析表明,GDH的核苷酸序列在相同血清型之间同源性高达99%以上,而在不同血清型之间的同源型也达到了96%以上,而其氨基酸序列同源性则都在99%以上,且都具有GDH1型家族的功能区域。说明猪链球菌GDH基因及其蛋白具有高度的保守性,为进一步研究与应用提供了重要依据。  相似文献   

16.
为了解山东地区副猪嗜血杆菌病的流行情况和流行菌株的生物学特性及致病性,将2016-2018年山东省12个地区送检的103个发病猪的病料进行细菌分离,并对疑似菌株进行形态学观察、PCR鉴定及血清型鉴定,对两株流行菌株进行了培养特性观察、生化特性鉴定、药敏试验及致病性研究。最终分离获得29株副猪嗜血杆菌,分离率为28.16%,其中血清型4型和5型最为流行,其次是1型和12型。该病多发于春秋两季,31~50日龄的仔猪感染率最高。两株流行菌株LZ株和LC株均对青霉素类、头孢类等药物高度敏感,LZ株对庆大霉素、卡那霉素等中度敏感,对林可霉素、链霉素不敏感,LC株对庆大霉素、林可霉素等中度敏感,对卡那霉素、链霉素不敏感;生化特性试验结果显示,LC株和LZ株的硝酸盐还原试验、接触酶试验、葡萄糖发酵试验以及果糖发酵试验的结果均为阳性,吲哚试验、氧化酶试验、甘露醇发酵试验的结果均为阴性;动物致病性试验表明LZ株和LC株均具有较强的毒力,最小发病剂量分别为4.5×10^9 CFU和6.0×10^9 CFU。该研究为副猪嗜血杆菌病的防治提供了重要的参考依据。  相似文献   

17.
Most isolates of Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) from human patients and from environmental materials in Japan have been identified as serotype A mating type a by the seroagglutination test and mating experiments. A PCR method using the mating type alpha allele-specific primer of the STE12 gene and the serotype- and mating type-specific primers of the STE20 gene for identification of C. neoformans has been developed. Using the PCR method, conserved strains and clinical isolates from feline cryptococcosis were examined for serotype and the mating type. The results showed that all clinical isolates examined were identified as serotype A, MATalpha, indicating that feline cryptococcsis cases in Japan are caused by C. neoformans serotype A, MATalpha, as is the case in humans.  相似文献   

18.
猪源链球菌的分离鉴定及生物学特性研究   总被引:10,自引:0,他引:10  
通过生化试验、药敏试验、PCR分型、毒力基因的PCR检测及动物试验对分离的猪源链球菌进行药物敏感性、血清型和分子流行病学初步研究。从不同省份猪链球菌病疑似病例猪的心血、肝、淋巴结、脑和关节液组织分离出97株链球菌,药敏试验结果表明各菌株对13种抗菌素的耐药谱不同,但对先锋霉素V和环丙沙星的耐药率均低于5%。通过对分离菌株进行PCR鉴定和分型,确认26株为猪链球菌,其中1株为1型,16株为2型,4株为7型,没有9型,另5株为其它型。进一步对1型、2型和7型猪链球菌mrp、epf和sly3种毒力基因的分布情况进行了PCR检测。动物试验表明能100%致死小白鼠的猪链球菌基因型均为epf^+mrp^+sly^+2型猪链球菌,1株2型和1株7型猪链球菌均能复制出典型的猪链球菌病例。  相似文献   

19.
Ribonuclease T1 oligonucleotide maps of strains of 4 of the endemic serotypes of foot-and-mouth disease virus isolated in Kenya between 1964 and 1982 have been compared with data obtained in complement-fixation and neutralization tests. There was a continual change in the oligonucleotide maps obtained for all the serotypes examined. This genetic heterogeneity was generally associated with antigenic variation. Viruses isolated during the 12-month course of an epidemic of the SAT 1 serotype showed few changes in their oligonucleotide fingerprints, and were serologically related. These maps form a data base that will be useful in future epidemiological studies on the maintenance and spread of foot-and-mouth disease virus in this region.  相似文献   

20.
The growth of five strains of infectious bursal disease virus--three strains of serotype I (SAL, D-78, 2512), one of serotype II (OH), and one variant strain (Variant-A)--were compared in Vero and chicken embryo fibroblast (CEF) cell cultures in order to characterize the replication of different strains of IBDV in Vero cells. For all five virus strains, the latent period in Vero cells ranged from 12 to 18 hr, which was longer than the 4-to-6-hr latent period observed in CEF cultures for strains SAL, D-78, and OH. Virus strains SAL, D-78, and OH, which were examined in both Vero and CEF cultures, also had a more extensive maturation phase and higher yields of virus in Vero than in CEF cultures. Total titers of these viruses of 5.35 to 6.10 log10 TCID50/ml in CEFs occurred 24 to 30 hr postinoculation (PI), although the cytopathic effect (CPE) was not seen until 72 hr PI. By comparison, their total infectious virus titers of 6.85 to 8.35 log10 TCID50/ml in Vero cells occurred from 48 hr PI, coinciding with the appearance of CPE. The growth curve of Variant-A in Vero cells differed from the other viruses by showing steadily rising extracellular and cell-associated virus titers throughout the 72-hr observation period. Only very low titers of Variant-A were obtained in CEF cultures, and thus no growth curve in CEFs was performed.  相似文献   

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