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为了查明河北省唐山市某豚鼠养殖场发病豚鼠感染的病原菌,本试验采集发病豚鼠的眼角脓性分泌物,采用细菌分离纯化、革兰染色、生化鉴定、16S rRNA基因PCR扩增方法对分离菌进行鉴定,通过K-B试纸法检测分离菌株对24种抗菌药的敏感性,试管二倍稀释法测定19种中药对该分离菌株的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),并通过动物回归试验检测该分离菌株的致病性。结果显示,分离菌株在血琼脂培养基培养24 h生长为针尖状透明菌落、呈β溶血,革兰染色为阳性圆球状、3~5个连接呈短链状,生化特性与马链球菌兽疫亚种相符,将其命名为MLQJ-1;16S rRNA序列比对结果显示,MLQJ-1与马链球菌兽疫亚种同源性高达99.72%,在系统进化树中处于同一分支,鉴定为马链球菌兽疫亚种;MLQJ-1对24种抗菌药的敏感性不同,对青霉素、头孢曲松和阿莫西林等8种抗菌药敏感,对阿米卡星、新霉素和卡那霉素等10种抗菌药耐药;苦地丁、苦参、黄芩和千里光对MLQJ-1的MIC值介于1.9~15.6 mg/mL,MBC值介于1.9~31.2 mg/mL;小鼠腹腔接种MLQJ-1菌液后24 h内全部死亡。结果表明,... 相似文献
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为了解马源马链球菌兽疫亚种(S.zooepidemicus)新疆分离株马链球菌兽疫亚种类M蛋白(SzM)基因的分子进化与变异情况,为该菌引起的感染性疾病的防控提供依据,本试验对新疆地区某马场采集的病马淋巴结样品进行病原菌的分离培养和生化鉴定,并对分离菌株进行药物敏感性试验。根据已发表的马源SzM基因序列设计引物,对其SzM基因进行PCR扩增及序列测定。将获得的SzM序列与GenBank中不同动物源马链球菌兽疫亚种分离株序列进行同源性比对和遗传进化分析。结果显示,分离得到了一株革兰氏阳性链球菌,将其命名为马链球菌兽疫亚种ZMSY15-1。药物敏感性试验结果表明,分离菌株对青霉素、磺胺嘧啶钠耐药,对其他14种药物均敏感。序列分析结果显示,马链球菌兽疫亚种ZMSY15-1与国内外不同动物源分离株SzM基因氨基酸同源性为56.0%~70.0%。遗传进化分析结果显示,这些菌株可分为4个群。马链球菌兽疫亚种ZMSY15-1与猪源分离株SzM蛋白的氨基酸同源性为59.9%,分别属于2个不同的群,其与美国马源分离株NH55426亲缘关系最近。本试验结果可丰富国内马源马链球菌兽疫亚种SzM基因的信息数据,为马链球菌兽疫亚种的致病机制研究和预防控制提供参考依据。 相似文献
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本试验旨在建立一种快速、特异、敏感的双重PCR鉴定猪链球菌和马链球菌兽疫亚种病原检测方法。根据猪链球菌GDH蛋白和马链球菌类 M 蛋白的基因保守区分别设计引物,优化了该双重PCR检测方法的引物浓度及比例,并筛选了其最佳退火温度;用该双重PCR反应体系以其他几株阴性菌株为对照,检测了该反应体系的特异性。以新鲜培养的猪链球菌倍比稀释后进行菌落计数,对该检测方法的敏感性进行了鉴定。M-like和GDH引物的加入量均为1 μL(20 pmol/L),最佳退火温度为52.3℃;该双重PCR反应体系有较高敏感性,检测马链球菌兽疫亚种和猪链球菌的敏感度分别达100和10 CFU;特异性试验结果显示,常见的5种病原菌在该双重PCR体系中无特异性条带出现;临床应用该方法分离鉴定了1株猪链球菌和2株马链球菌兽疫亚种。本试验建立了一种能同时检测猪链球菌和马链球菌兽疫亚种的双重PCR方法,且该方法应用快速、特异且敏感。 相似文献
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新疆伊犁某地区一些马群中不断出现以全身多发性脓肿为特征的患病马匹,为了查明其是否为马腺疫,以及由何种马链球菌感染所致,无菌采集典型患病马匹脓汁样品,常规THB增菌,绵羊鲜血平板划线分离单菌落后,用透射电镜对分离菌的微观形态进行观察,PCR对其fneB基因进行克隆与序列分析,最后用生化试验进行验证。结果表明,从12匹患病马的脓肿组织中,共采集了25份样品,从中分离鉴定出来源于不同患病马的2株马链球菌马亚种菌株,这2株菌的形态均符合链球菌的特点;基因克隆测序结果表明,其中1株与英国源的兽疫亚种H70和马亚种4047核苷酸同源性较高,均达97%~98%,另1株与英国源的马亚种4047和瑞典源某马亚种核苷酸同源性较高,均达98%~99%。这2株菌之间核苷酸序列同源性达98.3%,氨基酸同源性则达99.3%。马链球菌马亚种是引起伊犁某地区这些患马全身多发性脓肿的主要致病菌,它们可能来源于之前从欧洲和美洲引进的马匹,提示在进行疫病防控工作时,要特别注意海关检疫,积极防止疫病的传入。 相似文献
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1998年从病死猪分离了数株链球菌(其中2株编号为9801,9802),经鉴定,9801株为猪链球菌2型,9802株为马链球菌兽疫亚种。9801株系从暴发流行地区的病猪中分离,同时在该流行地区还发生畜禽从业人员感染相同血清型的链球菌而死亡的病例;9802株系从散发病猪中分离。临床有共同点,又有各自特点。病理变化均呈现典型的败血症变化,但肝、脾、肺等病变上有一些差别,而染色镜检基本一致。对兔均敏感,对小鼠和猪的敏感性有所不同。9801株菌落灰色,中心浑浊,直径0.5mm,α溶血,有草绿色色素沉着;9802株菌落湿润,半透明,直径1.5-2mm,β溶血。9801株对氧哌嗪青霉素、头孢他啶、氨苄青霉素最敏感;9802株对氧哌嗪青霉素、复方磺胺、头孢哌酮、呋喃妥因最敏感。 相似文献
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从我区某羊场流产波尔山羊的胎儿与羊水中分离到一株球菌。将该菌人工感染试验动物,证实为致病菌。通过对该菌的形态特征、培养特性、生化特性及致病特性等进一步进行检测,证明该分离菌为羊链球菌。同时对该细胞的耐药性进行了检测,结果表明,该菌对庆大霉素、卡那霉素高敏,而对青霉素产生了极强的耐药性。 相似文献
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猪链球菌2型与马链球菌兽疫亚种生物学特性的初步比较 总被引:1,自引:0,他引:1
1998年从病死猪分离了数株链球菌 ,其中 2株编号为 980 1,980 2。经鉴定 ,980 1株为猪链球菌 2型 ,980 2株为马链球菌兽疫亚种。 980 1株系从暴发流行地区的病猪中分离 ,同时在该流行地区还发生畜禽从业人员感染相同血清型的链球菌而死亡的病例 ;980 2株系从散发病猪中分离。临床有共同点 ,又有各自特点。病理变化均呈现典型的败血症变化 ,但肝、脾、肺等病变上有一些差别 ,而染色镜检基本一致。对兔均敏感 ,对小白鼠和猪的敏感性有所不同。 980 1株菌落灰色 ,中心浑浊 ,直径 0 5mm ,α溶血 ,有草绿色色素沉着 ;980 2株菌落湿润 ,半透明 ,直径 1 5~ 2mm ,β溶血。 980 1株对氧哌嗪青霉素、头孢他啶、氨苄青霉素最敏感 ;980 2株对氧哌嗪青霉素、复方磺胺、头孢哌酮、呋喃妥因最敏感 相似文献
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马链球菌兽疫亚种类M蛋白亚单位疫苗小鼠免疫试验 总被引:2,自引:0,他引:2
为获得控制猪链球菌病安全高效的疫苗,本试验进行了马链球菌兽疫亚种类M蛋白亚单位疫苗的研制。应用热酸法提取马链球菌兽疫亚种ATCC35246株类M蛋白,并分别采用羟基磷灰石(HAT)层析和冷酒精沉淀法进行纯化,结果得到较纯的类M蛋白。用热酸提取的粗制类M蛋白、2种纯化的类M蛋白及全菌苗免疫小鼠,结果保护率分别为92%(11/12)、100%(12/12)、100%(12/12)和83%(10/12),表明类M蛋白亚单位疫苗有进一步开发的必要。 相似文献
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马链球菌兽疫亚种 (Streptococcusequisubsp.zooepidemicus) 是引起我国猪链球菌病的主要病原之一 ,其类M蛋白是一重要的毒力因子和保护性抗原。用热酸提取的马链球菌兽疫亚种ATCC352 4 6株类M蛋白 ,作SDS PAGE电泳 ,在电泳图谱中出现相对分子质量为 43 0 0 0、 34 0 0 0、33 0 0 0、31 0 0 0和 30 0 0 0等 5条主蛋白带。免疫印迹显示 ,这 5条主蛋白带都能被ATCC352 4 6的全菌兔血清识别 ,表明它们具有有效的抗原表位 相似文献
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Alber J El-Sayed A Lämmler C Hassan AA Weiss R Zschöck M 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2004,51(10):455-458
The closely related streptococcal species Streptococcus equi subsp. zooepidemicus and S. equi subsp. equi were identified by polymerase chain reaction using oligonucleotide primers designed according to species-specific parts of the superoxide dismutase A encoding gene sodA. A further differentiation of both subspecies could be performed by amplification of the genes seeH and seeI encoding the exotoxins SeeH and SeeI, respectively, which could be detected for S. equi subsp. equi but not for S. equi subsp. zooepidemicus. A further simplification of the identification and differentiation of both subspecies was conducted by sodA-seeI multiplex polymerase chain reaction. 相似文献
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Real-time PCR for detection and differentiation of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus 总被引:1,自引:0,他引:1
Strangles is a contagious equine disease caused by Streptococcus equi subsp. equi. In this study, clinical strains of S. equi (n=24) and Streptococcus equi subsp. zooepidemicus (n=24) were genetically characterized by sequencing of the 16S rRNA and sodA genes in order to devise a real-time PCR system that can detect S. equi and S. zooepidemicus and distinguish between them. Sequencing demonstrated that all S. equi strains had the same 16S rRNA sequence, whereas S. zooepidemicus strains could be divided into subgroups. One of these (n=12 strains) had 16S rRNA sequences almost identical with the S. equi strains. Interestingly, four of the strains biochemically identified as S. zooepidemicus were found by sequencing of the 16S rRNA gene to have a sequence homologous with Streptococcus equi subsp. ruminatorum. However, they did not have the colony appearance or the biochemical characteristics of the type strain of S. ruminatorum. Classification of S. ruminatorum may thus not be determined solely by 16S rRNA sequencing. Sequencing of the sodA gene demonstrated that all S. equi strains had an identical sequence. For the S. zooepidemicus strains minor differences were found between the sodA sequences. The developed real-time PCR, based on the sodA and seeI genes was compared with conventional culturing on 103 cultured samples from horses with suspected strangles or other upper respiratory disease. The real-time PCR system was found to be more sensitive than conventional cultivation as two additional field isolates of S. equi and four of S. zooepidemicus were detected. 相似文献
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Abdulmawjood A Lämmler C 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1999,46(10):723-729
In the present study the soft agar technique was used to isolate phase variants of S. equi subsp. zooepidemicus-cultures isolated from infections of horses. The phase variants were characterized by a compact or diffuse colony morphology in this media. The variants could be cultivated separately and further characterized genotypically by RAPD analysis and by macrorestriction analysis of their chromosomal DNA by pulsed-field gel electrophoresis, indicating the identity of both strains of each pair. The diffuse colony variants grew uniformly turbid after cultivation in fluid media, did not haemagglutinate rabbit erythrocytes, and displayed a reduced surface hydrophobicity in hexadecane and phenyl-sepharose adherence tests. The compact colony variants generally grew as sediment with clear supernatant in fluid media, haemagglutinated rabbit erythrocytes and showed an enhanced surface hydrophobicity in both hydrophobicity tests. The presented soft agar technique allowed a demonstration of phase variation of S. equi subsp. zooepidemicus and a subsequent isolation of the variants. This might be an important prerequisite to understanding the pathogenic importance of phase variation among isolates of this bacterial species. 相似文献
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J Hewson C K Cebra 《The Canadian veterinary journal. La revue veterinaire canadienne》2001,42(6):465-467
A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp. zooepidemicus. Clinical findings, medical treatment, and case outcome are described. Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case. Possible transmission of the organism through contact with horses is discussed. 相似文献
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Alber J El-Sayed A Estoepangestie S Lämmler C Zschöck M 《Veterinary microbiology》2005,109(1-2):135-141
Bacterial superantigens are one of the major virulence factors produced by Streptococcus pyogenes and Staphylococcus aureus. The two novel superantigen encoding genes seeM and seeL were described for S. equi subsp. equi which is known as the causative agent of strangles in equids. In the present study previously characterized S. equi subsp. equi strains and strains of various other animal pathogenic streptococcal species and subspecies were investigated for the presence of the superantigen encoding genes seeM and seeL by polymerase chain reaction. According to these studies seeL and seeM appeared to be a constant characteristic of all investigated S. equi subsp. equi strains. Surprisingly, one S. equi subsp. zooepidemicus strain (S.z. 122) was also positive for both genes. The species identity of this S. equi subsp. zooepidemicus strain could additionally be confirmed by sequencing the 16S rRNA gene and the 16S-23S rDNA intergenic spacer region. The superantigen encoding genes could not be found among additionally investigated S. equi subsp. zooepidemicus strains or among strains of seven other streptococcal species. The seeL and seeM genes of the S. equi subsp. equi strain S.e. CF32 and the genes szeL and szeM of the S. equi subsp. zooepidemicus strain S.z. 122 were cloned and sequenced. A sequence comparison revealed a high degree of sequence homology between seeL, szeL, speL and seeM, szeM and speM, respectively. The superantigenic toxins L and M seemed to be widely distributed virulence factors of S. equi subsp. equi, rare among S. equi subsp. zooepidemicus but did not occur among a number of other animal pathogenic streptococcal species. 相似文献
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Streptococcus equi subsp. zooepidemicus (SEZ) is an opportunistic and zoonotic pathogen of horses. In this study, genetic intraspecies variability of SEZ obtained mainly from respiratory and genital samples of horses was investigated by analysis of the 16S–23S rRNA intergenic spacer region (ISR) and of the 16S rRNA gene. 16S–23S ISR rRNA type A1 was predominant, although a high rate of multiple products (30.5%) was obtained. Phylogenetic analysis of the 16S rRNA gene detected three genogroups (I, II and III). 16S rRNA variable regions V1 and V2 are the most important regions for evaluating SEZ intraspecies variability, but at least V1-V5 regions should be considered to avoid mistakes. Analysis of all 16S rRNA sequences available in databases assigned human SEZ to groups I and III but not to group II. These results show a high genetic variability in SEZ collected from different specimens of horses from various regions of Italy. 相似文献