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1.
从健康仔猪鼻腔分离到1株血清8型猪链球菌,基因型为mrp-efsly+,命名为RC4726。对青霉素类、头孢菌素类、磺胺类、喹诺酮类药物敏感,对红霉素、阿齐霉素、克林霉素和四环素耐药,MIC值分别为0.5 mg/mL,2 mg/mL,0.5 mg/mL,4 mg/mL。4×108CFU不能使C57BL6小鼠发病。MLST结果显示,RC4726的7个管家基因的座位分别为18,34,24,12,1,1,4。试验结果表明RC4726是一个新ST型,属于非致病性猪链球菌。  相似文献   

2.
为分析猪链球菌4型(Streptococcus suis type 4,SS4)分离株的病原生物学特性,试验对来自中国不同地区的10株猪链球菌4型进行了多位点序列分型,通过PCR方法对猪链球菌7个保守管家基因aroA、gki、dpr、mutS、recA、thrA、cpn60进行扩增,测序后将结果上传至MLST数据库查找序列型,然后制作聚类分析图来阐明菌株之间的亲缘关系;采用PCR方法对7种主要的毒力基因gdh、mrp、epf、sly、fbps、gapdh与orf2进行鉴定,通过毒力因子谱来分析猪链球菌4型毒力因子的分布;以纯化的10株细菌对BALB/c小鼠进行动物致病性试验,根据小鼠致死数量筛选出最强毒株,并进行新西兰兔致病性试验。结果显示,10株猪链球菌4型经多位点序列分型,6株为ST850型,3株为ST1006型,1株为ST94型;结合菌株分离地区分析发现,广东和江苏地区菌株有较高的同源性,江沪地区菌株出现分化现象,表现为遗传多样性;10株菌株均检测到了gdh、gapdh和orf2毒力基因,7株检测到sly基因,4株检测到fbps基因,根据毒力因子谱发现共有3个毒力基因型,gdh+sly+gapdh+orf2+型有6株,gdh+fbps+gapdh+orf2+型有3株,gdh+sly+fbps+gapdh+orf2+型仅有1株。动物致病性试验表明,10株细菌均能使BALB/c小鼠死亡,其中SH1510的半数致死量低至1×108 CFU,对小鼠的致病性最强,将纯化的SH1510菌液接种新西兰兔,可使新西兰兔出现典型的神经症状并死亡。以上结果为猪链球菌的遗传进化、毒力研究提供了新的数据,丰富了猪链球菌病的研究。  相似文献   

3.
为分析猪链球菌4型(Streptococcus suis type 4,SS4)分离株的病原生物学特性,试验对来自中国不同地区的10株猪链球菌4型进行了多位点序列分型,通过PCR方法对猪链球菌7个保守管家基因aroA、gki、dpr、mutS、recA、thrA、cpn60进行扩增,测序后将结果上传至MLST数据库查找序列型,然后制作聚类分析图来阐明菌株之间的亲缘关系;采用PCR方法对7种主要的毒力基因gdh、mrp、epf、sly、fbps、gapdh与orf2进行鉴定,通过毒力因子谱来分析猪链球菌4型毒力因子的分布;以纯化的10株细菌对BALB/c小鼠进行动物致病性试验,根据小鼠致死数量筛选出最强毒株,并进行新西兰兔致病性试验。结果显示,10株猪链球菌4型经多位点序列分型,6株为ST850型,3株为ST1006型,1株为ST94型;结合菌株分离地区分析发现,广东和江苏地区菌株有较高的同源性,江沪地区菌株出现分化现象,表现为遗传多样性;10株菌株均检测到了gdh、gapdh和orf2毒力基因,7株检测到sly基因,4株检测到fbps基因,根据毒力因子谱发现共有3个毒力基因型,gdh+sly+gapdh+orf2+型有6株,gdh+fbps+gapdh+orf2+型有3株,gdh+sly+fbps+gapdh+orf2+型仅有1株。动物致病性试验表明,10株细菌均能使BALB/c小鼠死亡,其中SH1510的半数致死量低至1×108 CFU,对小鼠的致病性最强,将纯化的SH1510菌液接种新西兰兔,可使新西兰兔出现典型的神经症状并死亡。以上结果为猪链球菌的遗传进化、毒力研究提供了新的数据,丰富了猪链球菌病的研究。  相似文献   

4.
为了解我国猪链球菌2型毒力因子和MLST型的特点,对四川、重庆和山东等地的22株猪链球菌2型进行毒力因子检测和ST型分析。结果显示,78%(17/22)的分离株为sly+mrp+epf+基因型的强毒株,仅山东省的5株分离株为弱毒力型。四川省和重庆市89%(15/17)的分离株为毒力较强的ST7型,而山东省的分离株均为毒力稍弱的ST1型。结合这些地区的猪链球菌病发病情况,分析提示猪链球菌2型的毒力因子和ST分型与菌株的致病性有一定的相关性。  相似文献   

5.
《中国兽医学报》2014,(6):936-941
通过对山西某猪场发病猪组织器官分离的病原菌鉴定以确定病原微生物以及其毒力强弱。主要从发病猪的关节液、肝脏、心脏、脾脏等器官内采集病原菌经过培养、镜检、生化试验初步鉴定后,用PCR方法通过扩增gdh基因和16SrDNA基因,16SrDNA测序后与国内外一些地区分离株的同源性比对,MLST(aroA、cpn60、dpr、gki、mutS、recA、thrA)分型研究、血清学分型以及mrp,ef,sly 3对毒力基因的扩增分析,结果鉴定为猪链球菌2型(SXZ-1株),16SrDNA序列与国内外部分地区菌株序列的同源性达到99%100%。MLST型为ST7型,具有强致病性(mrp+ef+sly+),表明山西存在ST7型强毒2型猪链球菌。  相似文献   

6.
猪链球菌扁桃体分离株的毒力因子分布特征与致病性   总被引:6,自引:0,他引:6  
本研究设计并合成7对引物,用PCR方法对猪链球菌7种主要毒力因子,包括谷氨酸脱氢酶(gdh)、溶血素(sly)、胞外蛋白因子(ef)、溶茵酶释放蛋白(mrp)、纤连蛋白/血纤蛋白原结合蛋白(fpbs)、三磷酸甘油醛脱氢酶(gadph)和毒力相关序列orf2进行检测,分析了29株猪链球菌扁桃体分离株的毒力因子分布特征.在被检的25株2型菌株中,共检测出7个基因型.其中10株(40%)的基因型为cps2/gdh+/sly+/ef+/mrp+/fbps+/gadph+/orf2+,7株(28%)表现为cps2/gdh+/sly-/ef-/mrp+/fbps+/gadph+/orf2+,4株(16%)表现为eps2/gdh+/sly-/ef+/mrp+/fbps+/gadph+/orl2+,基因型表现为eps2/gdh+/sly+/ef+/mrp+/fbps+/gadph+/orf2-、cps2/gdh+/sly-/ef-/mrp+/fbps+/gadph+/orf2-、eps2/gdh+/sly-/ef-/mrp*/fbps+/gadph+/orf2+、eps2/gdh+/sly-/ef-/mrp-/fbps-/gadph+/orf2-各1株;1株猪链球菌7型(SS7)分离株,基因型表现为cps7/gdh+/sly+/ef-/mrp-/fbps+/gadph+/orf2+;3株猪链球茵9型(SS9)扁桃体分离株,均表现为cps9/gdh+/sly-/ef-/mrp-/fbps+/gadph+/orf2+.可见我国SS分离株的毒力基因分布较为复杂,而且SS2的优势流行菌株是同时具有多种毒力因子的高致病茵株.通过对不同毒力基因型毒株对西藏小型猪的感染试验发现:毒力基因型为cps2/gdh+/sly-/ef+/mrp+/fbps+/gadph+/orf2+的健康扁桃体分离株SH06-21D对西藏小型猪有较强的致病性,仅次于标准株HA9801,而基因型为sly-/ef-/mrp-/fbps-/gadph+/orf2-的健康扁桃体分离株GZ06-122B对西藏小型猪不表现明显的致病性.  相似文献   

7.
《中国兽医学报》2017,(2):272-277
为了解吉林省猪链球菌的流行情况,从屠宰场采集的猪咽拭子和鼻拭子样品中分离鉴定猪链球菌,并进行菌株血清型、基因型和毒力表型的鉴定,以及致病性和耐药性的分析。结果表明,从100份样品中共分离鉴定猪链球菌104株,其中29株鉴定为血清2型、9型和1型等几种常见的致病性血清型,其他75株不属于常见的致病性血清型。血清2型的菌株中,2株经鉴定为ST1基因型和mrp+epf+sly+毒力型,并经动物试验鉴定为强毒菌株;其他菌株均为ST28基因型和mrp+epf-sly-毒力型,具有中等毒力。血清9型的菌株,均为mrp-epf-sly-毒力型,动物试验鉴定均为强毒菌株。根据Kirby-Bauer纸片扩散法的药敏试验结果,98%的猪链球菌分离株对四环素耐药;对大环内酯类、克林霉素和链霉素的耐药率都在50%以上;对β-内酰胺类、氯霉素和喹诺酮类抗生素的耐药率小于20%。总体分析,从屠宰场分离的猪链球菌强毒菌株所占的比例并不高,但是菌株多重耐药的情况非常严重。  相似文献   

8.
为了解安徽地区猪链球菌2型(SS2)临床分离株毒力基因分布、分子分型特征及其与致病性的相关性,本研究收集了58株源自临床患病猪的SS2,应用PCR技术检测其毒力基因:胞外蛋白因子基因(epf)、溶菌酶释放蛋白基因(mrp)和溶血素基因(sly),应用多位点序列分型方法(MLST)对其进行基因分型,并分析其与动物致病性之间的相关性。结果显示,58株分离菌,epf^+/mrp^+/sly^+型为44.83%(26/58),epf^+/mrp^-/sly^+型为25.86%(15/58),epf^-/mrp^+/sly^+型为18.97%(11/58),epf^-/mrp^-/sly^+和epf^-/mrp^-/sly^-型均为5.17%(3/58)。共分出10种ST型,其中ST1 26株(44.83%),ST7 18株(31.03%),ST28 6株(10.34%),ST117、ST156和ST308均为1株,各占1.72%,另有4种新发现的ST型,即ST958 2株(3.45%),ST957、ST959、ST970各1株(1.72%);分为4种克隆复合物(ST1 complex、ST27 complex、ST87 complex、ST188 complex)。ST1以epf^+/mrp^+/sly^+型为主(69.23%),其中23株(88.46%)分离自全身感染病猪;ST7以epf^+/mrp^-/sly^+型为主(55.56%),其次为epf^+/mrp^+/sly^+型(38.89%),其中13株(72.22%)分离自全身感染病猪。通过对斑马鱼、昆明鼠感染试验和LD50的测定,筛选出的6株强毒菌株分为ST1型(2株)和ST7型(4株)。结果表明,epf^+/mrp^+/sly^+是安徽地区SS2的优势毒力基因型,其次为epf^+/mrp^-/sly^+型。获得10种ST型,其中4种为首次发现,并且ST1和ST7为优势型。具有ST1/epf^+/mrp^+/sly^+遗传特征的菌株在临床上均倾向于导致全身感染。本研究结果揭示了安徽地区SS2毒力基因型、MLST遗传特征与毒力、动物临床症状之间的相关性,为开展SS2流行病学研究,区分不同菌株间毒力差异的研究提供参考依据。  相似文献   

9.
为了解人源、牛源及兔源无乳链球菌(S.agalactiae)的基因型差异及分子分型的总体结构特征,本实验对30株2012年~2015年分离于人、牛及兔的S.agalactiae在cfb基因鉴定的基础上,通过荚膜多糖分子血清分型(CPS)、多位点序列分型(MLST)和脉冲场凝胶电泳(PFGE)进行基因型的确定和分子特征差异性比较。CPS结果显示,牛源和兔源S.agalactiae均为Ia血清型,而人源无乳链球菌包括Ia和III型两种血清型,以Ia血清型为主要血清类型;MLST分型获得8个STs序列型(ST7、ST10、ST19、ST61、ST103、ST199、ST486、ST651)和6个克隆群(CC7、CC10、CC19、CC23、CC61、CC103),其中人源S.agalactiae拥有最多的STs和克隆群分型;PFGE聚类分析可以将其分为18个PFGE带型,且不同动物源性S.agalactiae之间基因分型差异性较大。血清分型、STs序列型、PFGE带型与宿主的来源之间无明显的相关性。不同来源菌株可以同属于血清型Ia,且所有Ia血清型菌株可以分布于不同的STs序列型和PFGE带型中。但分型为Ia/ST651的人源S.agalactiae在MLST的克隆分群中是分型为Ia/ST103牛源S.agalactiae的克隆衍生物,同属克隆群CC103,Ia/ST651只是Ia/ST103单一位点变化的产物,从分子水平的相似性推测二者存在交叉感染的可能性。本研究为奶牛乳房炎的有效预防和治疗提供依据。  相似文献   

10.
为了探明2018年3月江苏省某规模化猪场发病猪的病原,本研究对发病猪进行采样和细菌分离,并对细菌进行了PCR鉴定、多位点序列分型(MLST)及小鼠致病力分析。结果发现:在该场4份发病猪组织中分离到猪链球菌2型(SS2)、猪链球菌9型(SS9)菌株各2株,MLST分析显示分离的SS2菌株属于ST7,SS9菌株属于ST243,SS2、SS9分离菌株的毒力基因表现型分别为mrp^+epf^+sly^+orf2^+sao^+fbps^+gdh^+和mrp^-epf^-sly^-orf2^+sao^-fbps^+gdh^+。动物试验显示其中3株猪链球菌分离菌株对BALB/c小鼠有较强致病力。研究结果为该养殖场猪链球菌感染防控措施的制定提供了重要参考。  相似文献   

11.
Streptococcus suis is an important agent of swine and human meningitis. Sequence type (ST) 7 emerged in China and was responsible for the human epidemic caused by S. suis in 2005. The virulence of S. suis ST7 is greater than the wild type pathogenic S. suis, ST1; however, the mechanisms for this increased pathogenicity are unknown. The aim of this study was to determine the role of different toll-like receptors (TLRs) involved in regulating the host response to the S. suis infection and to speculate on differing mechanisms used by ST7 strains to induce disease. Here we compared two ST7 strains isolated in the 2005 Sichuan outbreak to two ST1 strains. Our data show TLR2, 6 and 9 are involved in the recognition of heat-killed S. suis independent of the ST type. We found the TLR-dependent cytokine production differed between the two types of strains using whole cell lysate proteins. TLR6 played a greater role in cytokine production induced by the whole cell lysate proteins from the ST7 strain than in that induced by the ST1 strain lysates. The data suggest that mechanisms of inflammation induced by S. suis strains differ where this will be useful in designing efficient strategies in combating streptococcal toxic shock-like syndrome caused by the S. suis ST7 strains.  相似文献   

12.
A total of 142 strains from different serotypes of Streptococcus suis isolated in Spain from diseased pigs (88 strains) and healthy carrier pigs (54 strains) were studied for the presence of a muramidase released protein (MRP) and an extracellular factor (EF). The following five phenotypes: MRP+EF+, MRP+EF-, MRP-EF+, MRP+EF* and MRP*EF- were detected. A high percentage of S. suis serotype 2 strains isolated from diseased pigs (84 per cent) belonged to phenotype MRP+EF+, but this phenotype has also been noticed in other serotypes (serotypes 1, 1/2 and 14). Both proteins were detected in S. suis serotype 2 strains (26%) isolated from healthy carrier pigs and one of both proteins in serotypes 1 and 14 (phenotype MRP+EF*). The isolation of S. suis strains from healthy pigs which have shown both proteins may support the epidemiological significance of these carriers in the maintenance, transmission and distribution of virulent strains within and between swine farms.  相似文献   

13.
A molecular analysis of strains of Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and Mycoplasma mycoides subsp. mycoides, small colony type (M. mycoides SC) isolated from goats was performed using the amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) fingerprinting techniques. Among the 11 field strains of M. capripneumoniae from Tanzanian goats, two AFLP patterns were demonstrated, with 10 of the strains showing indistinguishable patterns. Five Kenyan strains of M. capripneumoniae produced three AFLP patterns, with two of them being indistinguishable from the 10 identical Tanzanian and one Ugandan strain (M74/93) isolated from sheep. The AFLP pattern of the type strain (F38(T)) was identical to two Kenyan strains (Baringo and G183/82). On PFGE analysis, all the examined M. capripneumoniae strains exhibited identical PFGE profiles.Five field strains of M. mycoides SC isolated from goats displayed identical AFLP patterns except for one strain which differed from others at only one position. The AFLP pattern of the type strain of M. mycoides SC (PG1(T)) was different from the field strains. The five field strains of M. mycoides SC produced identical PFGE profiles, which were, however, different from the type strain. The AFLP and PFGE profiles of M. mycoides SC strains from goats were identical to those of six strains isolated from cattle affected with contagious bovine pleuropneumonia (CBPP) in the same areas. The results of this study suggest a close epidemiological linkage between strains of M. capripneumoniae and between M. mycoides SC type, respectively, isolated from goats in Tanzania.  相似文献   

14.
Experimental infections of mice and pigs with Streptococcus suis type 2.   总被引:6,自引:0,他引:6  
Five inbred strains of mice were tested for their susceptibility to Streptococcus suis type 2 including the type strain, two isolates from meningitis in pigs and two isolates from tonsils of clinically healthy pigs. C57BL/6, ICR and ddY strain mice showed lower susceptibility to all strains of S. suis type 2 than BALB/c and SS strain mice. The type strain and the isolates from diseased pigs produced septicaemia and meningitis in BALB/c and SS mice inoculated with 10(8) colony forming unit of the bacteria and 60 to 100% of these infected mice died. On the other hand, mice inoculated with the isolates from healthy pigs showed mild clinical signs but none of them died. In BALB/c mice which died or developed nervous signs, the purulent meningo-encephalitis, myocarditis, ophthalmitis, labyrinthitis and otitis media were observed. S. suis type 2 antigen was demonstrated in these lesions by immunoperoxidase staining using rabbit S. suis type 2 antiserum. These results were similar to those in the experimentally infected pigs with these virulent and avirulent strains against mice. These results indicate that BALB/c and SS strains of mice are useful as an experimental model of S. suis type 2 infections in pigs, and that there are virulent and avirulent strains against mice and pigs among the strains of S. suis type 2.  相似文献   

15.
The efficacy of triclabendazole in sheep experimentally infected with Fasciola hepatica was studied. Two groups of 12 lambs were infected with a susceptible (S) or a resistant (R) strain of F. hepatica. Eight weeks after infection, six lambs of each group (ST and RT) were treated with triclabendazole (10mg/kg). The other lambs were used as untreated controls (SC and RC). The parameters studied were: GLDH, gamma-GT, ELISA measuring antibodies against recombinant cathepsin-L(1) and eggs per gram faeces (epg). The lambs were slaughtered 16 weeks after infection and the number of flukes counted.The GLDH, gamma-GT levels and the OD value of the ELISA decreased as a result of the treatment in group ST. Patent infections were observed in all animals of groups SC, RT and RC. In group ST, occasionally a few eggs were found in five lambs. The percentage of flukes was 31.3 in SC and 37.6 in RC. In the treated groups ST and RT, the percentage of flukes was 0.06 and 33.6, respectively. These results corresponded to efficacies of 99.8% in the susceptible and 10.8% in the resistant strain. Since the resistant strain was isolated from a mixed cattle and sheep farm, it confirms the presence of triclabendazole resistance in the Netherlands.  相似文献   

16.
The production of muramidase-released protein (MRP), extracellular protein factor (EF) and hemolysin (suilysin) by 101 Canadian field strains of Streptococcus suis capsular type 2 is described. Most strains (72%) isolated from diseased pigs were MRP-EF- and only 1 strain was MRP+EF+. This strain was also the only 1 to produce the hemolysin. Thirteen strains (15%) were MRP+ EF- and only 3 strains were MRP* EF-. All the strains isolated from clinically healthy pigs as well as a bovine and 2 human isolates had a MRP-EF- phenotype. In addition, 7 strains (8%) had a MRPS phenotype, which had so far been described for S. suis capsular type 1. In conclusion, most Canadian field isolates of S. suis capsular type 2 tested in this study do not produce the virulence-related proteins described so far for this bacterial pathogen.  相似文献   

17.
We developed a PCR assay for the rapid and sensitive detection of virulent Streptococcus suis type 2 and highly virulent S. suis type 1 in tonsillar specimens from pigs. The PCR primers were based on the sequence of the gene encoding the EF-protein of virulent S. suis type 2 strains (MRP+EF+) and highly virulent S. suis type 1 strains (MRP(s)EF+) and of the EF protein of weakly virulent S. suis type 2 strains (MRP+EF). The latter strains give rise to larger PCR products than the virulent strains of S. suis type 1 and 2. A positive control template was included in the assay to identify false negative results. The PCR was evaluated using tonsillar specimens from herds known (or suspected) to be infected and herds without an S. suis history. The results obtained with the PCR assay were compared with the results obtained with a newly developed bacteriological examination. In this bacteriological examination we were able to identify the EF-positive strains directly in the tonsillar specimens. From the 99 tonsils examined, 48 were positive in the PCR and 51 negative. All specimens from which EF-positive S. suis strains were isolated were also positive in the PCR assay. Three samples were positive in the PCR, but negative by bacteriological examination. The results demonstrated that the PCR is a highly specific and sensitive diagnostic tool for the detection of pigs carrying virulent strains of S. suis type 2 and highly virulent strains of type 1. Application of the assay may contribute to the control of S. suis infections.  相似文献   

18.
对分离鉴定的鸽源冠状病毒PSH株纤突蛋白S1基因进行RT-PCR法扩增、克隆和序列测定分析.结果表明其S1基因由1 626个核苷酸组成,编码541个氨基酸,S蛋白的切割识别位点为精氨酸-精氨酸-苯丙氨酸-精氨酸-精氨酸(RRFRR),与常见的鸡传染性支气管炎病毒(IBV)S蛋白切割识别位点相似(RRF/SRR).该病毒与火鸡蓝冠病病毒(TCV)Gh、G1株S1基因推导的氨基酸同源性仅为24.7%、25%,而与IBV H52、H120、M41、Beau、Conn、Gray、Hotel、SH1、SH2、SH5、SH6基因推导的氨基酸同源性在75.0%~99.6%,其中与SH2、SH5的氨基酸同源性更是达到了99.6%,进一步证明该冠状病毒可能来源于IBV.  相似文献   

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