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1.
为了解引起山羊皮下脓肿的主要病原,本研究从四川乐至县、金堂县和蓬溪县3个山羊养殖场无菌采集患皮下脓肿病羊脓液样本19份,通过细菌分离培养、生化试验及特异性PCR方法对其病原进行检测和鉴定,并通过药敏试验测定分离病原的药物敏感性。结果显示,PCR扩增法共检测出病原菌19株,其中伪结核棒状杆菌、金黄色葡萄球菌及化脓隐秘杆菌分离率分别为57.9%(11/19)、26.3%(5/19)及15.8%(3/19);采用细菌分离培养法分离到3种形态不同的病原菌共计17株,其中革兰氏阳性短杆菌经鉴定为伪结核棒状杆菌,分离率57.9%(11/19),革兰氏阳性球菌经鉴定为金黄色葡萄球菌,分离率为21.1%(4/19),革兰氏阳性杆菌经鉴定为化脓隐秘杆菌,分离率为10.5%(2/19),PCR鉴定较细菌分离培养法灵敏度更高。3种病原菌对丁胺卡那、氟苯尼考等多种药物敏感。结果表明,伪结核棒状杆菌是引起该地区山羊皮下脓肿的主要病原,金黄色葡萄球菌和化脓隐秘杆菌同样可引起山羊皮下脓肿,特异性PCR方法较细菌分离培养更为高效、准确。本研究结果可为后续进一步分析其生物学特性,以及四川地区山羊皮下脓肿的综合性防制提供依据。  相似文献   

2.
为了分析引起湖南省永州市山羊皮下脓肿的病原种类,从3个规模化养殖场的患病山羊采集病料,同时采用细菌分离鉴定和PCR检测两种方法,对引物山羊皮下脓肿的主要病原进行鉴定,并对分离菌株对药物的敏感性进行了评价。结果显示,主要病原为伪结核棒状杆菌和金黄色葡萄球菌,且发现两种病原混合感染现象,未发现其他病原。伪结核棒状杆菌分离株对四环素和头孢噻吩敏感性最高;金黄色葡萄球菌分离株对苯唑西林、丁胺卡那和头孢噻吩的敏感性最高。研究结果为该地区山羊皮下脓肿的综合性防制提供了依据。  相似文献   

3.
旨在对重庆地区山羊体表型淋巴结炎病原菌进行分离鉴定,并建立相应检测方法。无菌采集山羊患病部位脓液,进行病原菌分离鉴定,依据菌株16SRNA基因序列构建系统进化发育树,同时设计特异性引物,建立PCR检测方法。结果表明,从病料中主要分离到3种病原菌,分别为伪结核棒状杆菌、化脓隐秘杆菌和金黄色葡萄球菌;依据伪结核棒状杆菌磷脂酶D、化脓隐秘杆菌16SRNA和金黄色葡萄球菌nuc保守核苷酸序列设计引物,建立的多重PCR检测方法,可以从3种菌株混合基因组中扩增出大小分别为507、1 378和278bp的对应菌株特异性序列,该方法最低检出浓度为103cfu·mL-1;对采集的80份临床样品进行多重PCR检测,发现伪结核棒状杆菌与金黄色葡萄球菌混合感染检出率为12.5%,伪结核棒状杆菌与化脓隐秘杆菌混合感染检出率为3.75%,三种菌混合感染检出率为7.5%。成功分离到山羊淋巴结炎的3种主要病原菌,并建立多重PCR检测方法,为该病的防控及治病机制研究奠定了基础。  相似文献   

4.
伪结核棒状杆菌的药物敏感性试验   总被引:1,自引:1,他引:0  
山羊皮下、淋巴脓肿病可由多种病原菌引起,经采集病料分离鉴定,其主要病原主要是伪结核棒状杆菌,该病是一种人畜共患的慢性传染病,也是世界上公认难以防治的传染病之一。该病主要引起患病羊发生一处或多处脓肿,消瘦、生产性能低下,严重者发生死亡,该病一旦侵入羊群则很难彻底消除。近年来,随着养羊业的发展,大量从省内外引进种羊,加之商品羊的频繁流动,我省山羊群发生皮下、淋巴脓肿病较普遍,给养羊业发展造成了较大的危害。本试验在本省分离鉴定的伪结核棒状杆菌中,选取不同地区、不同品种山羊病料分离的5株伪结核棒状杆菌,采用圆滤纸片法,…  相似文献   

5.
波尔山羊干酪样淋巴结炎的病原分离与初步鉴定   总被引:1,自引:0,他引:1  
某波尔山羊场羊群患有以淋巴结肿 大、化脓为主要特征的疾病,怀疑为山羊干酪 样淋巴结炎。从一患病羊只的淋巴结脓肿中 分离得到1株病原菌,对该病原菌分别进行 病原形态、培养特性、生化特性以及致病性等 方面的鉴定,初步确定为致病性假结核棒状 杆菌。  相似文献   

6.
为了确诊自然感染疑似化脓隐秘杆菌引起的山羊淋巴结炎病例病原,进行了病原菌分离培养、染色镜检、生化试验和16SrRNA鉴定、动物试验、病理组织切片观察。该病原菌染色镜检为革兰阳性的丝状、杆状或棒状细菌,VITEK2Compact全自动微生物分析系统鉴定为化脓隐秘杆菌,可信度为88%,16SrRNA鉴定与化脓隐秘杆菌相似性高于99%,判定该病原菌为化脓隐秘杆菌;药敏试验结果显示该菌对β-内酰胺类药物和氟喹诺酮类药物有高度的敏感性,对磺胺类药物有一定的耐药性;病理切片观察到病羊心肌纤维变性,肺和肾小管管腔内有浆液-纤维素性变性,肝有炎性细胞浸润形成肝炎,肩前淋巴结脂肪变性,脾呈浆液-化脓性炎症。结果表明,本次致使山羊淋巴结炎的病原菌为化脓隐秘杆菌,且病羊病理组织学变化为主要脏器的化脓性炎症。  相似文献   

7.
贵州山羊饲养场常发生皮下、淋巴脓肿病,该病不分品种、性别、年龄,多发于春秋两季,带病羊场占调查数的80%以上,群内发病率为6%左右(3%~30%).场内一旦发生本病,则经年不断,难以根除,给养羊业造成不小的经济损失.我们在对山羊皮下、淋巴脓肿病作流行病学调查时发现,该病主要由创伤感染引起,可直接接触感染,也会间接接触感染,还会因人为因素(如消毒不严的预防注射等)引起.为弄清病原,采集病料进行分离鉴定.结果表明,由于产生的原因不同,病原也不尽一致,但其主要病原为人畜共患的伪结核棒状杆菌(Corynebacterium pseudotuberculosis),其次有松鼠葡萄球菌及链球菌、四联球菌等.现将其主要病原"伪结核棒状杆菌"的分离鉴定报道如下:  相似文献   

8.
参照已发表的伪结核棒状杆菌16S rRNA序列,设计合成了1对引物,用此引物进行PCR扩增,建立伪结核棒状杆菌的PCR检测方法。用此方法对羊链球菌、马尔他布鲁氏菌、结核分支杆菌进行PCR扩增结果对比,确证该方法的特异性。同时,用建立的PCR方法检测病料,检测结果与病原菌的分离鉴定结果进行比较,以确证其准确性。结果表明,分离菌株与参考株的同源性分别为96.8%和97.1%;检测17份病料,阳性率为70.6%,与病原菌的分离鉴定结果完全一致。  相似文献   

9.
通过对重庆某羊场送检病死羊只进行病原菌的分离培养、形态学鉴定、生化鉴定、药敏试验、致病性试验及16S r RNA序列分析。结果从病变组织中分离到1株革兰阳性短棒状杆菌,其对多种抗生素敏感,能够引起小鼠肝脏和肺脏发生病变,PCR扩增出长为1 434 bp的目的序列,已被Gen Bank数据库收录,序列号为KJ842125,将该序列与NCBI中公布的序列进行比对,发现所得序列与化脓隐秘杆菌(Arcanobacterium pyogenes,登陆编号为HQ712123)同源性高达98%,将该序列与其相关性较高菌株的16S r RNA构建系统发育进化树,与化脓隐秘杆菌聚为一簇。本研究为化脓隐秘杆菌致病机理、诊断技术和疾病控制奠定了基础。  相似文献   

10.
为探明贵州省某猪场引起猪体表脓肿的原因,本研究对该猪场脓肿部位的脓汁进行细菌分离培养,并对分离所得的细菌进行革兰氏染色镜检、生化试验、药敏试验、16S rDNA序列分子分析及动物感染试验。结果显示,从脓汁中成功分离到了3株菌落形态不一的菌株,分别为金黄色葡萄球菌、化脓隐秘杆菌、停乳链球菌类马亚种,根据分离地点和时间将其分别命名为GZGP2018-1、GZGP2018-2和GZGP2018-3;GZGP2018-1菌株与NCBI上金黄色葡萄球菌的同源性高达99.9%,GZGP2018-2菌株与NCBI上化脓隐秘杆菌的同源性高达99.9%,GZGP2018-3菌株与NCBI上停乳链球菌类马亚种的同源性高达100%;3株分离菌株对头孢拉定、环丙沙星、磺胺间甲氧嘧啶和氟苯尼考较敏感,对青霉素类药物和红霉素耐药;3株分离菌株对试验小鼠均具有致死性。本研究为该猪场猪体表脓肿的发病原因、实验室诊断方法及日常防控提供了理论参考。  相似文献   

11.
Accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. Here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time PCR amplification of 16S rRNA gene fragment, spanning the variable region V5 and V6 was performed with a resulting amplicon of 290bp. First, a library was generated of melt curves of 9 common pathogens that are implicated in bovine mastitis. Six of the isolates, Escherichia coli, Streptococcus agalactiae, Klebsiella pneumoniae, Streptococcus uberis, Staphylococcus aureus and Mycoplasma bovis, were type strains while the other 3, Arcanobacterium pyogenes, Corynebacterium bovis and Streptococcus dysgalactiae, were bovine mastitis field isolates. Four of the type strains, E. coli, S. agalactiae, K. pneumoniae and S. aureus, were found to be of human origin, while the other 3 type strains were isolated from bovine infections. Secondly, the melt curves and corresponding amplicon sequences of A. pyogenes, E. coli, S. agalactiae, S. dysgalactiae, K. pneumoniae, S. uberis and S. aureus were compared with 10 bovine mastitis field isolates of each pathogen. Based on the distinct differences in melt curves and sequences between human and bovine isolates of E. coli and K. pneumoniae, it was deemed necessary to select a set of bovine strains for these pathogens to be used as reference strains in the HRMA. Next, the HRMA was validated by three interpreters analyzing the differential clustering pattern of melt curves of 60 bacterial cultures obtained from mastitis milk samples. The three test interpreters were blinded to the culture and sequencing results of the isolates. Overall accuracy of the validation assay was 95% as there was difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of S. uberis. The present study revealed that broad-range real-time PCR with HRMA can be used as a powerful, fast and low-cost tool for the differentiation of clinically important bacterial mastitis pathogens.  相似文献   

12.
The aim of the study was to identify the species of microorganisms isolated from the uterus of healthy cows (control group) and cows affected with puerperal metritis (PM) before and after an experimental therapy with an immunomodulator and antibiotics versus commonly applied methods (antibiotic + beta-blocker or antibiotic + PGF2alpha). Examinations were carried out on 110 cows with PM in three farms with similar system of rearing and nutrition. The control group consisted of 21 cows without postpartum disturbances. Smears from the uterus were taken before treatment and then at 21st day of observation. Escherichia coli and other species of Enterobacteriaceae family were isolated from 48.2% of PM cows and 47.6% of healthy cows. The degree of Arcanobacterium pyogenes infection was statistically lower in healthy than in sick cows (9.5% versus 30.0%). Streptococcus sp. was isolated from 13.6% of PM cows and from 16% of control ones. Staphylococci were isolated from 10% of PM and from 30% of control cows. Other bacteria species were isolated from about 10% of the examined cows. The best elimination of infections (66%) was noted in cows treated with the intrauterine antibiotic in combination with PGF2alpha i.m. injection. Examination showed that species of bacteria in the postparturient uterus were similar in healthy and sick cows. However, Arcanobacterium pyogenes was isolated 3 times more often from the sick animals. This pathogen was identified in 11 cows out of 19 (57.9%) culled subsequently because of infertility after the metritis puerperalis was clinically cured.  相似文献   

13.
金黄色葡萄球菌是引起人和动物感染及食物中毒的重要病原体之一。为了解四川省山羊金黄色葡萄球菌的健康带菌情况及其耐药性,采集了省内4个养殖场共32只山羊的鼻腔棉拭子样本,进行金黄色葡萄球菌的分离、鉴定及药敏试验。共从11只羊的鼻腔棉拭子样本中分离出金黄色葡萄球菌,检出率为34.4%。对临床分离株进行了苯唑西林、头孢西丁、万古霉素等11种抗菌药物的药敏试验,结果表明,分离株对氨苄西林的耐药率为72.7%,对其它抗菌药物没有耐药性,未发现耐甲氧西林金黄色葡萄球菌(MRSA)。本研究结果表明,四川地区山羊的金黄色葡萄球菌的带菌情况较普遍,因此,应该规范屠宰过程的卫生操作,并进行严格监控,减少金黄色葡萄球菌的污染。  相似文献   

14.
从天津地区某猪场病猪肺脏脓肿中分离到一株具有β溶血特性的细菌,经形态学与生化特性分析,初步确定为化脓隐秘杆菌(Arcanobacterium pyogenes),并命名为TJjh1913.利用PCR扩增16S rRNA基因,经Blast序列分析,其序列与GenBank数据库中猪源化脓隐秘杆菌的核苷酸序列同源性高达99....  相似文献   

15.
One hundred and one pig abscesses of different localisation were examined. Bacteria were re-covered from all abscesses. In 6’8 abscesses a mixed aerobic and anaerobic flora was found. In 22 abscesses solely aerobes and in 11 solely anaerobes were found. Gorynebaoterium pyogenes and Fuso-bacterium necrophorum were found in 61 and 45 abscesses, respectively. Hemagglutination litres of G. pyogenes strains were higher with pig erythrocytes than with sheep erythrocytes. Three of 4 isolates of Staphylococcus aureus produced enterotoxin type A.Key words: pig abscess, Corynebacterium pyogenes, Fuso-bacterium necrophorum, hemagglutination  相似文献   

16.
为检测江苏省某奶牛场中乳房炎是否为金黄色葡萄球菌所致,采取4份患病乳汁样本进行试验分离和鉴定,并针对金黄色葡萄球菌采用分离后鉴定,结果显示,分离到的2株致病菌为金黄色葡萄球菌,其他菌株还包括大肠杆菌、肠球菌等,为多种致病病原菌同时引起,主要为金黄色葡萄球菌感染。  相似文献   

17.
为指导江苏地区奶牛场奶牛乳房炎的临床用药,本研究对2015-2018年江苏地区部分奶牛场的牛奶样品进行细菌分离,使用基质辅助激光解析电离飞行时间质谱(MALDI-TOFMS)方法和纸片法对分离菌株进行鉴定和药物敏感性分析,对分离得到的葡萄球菌进行了葡萄球菌D环实验,并且使用PCR技术检测了D环实验阳性菌株携带的耐药基因。结果显示:218株细菌鉴定为葡萄菌属细菌,其中金黄色葡萄球菌58株,凝固酶阴性葡萄球菌(CNS)160株;药敏试验表明,能耐三种以上药物的菌株占64.22%,耐药率最高的药物为青霉素,有82.11%分离菌株耐药,耐药率最低为头孢西丁,无分离菌株对其耐药;共有6株葡萄球菌分离株D环实验阳性,同时PCR结果显示,耐药基因D环实验阳性菌株携带的均为ermA基因。本研究为临床用药与研究菌株流行趋势提供可靠数据,对临床选择合适药物和奶牛乳房炎监测有重要意义。  相似文献   

18.
猪化脓隐秘杆菌的分离与鉴定   总被引:3,自引:1,他引:2  
从一例肝、肺化脓性结节,脾出血性肿大的病猪体内分离到一株革兰氏阳性、细长、形态不规则的杆菌,该菌在TSA平板、普通绵羊血琼脂、巧克力平板上生长缓慢,在普通绵羊血琼脂上呈α-溶血。对该菌的16S rDNA进行PCR扩增、测序、比较,结果显示该菌与化脓隐秘杆菌的16S rDNA有99%的同源性,进一步用API生化鉴定,结果也与其相符。该菌在24~52 h内100%致死小白鼠,腹腔接种新西兰兔,接种兔72 h内死亡,表明该菌为致病性细菌。  相似文献   

19.
The population under study included young calves with pneumonia (group A, n = 13) and their controls (group B, n = 9), as well as older calves from which the lungs with (group C, n = 90) or without (group D, n = 10) lesions were collected after slaughter. Arcanobacterium pyogenes was the organism most commonly isolated from calves in group A (46%), followed by Haemophilus somnus (23%), Mannheimia haemolytica (15%), Streptococcus suis and Pasteurella multocida (7.7% each). Only S. suis (22%) and P. multocida (11%) were found in group B. P. multocida was isolated from 32% group C calves, H. somnus from 11%, A. pyogenes from 7.8%, M. haemolytica from 2.2% and S. suis from 1.1%. No specific pathogens were isolated in group D. Prevalence of Mycoplasma bovis infection was 69% in group A and 37% in group C. Ninety-eight strains were tested for resistAnce to antibiotics. Resistance to penicillin and ampicillin was present only in M. haemolytica (46%). High percentages of resistant strains were observed for streptomycin (48-100%), tetracycline (15-43%), sulfonamides alone (14-100%) or in combination with trimethoprim (0-100%). Therapeutic approaches to bacterial calf pneumonia in the area under study should be modified according to the isolated bacterial population, the observed antimicrobial resistances and the growing importance of Mycoplasma bovis.  相似文献   

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