共查询到19条相似文献，搜索用时 125 毫秒
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青海省互助县某羊场藏系绵羊患病死亡,伴有腹泻、神经症状,为快速确诊发病原因,及时防控和治疗,采集病羊样品5份,通过镜检、细菌分离培养、分子生物学试验及致病性试验进行病原鉴定分析。结果证明此次病原菌为羊致病性D型魏氏梭菌,毒素基因分析显示该菌同时含有α和ε两种毒素基因,动物致病性试验结果表明该菌对昆明系小鼠具有较强的致病性,并从试验致死的小鼠中分离到与藏羊病原相一致的细菌。三种基因的遗传进化树显示,该菌具有较强的多样性。研究确定了本次藏羊致死的主要病原为D型魏氏梭菌,结果可为该羊场梭菌病的治疗和预防提供科学依据。 相似文献
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无菌采取内蒙古通辽市某羊场病死羊肠道内容物、肝脏和肺脏,进行细菌的分离培养。将从十二指肠内分离到的1株疑似致病菌株进行生化试验、小鼠致病性试验,再将其通过魏氏梭菌多重PCR试验、魏氏梭菌ELISA试验及16S rRNA PCR试验进行鉴定。将PCR产物进行测序并进行了16S rRNA基因的进化树分析。结果显示,经细菌生化试验、多重PCR试验、ELISA试验和16S rRNA试验均证实此分离株为A型产气荚膜梭菌;进化树分析显示该菌与序列号为HQ808749.1(美国)的A型产气荚膜梭菌遗传距离最近。结果表明,该羊病例所分离的致病菌为A型产气荚膜梭菌。 相似文献
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犊牛魏氏梭菌的分离鉴定及致病性研究 总被引：1，自引：0，他引：1
2017年1月,山东省某规模化奶牛场10～20日龄犊牛出现腹泻、拉血便、呼吸困难等症状,部分犊牛突然死亡。通过流行病学调查和尸体剖检,初步怀疑为魏氏梭菌感染。采集病料进行病理组织学检查、细菌分离鉴定、生化试验、16S及毒素基因检测等,证明疫情由A型魏氏梭菌所致。对死亡犊牛的肠内容物及分离菌株进行致病性研究,发现肠内容物及分离菌株具有较强的致病性。药敏试验结果显示,该菌对青霉素、头孢噻呋、恩诺沙星等表现极敏感。根据病原确诊情况及药敏试验结果,指导临床用药,及时控制了牛场疫情。 相似文献
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笔者从某猪场疑似猪魏氏梭菌病病例中分离到一株魏氏梭菌,为了研究其生物学特性及其发病机理,对分离菌进行了形态培养、生化试验、动物实验等鉴定,鉴定结果为C型魏氏梭菌.药敏试验表明本株魏氏梭菌对青霉素、磺胺药敏感性较高. 相似文献
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Isolation,Identification and Pathogenicity Research of a Strain of Clostridium perfringens from Goat
A pathogenic bacteria was isolated from a dead goat,and it was identified as goat-pathogenic type A Clostridium perfringens by differential culture,biochemical test,molecular biological technology and animal pathogenicity test.Analysis results of toxin genes showed that this bacteria contained both α and β2 toxin genes.Animal pathogenicity test result showed that this bacteria was highly pathogenic to mice,and the same bacteria was isolated from dead mice as which was isolated from the dead goat,so it was certain that type A Clostridium perfringens was the main pathogenic bacteria which caused the goat dead.The results would provide scientific basis for the treatment and prevention of the disease caused by Clostridium perfringens in this goat farm. 相似文献
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旨在获得产气荚膜梭菌β毒素（CPB）的重组突变体,并评价其毒力及免疫保护性。对已知的产气荚膜梭菌CPB编码基因进行优化设计,同时引入4个氨基酸突变位点,分别是第212位的精氨酸突变为谷氨酸,第268位的亮氨酸突变为甘氨酸,266位的酪氨酸和275位的色氨酸突变为丙氨酸。此外,在该基因5'端添加Th细胞表位（T）和鞭毛蛋白（flagellin）N末端的编码序列,经人工合成获得重组基因片段（GTFNCPBm4）。将GTFNCPBm4克隆至原核表达载体pET-30a（＋）中进行表达与纯化,获得重组蛋白rTFNCPBm4。利用Western blot方法检测rTFNCPBm4与C型产气荚膜梭菌毒素抗血清的反应性,并检测rTFNCPBm4对小鼠的毒力。随后,以rTFNCPBm4免疫家兔,按照《中华人民共和国兽药典》（2015年版）规定的方法检测家兔血清对C型产气荚膜梭菌毒素的中和抗体效价。结果表明,rTFNCPBm4主要以包涵体的形式表达且能与C型产气荚膜梭菌毒素抗血清反应。小鼠安全性试验显示,50 μg的rTFNCPBm4对小鼠仍无致死性;免疫rTFNCPBm4后,每毫升家兔二免抗血清可中和10~20个小鼠最小致死量（MLD）的C型产气荚膜梭菌毒素;1个家兔MLD的C型产气荚膜梭菌毒素攻毒后,对照组家兔4/4死亡,免疫组家兔得到了100%（4/4）的保护。以上结果说明,rTFNCPBm4在丧失毒力的同时保留了良好的免疫原性,从而为C型产气荚膜梭菌病基因工程疫苗的研制提供了重要的数据。 相似文献
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DU Jige ZHU Zhen XU Zhongqing LI Qianlin YAO Wensheng LI Qihong YIN Chunsheng YANG Liu FU Lizhi CHEN Xiaoyun LIU Ying XUE Qi 《畜牧兽医学报》1956,51(11):2794-2801
This study was conducted to obtain the Clostridium perfringens β toxin (CPB) derivative and to evaluate its virulence and immunoprotection. Based on the known sequence, four amino acid mutations, R212E, L268G, Y266A and W275A, were introduced into the gene of Clostridium perfringens CPB. Meanwhile, genes of Th cell and N-terminal of flagellin were added to 5' of the CPB gene, respectively. Then the gene GTFNCPBm4 was optimized and synthesized, and was subsequently cloned into the prokaryotic expression vector pET-30a (＋) for expression and purification to get the recombinant protein, rTFNCPBm4. Reactivity of rTFNCPBm4with antiserum of Clostridium perfringens type C crude toxins was detected by Western blot. Meanwhile, the toxicity of rTFNCPBm4 to mice was evaluated. According to the method prescribed in Chinese Veterinary Pharmacopoeia (2015), rabbits were immunized with rTFNCPBm4 to prepare antiserum and detect the neutralizing titer against Clostridium perfringens type C crude toxins. Results showed that rTFNCPBm4 was presented predominantly in an insoluble form (inclusion bodies), and it could react with the antiserum of Clostridium perfringens type C crude toxins. rTFNCPBm4 with an injection volume of 50 μg was still not fatal to mice. Sera from rabbits immunized with rTFNCPBm4can neutralize 10-20 mouse minimum lethal doses (MLD) of Clostridium perfringens type C crude toxins after twice immunization. Moreover, rabbits immunized with rTFNCPBm4 can fully resist 1 rabbit MLD of Clostridium perfringens type C crude toxins challenge, whereas all of the rabbits died (4/4) in the control groups. These data suggest that rTFNCPBm4 is a potential vaccine candidate for the subunit vaccine of Clostridium perfringens type C. 相似文献
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circRNAs在病原菌感染宿主的肠道疾病发展中具有重要的调控作用。C型产气荚膜梭菌（C.perfringens type C）是引起仔猪腹泻及相关肠道炎症的主要细菌之一,对产业造成了严重的经济损失。然而,目前有关circRNAs如何调控仔猪C型产气荚膜梭菌性腹泻的全面且系统的研究未见报道。本研究通过RNA高通量测序研究分析了C型产气荚膜梭菌感染的7日龄仔猪回肠组织circRNAs的表达谱,筛选差异表达circRNAs,并进行差异表达基因GO和KEGG功能富集分析,利用miRanda等软件预测circRNA的microRNA靶点,构建circRNA-miRNA-mRNA的互作网络,并进行qPCR验证。结果显示,在C型产气荚膜梭菌感染的处理组（TI组）和对照组（CI组）中共鉴定出3 162个circRNAs,其中差异表达circRNAs有694个,上调表达circRNAs有404个,下调表达circRNAs有290个。功能富集分析结果表明,差异表达circRNAs的亲本基因主要富集在细胞周期、TGF-β信号通路、赖氨酸降解、Wnt信号通路、T细胞受体信号通路、MAPK信号通路等,调节仔猪对产气荚膜梭菌感染的抗性反应。此外,本研究构建了与C型产气荚膜梭菌感染致仔猪腹泻相关的circRNA-miRNA-mRNA互作网络,发现8 circRNAs-5 miRNAs-12 mRNAs组成的ceRNA网络与产气荚膜梭菌感染性疾病密切相关。本试验可为深入研究circRNA调控仔猪C型产气荚膜梭菌性腹泻疾病及猪抗腹泻病品系培育提供参考。 相似文献
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本研究自陕西省富平县某规模化关中奶山羊养殖场腹泻奶山羊肛门拭子中分离到5株产气荚膜梭菌,命名为21-D-1~21-D-5。毒素基因检测表明,其均为携带etx的D型产气荚膜梭菌,且21-D-5携带食源性致病毒素基因cpe。全基因组序列测定显示,5株D型产气荚膜梭菌基因组大小、GC含量和基因数量稳定;单核苷酸多态性（SNPs）分析显示,21-D-1和21-D-2以及21-D-3和21-D-4之间的SNPs差异极低（<25）,表明其大概率属于相同的产气荚膜梭菌菌株。分离的D型产气荚膜梭菌基因组中共检测到15种毒素基因,其中,毒素基因etx在分离的D型菌株中高度保守、基因环境相似,且在菌株21-D-5中毒素基因cpe位于etx下游。此外,包括噁唑烷酮类耐药基因optrA在内的9种耐药基因也在分离株中被检测到,并且erm（A）、optrA和fexA具有共同传播的可能性。本研究为国内首次对D型产气荚膜梭菌进行全基因组序列测定分析,结果对产气荚膜梭菌疾病的防治和基因组的进一步研究具有参考价值。 相似文献
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【目的】 查明引起福建省龙岩市某豪猪养殖场致豪猪腹泻死亡的病原菌及其特征,为该病的科学防控及合理用药提供参考依据。【方法】 分离腹泻死亡豪猪的病原,并结合形态特征、生理生化试验、16S rRNA基因扩增、种系发育分群鉴定分离菌株;通过毒力基因检测、动物致病性试验及药敏试验对分离菌株的致病性和耐药性进行研究。【结果】 从发病死亡豪猪肝脏组织中分离到1株大肠杆菌,命名为Fj/Porcupine2018,该分离菌株与22株不同来源的参考菌株16S rRNA序列之间的相似度为97.4%~99.8%,其中与禽源分离株（登录号：MN022583）和人源分离株（登录号：MW881377）的序列相似度高达99.8%;种系发育分群证实该分离菌株属于B1群。毒力基因检测结果显示,该分离菌株除检出肠侵袭型大肠杆菌毒力基因EinV外,还检出papC、iroC、Afa、luxs、stx2f及ompA 6种毒力相关基因。动物致病性试验显示,该分离菌株对小鼠具有较强的致病性,小鼠在攻毒后31 h内全部死亡,且肝脏、脾脏、肺脏及肠道等器官组织均有明显的病理损伤;在对常见抗菌药物的药敏试验中,该分离菌株对大环内酯类、四环素类、喹诺酮类、磺胺类、头孢类、氨基糖苷类、青霉素类7类11种药物表现出不同程度的耐药,耐药率为25%~100%,仅对头孢曲松和头孢西叮2种药物表现敏感。【结论】 本研究分离获得了1株腹泻死亡豪猪的病原菌大肠杆菌,通过一系列生物学特性研究证实该分离菌为1株具有较强致病力且呈现多重耐药性的B1群肠侵袭型大肠杆菌,为豪猪等野生动物大肠杆菌病的防控提供了重要的参考依据。 相似文献
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PENG Xiao-bing TIAN Dong-qing LI Xu-ni PENG Guo-rui WANG Meng JIANG Yu-wen 《中国畜牧兽医》2015,42(8):1950-1955
One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared. 相似文献
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本试验通过设计并合成1对引物,PCR扩增B型产气荚膜梭菌C58-2株α毒素完整成熟肽序列,并将其插入到pGEM-T Easy载体中,构建克隆载体pGEM-T-α。对克隆载体pGEM-T-α进行EcoRⅠ和Hind Ⅲ的双酶切,将得到的1125 bp片段以正确的阅读框架定向克隆于pET-28a(+)中,然后将重组质粒转化大肠杆菌BL21(DE3)plys宿主菌中,37 ℃、1.0 mmol/L IPTG诱导该片段获得良好表达。经SDS-PAGE分析,其表达的蛋白约为46.1 ku,与预期大小一致。Western blotting结果显示,该重组α毒素蛋白可被A型产气荚膜梭菌定型血清识别,表明该重组α毒素蛋白具备与天然毒素相似的反应原性。重组α毒素蛋白在菌液上清、超声波裂解上清和超声波裂解沉淀中均有分布,且以包涵体表达为主,表明重组α毒素蛋白可同时在胞外、周质和胞浆表达。小鼠毒力试验结果表明,重组α毒素蛋白不具有毒性。毒素—抗毒素中和试验结果表明,该抗血清具有α毒素特异性。以重组α毒素蛋白作为抗原免疫家兔制备血清,效价测定结果表明每1 mL重组α毒素蛋白抗血清可以中和100 MLD的A型毒素。 相似文献
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为探究羊源尸毒梭菌的生物学基本特性，进行生理生化分析、16S rDNA基因扩增、动物回感试验、耐药表型和耐药基因检测等试验。结果显示，菌株LJH-1为革兰阳性菌。PCR扩增获得长度为1 023 bp的片段。LJH-1与尸毒梭菌C4菌株聚类，从而判定LJH-1为尸毒梭菌。试验组小鼠全部死亡。小鼠肝中央静脉充盈大量红细胞，炎性细胞浸润，肾间质血管充血，肠黏膜上皮细胞脱落。LJH-1对头孢噻肟、氨苄西林等敏感，对头孢拉定、阿莫西林等耐药，并检出ant（3″）-Ia、aac（3）-IIa、aph（3'）-IIa、Sul2、Sul3和TEM耐药基因。这为该细菌性疾病的检测和防治提供了科学资料。 相似文献