鸡源肠球菌的分离与鉴定

本研究对１９５份死鸡胚和１日龄弱死雏鸡的病样,进行了链球菌和肠球菌的分离,并对所分离菌的某些生物学特性进行试验研究。结果共分离到１２０株目的菌,随机取４５株进行鉴定,皆属肠球菌,其中４３株能确定为肠球菌属中的已知菌种。     

狐狸源小肠肠球菌分离鉴定与药敏试验

为确定某养殖场患病死亡狐狸的病原菌,本试验对病死狐狸进行剖检和细菌分离培养、生化鉴定以及16S rDNA鉴定、动物致病性试验和药敏试验。结果显示,分离菌株为革兰阳性球菌,经过生化鉴定和16S rDNA序列分析,确定分离菌株为小肠肠球菌,并命名为E.hirae QHD-1;动物回归试验结果显示,分离菌株E.hirae QHD-1对小鼠具有较强的致病性,LD_(50)为1.26×10~4 CFU;药敏试验结果显示,分离菌株E.hirae QHD-1对氨苄西林和恩诺沙星等药物敏感,对替米考星、阿米卡星等耐药;毒力因子检测结果显示,该分离株具有较强的致病力。毛皮动物小肠肠球菌的感染病例尚未见有报道,本试验丰富了狐狸病原菌相关的研究,并为防控该菌引起的疾病提供了重要参考资料。     

一株黑叶猴源鸟肠球菌的分离鉴定及耐药分析

黑叶猴是我国一级重点保护动物,2018年11月贵州某动物园中黑叶猴出现死亡,为调查其原因,本研究采集病死黑叶猴的肺脏样品进行细菌分离纯化,并对分离菌进行革兰氏染色镜检、生化鉴定、16SrRNA基因序列鉴定、小鼠致病性试验及分离株的药敏试验和耐药基因检测.结果显示:经形态观察、培养特性鉴定以及16SrRNA的PCR鉴定,...     

1株羊源致病性屎肠球菌的分离与鉴定

鄂尔多斯市某羊场发生羊不明原因死亡的疫情,为了寻找病因并制定治疗方案和防治措施,本研究通过临床检查和实验室病理及分子生物学诊断方法确诊并提供治疗方案。试验以病死羊为试验材料,首先对其进行临床检查,包括尸体剖检和病理组织学采样。随后,采用常规无菌操作方法取回其脑组织至实验室,进行触片、染色镜检和分离细菌。病死羊剖检发现,肺脏淤血实变、心外膜点状出血、肾脏质地变软如泥、脑严重水肿等。选取脑组织制备病理切片,在镜下可观察到大脑皮质中性粒细胞浸润及中性粒细胞性血管管套现象;深层脑组织进行触片染色后,在镜下可见单个或成对排列的革兰氏阳性球菌,并分离出1株可疑细菌,命名为NEF1;采用Mega 6.0生物软件依据该分离株的16S rRNA基因而绘制的遗传进化树分析结果显示,分离株NEF1与屎肠球菌的国内分离株（MT197247.1）和伊朗分离株（KM495939.1）聚类在同一分支上,而与GenBank中其他屎肠球菌参考菌株遗传距离较远;药物敏感性试验结果表明,分离株NEF1只对环丙沙星呈现中度敏感,而对青霉素等其他选定的抗生素类药物均呈现明显的耐药性。本试验从病死羊脑组织中成功分离并鉴定了1株屎肠球菌NEF1,同时通过药物敏感性试验方法筛选出来的敏感药物有效控制了该病在发病羊场的进一步蔓延,为该地区羊细菌性疾病的有效防治提供了行之有效的试验数据。     

一株母猪产道益生性粪肠球菌的分离鉴定

产道作为母猪生殖系统的重要器官组织,与繁殖性能密切相关.本试验无菌采集江西省某养殖场窝产14头母猪产道中段阴道分泌物进行传统细菌分离培养,初步筛选出4株细菌,挑选其中一株通过生化反应和16S rDNA序列分析方法进行鉴定,同时进行抗生素抵抗能力试验和牛津杯法体外抑制试验.生化鉴定结果表明该菌可降解果糖、乳糖、半乳糖等,...     

动物源性肠球菌的分离鉴定及耐药性分析

了解肠球菌的耐药性,对控制该菌引起的感染,研制新一代抗菌药物具有重要价值。本试验从84种样品中分离鉴定出69株肠球菌,通过抗生素耐药性试验,发现肠球菌对多数抗生素表现出不同程度的耐药性,如青霉素耐药达52.2%、四环素达71.0%、万古霉素达85.5%。本文对肠球菌的分离与鉴定,药敏试验及风险分析三个方面作了分析和阐述。     

动物微生态制剂猪肠源肠球菌的分离与鉴定

肠球菌属细菌在自然界中分布很广,也是人和动物肠道微生物群落中的主要细菌。利用无毒害、兼性厌氧的肠球菌(Enterococcus),配合其他益生菌制成的动物微生态制剂(Animal Micro-ecological Agent,AMEA),可替代抗生素预防和治疗动物的消化道疾病。试验的目的是从基本自然生长的健     

致羔羊脑炎粪肠球菌的分离鉴定

为确定某羊场导致羔羊脑炎的病原,对从3只病死羔羊脑、肝、脾、淋巴结等组织中分离到革兰阳性球菌,采用生化试验、PCR检测、16SrRNA序列分析、进化树分析、药物敏感性及小鼠致病性测定进行鉴定。结果表明,分离菌符合粪肠球菌的生物学特性;对小鼠致病性较强;对庆大霉素、红霉素、克林霉素、万古霉素等有较强的耐药性,对呋喃妥因极敏感,对青霉素、氨苄西林、环丙沙星等敏感;应用DNA Man软件对分离株WS1、WS2、WS3、WS4、WS5 16SrRNA基因序列进行同源性比对,结果与GenBank数据库中粪肠球菌(NR-040789.1)的同源性分别为99.2%、99.0%、98.5%、98.5%和98.8%;经MEGA4.0软件分析的分子系统进化树(N-J法)表明5株分离菌与粪肠球菌之间的亲缘性最近。说明该羊场导致羔羊脑炎并死亡的病原为粪肠球菌。     

牦牛源肠球菌的分离鉴定及耐药性分析

试验结合革兰氏染色及16S rRNA分子鉴定,对分离自红原县的17份牦牛粪便样中的肠球菌进行鉴定。结果显示,通过细菌分离纯化及PCR扩增,从牦牛粪便样品中分离出8株疑似肠球菌,分离菌的扩增产物经凝胶电泳后均产生1 500 bp特异性条带。16S rRNA测序结果显示,8株疑似肠球菌中,6株为粪肠球菌,2株为屎肠球菌。同源性比对分析显示,粪肠球菌与参考序列同源性为99.7%~100.0%,屎肠球菌与参考序列同源性为98.2%~99.2%,说明牦牛源肠球菌在遗传进化过程中高度保守。运用K-B纸片法进行药敏试验,结果显示,分离株对氨基糖苷类抗生素耐受性较高,2株屎肠球菌中,11-1-2菌株表现为5重耐药,且该菌株耐万古霉素,粪肠球菌主要表现为3重耐药,药敏结果提示牦牛源肠球菌耐药较严重,应引起高度重视。     

一株致病性粪肠球菌的分离与鉴定

从1羽发病雏鸡中分离到1株溶血性革兰氏阳性球菌。初步分离鉴定后经攻毒试验能够造成小白鼠和雏鸡死亡,具有致病性。药敏试验表明,该菌对多种药物均表现耐药性,仅对庆大霉素、青霉素、氧氟沙星敏感。通过BLAST软件分析该菌株的16S rRNA序列,确定该致病性细菌为粪肠球菌。     

致鹅败血症粪肠球菌的分离鉴定

为确定导致通辽某鹅场雏鹅败血症的病原,本研究将从病鹅心、肝、淋巴结和脑组织液中分离的革兰氏阳性球菌纯化培养,进行形态学和生长耐受性观察,菌属区别、分群和菌种鉴定试验,16S rDNA检测、系统进化分析和致病性试验。结果表明,分离菌株(HQ831431)为中等致病力粪肠球菌,具有β型溶血特性,可致小鼠急性败血症。分离菌株与参考菌株ATCC29212及国内外分离的其他15株粪肠球菌16S rDNA的同源性均在99.0%以上,位于系统发育树的同一分支。     

Isolation and identification of Mycobacterium avium subspecies silvaticum from a horse

Routine cultivation methods are able to distinguish between isolates of the Mycobacterium avium and the Mycobacterium tuberculosis complex. However, molecular tools are needed to further identify the several subspecies in the M. avium complex, especially for the subspecies avium and silvaticum. A rapid technique using HhaI restriction digestion of a 349 bp amplification product of the 85B antigen (α-antigen) gene was used for the identification of M. avium subsp. silvaticum in a three-year-old gelding presenting with caseous, necrotizing, granulomatous lesions. The result was confirmed by sequencing of the 85B antigen gene.     

Isolation and characterization of Bordetella avium plasmids

Experiments were conducted to study the plasmids of Bordetella avium, B. avium-like, and B. bronchiseptica isolates from turkeys and the plasmids of the Art-Vax commercial vaccine strain. Plasmids were observed in 6 of 20 B. avium isolates, in 6 of 20 B. avium-like isolates, in all 5 B. bronchiseptica isolates, and in the Art-Vax strain. Plasmids of B. avium correlated with resistance to antibiotics but not with pathogenicity, hemagglutination of guinea pig erythrocytes, or expression of pili.     

Isolation of Bordetella avium from poultry

Four Australian isolates of Gram-negative nonfermentative bacteria obtained from poultry were compared with reference strains of Bordetella avium, Bordetella bronchiseptica and Alcaligenes faecalis. The Australian isolates were identified as Bordetella avium. A routine procedure for the identification of this recently recognised poultry pathogen is described.     

青贮料中肠球菌的分离与筛选

本试验从青贮料中分离肠球菌并对分离菌株进行初步鉴定与筛选,通过正交试验设计研究不同温度、初始pH和瘤胃液含量对3株分离菌生长的影响,筛选出适合作为奶牛饲用微生物的菌株。结果表明,3株分离菌的最佳培养条件相同为：培养基含葡萄糖2%、蛋白胨1%、瘤胃液20%、初始pH为5.8,接种量为3%,装液量为40 ml, 36℃培养24 h。 在该条件下,3株分离菌的生物量分别为1.9907、1.2963和1.2752。选择生物量最高的1号菌送中科院微生物所鉴定,结果为屎肠球菌。     

Isolation and Identification of Enterococcus faecalis from Fur-bearing Animals

In order to obtain Enterococcus faecalis from fur animals and evaluate its prebiotic properties,in this study,Enterococcus faecalis was isolated from the feces of healthy adult fur-bearing animals (mink,fox,raccoon dog),identified by morphological observation,biochemical test and 16S rRNA sequence analysis.The growth curve,acid production capacity and antibiotic sensitivity of the Enterococcus faecalis isolates were measured to evaluate their probiotic properties.Some strains were selected to determine their tolerance to temperature,artificial gastric juice and artificial bile salt.The results showed that five strains were Gram-positive,and their biochemical characteristics were basically consistent with the standard strains of Enterococcus faecalis,and they were identified as Enterococcus faecalis by 16S rRNA sequence analysis.The five strains all entered the logarithmic phase at 2 h after culture,and entered the stable phase at 8-10 h,and had weak acid production capacity.The resistance rate of the isolates to tetracycline and levofloxacin was 100%,followed by penicillin (80%),erythromycin (80%),gentamicin (80%) and chloramphenicol (40%).All the isolates were sensitive to ampicillin and vancomycin.Enterococcus faecalis from mink,fox and raccoon dog had strong tolerance to temperature below 60 ℃,artificial gastric juice with pH>3.0 and 0.3%-0.5% concentration of bile salt,but poor tolerance to temperature above 70 ℃,and artificial gastric juice with pH<3.0.In conclusion,five strains of Enterococcus faecalis from fur animals (mink,fox,raccoon dog) were obtained in this study.The isolated strains propagated rapidly,which were suitable for colonization and played a prebiotic role in fur animals' intestines,and had good prebiotic characteristics and stress resistance.They could be used as candidate strains for animal microbiological agents for further study.     

毛皮动物源粪肠球菌的分离鉴定

为获得毛皮动物源粪肠球菌,并评价其益生特性,本研究从健康成年毛皮动物（水貂、狐狸、貉）粪便中分离粪肠球菌,通过形态学观察、生化试验和16S rRNA序列分析等方法进行种属鉴定。测定分离菌株生长曲线、产酸能力、抗菌药敏感性,并挑取部分菌株测定其对于温度、人工胃液、人工胆盐的耐受能力。结果表明,分离菌株中共5株为革兰氏阳性菌,生化特性与粪肠球菌标准株基本相符,且经16S rRNA序列分析鉴定为粪肠球菌。5株菌均于培养后2 h进入对数期,8~10 h进入稳定期,且具有弱产酸能力。分离菌株对四环素、左氟沙星耐药性强,耐药率为100%,其次为青霉素（80%）、红霉素（80%）、庆大霉素（80%）、氯霉素（40%）,对氨苄西林和万古霉素敏感。水貂、狐狸和貉源的粪肠球菌对于60 ℃以下温度处理、pH>3.0的人工胃液、0.3%~0.5%浓度的胆盐耐受能力较强,对70 ℃以上高温和pH<3.0的人工胃液耐受性差。综上所述,本研究共获得5株毛皮动物源（水貂、狐狸、貉）粪肠球菌,分离菌株繁殖较快,适合在毛皮动物肠道中定植并发挥益生作用,且具有较好的益生特性和抗逆性,可作为动物用微生态制剂的候选菌种进一步研究。