乌鲁木齐市宠物源粪肠球菌和屎肠球菌耐药性及耐药基因检测

为了解乌鲁木齐市宠物源粪肠球菌和屎肠球菌对临床常用抗菌药物的耐药性及其耐药基因携带情况,在乌鲁木齐市多家宠物医院、警犬基地、犬养殖基地采集宠物(犬和猫)直肠粪样449份进行粪肠球菌和屎肠球菌的分离鉴定,通过琼脂稀释法测定10种抗菌药物的最小抑菌浓度,利用PCR方法检测11种耐药基因.结果分离鉴定出宠物源粪肠球菌和屎肠球...     

鸡源和兔源屎肠球菌的耐药性及相关基因分析

2020年4月,山东省某鸽场及某兔场发生腹泻情况,发病率较高.试验从病鸽及病兔肝脏中各分离到一株革兰氏阳性球菌,通过16S rRNA测序鉴定均为屎肠球菌.两株分离菌均未检测到cylA、gelE、ace等8种毒力基因.药敏试验和耐药基因结果显示,两个菌株都对β-内酰胺类耐药,耐药基因检测结果也一致.这些结果提示屎肠球菌或...     

一株羊源致病性屎肠球菌的分离鉴定及耐药性分析

试验从甘肃肃南县某羊场死亡羊中分离出疑似某球菌菌株XCP,为进一步确定该致病菌株及其耐药性情况,进行了病原菌分离、革兰氏染色、生化试验、16S rRNA PCR扩增鉴定、小鼠致病力试验、药敏试验、毒力基因和耐药基因扩增试验研究。结果发现,该菌革兰氏染色阳性,高盐、蜜二糖、蔗糖等生化反应阳性,VP、马尿酸盐、阿拉伯糖等生化反应阴性;16S rRNA基因PCR扩增后,进行BLAST比对,结果与GenBank中的屎肠球菌16S rRNA基因序列同源性为100%;小鼠致病力试验发现,该菌株具有很强的致病性;该菌株对β-内酰胺类抗生素苯唑西林、青霉素G,喹诺酮类抗生素诺氟沙星、环丙沙星、左氟沙星,以及四环素等高度耐药,对红霉素、万古霉素、克林霉素等抗生素敏感;毒力基因Asal、cylA、acm和耐药基因TetM、ant（6）-Ⅰ、aac（6'）-aph（2"）、ermB扩增均为阳性。结果表明,该菌为屎肠球菌（Enterococcus faecium）,具有较强的致病性和耐药性,可能与毒力基因和耐药基因的高阳性率有关。     

一株鸡源屎肠球菌的分离及鉴定

为丰富可用作禽用微生态制剂的肠道益生菌株,研究选择7日龄健康雏鸡肠道内容物,利用KF链球菌琼脂培养基分离培养目标分离菌株,通过形态特征和培养特性、生理生化及16S r DNA分子鉴定法,最终确定1株分离菌为屎肠球菌,命名为HEW-A527,为后续禽类益生菌的益生功能及微生态制剂研究提供理论依据。     

屎肠球菌益生作用研究概述

屎肠球菌是一种益生菌,可以通过吸收动物体内的养分来维持体内肠道菌群的平衡,同时也可以调节动物小肠绒毛的发育,增强其抗病能力等.本文对屎肠球菌的生物学特性、调节胃肠道菌群作用、营养调控作用、益生作用机理、作为饲料添加剂的独特优势以及在药品和食品中的应用等进行了综述,以期为广大同行提供有益参考.     

屎肠球菌新序列型ST989菌株的分离鉴定及多重耐药性分析

试验从青海民和县某羊场致病性感染半月龄羔羊中分离出革兰氏阳性菌,感染羔羊临床症状有精神萎靡,采食减少,发病1~2 h后死亡。两只濒死羔羊剖检结果发现,其中一只出现肝脏肿大及肝包膜脱落,其余脏器无明显病理变化;另一只各脏器均未发现病理变化。分离病料,革兰氏染色发现,从病料中分离到的细菌均为革兰氏阳性球菌。对分离菌株进行Tuf基因PCR扩增,BLASTn比对结果显示与屎肠球菌AUS0085核苷酸序列相似性为99.8%,据此初步确定这些菌株为屎肠球菌。药敏试验结果表明,这些菌株可分为两类,第1类对β-内酰胺类、氨基糖苷类、大环内酯类、四环素类、喹诺酮类、利福霉素类抗生素及林可霉素和克林霉素耐药,对氯霉素和万古霉素敏感;第2类对β-内酰胺类、氨基糖苷类、四环素类、喹诺酮类、利福霉素类抗生素耐药,对红霉素和罗红霉素中度耐药,但对克拉霉素、阿奇霉素、克林霉素、林可霉素、氯霉素和万古霉素敏感。对第1类耐药性代表菌株lby1和第2类耐药性代表菌株lbg3进行多位点序列分析(MLST),结果发现这些菌株为一个屎肠球菌的新序列型,为ST989。系统进化树结果显示屎肠球菌lbg3与屎肠球菌ST468亲缘关系最近,致病力试验结果表明屎肠球菌lby1和lbg3均具有致病性。     

屎肠球菌自溶的基因表达差异分析

为探讨屎肠球菌自溶状态下基因表达变化,利用高通量测序技术进行RNA-Seq分析。差异基因分析显示,自溶状态下共有783个基因表达出现差异,其中在自溶状态下上调和下调的基因分别为689和94个。GO功能聚类分析显示,差异基因主要富集在细胞、细胞组分、生物合成、有机物质生物合成几个方面。KEGG富集分析发现,差异基因主要参与了核糖体、氨基酰基-tRNA生物合成、嘌呤代谢、肽聚糖生物合成、嘧啶代谢、同源重组、氨基酸合成等通路,且下调的差异基因主要富集在碳代谢、糖酵解/糖异生、丙酮酸代谢等能量代谢相关通路。说明屎肠球菌自溶状态下代谢出现了明显改变,对碳源的利用不足和合成能力的下降可能是导致屎肠球菌自溶发生的重要原因。通过分析屎肠球菌自溶状态下基因的表达情况,得到了充分的基因数据,为进一步研究屎肠球菌的自溶发生机制提供了基础。     

一株仔猪屎肠球菌的分离及其种属鉴定

为分离出可用作猪用微生态制剂的肠道益生菌株,本研究选择锦州市凌海大业乡生态猪场为试验采样点,采集10日龄左右健康仔猪肠道粪样12份。利用MRS琼脂培养基分离培养初筛目标分离菌株的形态特征和培养特性,再通过生理生化鉴定法和16SrRNA基因分子鉴定法,最终确定1株分离菌为屎肠球菌,并命名为LN／EFl312株。为后续肠道益生球菌的益生功能研究奠定了物质基础。     

广东省养禽场肠球菌耐药性及毒力因子流行分布特征

旨在调查和分析广东省养禽场肠球菌的亚型屎肠球菌和粪肠球菌耐药性及其毒力因子流行分布特征,为控制禽源肠球菌耐药性传播、保障公共卫生安全提供理论依据。作者于2018年从广东省4个养禽场采集肠道样品493份,进行屎肠球菌和粪肠球菌的分离鉴定;采用琼脂二倍稀释法测定肠球菌的最小抑菌浓度（MIC）;PCR方法检测肠球菌的耐药基因和毒力基因。结果显示：1）共分离到125株肠球菌,其中粪肠球菌84株（鸡源66株,鸭源18株）;屎肠球菌41株,均来自鸡肠道样本。2）菌株对四环素、多西环素、红霉素几乎全部耐药,对氟苯尼考和氯霉素的耐药率高达89.60%和74.40%。屎肠球菌耐药率普遍高于粪肠球菌,而粪肠球菌对环丙沙星和利奈唑胺的耐药率高于屎肠球菌;鸭源粪肠球菌对利奈唑胺的耐药率（94%）显著高于鸡源粪肠球菌（39.4%）,屎肠球菌对利奈唑胺均敏感。从鸡分离的1株粪肠球菌对万古霉素耐药。3）耐药基因在屎肠球菌中的检出率高于粪肠球菌,鸭源分离株检出率高于鸡源。耐药基因tetL、fexA、ermB最为流行,检出率均高于90%。其次是optrA基因,检出率为73.60%,poxtA和fexB的检出率均低于20%。在3株鸭源粪肠球菌中检测出cfr基因。4）已检测的毒力基因中efaA的携带率最高,为63.04%（58/92）,其他依次为gelE（54.35%,50/92）、ace（47.83%,44/92）、asa1（44.57%,41/92）。对环丙沙星及高浓度氨基糖苷类耐药的菌株及携带cfr基因的菌株,大多携带agg、asal、gelE或ace。本研究显示养殖场禽源肠球菌耐药严重,鸭源肠球菌对利奈唑胺耐药率高,耐药基因和毒力基因流行且多样,且检测出人医临床重要抗生素耐药基因,应加强对养禽场肠球菌耐药性监测。     

一株猪源屎肠球菌的分离及其种属鉴定

为分离出可用作猪用微生态制剂的肠道益生菌株,本研究选择锦州市凌海大业乡生态猪场为试验采样点,采集10日龄左右健康仔猪肠道粪样12份。利用MRS琼脂培养基分离培养初筛目标分离菌株的形态特征和培养特性,再通过生理生化鉴定法和16SrRNA基因分子鉴定法,最终确定1株分离菌为屎肠球菌,并命名为LN/EF1312株,为后续肠道益生球菌的益生功能研究奠定了物质基础。     

家养林麝电刺激采精及精液品质研究

用电刺激采精法对20头次家养林麝进行电刺激采精及精液品质的分析。结果表明：电刺激射精电压为（4．6±0．5982）V;电刺激至射精的时间为（9．6667±6．3152）min;采精量为（0．369±0．242）mL;pH值为6．475±0．2552;精子密度为（6．935±7．0173）亿／mL;精子活力平均为0．67±0．1081。电刺激采精麻醉保定药品用量,氯胺酮（24．5075±10．0285）mg/kg体重,或陆眠宁（0．0225±0．005）mL/kg体重,均能达到满意的保定效果。     

Isolation,Identification and Multidrug-resistant Analysis of the New Sequence Type ST989 Strain of Enterococcus faecium

Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity.     

Genetic Diversity Analysis of Shaanxi Moschus berezovskii Based on mtDNA Cytb Gene and D-loop Region Sequence

【Objective】 This study was aimed to explore the genetic diversity of Shaanxi Moschus berezovskii population,and understand the genetic information of Moschus berezovskii.【Method】 The hair of Moschus berezovskii was collected to extract DNA,the mitochondrial DNA(mtDNA) cytochrome b(Cytb) gene and D-loop sequences of 43 Moschus berezovskii individuals were determined,and the base composition was counted.All sequences were integrated and compared using ClustalX 2.0 software to obtain nucleotide polymorphic sites (SNPs) in the population.The nucleotide diversity (Pi),number of haplotype (H),haplotype diversity (Hd) and average number of nucleotide differences (K) were calculated by DNASP 5.10 software.The genetic distance among different haplotypes of Cytb gene and D-loop sequences was calculated by Mega 7.0 software,and Neighbor-Joining (NJ) phylogenetic tree was constructed.【Result】 The AT content of Cytb gene and D-loop region were higher than GC content,indicating there was bias in base composition.There were 241 and 383 SNPs of Cytb gene and D-loop region,respectively.The nucleotide diversity of Cytb gene and D-loop region were 0.28343 and 0.07707,and the haplotype diversity was 0.983 and 0.975,respectively,indicating that the population genetic diversity was rich.The genetic distances of 35 haplotypes of Cytb gene ranged from 0.002 to 0.831,and 29 haplotypes of D-loop region ranged from 0.006 to 1.342.The phylogenetic tree showed that there were two mitochondrial lineages,indicating that there were two mitochondrial maternal origins.The evolutionary analysis of D-loop region also supported this conclusion.【Conclusion】 The nucleotide diversity and haplotype diversity of Moschus berezovskii population were high,and the genetic diversity was rich.At the same time further supported the view of Moschus berezovskii and Moshus moschiferus belonged to a branch of the view.     

粗放式管理条件下饲料营养对林麝麝香品质影响的研究

为了了解粗放式管理条件下饲料营养水平与麝香品质的关系,2008--2010年间对山西省宝鸡市凤县一养麝场泌香期雄麝饲料调整前后麝香的品质进行了比较研究。结果表明,调整后泌香期雄麝饲料主要营养成分为粗蛋白22．5％、粗脂肪4．8％、粗纤维9．1％、灰分5．7％、无氮浸出物56．7％,Ca：P为1：1,饲喂调整饲料后,麝香的麝香酮含量为3．2％（标准〉12．0％）、总灰分为4．2％（标准≤6．5％）、含水量为51．4％（标准≤35。0％）,麝香性状符合中国药典中的麝香标准指标,表明调整后饲料营养水平符合泌香期雄麝的营养需要。     

林麝胚胎移植研究初报

用獐子作受体进行林麝的胚胎移植试验。应用PMSG和PGF24诱导供体林麝和受体獐进行同期发情试验，供体林麝在发情配种后8～10天，手术法进行胚胎收集，结果表明，单独应用PMSG或PGF24使供体林麝与受体獐达到了预期同期发情效果，对2头林麝进行手术，发现卵巢上各有数枚黄体存在，在冲卵液中未检到胚胎。研究揭示，手术法可能是适合林麝的胚胎收集方法之一。     

林麝养殖中活体取香的方法及步骤

人工养殖林麝活体取香是有效保护林麝野生资源的重要途径。陕西凤县有着悠久的林麝人工养殖历史,在实践经验基础上,就人工养殖林麝活体取香的时间、工具、取香技术与操作方法步骤等方面进行了总结与阐述。     

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。     

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8 μg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256 μg/ml (62 strains) and 512 μg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.     

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。