五种动物源肠球菌耐药性监测

为了解不同动物源肠球菌耐药性,试验选用10种常用抗生素,采用统一材料、方法(KB法)和判断标准[美国临床实验室标准化协会(CLSI)2007年版]对从5种不同动物源(人、狗、鸡、牛、猪)粪便中分离得到的肠球菌进行耐药性检测。结果表明:共分离得到肠球菌646株,包括人源肠球菌146株、狗源肠球菌138株、鸡源肠球菌128株、牛源肠球菌111株、猪源肠球菌123株;各动物源肠球菌对四环素和利福平耐受性偏高,对青霉素G、万古霉素和高浓度庆大霉素耐受性偏低,其中耐万古霉素肠球菌47株,平均耐药率为7%;各动物源肠球菌的耐药性表现为鸡源肠球菌最强,猪源肠球菌、狗源肠球菌、人源肠球菌次之,牛源肠球菌最低。说明滥用抗生素已造成人和畜禽动物肠球菌较为普遍的耐药,应引起临床及畜禽业高度重视。     

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。     

2017-2021年上海市动物源粪肠球菌、屎肠球菌耐药性情况变迁分析

为了解上海地区动物源粪肠球菌、屎肠球菌对常见抗菌药物的耐药及其最小抑菌浓度(minimal inhibitory concentration, MIC)变迁情况,采用微量肉汤稀释法对近五年采集的粪肠球菌、屎肠球菌进行10种常见抗菌药物敏感性测试。结果表明,458株粪肠球菌及283株屎肠球菌对头孢西丁、头孢噻呋及氧氟沙星耐药率较高(均高于60%),对青霉素、阿莫西林/克拉维酸及万古霉素的耐药率较低(均低于11%),粪肠球菌耐药率整体高于屎肠球菌,两者对氧氟沙星、头孢噻呋、氟苯尼考、庆大霉素、利奈唑胺、恩诺沙星的耐药率存在较大差异(耐药率差异为15%～44%)。五年间,粪肠球菌对阿莫西林/克拉维酸、氧氟沙星的MIC₅₀及MIC₉₀均呈下降趋势;屎肠球菌对阿莫西林/克拉维酸、头孢西丁及氟苯尼考的MIC₅₀及MIC₉₀均呈下降趋势。研究表明,近年来上海地区动物源粪肠球菌、屎肠球菌的耐药情况有好转趋势,仍需继续加强肠球菌耐药性监测。     

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。     

万古霉素类耐药性肠球菌

据国际商报 ( 1998,7,16 )报道 ,最近日本官方抽查了来自欧洲和亚洲 6个国家的进口鸡肉 ,在德国、挪威和泰国的 6个鸡肉检样中发现含有万古霉素耐药性肠球菌( Vancomycin- resistant Enterococci,VRE)。VRE可引起体弱患者消化道及尿道感染并可致败血症引起死亡。鸡肉一直是我国     

动物源肠球菌对饲用抗菌促生长剂流行病学临界值的建立

【目的】建立动物源肠球菌对抗菌促生长剂的流行病学临界值,并基于此了解2019-2021年动物源肠球菌对饲用抗菌促生长剂的耐药情况。【方法】测定2019-2021年从北京、河北、四川、山西、陕西和内蒙古养殖场动物样本中分离得到的肠球菌对7种抗菌促生长剂（吉他霉素、黄霉素、恩拉霉素、那西肽、阿维拉霉素、维吉尼亚霉素和杆菌肽）的最小抑菌浓度（MIC）值。根据CLSI-VET中规定的方法,利用统计软件ECOFFinder对MIC分布数据进行非线性回归模拟,以95%置信区间下的流行病学临界值计算动物源肠球菌对7种抗菌促生长剂的耐药率。【结果】确定了动物源肠球菌对吉他霉素、黄霉素、恩拉霉素、那西肽、阿维拉霉素、维吉尼亚霉素和杆菌肽的流行病学临界值分别为8、8、8、0.25、8、8和32 μg/mL。由此计算的药物耐药率为64.38%、34.48%、80.06%、14.69%、19.06%、50.69%和24.96%,其中动物源肠球菌对恩拉霉素、吉他霉素和维吉尼亚霉素的耐药情况最为严重,而对那西肽最敏感。在2019-2021年的耐药率变化情况统计中,黄霉素和杆菌肽的耐药率下降趋势较为明显。【结论】本研究制定了动物源肠球菌对7种养殖业常用抗菌促生长剂的流行病学临界值,可作为动物源肠球菌对饲用抗菌促生长剂敏感和耐药的初步判定标准,为进一步制定耐药折点,监测动物源肠球菌对抗菌促生长剂的耐药情况变化提供数据基础和科学依据。     

Research on Antimicrobial Resistance Characteristics of Enterococcus spp. from Companion Animals in Beijing

In order to study the tolerance of Enterococcus spp.from companion animals to common antibiotics in Beijing,320 samples were collected from dogs and cats in a pet hospital in Beijing from May 2015 to January 2016.The strains were isolated by selective culture medium and identified by VITEK-2 Compact 60.The sensitivity test of 11 kinds of common antibiotics in 8 categories was carried out for the isolated and identified strains.At the same time,the high-level gentamicin and high-level streptomycin resistance test were carried out to analyze the drug resistance and multiple drug resistance of Enterococcus.A total of 318 strains of non repetitive enterococci with different colony morphology and biochemical characteristics were obtained.The isolation rates of Enterococcus faecalis (49.06%) and Enterococcus faecium (29.87%) were higher.The resistance rates of Enterococcus to high-level gentamicin and high-level streptomycin were 39.94% and 43.40%,respectively.The resistance rates to tetracycline (78.62%),erythromycin (67.30%) and quinoproptin dapfoptin (43.71%) were very high.All of them were sensitive to tigecycline,and the non-resistant rate to vancomycin and nitrofurantoin was also high (98.74%).Enterococcus tested showed different degrees of resistance to class 1 to class 7 antibiotics,and the multidrug resistance was common,and the multidrug resistance rate was as high as 57.23%.The number of resistant antibiotics was the highest (n=65).A total of 44 different drug resistance profiles were obtained.Among them,erythromycin-quinoptin-daptoptin-tetracycline-high level aminoglycoside resistant spectrum accounted for the highest proportion (45/318,14.15%).This study confirmed that Enterococcus spp.isolated from companion animals in Beijing was highly resistant to common antibiotics,and multidrug resistance was prevalent.Therefore,it was necessary to strengthen the monitoring of drug resistance.     

粪肠球菌动物致病的研究进展

粪肠球菌又称粪链球菌,能在不同环境中生长存活,对外界环境抵抗力较强,部分属于动物肠道中的正常茵群,还有部分在长期的进化中出现了毒力基因。近年来,粪肠球菌感染人和动物的报道日益增加,逐渐得到人们的广泛关注。论文综述了猪、牛、羊、禽和犬粪肠球菌的主要致病性、毒力基因及耐药性,为防制动物粪肠球菌感染提供参考。     

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...     

猪源粪肠球菌和屎肠球菌多重PCR快速鉴定方法的建立

粪肠球菌和屎肠球菌是引起猪感染发病的优势肠球菌种,以肠球菌的16 S rRNA基因设计属特异性引物,利用SodA基因多态性设计种特异性引物,同时优化反应条件,建立了能同时测定猪源粪肠球菌和屎肠球菌多重PCR方法。通过对来源于猪的临床菌株、粪便菌株和鲜猪肉菌株进行测定,均能成功扩增出属特异性片段和种特异性片段。经过与快速生化鉴定试剂盒(Vitek-32)和16 S rRNA测序方法比较,多重PCR与16 S rRNA测序方法对猪的临床菌株、粪便菌株和鲜猪肉菌株的鉴定符合率100%;与Vitek-32鉴定符合率为62.3%,其中,与分离于感染猪的临床菌株符合率仅有46.7%,特别是感染猪的屎肠球菌,符合率仅为22.3%。     

伴侣动物源肺炎克雷伯氏菌的分离鉴定及毒力和耐药性分析

为探究南宁伴侣动物源肺炎克雷伯氏菌的毒力和耐药情况,本研究采集犬、猫粪便拭子,通过分离培养、形态学观察、药敏试验及PCR扩增16S rRNA、khe基因、毒力基因及耐药基因等方法对细菌特性进行分析。结果显示,分离的菌株中有4株能在麦康凯培养基上形成液状菌落,轻挑拉丝且镜检为短杆状革兰氏阴性菌,疑为肺炎克雷伯氏菌。16S rRNA测序结果显示,分离菌与肺炎克雷伯氏菌同源性达99%,同时肺炎克雷伯氏菌特异性基因（khe）阳性。其中,分离株GXKP-C14、GXKP-D15对大部分临床常用抗菌药物敏感,分离株GXKP-D1则表现出高水平多重耐药,分离株GXKP-D4耐药性稍低于GXKP-D1,对临床常用药氨苄西林、哌拉西林、头孢呋辛、四环素、多西环素、呋喃妥因、复方新诺明表现耐药,对头孢吡肟、美罗培南、亚胺培南、阿米卡星、庆大霉素、链霉素、多黏菌素等敏感。部分菌株携带tet（A）、QnrS、bla_SHV、sul2、mcr-1等耐药基因和WabG毒力基因。本研究结果为犬、猫源肺炎克雷伯氏菌病的检测、诊断及治疗提供了试验依据。同时,黏菌素耐药基因mcr-1的检出将为多黏菌素的耐药性控制和合理使用提供参考。     

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.     

宠物源大肠埃希菌的分离鉴定和耐药性研究

为调查长春地区宠物源大肠埃希菌耐药流行情况,从2家宠物医院采集135份宠物肛拭子样品,分离和鉴定大肠埃希菌并进行多重PCR分群。测定大肠埃希菌分离株对19种抗菌药物的耐药性,并鉴定超广谱β-内酰胺酶(ESBLs)表型。共分离鉴定获得95株宠物源大肠埃希菌,它们对氨苄西林和哌拉西林的耐药率最高(78.9%和76.8%);其次是四环素(61.6%);对头孢唑林、头孢噻肟、头孢吡肟、庆大霉素、复方新诺明、环丙沙星和左氧氟沙星的耐药率均超过了50%;所有分离株均对亚胺培南和美罗培南敏感。其中15个分离株对受试的抗菌药物全部敏感(15.8%),63株呈多重耐药表型(66.3%)。ESBLs型菌株占53.7%。本研究探明了长春地区宠物源大肠埃希菌的耐药流行情况,对宠物临床上用药具有指导价值。     

Classification and Antimicrobial Susceptibilities of Enterococcus Species Isolated from Apparently Healthy Food‐Producing Animals in Japan

Summary A total of 1024 enterococci were recovered from faeces of healthy animals from 178 cattle, 178 pig and 156 broiler farms. Enterococcus faecium was the predominant species recovered (35.8%), followed by E. faecalis (31.3%) and E. hirae (25.6%). Oxytetracycline resistance was most frequently found among E. faecalis (85.9%), E. faecium (58.8%) and E. hirae (48.1%). Resistance rates to almost all antimicrobials were higher in E. faecalis than E. faecium and E. hirae. Isolates from cattle were more susceptible to the antimicrobials studied than those from pigs and broilers. VanA‐ or VanB‐type vancomycin‐resistant enterococci have not been found since the ban of avoparcin use 5 years ago.     

河南省某地区健康人源及动物源肠球菌种属分布及耐药性差异研究

为了解河南省某地区社区健康人源及动物(鸡、猪)源肠球菌种属分布及对多种抗菌药物的耐受情况,以及研究不同来源肠球菌的流行分布和抗菌药物耐药表型特点,本试验应用肠球菌选择性培养基对220份人源及动物源粪便样本进行肠球菌的分离培养,并对分离菌株采用16S rDNA序列分析结合API生化板条进行种属鉴定,纸片扩散法对分离肠球菌进行9种抗菌药物的敏感性检测.肠球菌分离及鉴定结果显示肠球菌总分离率、粪肠球菌分离率及屎肠球菌分离率在人源、鸡源、猪源3种来源间均差异显著(P<0.05):肠球菌总分离率为70.91%(156/220),猪源肠球菌分离率最高(86.00%),人源肠球菌分离率最低(62.63%),且人源与猪源肠球菌分离率差异显著(P<0.018);人源粪便样本中分离率最高的为屎肠球菌(31.36%),鸡源、猪源肠球菌中粪肠球菌分离率最高,分别为28.17%和32.00%.抗菌药物敏感性结果显示肠球菌对多种药物的耐药率在人源、鸡源、猪源3种来源间差异显著(P<0.05),且3种来源肠球菌的多药耐药率差异有统计学意义(P<0.05).人源肠球菌对红霉素(69.35%)、环丙沙星(37.10%)、氨苄西林(19.35%)等抗菌药物耐药率较其他来源的肠球菌要高;鸡源肠球菌对四环素(88.24%)、氟苯尼考(11.76%)、氯霉素(21.57%)等抗菌药物耐药率较其他来源的肠球菌要高;猪源肠球菌对抗菌药物耐药率总体较低,且其多药耐药率(7.84%)也低于人源(35.48%)及鸡源肠球菌(30.19%).提示,健康人及动物粪便样本中肠球菌种属分布不同及对抗菌药物耐药率有差别,并对多种常见抗菌药物耐药率较高,有关部门应加强社区人群及动物等非临床来源肠球菌耐药检查、监测,进而更好的了解中国耐药肠球菌的流行现状,更有效的控制耐药肠球菌的传播.     

宠物源菌株的分离鉴定及耐药性分析

为了解河南郑州市常见宠物携带菌株组成以及耐药情况,本试验采集了郑州地区不同宠物医院患病及健康犬、猫共计76个样本的肛拭子、眼拭子、耳拭子。试验通过细菌分离培养、形态学观察、生化试验、16s rRNA基因测序完成细菌鉴定,确定分离菌株种类,采用微量肉汤稀释法测定分离菌株耐药表型。结果共分离得到大肠杆菌50株、肠球菌27株、奇异变形杆菌24株、葡萄球菌46株、绿脓杆菌4株。奇异变形杆菌耐药最为严重,对多西环素、替米考星两种药物耐药率高于90.0%;大肠杆菌对替米考星的耐药率高达94.0%,对阿莫西林耐药率为62.0%,对多西环素、多粘菌素E、庆大霉素、环丙沙星、恩诺沙星、头孢噻呋的耐药率在4.0%~24.0%之间。肠球菌对多西环素、庆大霉素、头孢噻呋三种药物的耐药率在29.6%~40.7%之间;对阿莫西林、恩诺沙星、环丙沙星三种药物的耐药率在7.4%~14.8%之间,对替米考星的耐药率为66.7%。葡萄球菌对替米考星的耐药率为39.1%,对阿莫西林、多西环素、庆大霉素、环丙沙星、恩诺沙星、头孢噻呋的耐药率均低于20.0%。上述试验结果表明:大肠杆菌和葡萄球菌在宠物菌群组成中占比较大;大肠杆菌和奇异变形杆菌的耐药情况较为严重。