Transmission of opportunistic pathogens in a veterinary teaching hospital

Acinetobacter baumannii, Enterococcus faecalis, Enterococcus faecium, and Staphylococcus intermedius isolates from infected surgical wounds and other types of infections in a veterinary teaching hospital were typed by pulsed field gel electrophoresis. A first cluster of infections with a multiresistant A. baumannii strain was observed in dogs and cats in 1998-1999. This strain disappeared after cleaning and disinfection of the companion animals intensive care unit. It was followed in 2000 by a second multiresistant strain, which caused another cluster of infections and was also transmitted to a patient in the nearby horse clinic. Transmission of a multiresistant E. faecium strain between two cats with surgical wound infections was also observed during the same period. No multiresistant S. intermedius strain was observed during this time and transmission of this organism between patients did not seem to occur. The present study demonstrates the potential for hospital nosocomial resistance problems in veterinary medicine similar to those encountered in human medicine. The results suggest that the epidemiology of nosocomial infections with A. baumannii and E. faecium may differ from that of S. intermedius infections and that preventive measures may have to be adapted to the problem organism.     

Towards the establishment and standardization of a veterinary antimicrobial resistance surveillance and monitoring programme in South Africa

The objective of this study was to establish a repeatable, standardized laboratory procedure for monitoring the development of antimicrobial resistance in bacteria isolated from animals and food of animal origin in South Africa, with reagents prepared in-house. The emergence of resistance and the spread of resistant bacteria can be limited by implementing a veterinary antimicrobial drug policy, in which inter alia systematic monitoring and prudent use play essential roles. The bacteria included in this study represented three different categories, namely zoonotic bacteria (Salmonella), indicator bacteria (Escherichia coli, Enterococcus faecalis and Enterococcus faecium) and veterinary pathogens (Mannheimia haemolytica). Thirty isolates of each species were collected with the aim of standardizing the laboratory methodology for a future national veterinary surveillance and monitoring programme. Susceptibility to ten selected antimicrobial drugs was determined by means of minimum inhibitory concentrations (MICs) using the microdilution method. The method according to the National Committee for Clinical Laboratory Standards was used as the standard. Multi-well plates containing varying dilutions of antimicrobial drugs and prepared in-house for MIC determinations, yielded repeatable results. Storage of plates for 2 months at -70 degrees C did not influence results meaningfully. Within this limited sample of bacteria, MIC results did not indicate meaningful resistance against any of the ten selected antimicrobial drugs. The findings of the study will be used to establish a national veterinary antimicrobial resistance surveillance and monitoring programme in South Africa. To allow for international comparison of data, harmonisation of the surveillance and monitoring programme in accordance with global trends is encouraged. Ideally it should be combined with a programme monitoring the quantities of antimicrobial drugs used. The aim is to contribute to slowing down the emergence of resistance and the problems associated with this phenomenon by means of the rational use of antimicrobial drugs.     

Vancomycin for multi-drug resistant Enterococcus faecium cholangiohepatitis in a cat

A 12-year-old, neutered male domestic shorthair cat was evaluated with a life-long history of intermittent, predominantly small bowel diarrhea and a 3 day history of hematochezia. At presentation, the cat had increased liver enzyme activities and an inflammatory leukogram. Histopathology demonstrated inflammatory bowel disease (IBD), cholangiohepatitis and pancreatitis. The cholangiohepatitis was associated with a multi-drug resistant Enterococcus faecium. Gallbladder agenesis was also documented. Treatment with vancomycin was safely instituted for 10 days. Clinical signs resolved, however, cure of the bacterial cholangiohepatitis was not achieved. The risk of vancomycin resistant enterococci (VRE) in human and veterinary medicine is discussed.     

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。     

福州动物园动物高水平耐庆大霉素肠球菌耐药表型检测

为了解福州动物园动物细菌耐药现状,选用肠球菌为指示菌,采用琼脂平板筛选法(ADSP法)筛选高水平耐庆大霉素肠球菌(HLGRE)。根据美国临床实验室标准化协会(CLSI)推荐的微量肉汤稀释法测定耐药菌株对6种抗菌药物的耐药性,用PCR法对分离出的不同来源的高水平耐氨基糖苷类肠球菌进行菌种鉴定。从动物园圈养野生动物共分离出78株HLGRE,不同动物来源的分离率不同,其中草食动物HLGRE分离率最低,占55.56%;肉食动物HLGRE分离率最高,占84.38%。药敏试验结果显示,不同动物来源的HLGRE均对红霉素和土霉素的耐药率最高,分别为100%和96.1%,对氨苄西林和万古霉素的耐药率最低。     

福州动物园工作人员及普通人员高水平耐庆大霉素肠球菌耐药表型检测

为了解福州动物园饲养员和兽医以及普通人群的细菌耐药现状,选用肠球菌为指示菌,采用琼脂平板筛选法（ADSP法）筛选高水平耐庆大霉素肠球菌（HLGRE）;根据美国临床实验室标准化协会（CLSI）推荐的微量肉汤稀释法测定耐药菌株对6种抗菌药物的耐药性;用PCR法对分离出的不同来源的高水平耐氨基糖苷类肠球菌进行菌种的鉴定.从所有37个样本中共筛选出15株HLGRE.其中动物园饲养员及兽医的分离率为62.50％,而普通人群的分离率为23.81％,差异显著.分离到的肠球菌中,以粪肠球菌为主要,根据药敏实验结果显示,不同人群的HLGRE均以红霉素和土霉素的耐药率最高,均达到100％;以氨苄西林和万古霉素的耐药率最低.     

Susceptibility to vancomycin and other antibiotics of 165 Enterococcus strains isolated from dogs in Italy

The prevalence of antibiotic resistance, with special attention to vancomycin, in 165 Enterococcus strains isolated from dogs subjected or not to previous antibiotics treatment(s) was determined. For each strain, the minimum inhibitory concentration (MIC) to 9 antibiotics was assessed. All strains were sensible to vancomycin. High frequencies of resistance to erythromycin, tetracycline, rifampicin, and enrofloxacin were detected. E. faecium strains isolated from dogs subjected to antibiotic treatment were more resistant to tetracycline with respect to control dogs. Although enterococci from dog show a high degree of antibiotic resistance, they are sensitive to vancomycin. Therefore, the risk of transmission of vancomycin-resistant enterococcal strains from dogs to man is close to zero.     

Isolation，Identification and Antibiotics Resistance Analysis of Enterococcus from Yak

17 feces samples of yak which were collected in Hongyuan county were measured with Gram staining method and 16S rRNA molecular identification in this study.8 suspected Enterococcus were separate from feces samples by bacteria purification and PCR amplification with 1 500 bp specific band. 6Enterococcus faecalis and 2Enterococcus faecium were identified through 16S rRNA sequencing.The homology analysis of the strains revealed that the homology between Enterococcus faecalis and reference strains sequence were 99.7% to 100%,that of Enterococcus faecium and reference sequence were 98.2% to 99.2%,indicating that the yak Enterococcus was highly conserved in the process of genetic evolution.The drug sensitive test results showed that the isolated strains were highly resistance to aminoglycoside antibiotics.Enterococcus faecium 11-1-2 strain was not only 5 multi-resistant,but also showed resistence to vancomycin.Enterococcus faecalis strains was most 3 multi-resistant.The antibiotics resistance results revealed that the resistance of yak Enterococcus was serious and should be taken seriously.     

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。     

屎肠球菌的特性及其在畜牧生产中的应用研究进展

随着对畜产品需求的增加,人们越来越重视畜产品的安全性。屎肠球菌为常见乳酸菌,是近年来研究较多的益生菌,其代谢产物对病原菌的抑制、肠道菌群稳定和家畜生长具有重要意义,已广泛应用于畜牧生产中,但耐药性和致病性屎肠球菌的相继发现,引发了公众的担忧。因此,如何合理、规范应用屎肠球菌是其在畜牧生产中应用的关键。本文综述了屎肠球菌的生物学特性及其在畜牧生产中的应用研究进展,但抗生素耐药性屎肠球菌的发现,使得屎肠球菌的应用更为谨慎。     

我国18个省份猪源肠球菌耐药调查及分析

为研究我国猪源肠球菌的耐药情况,本研究采集18个省市猪养殖场的粪便样品836份进行了肠球菌的分离鉴定,并通过琼脂稀释法测定11种抗菌药物的最小抑菌浓度。结果显示:从836份样品中共分离出225株肠球菌,其中屎肠球菌106株,粪肠球菌56株,小肠肠球菌34株,鹑鸡肠球菌17株,铅黄肠球菌7株,坚忍肠球菌4株,Enterococcus thailandicus 1株。所有的肠球菌对被检抗菌药物耐药程度由高到低依次是四环素(98.2%)、阿米卡星(94.7%)、多西环素(89.3%)、米诺环素(88.9%)、氟苯尼考(88.4%)、红霉素(86.2%)、环丙沙星(63.1%)、氨苄西林(33.8%)、氯霉素(29.8%),未检测到万古霉素和利奈唑胺耐药菌株。除氨苄西林、万古霉素、利奈唑胺外,粪肠球菌对其余8种被检抗菌药物耐药率均高于屎肠球菌。小肠肠球菌对阿米卡星、红霉素、四环素、多西环素、米诺环素5种抗菌药物的耐药率高于屎肠球菌。肠球菌的多重耐药情况集中在6耐、7耐和8耐,占肠球菌总数的72.9%。本研究表明我国猪源肠球菌对常见抗菌药物耐药率较高,应引起更多关注。     

日粮中添加屎肠球菌对AA肉鸡生产性能、免疫器官指数和血液脂质代谢相关指标的影响

本试验旨在研究日粮中添加屎肠球菌对AA肉鸡生产性能、免疫器官指数和血液脂质代谢相关指标的影响，探讨屎肠球菌替代抗生素在肉鸡日粮中应用的可行性及其最适添加量。试验选取1日龄AA肉鸡公仔鸡600只（50 g/只），随机分成5组，每组6个重复，每个重复20只鸡。对照组（CON）饲喂基础日粮；抗生素组（ANT）饲喂基础日粮+0.1%金霉素；3个屎肠球菌试验组（LEF、MEF和HEF组）在基础日粮中分别添加50、100、200 mg/kg屎肠球菌。试验分2期，分别为1~21日龄（前期）和22~42日龄（后期）。结果显示：①各试验组肉鸡生长性能与CON组相比均无显著差异（P>0.05）；从饲养全期看，MEF组和HEF组AA肉鸡的死淘率最低。②在饲养后期，与CON组相比，MEF组胸腺指数显著提高（P<0.05），MEF组脾脏指数有提高的趋势（0.05 < P < 0.1）。③在饲养前期，与CON组和ANT组相比，LEF组和HEF组血清总胆固醇（TC）含量极显著降低（P<0.01）；与ANT组相比，LEF组和MEF组血清甘油三酯（TG）含量极显著降低（P<0.01）；与CON组相比，MEF组和HEF组血清高密度脂蛋白（HDL）含量极显著提高（P<0.01）；与ANT组相比，添加屎肠球菌试验组的低密度脂蛋白（LDL）含量极显著降低（P<0.01）。在饲养后期，与CON组和ANT组相比，LEF组肉鸡血清中TG含量极显著降低（P<0.01），LEF组和MEF组HDL含量极显著提高（P<0.01）。综上所述，在肉鸡饲养中，添加屎肠球菌可以在一定程度上降低死淘率，并有促进免疫器官发育的趋势，还具有一定的降低血脂的作用。综合考虑，建议在AA肉鸡日粮中添加100 mg/kg屎肠球菌。     

Detection of the esp gene in high-level gentamicin resistant Enterococcus faecalis strains from pet animals in Japan

We investigated the prevalence of the esp gene and the susceptibility to gentamicin in Enterococcus faecalis and E. faecium strains obtained from pet animals. Nine of 30 E. faecalis and 2 of 38 E. faecium strains from the pet animals had the esp gene. Three esp-positive E. faecalis strains, which were isolated from two dogs and a cat, showed gentamicin MICs of > or =256 microg/ml and harbored the high-level gentamicin resistance (HLGR) gene, aac(6')-Ie-aph(2')-Ia. Of the nine esp-positive E. faecalis strains, five, including the three strains with the HLGR gene, were closely related by numerical analysis of PFGE patterns. Longitudinal investigation needs to elucidate whether the HLGR gene was incorporated into a subpopulation of the esp-positive E. faecalis.     

中草药提取物对蜜蜂屎肠球菌(Enterococcus faecium)的抑制作用研究

为了观察中草药对蜜蜂屎肠球菌（Entorococcus faecium）的体外抑菌效果，选用14种中草药按常规法制备水提物，用纸片法和试管二倍稀释法检测中草药水提物对蜜蜂屎肠球菌的抑菌作用。试验结果表明，所选14种中草药中9种对蜜蜂屎肠球菌有中度以上抑菌作用，其中大青叶的抑菌作用最强，其次是黄连、金银花、黄芪、黄芩、大黄、板篮根、淫羊藿和青蒿。     

表达猪乳铁蛋白肽重组屎肠球菌对断奶仔猪生长性能的影响及其抗ETEC感染效果研究

本试验以表达猪乳铁蛋白肽的重组屎肠球菌（pNZ8112-PLFcin/Ef）饲喂断奶仔猪，研究其对仔猪生长性能的影响和抗产肠毒素大肠杆菌（enterotoxigenic Escherichia coli，ETEC）感染的效果。选取28日龄体重相近的健康断奶仔猪36头，随机分为3组（重组屎肠球菌组、空载体组和培养基组），每组3个重复，每个重复4头仔猪。重组屎肠球菌组和空载体组分别饲喂添加pNZ8112-PLFcin/Ef （6×10¹² CFU/kg）和pNZ8112/Ef （6×10¹² CFU/kg）的基础日粮，而培养基组饲喂含有相同体积的GM17液体培养基的基础日粮。试验期26 d。结果显示，与培养基组相比，重组屎肠球菌组断奶仔猪的平均日增重极显著提高（P<0.01）；料重比显著降低（P<0.05）；腹泻率明显降低，肠道菌群的均匀度和多样性指数均下降。为进一步探究表达猪乳铁蛋白肽的重组屎肠球菌对断奶仔猪抵抗ETEC感染的保护作用，在连续饲喂21 d后，每个重复中随机挑选1头体况相近的断奶仔猪灌服ETEC。结果发现，与培养基组相比，攻菌后重组屎肠球菌组断奶仔猪血清白细胞介素-2（IL-2）、免疫球蛋白G （IgG）含量、肠黏液中分泌型免疫球蛋白A （sIgA）水平均显著升高（P<0.05）；脾脏指数显著提高（P<0.05），但胸腺指数、肠段长度及重量则均无显著差异（P>0.05）。综上所述，表达猪乳铁蛋白肽的重组屎肠球菌能够起到促进断奶仔猪生长及抗ETEC感染的保护作用。     

屎肠球菌及其微生态制剂的研究进展

屎肠球菌作为一种潜在的新型饲用抗生素替代品,具有耐热、耐胃酸、耐胆盐、粘附力强、抑菌性和抗生素耐受性等优良的生物学特性,还能提高饲料的转化效率,增强动物机体的免疫力,降低腹泻率及死亡率,从而改善动物产品品质,屎肠球菌微生态制剂在畜禽养殖业中应用已十分广泛。其中微囊化屎肠球菌活菌制剂更是近几年的研究热点,其抗逆性强、稳定性高,可以作为饲用高活性微生态制剂应用于实际生产。本文主要阐述屎肠球菌的生物学特性及其在饲用微生态制剂中的研究进展。     

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16S 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。通过特异性和敏感性分析,该方法能扩增出肠球菌属特异性片段及粪肠球菌和屎肠球菌种特异性片段;对粪肠球菌敏感性为0...     

Extraction,Purification and Determination of Capsular Polysaccharide in Enterococcus faecium

In order to extract and purify the capsular polysaccharide of Enterococcus faecium and measure contents of polysaccharide, high pressure crushing methods and enzyme decomposition methods (RNase A (25 μg/mL), DNase (0.5 mg/mL), lysozyme (0.25 mg/mL) and protease K (1.25 mg/mL)) were used to remove the nucleic acids and proteins to obtain crude capsular polysaccharide, and centrifugal ultrafiltration with 30 and 100 ku ultrafiltration centrifuge tubes and DEAE Sepharose Fast Flow column chromatography were also adopted to obtain the purified capsular polysaccharides of Enterococcus faecium. Then the content of polysaccharide was determined by the phenol-sulfuric acid method. The results showed the capsular polysaccharide content was 75.21%. The results revealed that capsular polysaccharide ofEnterococcus faecium was extracted successfully. The method of extracting capsular polysaccharide in Enterococcus faecium was established initially.     

屎肠球菌荚膜多糖的提取纯化及其多糖含量的测定

为提取纯化屎肠球菌（E.faecium）荚膜多糖（CPS）并测定其多糖含量,试验采用高压法破碎菌体,加入核糖核酸酶（25 μg/mL）、脱氧核糖核酸酶（0.5 mg/mL）、溶菌酶（0.25 mg/mL）及蛋白酶K （1.25 mg/mL）等去除核酸和蛋白质,得到荚膜多糖粗提物,经过30 ku和100 ku超滤离心管超滤浓缩和DEAE Sepharose Fast Flow层析柱层析后,得到纯化的屎肠球菌荚膜多糖,再利用苯酚-硫酸法测定其含量。结果表明,成功从屎肠球菌中提取纯化出荚膜多糖,其含量为75.21%,初步确定了屎肠球菌荚膜多糖的提取纯化方法,为今后相关疫苗的研制奠定了基础。     

Genetic relatedness of high-level aminoglycoside-resistant enterococci isolated from poultry carcasses

Approximately 46% (75/162) or poultry enterococci collected between 1999 and 2000 exhibited high-level resistance to gentamicin (minimum inhibitory concentration [MIC] > or = 500 microg/ml), kanamycin (MIC > or = 500 microg/ml), or streptomycin (MIC > or = 1000 microg/ml). Forty-one percent of the isolates were resistant to kanamycin (n = 67), whereas 23% and 19% were resistant to genramicin (n = 37) and streptomycin (n = 31), respectively. The predominant species identified was Enterococcus faecium (n = 105), followed by Enterococcus faecalis (n = 40) and Enterococcus durans (n = 8). Using polymerase chain reaction, the isolates were examined for the presence of 10 aminoglycoside resistance genes [ant(6)-Ia, ant(9)-Ia, ant(4')-Ia, aph(3')-IIIa, aph(2")-Ib, aph(2")-Ic, aph(2")-Id, aac(6')-Ie-aph(2")-Ia, and aac(6')-Ii]. Five aminoglycoside resistance genes were detected, most frequently aac(6')-Ii and ant(6)-Ia from E. faecium. Seven E. faecalis isolates resistant to gentamicin, kanamycin, or streptomycin were negative for all genes tested, indicating that additional resistance genes may exist. Phylogenetic analysis revealed that the isolates were genetically different with little clonality. These data indicate that enterococci from poultry are diverse and contain potentially unidentified aminoglycoside resistance genes.