中国部分地区犬源伪中间葡萄球菌的流行特征分析

旨在调查犬源伪中间葡萄球菌的耐药性和分子特征，本研究自2017年11月—2019年4月收集601份送检至中国农业大学动物医院检验科和第三方检测机构的犬源临床样本，分离鉴定伪中间葡萄球菌；采用琼脂稀释法检测伪中间葡萄球菌对14种抗菌药物的敏感性；所有分离菌株均进行全基因组测序（WGS），并从WGS结果中获取耐药基因、多位点序列分型（ST）和葡萄球菌染色体基因盒（SCCmec）分型。结果显示，共分离到75株伪中间葡萄球菌（分离率为12.5%），其中包括42株（56%）耐甲氧西林伪中间葡萄球菌（MRSP）和33株（44%）甲氧西林敏感伪中间葡萄球菌（MSSP）；所有菌株均对青霉素、阿奇霉素、克林霉素、多西环素、环丙沙星、恩诺沙星、苯唑西林和氯霉素高度耐药，对利奈唑胺、万古霉素、阿米卡星和利福平敏感，多重耐药率达90.7%；所有MRSP均具有多重耐药性；75株伪中间葡萄球菌中检测到19种耐药基因，blaZ和aac（6'）-aph（2″）检出率最高，均为92%（69/75）；共发现70种ST型，54种为新ST型，无优势ST型；23株（54.7%） MRSP为SCCmec V型。综上表明，犬源伪中间葡萄球菌存在严重的多重耐药性；MRSP和MSSP均具有较大的遗传多样性。     

我国部分省区牛源金黄色葡萄球菌抗菌药物敏感性调查

为调查我国奶牛乳房炎奶样中金黄色葡萄球菌的药物敏感性情况,以便为该病的临床用药提供参考,本试验采用纸片扩散法对从北京、山西、内蒙古、新疆、浙江5省区奶牛乳房炎奶样中分离的148株金黄色葡萄球菌进行了抗菌药物敏感性检测。北京、山西、内蒙古分离株的临床药敏试验显示,所有菌株对头孢西丁、多西环素、四环素敏感或中介,对红霉素、克林霉素、青霉素、复方新诺明严重耐药,对氯霉素、环丙沙星和庆大霉素的敏感性有地域性差异,66%～100%的菌株同时对3种以上的抗菌药物耐药;新疆86株分离株和浙江20株分离株的临床药敏试验显示,所有菌株除对红霉素、克林霉素、青霉素、复方新诺明耐药外,对多西环素和四环素完全耐药,86%以上的分离株对5种以上抗菌药物耐药,同时还发现头孢西丁耐药株。调查结果显示,5省区乳房炎奶样中的金黄色葡萄球菌除对头孢类(头孢西丁)敏感外,对其他类型抗菌药物均存在一定程度的耐药,尤其对大环内酯类(红霉素)、林可胺类(克林霉素)、青霉素类(青霉素)、磺胺类(复方新诺明)抗菌药物严重耐药,对3～5种以上不同类型抗菌药物的多重耐药也较为普遍。调查表明,牛源性金黄色葡萄球菌耐药性的广泛存在将对金黄色葡萄球菌性奶牛乳房炎的抗菌药物治疗形成严重挑战。     

长春部分地区犬猫源葡萄球菌的高通量测序分析

为了解长春地区犬猫源金黄色葡萄球菌(Staphylococcu saureus,S.aureus)和伪中间型葡萄球菌(Staphylococcus pseudintermedius,SP)耐药基因和毒力基因携带情况,利用高通量测序技术对17株S.aureus与38株SP所携带的耐药基因与毒力基因进行分析,根据Kirby-Bauer纸片扩散法的结果分析耐药基因型与表型的关系,同时进行了MLST分型。结果表明S.aureus和SP对常规抗菌药物表现中介或耐药,但对呋喃妥因敏感率为100%;β-内酰胺类耐药基因bla_Z检出率较高,分别为100%和57.9%;各有1株同时携带bla_Z和bla_NDM;S.aureus中检出2株携带磷霉素类耐药基因fosA;SP中检出21株携带多药耐药基因cfr;各检出3株携带多药外排耐药基因mdfA;共检出4株MRSA和11株MRSP;优势ST型分别为ST25和ST1430;在大多数分离菌中携带有γ-溶血素hlgB、黏附素clf、fnb、杀白细胞素LukF-PV、LukS-PV等毒力基因。分离株耐...     

北京地区警犬皮肤伪中间型葡萄球菌药敏试验及耐药基因筛查

采用纸片扩散法对北京地区警犬皮肤分离出的23株伪中间型葡萄球菌进行药物敏感性试验,并用PCR扩增技术进行相关耐药基因筛查。结果显示:23株伪中间型葡萄球菌对头孢西丁、阿米卡星、阿莫西林-克拉维酸、头孢曲松、氟苯尼考全部敏感,对青霉素G耐药性最高,为52.17%;耐药基因筛查中,3株未检出耐药基因,占13.04%;耐药基因中,linA/linA’检出率最高,为47.83%;耐药基因谱型以aacA-aph D、linA/linA'为主,占17.39%。     

猫源临床样品的猫葡萄球菌耐药性调查

为了解猫葡萄球菌(Staphylococcus felis)在临床患猫中的携带和耐药情况，本调查使用选择性培养基从1 990份猫源临床样本中筛选葡萄球菌，并采用16S rDNA测序鉴定猫葡萄球菌；采用琼脂稀释法检测猫葡萄球菌对14种抗菌药的敏感性，并运用PCR检测耐药基因mecA。结果显示，本调查共分离到猫葡萄球菌48株，对青霉素耐药率最高(83.3%),对阿奇霉素、环丙沙星、恩诺沙星、克林霉素、氯霉素和氟苯尼考的耐药率均大于10.0%,对苯唑西林、利福平和多西环素的耐药率均为2.1%,对利奈唑胺、万古霉素、阿米卡星和庆大霉素均敏感；40株(83.3%)菌株对至少1种抗菌药耐药；16株(33.3%)菌株呈多重耐药；未检测到mecA基因。结果表明，猫葡萄球菌的耐药状况严峻，应加强小动物临床感染耐药性的监测，规范临床诊疗程序，合理使用抗菌药。     

猪肉和牛奶源葡萄球菌耐药性调查

2010年从贵州省猪肉和牛奶样品中分离鉴定出73株葡萄球菌,其中金黄色葡萄球菌50株,凝固酶阴性葡萄球菌23株。采用琼脂稀释法检测其对18种抗菌药物的敏感性,运用统计学方法比较分析金黄色葡萄球菌与凝固酶阴性葡萄球菌的耐药性差异。结果显示,动物性食品源葡萄球菌耐药较严重,对临床上常用药物耐药率较高,且为多重耐药,其中乳源葡萄球菌的多重耐药现象较猪肉源葡萄球菌严重;猪肉源金黄色葡萄球菌主要对青霉素类、四环素和大环内酯类耐药,乳源金黄色葡萄球菌主要对青霉素类、头孢菌素类、大环内酯类、四环素、克林霉素、泰妙菌素及利福平耐药;猪肉源凝固酶阴性葡萄球菌主要对氨苄西林、苯唑西林和泰妙菌素耐药,乳源凝固酶阴性葡萄球菌的耐药谱与乳源金黄色葡萄球菌类似。     

夏、冬两季猪源葡萄球菌耐药情况比较

本研究旨在比较夏、冬两季某规模化养猪场猪源葡萄球菌的耐药情况。分别在夏、冬两季对猪场同一猪采集其直肠肛拭子样和鼻腔鼻拭子样,采用微量肉汤稀释法对分离出的葡萄球菌进行临床9种常用抗菌药物的药敏试验。结果显示,夏季鼻腔分离的葡萄球菌对阿米卡星、氟苯尼考、氧氟沙星、苯唑西林和克林霉素5种抗菌药物的耐药率均高于冬季分离的鼻腔菌。夏季直肠分离的葡萄球菌对阿莫西林/克拉维酸、苯唑西林、庆大霉素、氧氟沙星、四环素、克林霉素和利福平7种抗菌药物的耐药率低于冬季分离的直肠菌。且夏、冬两季鼻腔分离的葡萄球菌多药耐药种数在0~9耐均有分布,夏、冬两季直肠分离的葡萄球菌多药耐药种数在1~9均耐有分布。冬季鼻腔分离的葡萄球菌6耐的多药百分率显著高于夏季分离菌的多药百分率（P＜0.05）,冬季直肠分离的葡萄球菌8耐的多药百分率显著高于夏季分离菌的多药百分率（P＜0.05）。结果表明,夏、冬两季猪不同部位分离的葡萄球菌对大多数被检抗菌药物的耐药率情况不同,其耐药率与养殖场季节性用药呈现相关性。     

山羊源伪中间葡萄球菌的分离鉴定及耐药性检测

伪中间葡萄球菌是一种常见的致病菌,多引起宠物不同程度感染。2016年3月初,吉林省某羊场育肥羊出现零星死亡,病羊主要表现为呼吸系统症状。采集病羊呼吸系统各器官,以进行细菌的分离、培养。分离菌株经培养特性、染色镜检、生化试验及16S r DNA序列分析,确定为伪中间葡萄球菌,其对小鼠有致病性。对分离到的菌株通过微量稀释法进行药物敏感性测定,结果显示,对庆大霉素、链霉素、红霉素、阿莫西林、头孢氨苄、磺胺间甲氧嘧啶、恩诺沙星、强力霉素耐药;对阿米卡星、氟苯尼考、环丙沙星敏感。     

湖南部分地区乳源金黄色葡萄球菌耐药性及溶血性分析

为调查湖南省部分地区奶牛场生鲜乳源金黄色葡萄球菌的耐药性及耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)的检出情况，同时调查金黄色葡萄球菌溶血素基因型与溶血表型的分布情况及二者的关系。采用肉汤稀释法对分离自湖南省部分地区奶牛场生鲜乳的54株金黄色葡萄球菌进行18种常见抗菌药物耐药性检测，采用绵羊血琼脂平板检测细菌溶血性，并采用PCR方法扩增分离菌株mecA基因及5个溶血素基因。结果显示，54株金黄色葡萄球菌均表现不同程度的耐药，耐药谱型达21种；21株金黄色葡萄球菌表现为多重耐药，最高耐药重数为9重；对青霉素耐药最严重，耐药率达85.19%,对头孢西丁、苯唑西林、克林霉素、庆大霉素及大环内酯类药物(红霉素、替米考星)耐药率在20%～30%之间，对多西环素与万古霉素敏感；从24株分离菌中检出mecA基因，鉴定为MRSA。54株分离株在羊血琼脂平板上主要以β和γ溶血为主，α、β及γ溶血率分别为12.9%、44.4%和42.6%;各溶血素基因检出率分别为98.1%(hla)、94.4%(hlb)、96.3%(hld...     

犬脓皮病耐甲氧西林伪中间型葡萄球菌抗菌药耐药性及消毒剂抗性研究进展

犬脓皮病是一类主要由耐甲氧西林伪中间型葡萄球菌（MRSP）感染而引起的化脓性皮肤病。葡萄球菌是一种人与动物均易感的细菌，常引起各种化脓性疾病，其中，MRSP作为一种动物源葡萄球菌还会成为耐药基因贮存库，可将耐药基因通过环境或食物链传给人类。近年来MRSP造成的皮肤疾病病例大幅上升，给感染的控制带来挑战。笔者综合了犬脓皮病致病菌的抗菌药耐药性及其消毒剂抗性的相关研究，从MRSP的致病机制出发，总结了MRSP通过破坏细胞免疫系统的功能导致感染发生的相关机制，简述了多个国家MRSP对抗菌药的显著耐药性与相关耐药基因，如mecA和cat基因等，介绍了MRSP对胍类消毒剂与季铵盐类消毒剂的抗性及抗性机制，对外排泵、基因调控与抗性基因的可转移性等多种机制进行了论述，同时为避免MRSP对抗菌药与消毒剂的共同耐药性对犬脓皮病的治疗造成干扰，笔者从移动遗传元件介导的获得性抗性与依赖于细菌细胞结构的固有抗性等方面系统地分析了MRSP对消毒剂抗性和抗菌药耐药性之间的争议与联系，以期寻找一种科学合理的治疗方案，为犬脓皮病的临床用药提供参考。     

Antimicrobial susceptibility of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from various animals

This study characterized the antimicrobial susceptibility of 221 Staphylococcus aureus isolated from various species, and 60 canine Staphylococcus pseudintermedius isolated from 1986 through 2000 at the Western College of Veterinary Medicine (WCVM). Resistance of S. aureus was most common to penicillin (31%) and tetracycline (14%); resistance of S. pseudintermedius to penicillin was present in 8% and to tetracycline in 34% of isolates. Resistance to trimethoprim/sulfamethoxazole was only seen among S. pseudintermedius, and there was no resistance to amoxicillin/clavulanate, ampicillin/sulbactam, cephalothin, amikacin, gentamicin, enrofloxacin, chloramphenicol, or rifampin among any isolate. Inducible clindamycin resistance was found in both S. aureus and S. pseudintermedius, highlighting the need for careful interpretation of culture and susceptibility test results. There were significant differences in the minimum inhibitory concentrations of penicillin, ciprofloxacin, enrofloxacin, clindamycin, erythromycin, chloramphenicol, and tetracycline between avian, bovine, equine, and porcine isolates.     

Carriage of methicillin-resistant Staphylococcus pseudintermedius in small animal veterinarians: indirect evidence of zoonotic transmission

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is increasingly reported in small animals and cases of human infections have already been described despite its recent emergence in veterinary practice. We investigated the prevalence of MRSP and methicillin-resistant Staphylococcus aureus (MRSA) among small animal dermatologists attending a national veterinary conference in Italy. Nasal swabs were obtained from 128 veterinarians, seven of which harboured MRSP (n = 5; 3.9%) or MRSA (n = 2; 1.6%). A follow-up study of two carriers revealed that MRSP persisted for at least 1 month in the nasal cavity. Methicillin-susceptible S. aureus (MSSA) was isolated from 32 (25%) conference participants, whereas methicillin-susceptible S. pseudintermedius (MSSP) was not detected, suggesting that MRSP may have a particular ability to colonize humans compared to MSSP. All isolates were characterized by spa typing. Methicillin-resistant isolates were further typed by antimicrobial susceptibility testing, SCCmec and multi-locus sequence typing. Two lineages previously associated with pets were identified among the five MRSP isolates; the European epidemic clone ST71-SCCmec II-III and ST106-SCCmec IV. One of the two MRSA isolates displayed a genotype (ST22- SCCmecIV) frequently reported in dogs and cats. MRSP isolates were resistant to more antimicrobial agents compared with MRSA isolates and displayed the typical multidrug resistance patterns of MRSP in pets. The 32 MSSA isolates belonged to 20 spa types and the most frequent types (t12, t15 and t166) were associated with common S. aureus lineages in humans (CC30 and CC45). Although low, the 3.9% MRSP carriage rate found among small animal dermatologists was surprising in consideration of the rare occurrence of S. pseudintermedius in humans, the lack of MSSP detection and the recent appearance of MRSP in Europe. As cases of human MRSP infection have been linked with pets, veterinarians should be aware of this zoonotic risk and proper preventative measures should be taken to avoid MRSP transmission from animal patients.     

Heterogeneity of methicillin-sensitive Staphylococcus pseudintermedius strains isolated from diseased dogs

Thirty nine canine S. pseudintermedius strains were examined for antibiotic susceptibility and genetic polymorphisms. All strains were methicillin-sensitive S. pseudintermedius (MSSP). Resistance to penicillin was most prevalent (66.6%), followed by resistance to neomycin (56.4%), erythromycin (53.8%), clindamycin (48.7%), chloramphenicol (48.7%), and tetracycline (46.2%). Pulsed-field electrophoresis (PFGE) showed a high genetic polymorphism in the investigated strains.     

Tandem repeat sequence analysis of staphylococcal protein A (spa) gene in methicillin-resistant Staphylococcus pseudintermedius

A putative staphylococcal protein A (spa) gene was discovered in the genome of Staphylococcus pseudintermedius and used for developing a species-specific spa typing protocol. Thirty-one clinical methicillin-resistant S. pseudintermedius (MRSP) isolates from dogs and cats in four countries were characterized by spa typing, pulsed-field gel electrophoresis (PFGE) and staphylococcal cassette chromosome (SCCmec) typing. The results indicated the occurrence of two MRSP clones that acquired distinct SCCmec elements in Europe (t02, PFGE type A, SCCmec type III,) and California (t06, PFGE type B, SCCmec type V). Sequence analysis of mecA revealed the occurrence of four alleles (mecA1 to mecA4), which correlated with the geographical origin of the isolates and enabled discrimination of two distinct subtypes within the European clone. The newly developed spa typing method appeared to be a promising tool for easy and rapid typing of MRSP, either alone or in combination with SCCmec and mecA typing for fine-structure epidemiological analysis.     

Emergence of methicillin-resistant Staphylococcus pseudintermedius in Switzerland: three cases of urinary tract infections in cats

Methicillin resistance has emerged in clinical isolates of Staphylococcus pseudintermedius from cats in Switzerland. Three cats suffering from urinary tract infections were infected with methicillin-resistant S. pseudintermedius (MRSP). Phenotypic and genotypic characterization of the resistance profile showed that the isolates displayed resistance to all beta-lactams and cephalosporins (blaZ, mecA), fluoroquinolones, tetracyclines [tet(K)], macrolides, lincosamides and streprogramins B [erm(B)], chloramphenicol (catpC221), trimethoprim [dfr(G)] and the aminoglycosides gentamicin [aac(6')-Ie-aph(2')-Ia], kanamycin and neomycin [aph(3')-III] and streptomycin [ant(6)-Ia]. They also harbor the leukocidin gene lukS-I. MRSP represents a new challenge for antibiotic therapy and this zoonotic bacteria may rapidly spread to animals and humans.     

Meticillin-resistant Staphylococcus pseudintermedius: clinical challenge and treatment options

Meticillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a major therapeutic challenge for small animal veterinarians over the past 10 years and continues to spread worryingly in many countries. This review focuses on the clinical aspects of MRSP infections seen in patients with skin disease and on currently available treatment options. In addition, it discusses the implications for in-contact people, other animals and the environment, because infection control strategies are likely to have a significant impact on treatment success and prevention of spread. There is currently no indication that MRSP is more virulent than meticillin-susceptible S. pseudintermedius, and reported infections have mostly been treated successfully, although possibly with a longer time to resolution than infections with more susceptible S. pseudintermedius. However, in vitro testing of MRSP isolates indicates resistance to most or all antibacterial agents licensed for use in pets. Based on susceptibility results, the most useful systemic antimicrobials may include chloramphenicol, rifampicin, amikacin, clindamycin and/or minocycline. Adverse effects of some of these medications may limit their usefulness. While in vitro susceptibility to vancomycin and linezolid is reported by some laboratories, use of these drugs in animals is strongly discouraged because of ethical considerations. Aggressive topical therapy has been effective as the only treatment in certain cases. Awareness, continued research and comprehensive management of infections are required by veterinary practitioners not only to help treat infected animals but also to limit the spread and prevent the establishment of this highly drug-resistant and zoonotic pathogen in veterinary facilities and in the community.     

Prevalence of resistance to 11 antimicrobials among Campylobacter coill isolated from pigs on 80 grower-finisher farms in Ontario.

We carried out a cross-sectional study to investigate antimicrobial resistance patterns of Campylobacter coli isolated from Ontario grower-finisher pigs. From January to June 2004, 1200 samples were collected from 80 farms by obtaining a constant number (15) of fecal samples per farm. Susceptibility of the isolates to 11 antimicrobial drugs was determined by the agar-dilution technique. The overall prevalence of resistance to 1 or more antimicrobials among the isolates was 99.2%. High levels of resistance were observed for azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline: 91.7%, 82.5%, 81.4%, 70.7%, and 63.7%, respectively. For sulfamethoxazole, ampicillin, and nalidixic acid, resistance was observed in 40.3%, 26.6%, and 22.7% of the isolates, respectively. Although at very low levels, resistance was observed for ciprofloxacin (a fluoroquinolone), chloramphenicol, and gentamicin: in 2.4%, 1.7%, and 0.2%, respectively. Many of the isolates (29.7%) were resistant to 5 antimicrobials, the most common being azithromycin, clindamycin, erythromycin, streptomycin, and tetracycline. Isolates from the same farm showed at least 5 patterns of resistance. Results from this study indicate high levels of resistance to the antimicrobial drugs most commonly used in the Canadian swine industry (macrolides, lincosamides, and tetracyclines) among C. coli isolated from grower-finisher pigs in Ontario. Macrolides and fluoroquinolones are the drugs most commonly used to treat severe human campylobacteriosis. Fortunately, at present, there is little resistance to fluoroquinolones among C. coli from pigs in Ontario.     

Transmission of methicillin-resistant Staphylococcus pseudintermedius between infected dogs and cats and contact pets, humans and the environment in households and veterinary clinics

The objective of this study was to investigate the prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in people, pets and the environment in households with a pet with a clinical MRSP-infection within the past year. Personnel and the environment at veterinary clinics were also screened. Nasal swabs (humans), nasal and perineal swabs (pets) and environmental wipes were examined using selective culturing. Twenty households were enrolled; 10/20 index cases still had clinical signs of infection at the start of the study and all were MRSP-positive. Of the remaining 10 index cases five were MRSP-positive in nasal and/or perineal samples. Five of 14 (36%) contact dogs and four of 13 (31%) contact cats were found MRSP-positive. In the households with an index case with clinical signs of infection 6/7 (86%) contact animals were MRSP-positive. MRSP was cultured from 2/45 (4%) human nasal samples. Domestic contamination was widespread as positive samples were found in 70% of the households and 44% of all environmental samples were MRSP-positive. In all but one of these MRSP-positive households the index case was still MRSP positive. Among the personnel in veterinary clinics 4/141 (3%) were MRSP-positive. MRSP was cultured from 31/200 environmental samples in 7/13 clinics at the first sampling and in 3/6 clinics the environment remained MRSP-positive after cleaning and disinfection indicating that current cleaning procedures often were unable to eliminate MRSP. These results show that transmission of MRSP between infected or colonized dogs and cats and healthy people does occur but is relatively uncommon, while transmission to contact pets occurs frequently, especially when the index case still has clinical signs of MRSP-infection.     

The In Vitro Activity of 15 Antimicrobial Agents against Bacterial Isolates from Dogs

The in vitro activity of 15 antimicrobial agents against clinical isolates of Staphylococcus pseudintermedius, Staphylococcus aureus, Escherichia coli, Pasteurella spp. and Streptococcus canis from dogs was investigated. For Staphylococcus spp., the highest frequency of resistance was observed for penicillin, followed by ampicillin, tetracycline and chloramphenicol. The highest frequency of resistance in E. coli isolates was recorded for tetracycline and streptomycin. Pasteurella spp. and S. canis had the highest resistance rate for tetracycline and chloramphenicol. Most isolates showed full susceptibility to low-level resistance to colistin, florfenicol and fluoroquinolones. Further studies using larger number of isolates from both healthy and diseased dogs would provide a broader picture of antimicrobial resistance at a national level and promote prudent use of antimicrobial agents in companion animals.