Isolation of methicillin-resistant Staphylococcus aureus from the environment in a veterinary teaching hospital

After recognition of a cluster of methicillin-resistant Staphylococcus aureus (MRSA) infection in horses and humans at the Ontario Veterinary College Veterinary Teaching Hospital, environmental contamination with MRSA was evaluated. A total of 260 environmental surfaces were sampled during periods when MRSA-infected horses were hospitalized, and MRSA was isolated from 25/260 sites (9.6%). The most commonly contaminated sites were stalls housing MRSA-positive horses, but other stalls, medical equipment, personal items, and equine restraint devices also were contaminated. The role of the environment in the transmission of MRSA infection to horses or humans is unclear. However, relatively widespread contamination of the hospital environment, as apparently occurs when infected horses are hospitalized, suggests that the environment may be an important source of MRSA infection. This possibility must be taken into consideration when designing infection control and disinfection protocols.     

Clindamycin-resistance in methicillin-resistant Staphylococcus aureus isolated from animals

Clindamycin is widely used in veterinary medicine to treat a variety of bacterial infections. Resistance to clindamycin amongst staphylococci may be due to modification of the ribosomal target site. Resistance due to this mechanism is encoded by the erm genes, which may be either constitutively or inducibly expressed. Inducible expression of resistance is not usually demonstrable by routine laboratory susceptibility testing, however, a relatively simple double disc agar diffusion test (D-test) may be used to detect the presence of inducible resistance. Two hundred and eighty-five isolates of methicillin-resistant Staphylococcus aureus (MRSA) isolated from infections in animals were tested using the D-test for the presence of inducible clindamycin resistance. One hundred and seven (71%) of the strains tested demonstrated inducible resistance to clindamycin, which were not detected on initial susceptibility testing. It has been shown that there is potential for staphylococci to develop resistance to clindamycin whilst on treatment when inducible resistance mechanisms are present. Therefore, routine screening for inducible resistance may be prudent.     

Characterization of methicillin-resistant Staphylococcus aureus isolated from dogs in Korea.

Twelve strains of the methicillin-resistant Staphylococcus aureus (MRSA) recovered from hospitalized dogs were analyzed for in vitro antimicrobial susceptibility and virulence, and were genetically characterized by pulsed-field gel electrophoresis (PFGE). Antibiotic susceptibility test showed that nearly all isolates were resistant to beta-lactam antibiotics tested and all the strains were fully susceptible to glycopeptides. There were no inhibitory activities among the aminoglycosides. The 50% lethal dose (LD50) was determined by intraperitoneal injection of cell suspensions and estimated by the Spearman-K?rber method. The mouse lethality of MRSA and methicillin-susceptible S. aureus (MSSA) was not significantly different in both normal and cyclophosphamide-treated mice (p>0.05), indicating that they were equally virulent. There was a great difference in the incidence of toxin production between the MRSA and MSSA group; 83.3% (10 of 12) of the MRSA and 14.3% (1 of 7) of the MSSA were toxin producers. The predominant types produced by MRSA was B. All the MRSA strains were capsular type 5 producers, while of 7 MSSA strains, four were type 5, one for type 8, and two were nontypeable. Based on the PFGE analysis, the 12 MRSA isolates generated 9 to 11 fragments in the size range of <48.5 to 630.5 kb, and yielded 6 different patterns. The results indicated that production of toxin and capsule type do not play a role in the pathogenicity to mouse and PFGE is a valuable tool for the characterization of MRSA. This report is the first such cases in the veterinary literature in Korea and may indicate the frequent emergence of MRSA in veterinary clinic hereafter.     

Isolation of methicillin-resistant Staphylococcus lugdunensis in a dog with endocarditis

A 4-year-old male castrated Borzoi dog presented on referral for evaluation of pleural effusion, ventricular arrhythmias, and suspected dilated cardiomyopathy. Echocardiogram identified several masses along the chordae tendineae, as well as a long the annulus of the mitral valve. A blood culture was positive for methicillin resistant Staphylococcus lugdunensis. The dog was also positive for Bartonella DNA on PCR testing. Aggressive antibiotic therapy was instituted. However, the dog continued to have recurrent pleural effusion and progressive azotemia. The dog was euthanized 39 days after diagnosis. Necropsy confirmed the presence of intracardiac abscesses, thrombosis and endocarditis. S. lugdunensis is a recently identified rare cause of endocarditis in humans characterized by intracardiac abscess formation, highly destructive valvular lesions preferentially affecting the mitral valve and a high mortality rate. This is the first reported case of S. lugdunensis isolation in a dog with endocarditis.     

兔金黄色葡萄球菌的分离鉴定

本研究从福建省某养殖场病兔分离到3株革兰氏阳性球菌,并对其进行了16S rRNA扩增、生化反应、药敏试验等检测,结果显示该病原菌为金黄色葡萄球菌。     

Carriage of methicillin-resistant Staphylococcus aureus by veterinarians in Australia

Objective To estimate the prevalence of carriage of methicillin‐resistant Staphylococcus aureus (MRSA) among Australian veterinarians. Methods Individuals attending veterinary conferences in Australia in 2009 were recruited to provide nasal swabs and complete a questionnaire about their professional activities. Swabs were processed by standard methods for detecting MRSA and questionnaire responses were used to group veterinarians according to their areas of major work emphasis (species and practice type). Prevalence was estimated for each of these grouping and contingency tables and regression tree analysis used to explain the variation in MRSA carriage. Results Among the 771 respondents ‘industry and government veterinarians’ (controls) had the lowest prevalence of MRSA carriage at 0.9%. Veterinarians with horses as a major area of work emphasis had a prevalence of 11.8% (13‐fold that of controls) and those whose only major emphasis was horses had a prevalence of 21.4% (23‐fold that of controls). Veterinarians with dogs and cats as a major activity had a 4.9% prevalence (5‐fold that of controls). Prevalence rates for other major activities (pigs, dairy and beef cattle, avian and wildlife) were also increased, but were estimated from smaller numbers of respondents. Regression tree analysis clearly isolated equine veterinarians and dog and cat practitioners as groups at increased risk of carriage of MRSA. Conclusion Carriage of MRSA is a notable occupational health issue for veterinarians in clinical practice in Australia, particularly those who work with horses.     

Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds.

Sixty-eight methicillin-resistant Staphylococcus aureus strains were isolated from mastitis milk samples originating from 20 Belgian dairyherds. All these strains appeared to be representatives of one single strain which was probably of human origin. Evidence is presented indicating a rapid in vivo evolutionary change in this strain. The following characteristics were found to be variable: the production of beta haemolysin inversely connected with fibrinolysin (staphylokinase) activity; the production of lipase, enterotoxin B and delta haemolysin; the resistance to neomycin, chloramphenicol, tetracyclines, methicillin and spectinomycin associated with constitutive or inducible macrolide resistance.     

Characterization of methicillin-resistant Staphylococcus aureus isolated from retail raw chicken meat in Japan

Two isolates of mecA-positive methicillin-resistant Staphylococcus aureus (MRSA) from retail raw chicken meat were characterized by phenotypic and genotypic methods. One isolate showed the human biovar, coagulase type III, phage group I * III, the lack of production of enterotoxins and TSST-1, and resistance to PCG/ABPC/EM/GM/KM. The other isolate showed the human biovar, coagulase type III, phage group III, production of enterotoxin C and TSST-1, and resistance to PCG/ABPC/CEZ. The biotyping results indicate that the two isolates showed characteristics of human S. aureus. They also harbored SCCmec type IV, which has prevalently been found in community-acquired MRSA isolates. This paper is the first publication regarding MRSA isolates from raw chicken meat in Japan.     

猪源耐甲氧西林金黄色葡萄球菌的耐药性及其SCCmec基因分型研究

为了解陕西猪场耐甲氧西林金黄色葡萄球菌(MRSA)的耐药表型及其SCCmec基因型分布情况,我们于2009年1月～7月在陕西地区6个规模化猪场,采集鼻腔拭子共计173份,利用选择性培养基分离金黄色葡萄球菌(SA),采用多重PCR方法扩增其中是否含有nuc基因和mecA基因进行MRSA鉴定;以琼脂稀释法进行药敏试验;并通过多重PCR方法对SCCmec进行基因分型。结果显示,调查的6个猪场中有2个猪场分离到MRSA,共分离到83株SA,其中有14株(16.9%)为MRSA,均为SCCmec-IVb型;药敏结果显示MRSA除对CFP、CHL、AMK和VAN敏感外,对CIP、OXA耐药率分别为57.1%和92.6%,对FOX、TET、ERY、甲氧嘧啶和卡那霉素的耐药率均为100%。结果显示了猪源SCCmec-IVb型MRSA株具有多重耐药特点,与人源SCCmec-IV型MRSA株耐药谱存在较大差异,这一结果提示猪源MRSA容易通过猪及其产品感染人。     

奶牛乳房炎金黄色葡萄球菌的分离鉴定

为检测江苏省某奶牛场中乳房炎是否为金黄色葡萄球菌所致,采取4份患病乳汁样本进行试验分离和鉴定,并针对金黄色葡萄球菌采用分离后鉴定,结果显示,分离到的2株致病菌为金黄色葡萄球菌,其他菌株还包括大肠杆菌、肠球菌等,为多种致病病原菌同时引起,主要为金黄色葡萄球菌感染。     

Dissimilarity of ccrAB gene sequences between methicillin-resistant Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus among bovine isolates in Korea

The sequences of the ccrAB genes from bovine-, canine- and chicken-originating methicillin-resistant Staphylococcus (S.) epidermidis (MRSE) and bovine methicillin-resistant Staphylococcus (S.) aureus (MRSA) were compared to investigate the frequency of intra-species horizontal transfer of the staphylococcal cassette chromosome mec (SCCmec) complex. Nineteen MRSE strains were isolated from bovine milk, chickens, and dogs, and their genetic characteristics were investigated by multilocus sequence typing and SCCmec typing. Among the animal MRSE strains, the most frequent SCCmec type was type IV, which consisted of the type B mec complex and ccrAB type 2. The ccrA2 and ccrB2 genes were sequenced from the bovine, chicken and canine MRSE strains and compared with those of the bovine MRSA strains. The sequences generally clustered as MRSA and MRSE groups, regardless of the animal source. Additionally, no bovine MRSE sequence was associated with the bovine MRSA groups. Although most of the bovine MRSE and MRSA isolates possessed SCCmec type IV sequences, our results suggest that the intra-species gene transfer of the SCCmec complex between bovine S. aureus and bovine S. epidermidis strains is not a frequent event.     

The prevalence of methicillin-resistant Staphylococcus aureus colonization in feedlot cattle

Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen and recent evidence has implicated food animals in the epidemiology of human infections in some regions. While the role of food in MRSA transmission and human health relevance are unclear, MRSA can be found in retail meat products internationally, including beef, yet there has been minimal investigation of MRSA in beef cattle. This study involved screening feedlot cattle for nasal and gastrointestinal colonization with MRSA shortly before the time of slaughter. Methicillin-resistant S. aureus was not isolated from any of 491 nasal swabs and 488 faecal samples. This finding is in contrast to studies that have isolated MRSA from retail beef in Canada, performed in the same laboratory using comparable culture techniques. The reason for this discrepancy is unclear but it demonstrates that further study of MRSA in livestock as well as slaughter, processing and retail environments is needed to elucidate the epidemiology of MRSA contamination of meat.     

Cluster of methicillin-resistant Staphylococcus aureus colonization in a small animal intensive care unit

CASE DESCRIPTION: Methicillin-resistant Staphylococcus aureus (MRSA) was isolated from the tracheostomy tube of an 18-month-old castrated male Golden Retriever in the intensive care unit (ICU) of the Ontario Veterinary College. This prompted an investigation of MRSA colonization in other animals in the ICU. CLINICAL FINDINGS: On day 1 of the investigation, MRSA was isolated from nasal swabs obtained from 2 of 10 animals (2/7 dogs and 0/3 cats), including the index case. Subsequently, MRSA was isolated from 3 of 12 animals on day 9; 3 of 9 animals on day 13; and none of 14, 5, and 6 animals on day 20, 27, and 78, respectively. Overall, MRSA was isolated from 6 of 26 (23%) animals during the outbreak period (4/22 dogs and 2/4 cats). The apparent incidence of MRSA acquisition in the ICU from days 1 through 13 was 20% (5/25 animals). No clinical signs of MRSA infections developed. All isolates were indistinguishable from one another. TREATMENT AND OUTCOME: Infection-control measures including active surveillance of all animals in the ICU, barrier precautions, and hand hygiene were used to control the apparent outbreak. CLINICAL RELEVANCE: Methicillin-resistant S aureus is an emerging problem in veterinary medicine. Intensive care units may be at particular risk for periodic outbreaks of colonization and disease. The outbreak of this report highlights the potential for clinically inapparent transmission of MRSA within a facility; infection-control measures that might facilitate MRSA eradication should be considered in ICU settings.     

Development of a new selective medium for isolation of methicillin-resistant Staphylococcus aureus

A new selective medium containing cephem antibiotics was developed for isolation of methicillin-resistant Staphylococcus aureus (MRSA). MRSA colonies on a medium containing ceftazidime (CAZ) were most easily identifiable and a medium containing cefoperazone (CPZ) was superior in suppressing the growth of other bacteria. With the medium containing a couple of CAZ and CPZ, MRSA and methicillin-resistant coagulase-negative staphylococci (MRCNS) were detected from 2 and 1 of 15 chicken meat samples respectively. The MRSA and MRCNS recovery test showed that the medium was effective for MRSA isolation, suppressing the growth of other bacteria efficiently. These results suggested that the medium containing a couple of CAZ and CPZ was useful for MRSA detection from foods and animals.     

Molecular characterization of methicillin-resistant Staphylococcus aureus strains from pet animals and veterinary staff in China

The aim of this study was to determine the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) isolates from pet animals and veterinary staff and the characteristics of these isolates. A total of 22 MRSA isolates were isolated from nasal swabs from dogs, cats and veterinary staff in six pet hospitals in six cities, and examined for antimicrobial susceptibility, the presence of resistance genes, Panton-Valentine leukocidin gene lukF-lukS, staphylococcal chromosomal cassette (SCC) mec typing, spa tying, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. Of 22 MRSA isolates, 21 were recovered from pet animals, and one was isolated from a member of sstaff. All 22 MRSA strains were resistant to penicillin, oxacillin, azithromycin, clindamycin and ceftriaxone, and harboured mecA, ermB and linA genes. The lukF-lukS gene was not detected in any of the MRSA isolates. Eighteen MRSA strains from Qingdao belonged to ST59-MRSA-IV-spa t437. Of four MRSA isolates from Beijing, one belonged to ST398-MRSA-V-spa t034, and three belonged to ST239-MRSA-III-spa t030 profiles. Two PFGE types (A and B) were identified. Two isolates originating from dogs and one isolate originating from a staff member in Beijing shared similar PFGE patterns. Our cumulative data suggested that cross-transmission of MRSA may have occurred between pet animals and veterinary staff.     

An outbreak of methicillin-resistant Staphylococcus aureus skin infections resulting from horse to human transmission in a veterinary hospital

There are increasing reports of methicillin-resistant Staphylococcus aureus (MRSA) infection and colonization in horses and evidence that MRSA can be transmitted between horses and humans. The objective of this study was to investigate reports of skin infection in personnel working with a foal with community-associated MRSA colonization and subsequent infection. Clinical diagnostic specimens were collected from individuals reporting skin lesions following contact with the affected foal. Nasal and groin screening swabs were collected from other veterinary personnel that attended a voluntary screening clinic. MRSA skin infections were identified in three neonatal intensive care unit personnel. Nasal colonization was subsequently identified in 10/103 (9.7%) other veterinary hospital personnel. Isolates were indistinguishable by pulsed field gel electrophoresis, classified as Canadian epidemic MRSA-5, possessed SCCmecIV, were negative for the Panton-Valentine leukocidin and were multidrug resistant. Transmission to veterinary personnel despite short-term contact with standard protective barriers highlights the potential importance of MRSA as an emerging zoonotic pathogen, and indicates that further evaluation of interspecies transmission of MRSA and means to prevent zoonotic infection are required.     

The first nosocomial outbreak of methicillin-resistant Staphylococcus aureus in horses in Sweden

Background

Methicillin-resistant Staphylococcus aureus (MRSA) in animals is a rare finding in Sweden. In horses, MRSA was first detected in a screening survey in 2007. In 2008, six clinical cases occurred in an equine hospital, indicating an outbreak.

Method

All MRSA isolates detected, 11 spa-type t011 and one t064 (n = 12), in infected horses (n = 10) and screening of horses (n = 2) in Sweden from December 2007 to March 2010 were retrospectively analysed with pulsed-field gel electrophoresis (PFGE) using Cfr9I and ApaI restriction enzymes, to study relationship between the isolates. Medical records of infected horses and outbreak investigation notes were scrutinised to monitor the clinical outcome and other aspects of the outbreak.

Results

Eight of the 10 infected horses were linked to one equine hospital and two to another hospital in the same region. The six horses infected with MRSA in 2008 underwent surgery during the period 22 May-7 July in one of the hospitals. Four more infections linked to the two hospitals were notified between 2009 and March 2010.Nine of the 11 spa-type t011 isolates had identical Cfr9I and ApaI PFGE pattern. All six infected horses from 2008 presented with this MRSA. Two t011 isolates differed in one and two bands, respectively, in PFGE.Nine horses suffered from surgical site infections (SSI). No antimicrobials were used following the MRSA diagnosis and the infections cleared. The time from surgery to MRSA diagnosis differed greatly between the horses (range 15-52 days).

Conclusions

Association in time and space of six horses infected with an identical MRSA strain of spa-type t011 confirmed an outbreak. Two isolates found in 2009 and 2010 in the outbreak hospital were closely related to the outbreak strain, indicating one circulating strain. Both spa-type t011 and t064 have been reported in horses in Europe prior to these findings. The observation that the infections cleared although antimicrobials were not used is encouraging for future prudent use of antimicrobials. The time from surgery to bacteriological diagnosis was not acceptable in most cases, as contagious spread was a risk. Sampling when symptoms of infection are noticed and accurate analysis are thus important.     

Isolation of Staphylococcus aureus from raw fish in relation to culture methods

Five hundred and fifty fish samples from various stages in the course of distribution in Hyogo Prefecture (209 retailed in super markets, 173 obtained from fishery cooperatives at a harbor, 91 caught by trawling and 77 caught by rod fishing) were examined for contamination with Staphylococcus aureus (S. aureus). S. aureus was detected in 41 (19.6%) of the retail fish samples and 46 (26.6%) of the samples from the fishery cooperatives. No S. aureus was isolated from the live fish (91 trawled and 77 fished by rod). With regard to the retail fish, the contamination rate of processed fish (26.0%) was significantly higher than that of unprocessed fish (14.2%). For 88 samples, the efficacy of the selective medium was compared using Baird-Parker agar and mannitol salt agar supplemented with egg yolk (MSEY agar) by the direct plate and enrichment culture methods. Using the direct culture method, the S. aureus positive rate with the Baird-Parker agar (30.7%) was significantly higher (P<0.01) than that with the MSEY agar (6.8%). The enrichment culture method remarkably raised the S. aureus detection rate. Seventy-eight (85.7%) of 91 isolates belonged to the human ecovar. Sixty-two (68.1%) of the 91 isolates had some enterotoxin genes, including 44 (48.4%) with the sea gene. These data showed that the fish were contaminated with S. aureus after landing and that Baird-Parker agar had an advantage in detecting S. aureus with a direct plate culture.