Clinical, microbiological, and molecular characterization of methicillin-resistant Staphylococcus aureus infections of cats

OBJECTIVE: To compare clinical information obtained from medical records of cats with methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S aureus (MSSA) infections, evaluate antibiograms of MRSA and MSSA for multiple-drug resistance (MDR), and characterize the strain type and staphylococcal chromosome cassette (SCC)mec type of each MRSA. SAMPLE POPULATION: 70 S aureus isolates obtained from 46 cats. PROCEDURES: Clinical information obtained from medical records, including signalment, clinical signs, histologic examination of affected tissues, and outcomes, was compared between the 2 groups. Composite antibiograms of MRSA and MSSA were compared statistically. The MRSA strains were characterized by use of pulsed-field gel electrophoresis and SCCmec typing. RESULTS: No statistical differences in signalment or subjective differences in clinical signs or outcomes were detected between groups with MRSA or MSSA infection. Significant differences in antimicrobial resistance were detected, with MRSA having complete resistance to fluoroquinolone and macrolide antimicrobials, whereas MSSA maintained a high frequency of susceptibility. Seven pulsed-field patterns were observed in 15 MRSA strains; all but 1 were highly related. All MRSA isolates contained a type II SCCmec element. CONCLUSIONS AND CLINICAL RELEVANCE: Because MDR cannot be predicted in staphylococcal infections in cats on the basis of clinical signalment, culture and susceptibility testing are recommended whenever initial empirical treatment is unsuccessful. Molecular characterization of MRSA strains suggests that there has been reverse-zoonotic transmission from humans. IMPACT FOR HUMAN MEDICINE: The SCCmec type II element is typically associated with nosocomial MRSA infections of people. Cats may serve as reservoirs for MRSA infections in humans.     

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.     

Antimicrobial susceptibility of canine and human Staphylococcus aureus collected in Saskatoon, Canada

Staphylococcus aureus is one of the most common causes of infection in people and is increasingly recognized in dogs. The increasing prevalence of methicillin resistant S. aureus (MRSA) is complicating the treatment of these infections. Panton Valentine leukocidin (PVL), a toxin involved in the pathogenesis of necrotic syndromes in people may be partially responsible for the rise of MRSA. Canine and human S. aureus from the same geographic area are genetically similar, indicating a common population and likely transmission. The implications of increasing antimicrobial resistance complicated by interspecies transmission, necessitates including both dogs and humans in S. aureus resistance surveillance studies. A collection of 126 S. aureus isolates from people (n = 99) and dogs (n = 27) were included, minimum inhibitor concentrations to a panel of 33 antimicrobials used in human and veterinary medicine were determined. No resistance to vancomycin, linezolid, daptomycin, quinupristin/dalfopristin or nitrofurantoin was found. A wide range of antibiograms were found; including resistance to 0-12 drugs (0-6 drug classes). Outstanding antibiograms included a canine MRSA resistant to rifampin and a human MRSA resistant to chloramphenicol. Inducible clindamycin resistance was found among 78% and 4% of canine and human MRSA and 17% and 25% of canine colonizing and human methicillin susceptible S. aureus (MSSA), respectively. Resistance to mupirocin was only found among human isolates including 20% of MRSA and 4% of MSSA. While no canine isolates were PVL positive, 39% of human MRSA and 2% of MSSA carried the gene. The bidirectional transmission of S. aureus between people and dogs necessitates the inclusion of isolates from both species in future studies.     

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from animals and veterinary personnel in Ireland

Reports of methicillin-resistant Staphylococcus aureus (MRSA) in animals have become more frequent in recent years. This paper documents the recovery of MRSA from animals with respiratory, urinary tract or wound infection and from animals subjected to surgical procedures following treatment in one veterinary hospital and 16 private veterinary clinics in different geographical locations throughout Ireland. MRSA was recovered from 25 animals comprising 14 dogs, eight horses, one cat, one rabbit and a seal, and also from 10 attendant veterinary personnel. Clinical susceptibility testing suggested that the 35 isolates fell into two different groups. One group of isolates (Group 1) was resistant to one or more of the following classes of antimicrobials: macrolides, lincosamines, tetracyclines and/or fluoroquinolones. The second group (Group 2) was resistant to macrolides, aminoglycosides, tetracyclines and trimethoprim/sulphamethoxazole and variably resistant to fluoroquinolones, lincosamines and rifampicin. One isolate in Group 2 was susceptible to trimethoprim. Epidemiological typing by antibiogram-resistogram (AR) typing, biotyping and by chromosomal DNA restriction fragment length polymorphism analysis using SmaI digestion followed by pulsed field gel electrophoresis (PFGE), confirmed these two major clusters. PFGE analysis showed that most isolates from non-equine animals were indistinguishable from each other and from the isolates from personnel caring for these animals. MRSA was isolated from eight horses which attended six different veterinary practices before referral to an equine veterinary hospital. Isolates from the eight horses and from their attendant personnel had PFGE patterns that were indistinguishable and were unlike the patterns obtained from the other isolates. Comparison of PFGE patterns of isolates from veterinary sources with patterns from MRSA recovered in human hospitals showed that the most frequently occurring pattern of MRSA from non-equine animals was indistinguishable from the predominant pattern obtained from the most prevalent MRSA strain in the human population in Ireland. However, the patterns of the isolates from horses were unlike any patterns previously reported in Irish studies of human isolates. This study shows that transmission of two strains of MRSA is occurring in veterinary practices in Ireland and that one strain may have arisen from human hospitals. The source of the second strain remains to be determined.     

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from small and exotic animals at a university hospital during routine microbiological examinations

Clinical specimens of small animals (n=869) were screened for the occurrence of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA; MRSA) during routine microbiological examinations, and results were confirmed by a multiplex PCR strategy. The genetic relatedness of all mecA-positive S. aureus isolates was further investigated by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), PCR for Panton-Valentine leukocidine genes (PVL) and staphylococcal cassette chromosome mec-typing (SCCmec). A total of 61 S. aureus isolates were found during a 20-month period of investigation, 27 (44.3%) of them harbouring the mecA gene for methicillin-resistance. The majority of MRSA were isolated in specimens from dogs (n=18) and cats (n=4). One guinea pig and one rabbit were found to be positive for an MRSA infected site. Similarly, three exotic animals, a turtle, a bat and a parrot, were found to be infected with MRSA. PFGE and MLST analysis revealed a certain genotype ("A" and "A-1") dominating the isolate collection (23 of 27). Furthermore, one isolate showed homologous PFGE pattern to the German epidemic strain Barnim ("BE") and another one ("BE-1") was considered to be closely related. A third genotype ("B") was detected in two cases. Two different sequence types (ST) were identified among the 27 MRSA isolates. PFGE type "A" and both strains related to the Barnim epidemic strain were assigned to ST22, whereas ST239 was associated to PFGE profile "B". The present data show that certain MRSA genotypes are capable of infecting a wide spectrum of small and exotic animals, especially in clinical facilities.     

胶东地区不同家禽MSSA和MRSA的流行分布与耐药性分析

通过了解胶东地区肉鸡、蛋鸡和水禽(鸭、鹅)中甲氧西林敏感/耐药金黄色葡萄球菌(MSSA/MRSA)的流行情况及耐药现状,为临床合理用药、有效控制耐药菌株的流行传播提供依据。采用细菌常规分离培养、质谱/PCR鉴定、微量肉汤稀释法和spa分型等方法,对2021年8月至10月在青岛地区采集的2730份禽源咽拭子进行MSSA及MRSA分离鉴定、spa分型及药敏试验,利用SPSS26.0软件和BioNumericsv7.6软件进行数据处理分析。2730份禽源咽拭子中共获得金黄色葡萄球菌742株(27.18%),其中MSSA 528株(19.34%),MRSA 214株(7.84%)。三种家禽菌株分离率依次为水禽(38.33%)、蛋鸡(28.45%)、肉鸡(20.33%)。351株代表性金黄色葡萄球菌共获得16种spa型别,MSSA(70.80%)和MRSA(95.30%)均是以t899为主。部分型别仅在蛋鸡(t010、t002、t3155)或肉鸡(t571、t011)或水禽(t1793、t5268、t267)中分离到。MSSA和MRSA对青霉素类、喹诺酮类等多种药物显示出高水平耐药,分别对8种和...     

Transmission of methicillin-resistant Staphylococcus aureus strains between different kinds of pig farms

The main objective of the present study was to investigate if different kinds of pig farms, like farrowing farms and rearing farms, play a role in the transmission of methicillin-resistant Staphylococcus aureus (MRSA) to Dutch finishing farms. Twelve farrowing farms, 11 finishing farms, 6 farrow-to finish farms, 1 rearing farm and 1 centre for artificial insemination were included. Screening of 310 pigs from these 31 farms showed 35 pigs (11%) to carry MRSA in their nares. On 7 of the 31 (23%) investigated farms colonized pigs were found, including 3 finishing farms, 3 farrowing farms and 1 farrow-to-finish farm. The use of standard antimicrobial medication of the pigs seemed to be a risk factor for MRSA carriage. Screening of the pigs on six farms supplying pigs for the MRSA positive farms revealed that the pigs on all but one farm were MRSA positive. Genotyping revealed that all MRSA strains were non-typeable by PFGE using the SmaI restriction enzyme and had multilocus sequence type (MLST) ST398. Different spa-types were found including t011, t108, t567, t899 and t1939, but the spa-types on epidemiologically related farms were identical indicating that MRSA are transmitted between farms through the purchase of colonized pigs. Two SCCmec types were found among the MRSA: type IV and type V. SCCmec type V was predominant. On two farms MRSA isolates with ST398, the same spa-type but with different SCCmec types (IV and V) were found, suggesting that different SCCmec elements have been inserted into MSSA with the same genotype. All MRSA strains were resistant to tetracycline, but additional resistances to erythromycin, lincomycin, kanamycin and gentamicin were also found. All MRSA isolates were negative for the exfoliative toxin genes (eta and etb), PVL toxin genes (lukF and lukS), toxic shock syndrome gene (tst-1), and the leukotoxin genes (lukE, lukD, lukM, lukF').     

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.     

乳房炎奶牛金黄色葡萄球菌毒素基因的检测及PFGE分型研究

作者针对临床及亚临床乳房炎奶牛乳汁中金黄色葡萄球菌分离株的毒素基因进行检测和脉冲场凝胶电泳(PFGE)基因分型,比较2种类型乳房炎金黄色葡萄球菌分离株的差异.无菌法采集奶样,采用国际标准方法从中分离金黄色葡萄球菌,用多重PCR方法扩增nuc基因和mecA基因以确证金黄色葡萄球菌(SA)和耐甲氧西林金黄色葡萄球菌(MRSA).进一步用PCR方法检测SA的各种毒素基因(SEs、ETs、TSST 1和PVL基因等).利用限制性内切酶Sma Ⅰ对SA基因组DNA进行酶切和PFGE分析,最后利用BioNumerics软件进行聚类分析.结果:19.3％(23/119)的临床乳房炎奶样和14.8％(26/176)的亚临床乳房炎奶样确定为金黄色葡萄球菌阳性样品,分别从中分离鉴定出43株和26株金黄色葡萄球菌,其中临床乳房炎分离株中有5株为mecA基因阳性.临床乳房炎奶牛奶样中检测到SA的SEA、SEB、SED、SEJ和PVL毒素基因,检出率分别为3.8％(1株)、11.5％(3株)、19.2％(5株)、7.7％(2株)和31.2％(10株);亚临床乳房炎奶牛乳样中仅检测到SA的SEA和PVL毒力基因,检出率分别为7.0％(3株)和84.1％(37株).表明临床与亚临床乳房炎奶牛乳汁中SA菌株携带的毒素基因不一样,SEs可能是临床乳房炎菌株的重要致病基因,PVL可能是亚临床乳房炎菌株的重要致病基因.69株SA使用Sma Ⅰ酶切分型后,可分为7个大簇、50个基因型,来源相同的SA分型后大部分位于同一簇内.临床乳房炎奶牛乳汁中检测到MRSA菌株,PVL基因在亚临床乳房炎中的检出率为临床乳房炎的2.7倍.PFGE方法能较好的区分临床乳房炎和亚临床乳房炎的SA分离菌株.     

Prevalence of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus Carriage in Three Populations

Background: A higher prevalence of methicillin-resistant Staphylococcus aureus (MRSA) colonization is reported in healthcare workers compared with nonhealthcare workers.
Hypothesis: The prevalence of MRSA colonization differed in people and pets in households with healthcare workers as compared with households without healthcare workers.
Subjects: A person and 1 dog or cat from 586 households defined as either a nonhealthcare (n = 213), veterinary healthcare (n = 211), or human healthcare (n = 162) worker household.
Methods: Prospective cross-sectional study. Samples from humans and pets were cultured in vitro. Staphylococcus aureus was identified as methicillin sensitive (MSSA) or MRSA with mec A polymerase chain reaction. Pulsed-field gel electrophoresis and spa -typing were used to characterize relatedness of S. aureus and MRSA and assign USA types.
Results: The prevalence of MSSA and MRSA in humans was 21.5% (126/586) and 5.63% (33/586), respectively, and 7.85% (46/586) and 3.41% (20/586), respectively, in pets. There were no differences in prevalences of either MSSA or MRSA between household types. The proportion of MRSA among all S. aureus isolates in humans and pets was 20.8% (33/159) and 30.3% (20/66), respectively. In <1.0% (4/586) of households, the same strain of MRSA was found in both a person and a pet.
Conclusions and Clinical Importance: There were no differences in the prevalences of MSSA or MRSA between healthcare worker and nonhealthcare worker households. Pets and people colonized with S. aureus were as likely to be colonized with MRSA. Colonization of a person and their pet with the same strain of MRSA was rare.     

Zinc resistance of Staphylococcus aureus of animal origin is strongly associated with methicillin resistance

This study was conducted to determine the occurrence of zinc and copper resistances in methicillin-resistant Staphylococcus aureus (MRSA) from swine and veal calves in a global strain collection. The test population consisted of 476 porcine MRSA isolates from ten European countries, 18 porcine MRSA isolates from Canada and seven MRSA from China, 92 MRSA and 60 methicillin-susceptible S. aureus (MSSA) isolates from veal calves in the Netherlands and 88 porcine MSSA isolates from four European countries. Most porcine MRSA (n=454) and all bovine MRSA belonged to clonal complex (CC) 398 whereas 37 of the pig MRSA from Europe and the seven Chinese isolates belonged to other CCs and 3 isolates were not classified into a CC. All isolates were tested for susceptibility to zinc chloride and copper sulphate using agar dilution and tested by PCR for the czrC gene encoding zinc resistance. Phenotypic zinc resistance (MIC>2mM) was observed in 74% (n=324) and 42% (n=39) of European MRSA CC398 from pigs and veal calves, respectively, and in 44% of the Canadian isolates (n=8), but not among the Chinese isolates. Almost all (99%) zinc-resistant MRSA carried czrC. Of the 37 European non-CC398 MRSA, 62% were resistant to zinc, but only 46% of them carried czrC. The MICs of the MSSA isolates to zinc chloride ranged from 1 to 4mM and none carried czrC. The MICs of copper sulphate were associated neither with methicillin resistance nor with the detection of czrC. This study showed that zinc resistance and the czrC gene are widespread among CC398 MRSA isolates. This suggests that the use of zinc in feed might have contributed to the emergence of MRSA.     

Staphylococcus aureus: study of genomic similarity of strains isolated in veterinary pathology using amplified fragment length polymorphism (AFLP)

Staphylococcus aureus is a widespread pathogen causing infections in different animal species. The extensive use of antibiotics, particularly methicillin, causes the rise of antibiotic-resistant strains (MRSA). In order to verify the epidemiology and genetic relatedness among MRSA and sensible strains (MSSA), an accurate fingerprinting technique, the amplified fragment length polymorphism (AFLP), was carried out. The isolates were cultured, subdivided on MRSA and MSSA and submitted for the genomic DNA extraction that was utilized for AFLP. The data were analysed for genetic similarity using the Dice coefficient. The results of genomic analysis among MRSA and MSSA and within them revealed that the major component of variation was due to variation within strains (82.12%), while variance among strains was lower (17.88%). The low level of genomic similarity found among S. aureus strains implies high level of genetic diversity. Different similarity was found as well in all strains independently of the source.     

Prevalence, antibiotic resistance, virulence traits and genetic lineages of Staphylococcus aureus in healthy sheep in Tunisia

Nasal swabs of 163 healthy sheep were obtained from two farms and one abattoir in Tunisia during 2010. Samples were inoculated in Baird Parker agar and ORSAB medium for Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) recovery, respectively. MRSA was detected in 5 of these 163 samples (3%) in ORSAB medium, and one isolate per sample was further studied. MRSA isolates were mecA-positive, typed as ST153-CC80-t044-agrIII, and contained blaZ, ant(6)-Ia, aph(3')-IIIa, erm(C), tet(K), and fusB genes encoding penicillin, streptomycin, kanamycin, erythromycin, tetracycline and fusidic acid resistance, respectively. These MRSA isolates showed indistinguishable or closely related PFGE-patterns and harboured the lukF/lukS gene encoding the Panton-Valentine leukocidin and the luk-ED, hla, hld, and hlg(v) genes. Methicillin-susceptible S. aureus (MSSA) were recovered in 68 of the 163 samples (41.7%) and one isolate per sample was characterized. Most of MSSA (82.4%) showed susceptibility to the tested antibiotics with exceptions: penicillin (6%, carrying blaZ gene), tetracycline (19%, carrying tet(K) gene) and fusidic acid (9%). The following toxin-genes were identified among MSSA: tst (53 isolates), luk-M (52), luk-ED, hla, hlb, hld and hlg(v) (67), hlg (1), sec (49), sel (52), and the egc-cluster-like sen-sem-sei-seo-seg (1). Ten spa-types (two of them new ones) and nine sequence types (six new ones) were detected among the 73 S. aureus isolates, and they were ascribed to agr types I and III. All MRSA and MSSA isolates were able to coagulate bovine plasma and MRSA harboured the immune-evasion-gene-cluster type E. Conclusions. Nares of healthy sheep could be a reservoir of PVL-positive community-associated-MRSA and also of TSST-positive S. aureus isolates, with potential implications in public health.     

Staphylococcus aureus (MSSA) and MRSA (CC398) isolated from post-mortem samples from pigs

There are many reports on the occurrence of Livestock Associated Methicilline resistant Staphylococcus aureus (LA-MRSA, CC398) in healthy pigs. There are however, very few reports of LA-MRSA being associated with pathological lesions in pigs. With this study we try to find the answers to the questions: (1) how often is S. aureus found in post-mortem material from pigs, (2) how many of these isolates are methicillin resistant, (3) are these equally distributed over the years? Here we report the isolation of MRSA and of methicillin sensitive S. aureus (MSSA) from samples derived from post-mortem examinations at the Animal Health Service in The Netherlands in the period from 2003 through October 2008. The MSSA and MRSA described here were isolated from 159 pathological lesions and from 7 submissions of aborted foetuses derived from a total of 116 animals, representing 103 submissions coming from 92 different herds. This is approximately 0.5% of all pigs submitted for post mortem examination in those years. The proportion of pigs from which S. aureus (both MSSA and MRSA) was isolated from, did not increase over the years. MSSA (N=97) and LA-MRSA CC398 (N=18) were present mainly in (peri)arthritis in over 30% of all cases, but were also isolated from internal organs such as lung, brain, spleen, kidneys, heart, indicating septicaemia. Remarkably, one non-CC398 MRSA (ST1) was isolated in a joint and a kidney of one pig. This isolate was resistant to 5 out of 6 antimicrobials tested. There was no significant difference in the type of lesions in which LA-MRSA was found compared to MSSA. The number of antimicrobials these isolates were resistant to, increased rapidly after 2004. LA-MRSA was isolated for the first time in 2005 and then again in 2007 and 2008, suggesting that this is an emerging pathogen. However, due to changes in the panel of antimicrobials used to test S. aureus for antimicrobial susceptibility in 2005 and 2007, the possibility exists that we may have missed some MRSA isolates. LA-MRSA isolates are resistant to at least three but sometimes five out of six antimicrobials tested. All isolates were susceptible to the combination of Trimethoprim/Sulfamethaxol.     

High occurrence of methicillin-resistant Staphylococcus aureus ST398 in equine nasal samples

Methicillin-resistant Staphylococcus aureus (MRSA) infections do occur in equine patients. Little is known, however, about their origin and the general equine MRSA colonization status. In West European horses in particular, neither the colonization rate nor the present strains or their antimicrobial susceptibility patterns are known. In the present study, a sample of 110 (Belgian, French, Dutch and Luxemburg) horses presented at a Belgian equine clinic was screened for nasal MRSA carriage. An indirect culturing protocol using a 0.001% colistin and nalidixic acid containing broth was compared to a direct agar method. Phenotypic identification following growth on a chromogenic MRSA screening agar (ChromID MRSA) was combined with genotypic analysis (PCR, PFGE, SCCmec, spa, and MLST typing). Antimicrobial susceptibility was tested through disk diffusion. Twelve (10.9%) horses carried MRSA, with the enrichment protocol resulting in a significantly higher isolation rate. None of the isolated strains were typeable through SmaI PFGE. They all harboured SCCmec type IVa or V and belonged to spa type t011 or t1451 of the ST398 lineage. All isolates were tetracycline resistant and sulfonamide and enrofloxacin susceptible. Macrolide, lincosamide, trimethoprim and aminoglycoside susceptibility varied and in total five different antimicrobial resistance patterns were distinguished. These results show that ST398 is certainly present in West European horses. Due to its known interspecies transmission and the structure of the equine industry, the presence of this clone in horses poses a substantial health hazard for both animals and humans.     

Biofilm production by Staphylococcus aureus associated with intramammary infection

Biofilm production by 221 Staphylococcus aureus isolates from 45 dairy herds was evaluated. Isolates were from composite milk of 117 cows, from teat skin of 70 cows, and from 34 milking machine unit liners. Of S. aureus from milk samples, 41.4% were biofilm producers, as compared to 24.7 and 14.7% of the isolates collected from skin and liners. Pulsed field gel electrophoresis (PFGE) best categorized S. aureus biofilm producers as compared to phage typing and binary typing. PFGE types that were significantly associated with isolation from milk as opposed to teat skin or liners, had isolates that were more likely to produce biofilm than PFGE types that were isolated from milk, skin and liners at similar frequencies. By contrast, PFGE type A was significantly associated with isolation from teat skin and had few biofilm producers. PFGE type Q, which is exclusively a milk, isolate produced more biofilm as evidenced by absorbance values. Given S. aureus that are associated with milk are more likely to produce biofilm as compared to extramammary sources (teat skin and milking unit liners), suggests that biofilm production is a risk factor for infection.     

Detection of methicillin-resistant Staphylococcus aureus (MRSA) in backyard pigs and their owners, Michigan, USA

Methicillin-resistant Staphylococcus aureus (MRSA) have been reported in commercially raised pigs and their human handlers, raising concerns of zoonotic transmission. To determine whether MRSA in backyard-raised pigs is commonly transmitted to their human owners, a matched study of this type of pigs and their owners was conducted in selected counties in Michigan. Nasal swabs from matched owner-pig pairs (n = 50 pairs) with a few unmatched pig (n = 3) and human (n = 4) samples were collected and processed using standard isolation and identification protocols. No matched owner-pig pair was found; however, MRSA was isolated from 1/54 (1.9%) human samples and 2/53(3.8%) of the pigs. The single human isolate was not strain type USA100-1100 by pulsed-field gel electrophoresis (PFGE), was sequence type (ST) 8 by multilocus sequence typing (MLST), possessed SCCmec type IVb and agr I and was negative for the Panton-Valentine leukocidin (PVL) toxin gene. The two pig isolates were indistinguishable by PFGE (not USA100-1100), and both isolates were ST5 by MLST, possessed SCCmec type III and agr II and were negative for the PVL gene. Persons raising backyard swine from the selected Michigan counties had MRSA carriage rates similar to that of the general US population, suggesting that their avocational pig exposure did not increase their risk of MRSA.     

Prevalence and Characteristics of Staphylococcus aureus in Connecticut Swine and Swine Farmers

This study explores the characteristics of Staphylococcus aureus (S. aureus) in swine and their human handlers in a convenience sample of 35 farms in Connecticut. Husbandry practices are clearly different from better‐known concentrated animal feeding operations (CAFOs) with less intensive rearing conditions. Nasal samples were collected from 263 pigs and nine humans on 35 farms during the 2010 rearing season. Samples were analysed using established microbiology methods, and resulting methicillin‐sensitive (MSSA) and resistant (MRSA) isolates were typed by pulsed‐field gel electrophoresis (PFGE) and spa typing. PCR was used to detect the presence of the Panton‐Valentine Leukocidin (PVL) gene, a cytotoxin usually associated with CA‐MRSA infection. A farm assessment form and questionnaire were used to obtain the information about husbandry practices and human exposure risk, respectively. Staphylococcus aureus colonized swine and humans were found in 51% (18/35) of the farms sampled at a rate of 30% (85/259) and 22% (2/9), respectively. Eight pigs and two humans were MRSA positive on five farms. MRSA in swine was related to healthcare‐associated (HA), community‐associated (CA) or livestock‐associated (LA) MRSA strains, whereas humans were colonized with HA‐MRSA. On the basis of spa typing, there was evidence of human–animal transmission thereby signifying humanosis/reverse zoonoses. The PVL gene was found in 88% (7/8) of MRSA swine isolates, the first time this gene has been seen in colonized pigs sampled on US farm. MSSA isolates belonged to six spa types: t337 (41%), t034 (12%), t334 (12%), t4529 (12%), t8760 (18%) and t1166 (6%) including LA strains. This is the first time spa type t8760 has been reported and the only MSSA with the PVL gene. In summary, MRSA including LA strains (LA‐MRSA) can be found on small farms with different husbandry practices from CAFOs, suggesting that preventive measures for zoonotic MRSA infection should address a range of animal production.     

A real-time PCR assay to detect the Panton Valentine Leukocidin toxin in staphylococci: screening Staphylococcus schleiferi subspecies coagulans strains from companion animals

Recent reports suggest that methicillin-resistant strains of Staphylococcus schleiferi subspecies coagulans are now commonly isolated from dogs. Given the association of a potentially mobile SCCmec type IV element with lysogenic phage-encoded Panton Valentine Leukocidin (PVL) toxin genes in community-acquired methicillin-resistant Staphylococcus aureus strains we hypothesized that methicillin-resistant S. schleiferi ssp. coagulans strains may also encode PVL toxin genes. Forty S. schleiferi ssp. coagulans strains isolated from companion animals were studied. Susceptibility to oxacillin was determined by broth microdilution and all isolates were screened by PCR for the presence of the mecA gene. SCCmec typing was performed on 14 isolates. A real-time PCR assay was developed for the detection of the PVL genes using a SmartCycler. Pulsed-field gel electrophoresis (PFGE) was performed to determine whether S. schleiferi ssp. coagulans strains were homogeneous. Twenty-eight of the 40 isolates (70%) were resistant to oxacillin and 26/28 possessed the mecA gene by PCR. SCCmec IV was identified in seven strains; the other seven isolates were not typable by this technique. All 40 strains were negative for the PVL toxin gene. PFGE showed a heterogeneous population and 13 different profiles were determined. In conclusion, this study showed that PVL toxin genes were not detected in a heterogeneous population of methicillin-resistant S. schleiferi ssp. coagulans strains isolated from companion animals.     

Study of methicillin resistant Staphylococcus aureus (MRSA) in Danish pigs at slaughter and in imported retail meat reveals a novel MRSA type in slaughter pigs

Methicillin-resistant Staphylococcus aureus (MRSA), especially CC398, have emerged in livestock worldwide. We investigated the occurrence of MRSA in pigs at slaughter and in retail meat. During 2009, nasal swabs (n=789) were taken from pigs at slaughter. Moreover, 866 meat samples [Danish: pork (153), broiler meat (121), beef (142) and; imported: pork (173), broiler meat (193), and beef (84)] were randomly collected in retail stores and outlets. MRSA was isolated from nasal swabs or from meat samples after preenrichment (Mueller Hinton broth with 6.5% NaCl), selective enrichment (tryptone soya broth with 4 mg/L cefoxitine and 75 mg/L aztreonam) and selective plating on Brilliance Chromogenic MRSA agar. The presence of mecA was confirmed by PCR and the MRSA isolates were spa typed. Novel MRSA spa types were characterized by MLST, PFGE and SCCmec typing. Thirteen percent (101/789) of the pigs had MRSA. Based on spa types 93% corresponded to CC398 (spa t011, t034, t1451, t2876, t2974), 4% to CC30 (t1333) and one isolate to CC1 (t0127). The spa type t1333 (CC30), which is common among methicillin susceptible S. aureus (MSSA) from pigs in Denmark, contained a SCCmec cassette type V and czrC zinc resistance gene. Imported broiler meat had the highest occurrence (18%) of MRSA, followed by imported pork (7.5%) and Danish pork (4.6%). MRSA ST398 was found for the first time in Danish beef (1.4%). The finding of MRSA CC30 (spa t1333) suggest possible spread of the SCCmec cassette normally associated with ST398 into another S. aureus lineage common in pigs.