Methicillin-resistant Staphylococcus pseudintermedius among dogs admitted to a small animal hospital

The aim of this study was to determine the frequency of carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) among dogs admitted to a small animal hospital during a 17-month period, to characterize these isolates and to initially screen for possible factors associated with MRSP carriage. Swabs were taken from the nose/pharynx and the perineum as well as from wounds and skin infections (if present) of 814 dogs before entering the small animal hospital. A questionnaire for background information was completed. The staphylococcal species and methicillin resistance were confirmed pheno- and genotypically. The identified MRSP isolates were characterized by SCCmec typing, testing for susceptibility to 25 antimicrobial agents and SmaI-directed pulsed-field gel electrophoresis. A first screening for possible risk factors for MRSP carriage was performed by means of unifactorial contingency tables and CART analysis. Sixty (7.4%) dogs were positive for MRSP. All MRSP isolates harboured a type II-III SCCmec cassette and showed extended resistance to antimicrobial agents. Fifteen different SmaI patterns were observed. The major factors that clustered with MRSP carriage were former hospitalization and antibiotic treatment within the last six months before sampling. This study showed that only a minor part of the sampled dogs carried multi-resistant MRSP isolates. The facts that prior hospitalization and/or antibiotic therapy are potential associated factors for MRSP carriage underline the necessity of a judicious use of antibiotics in small animal medicine.     

Species and staphylococcal cassette chromosome mec (SCCmec) diversity among methicillin-resistant non-Staphylococcus aureus staphylococci isolated from pigs

While methicillin-resistant Staphylococcus aureus (MRSA) ST398 is known to be widespread in pig farms, few studies have investigated the species diversity and SCCmec types of methicillin-resistant non-S. aureus staphylococci (MRNAS) residing in the nose of pigs. We examined nasal swab samples of 200 pigs originating from 10 Belgian pig farms previously found positive for MRSA ST398. Suspected staphylococcal isolates were subjected to a 16S rRNA-mecA-nuc PCR. Confirmed MRNAS were genotypically identified to the species level and investigated with a SCCmec typing PCR. MRNAS (n=72) were detected on all 10 farms and were carried by 29.5% of the pigs. Seven MRNAS species were found: Staphylococcus epidermidis (38.9%), Staphylococcus sciuri (18.1%), Staphylococcus pasteuri (18.1%), Staphylococcus rostri (12.5%), Staphylococcus warneri (8.3%), Staphylococcus haemolyticus (2.7%) and Staphylococcus hominis (1.4%). SCCmec cassettes were of type IVa (29.2%), type IVc (25%), type III (22.2%), type V (5.6%) or could not be assigned to any of the known types (NT types) (18.1%). Five distinct NT types were found. The predominance of methicillin-resistant S. epidermidis (MRSE) in our samples is remarkable, as MRSE is mainly associated with humans. The finding of three different SCCmec elements (IVa, V, NT type 1) in MRNAS that also prevail or predominate in MRSA ST398 shows that MRNAS might be an important SCCmec reservoir for MRSA in pigs. Yet, the occurrence of multiple other SCCmec types illustrates that further studies are required to understand the presence and spread of SCCmec in methicillin-resistant staphylococci from animals.     

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.     

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').     

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.     

Characteristics of methicillin resistant Staphylococcus aureus isolated from chicken meat and hospitalized dogs in Korea and their epidemiological relatedness

Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important pathogens in human and veterinary hospitals. The isolation of MRSA from animals and foodstuffs has been reported with an increased incidence. However, methicillin (oxacillin) is not used in animal husbandry or in animal hospitals in Korea. In this study, three pre-MRSA and one silent mecA-carrying methicillin susceptible S. aureus (smMSSA) were isolated from retail chicken meat, and three MRSA were isolated from hospitalized dogs in Korea. The three pre-MRSA isolates were determined to have a staphylococcal cassette chromosome mec (SCCmec) type III, and the smMSSA isolate was not classified. The animal hospital isolates were found to contain SCCmec type II. Seven and 15 S. aureus isolated from hospitalized humans and bovine milk, respectively, were also examined in this study in order to determine the epidemiological origins of MRSA. Multilocus sequencing typing (MLST) revealed that the chicken meat and bovine milk isolates were closely related to the animal hospital isolates. The SCCmec characteristics and MLST analyses indicated the possibility of the human to animal transmission of MRSA. These results highlight the importance of identifying MRSA carriers as well as intercepting MRSA transmission because MRSA is becoming increasingly widespread without any plausible relationship with the use of methicillin (oxacillin).     

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.     

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.     

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.     

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.     

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.     

Antimicrobial activity of gallium maltolate against Staphylococcus aureus and methicillin-resistant S. aureus and Staphylococcus pseudintermedius: An in vitro study

Gallium is a trivalent semi-metallic element that has shown antimicrobial activity against several important human pathogens. This antimicrobial activity is likely related to its substitution in important iron-dependent pathways of bacteria. The genus Staphylococcus, which includes human and animal pathogens that cause significant morbidity and mortality, requires iron for growth and colonization. In this study, gallium maltolate, at various concentrations between 50 and 200μM, inhibited the in vitro growth of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) at time-points between 8 and 36h after inoculation. The inhibitory activity of gallium maltolate against clinical isolates of MRSA and methicillin-resistant Staphylococcus pseudintermedius (MRSP) from a veterinary teaching hospital was determined.     

Prevalence of methicillin-resistant Staphylococci on a farm: staff can harbour MRS when animals do not

The aim of this work was to establish the prevalence of methicillin-resistant Staphylococci (MRS) in the animals and staff of a teaching and research farm. Samples of dairy cattle (36), beef cattle (26), sheep (19), horses (21), pigs (23), goats (23) and humans (13) were collected and screened for the presence of MRS. The detection of mecA gene was performed by PCR to determine the resistance of the samples to methicillin. Antimicrobial-resistance testing to penicillin, meropenem, ceftriaxone, cephalothin, oxacillin, levofloxacin, enrofloxacin, chloramphenicol, ciprofloxacin, gentamicin, clindamycin, erytromycin, linezolid, sulfamethoxazole/trimethoprim, tetracycline, doxycycline and vancomycin was performed on the mecA+ isolates. From the 161 samples, four methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were isolated from human beings (31%), whereas none was isolated from animals (0%). No methicillin-resistant Staphylococcus aureus (MRSA) were isolated. All of the MRCoNS isolates from this work presented different antimicrobial resistance patterns. MRCoNS may be present in humans associated with animals while not present in the animals. Selective pressure outside of the farm and a lack of MRCoNS transmission between humans and animals may be responsible for this lack of correlation.     

The prevalence of methicillin-resistant staphylococci in healthy horses in the Netherlands

Two hundred healthy horses housed at 23 different farms and one clinic and 42 persons in close contact with these horses were screened for the presence of methicillin resistant staphylococci. Samples were taken from the nose and the pastern of the horses and from the nose and throat of the humans and incubated in selective media. Isolates were identified by standard techniques and their susceptibilities were tested using an agar diffusion method. Methicillin-resistant strains were tested for the presence of the mecA gene by PCR. In 45 horses (22.5%) and 15 humans (35.7%) mecA positive staphylococci were found. All isolates were coagulase negative staphylococci, except for one methicillin-resistant Staphylococcus aureus isolated from a veterinarian. Staphylococcus sciuri was the predominant species found among the methicillin resistant staphylococci (MRS) in the horses, whereas S. epidermidis predominated in the humans. From the horses, often more than one species of MRS could be isolated, resulting in a total of 175 mecA positive equine isolates. The equine isolates were predominantly susceptible to most antimicrobials tested, whereas the human isolates showed more resistance. In conclusion, no methicillin-resistant Staphylococcus aureus was found in healthy horses in the Netherlands, but methicillin-resistant coagulase negative staphylococci were found frequently. Further studies are needed in order to investigate whether horses can be a reservoir for MRS or the mecA gene for humans.     

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.     

Molecular characterization of community‐associated methicillin‐resistant Staphylococcus aureus from pet dogs

Community‐associated methicillin‐resistant Staphylococcus aureus (MRSA) is a serious public health concern and in Australia, one that disproportionately affects Aboriginal people. Paralleling MRSA in human medicine, methicillin‐resistant S. pseudintermedius (MRSP) is an increasingly prevalent pathogen in veterinary medicine. We aimed to characterize the carriage of MRSA and MRSP in dogs and cats from predominantly Aboriginal communities in a very remote region of New South Wales (NSW), Australia. Pets (303 dogs and 80 cats) were recruited from six communities in western NSW. Three swabs were collected from each animal (anterior nares, oropharynx and perineum) and from skin lesions or wounds (if present) and cultured on selective media for methicillin‐resistant staphylococci. Human host‐adapted community‐associated MRSA representing four multilocus sequence types (ST1‐IV, ST5‐IV, ST72‐IV, ST93‐IV) were isolated from eight dogs (prevalence 2.6%, 95% confidence interval 1.3%–5.1%). Two ST5‐IV isolates from a single dog were phenotypically trimethoprim‐resistant, harbouring trimethoprim‐resistant gene dfrG within the SCCmec type IVo mobile genetic element. MRSA was not isolated from any cats and MRSP was not isolated from any dogs or cats. This study estimated a high prevalence of human host‐adapted community‐associated MRSA carriage in dogs despite an absence of MRSP. This suggests MRSA carried by dogs in remote NSW originate from human hosts. The cycle of transmission between people, dogs and common environmental sources warrants further investigation. To our knowledge, this is the first report of trimethoprim‐resistant ST5‐IV in eastern Australia and the first report of trimethoprim‐resistant ST5‐IV from a dog.     

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.     

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.     

Microarray based study on virulence-associated genes and resistance determinants of Staphylococcus aureus isolates from cattle

Staphylococcus aureus is a common pathogen which can colonise and infect not only man, but also domestic animals. Especially, infection of cattle is of high economic relevance as S. aureus is an important causal agent of bovine mastitis. In the present contribution, a DNA microarray was applied for the study of 144 different gene targets, including resistance genes and genes encoding exotoxins, in S. aureus isolated from cows. One hundred and twenty-eight isolates from Germany and Switzerland were tested. These isolates were assigned to 20 different strains and nine clonal complexes. The majority of isolates belonged either to apparently closely related clonal complexes 8, 25, and 97 (together 34.4%) or were related to the sequenced bovine strain RF122 (48.4%). Notable characteristics of S. aureus of bovine origin are the carriage of intact haemolysin beta (in 82% of isolates tested), the absence of staphylokinase (in 89.1%), the presence of allelic variants of several exotoxins such as toxic shock syndrome toxin and enterotoxin N, and the occurrence of the leukocidin lukF-P83/lukM (in 53.1%). Two isolates were methicillin-resistant S. aureus (MRSA). One of them was a clonal complex 8 MRSA related to the epidemic MRSA strain Irish 01. The other one belonged to ST398/spa-type 34 resembling a newly emerging MRSA strain which has been described to occur in humans as well as in domestic animals. The presence of these two strains highlights the possibility of transfers of S. aureus strains between different host species.