Occurrence and characterization of methicillin-resistant staphylococci from bovine mastitis milk samples in Finland

Background

Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods.

Results

Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates.

Conclusions

The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSA_mecC ST130. The study provides a baseline for further MRS monitoring.     

Genetic diversity and methicillin resistance of Staphylococcus aureus originating from buffaloes with mastitis in Iran

The aims of the present study were to investigate the genetic diversity and methicillin resistance in S. aureus isolates recovered from mastitis-affected buffaloes. Five hundred seventy-eight milk samples were obtained from buffaloes with mastitis in three provinces, Iran. Ninety-one of the 578 tested samples contained S. aureus (15.74%), in two cases were methicillin resistant S. aureus (MRSA). Isolates were typed by spa typing, followed by MLST on some representative isolates and SCCmec typing for MRSA strains. The presence of genes encoding Panton–Valentine leukocidin (PVL) was also tested by PCR. Eight spa types were identified, with t3576 (n = 18), t7311 (n = 18) and t937 (n = 17) were the most common, followed by t304 (n = 11), t7308 (n = 9), t521 (n = 7), t267 (n = 6), and t527 (n = 5). MLST revealed four different sequence types (STs) including ST97 (related to t521 and t527 spa types), ST352 (related to t267), ST291 (related to t304 and t937) and ST522 (related to t7338, t7311 and t3576). Two MRSA were identified as t304-ST291-SCCmecIV and t7311-ST522-SCCmecIV. No PVL-positive S. aureus were found. A significant difference in geographical distribution of genotypes was observed, with some types being prevalent in all studied provinces (P < 0.001). The results demonstrated genetic diversity among the S. aureus strains involved in mastitis in buffaloes. This study also provides evidence of the presence of MRSA belonging to genotypes which have been earlier reported in human infections, emphasizing the need for their epidemiological monitoring.     

Molecular Epidemiology and Antimicrobial Resistance Mechanisms of Methicillin‐Resistant Staphylococcus aureus Isolated from Bovine Milk

The aim of this study was to identify methicillin‐resistant Staphylococcus aureus (MRSA) strains gathered from 2002 to 2006 from milk samples in Aydin region in Turkey. Among 93 S. aureus strains isolated from bovine milk with mastitis, 16 were resistant to methicillin. Methicillin‐resistant S. aureus strains were studied further for their staphylococcal cassette chromosome mec (SCCmec) types, pulsotypes, spa and MLST types, antimicrobial susceptibilities, mechanisms of resistance and presence of Panton–Valentine leucocidin (PVL) toxin gene. The MRSA strains were multi‐drug resistant. The susceptibility rates to antimicrobials tested were 0%, 0%, 0%, 0%, 6.25%, 16.25% and 56.25% for erythromycin, clindamycin, chloramphenicol, gentamicin, tetracyclin, ciprofloxacin and vancomycin, respectively. All tetracycline and gentamicin resistant strains carried tet(M) and aac(6)‐aph(2) gene, respectively. Among macrolide‐resistant isolates, nine had erm(A), and seven had both erm(A) and erm(B) genes. The molecular characterization by pulsed‐field gel electrophoresis showed presence of three pulsotypes with their variants. The pulsotype B strains were type IV with SCCmec typing, and representative of pulsotype B was t190 by spa typing and ST8 by MLST typing. The strains with pulsotype A and C were SCCmec III, and representative of these pulsotypes was t030 by spa typing. The MLST type of pulsotype A was ST239 and pulsotype C was one allele variant of ST239. None of the isolates harboured the PVL gene. Presence of hospital‐related MRSA strains may indicate transmission of these strains between human and animals. In case of clonal spread beside the infected animals’ treatment of MRSA carrier, farm workers should also be considered. Hygienic measures and rational antibiotic use may avoid resistance selection, clonal dissemination of resistant strains and decrease losses because of mastitis in dairy herds.     

Low Rate of Methicillin‐resistant Coagulase‐positive Staphylococcal Colonization of Veterinary Personnel in Hong Kong

Elevated rates of methicillin‐resistant Staphylococcus aureus (MRSA) carriage have been reported in veterinary personnel, suggesting an occupational colonization risk. Hong Kong veterinary personnel (n = 150) were sampled for coagulase‐positive staphylococci (CPS) nasal colonization. Risk factors for colonization were assessed by questionnaire. Isolates were identified and antibiotic susceptibility determined. All CPS isolates were investigated for mecA carriage, SCCmec type and PVL genes. Two subjects were colonized with methicillin‐resistant CPS: one with MRSA (spa type t002 (CC5), SCCmec type II) and one with methicillin‐resistant Staphylococcus pseudintermedius (MRSP) (MLST type ST71, SCCmec type II‐III). MLST type ST71 S. pseudintermedius strain is the predominant MRSP clone circulating in dogs in Europe and in Hong Kong. The low MR‐CPS colonization rate may be associated with low levels of large animal exposure or low rates of MRSA colonization of companion animals in Hong Kong. Colonization with non‐aureus CPS, which may cause human infection, must also be considered in veterinary personnel.     

Methicillin-resistant Staphylococcus aureus in a population of horses in Australia

Objective To evaluate if methicillin‐resistant Staphylococcus aureus (MRSA) is present in the horse population in Australia. Design A two‐part retrospective study of laboratory submissions of microbial culture results from horses. Methods Part A: medical records of 216 horses that had MRSA screening performed on nasal swabs collected over a 30‐day period at admission to the Scone Equine Hospital Clovelly Intensive Care Unit were retrieved. Part B: laboratory records from 2004 to 2009 of culture submissions to the Scone Veterinary Laboratory were reviewed and cultures that grew MRSA were identified. The MRSA isolates from Parts A and B were genotyped over an 18‐month period. Results MRSA screening of 216 horses identified eight (3.7%) positive samples. MRSA was isolated from cultures of 80 (0.002%) clinical bacteriology samples over a 6‐year period. Genotypic analysis was performed on 36 isolates. All MRSA characterised had the same pulse field gel electrophoresis pattern (type 1), with eight closely related subtypes identified (subtypes A–F and H) and 66% of isolates classified as subtype D, which multilocus sequence and staphylococcal cassette chromosome mec typing analysis identified as ST612‐MRSA‐IVa, a clonal complex (CC) 8 S. aureus strain. Antimicrobial resistance to more than two classes of antimicrobials was common. Conclusions MRSA was present in a population of horses in Australia. Genotypic analysis of the isolates identified the MRSA strain as CC8 S. aureus. Further research needs to be undertaken to evaluate MRSA infection and colonisation of horses and personnel in Australia.     

Methicillin and aminoglycoside resistance in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolates from bovine mastitis and sequence analysis of their <Emphasis Type="Italic">mecA</Emphasis> genes

Although methicillin-resistant Staphylococcus aureus (MRSA) were generally isolated from human beings; these agents were recently isolated from various animal species. It has been shown that MRSA isolates are not only resistant to beta-lactam antibiotics, but can also be resistant to the other commonly used antibiotics. In this study, 18 phenotypic methicillin resistant S. aureus isolates from bovine mastitis cases were analyzed by PCR for the presence of mecA gene encoding methicillin resistance and aac(6′)/aph(2″), aph(3′)-IIIa and ant(4′)-Ia genes encoding aminoglycoside resistance. Out of 18 S. aureus isolates (oxacillin MICs, ≥4 μg/ml), 3 were positive for mecA gene. Only one from 3 mecA positive isolates was positive for genes encoding aminoglycoside-modifying enzymes and this isolate carried aac(6′)/aph(2″) in combination with aph(3′)-IIIa gene. The aph(3′)-IIIa gene was detected in 3 isolates. These three isolates carrying the aminoglycoside-modifying enzyme genes were resistant to gentamicin, kanamycin and neomycin. The mecA gene of 3 MRSA isolates was sequenced. All three mecA genes of these isolates were identical to that found in human MRSA strains, except a one-base substitution at nucleotide position 757. From the data presented in this study, it can be concluded that MRSA isolated from bovine mastitis may be originated from human beings, but further studies are needed to investigate the possibility of zoonotic transfer of MRSA.     

SCC <Emphasis Type="Italic">mec</Emphasis> typing and antimicrobial resistance of methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> (MRSA) from pigs of Northeast India

Staphylococcus aureus is one of the most important pathogens of both humans and animal. Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that causes serious infections both in hospitals and communities due to its multidrug resistance tendency. This study was undertaken to characterize the MRSA isolates from pigs and to determine the antimicrobial resistance of these isolates. Forty nine MRSA strains (one strain per positive pig) isolated from pigs of Northeast India were characterized by SCCmec typing and antimicrobial resistance. The overall prevalence of MRSA was 7.02 % with the highest prevalence recorded in pigs aged 1–3 months (P = 0.001) and in nasal samples (P = 0.005). Two SCC mec types (type III and V) were found in Indian pigs with predominance of type V. All isolates were resistant to penicillin. Seventeen resistance groups were observed where 87.75 % isolates showed multidrug resistance (showed resistance to three or more classes of antimicrobials). The most predominant resistance pattern observed was Oxytetracycline + Penicillin + Sulfadiazine + Tetracycline accounting 12.24 % of the isolates. The present study contributes to the understanding of characteristics and antimicrobial resistance of porcine MRSA isolates which in turn will help in devising strategy for the control of this pathogen. Findings of the study also throw light on multidrug resistance MRSA and emphasize the need for judicious use of antimicrobials in animal practice.     

Swine as reservoirs of zoonotic borderline oxacillin-resistant Staphylococcus aureus ST398

Methicillin resistance mediated by the mecA gene in Staphylococcus aureus, also known as “true MRSA”, is typically associated with high oxacillin MIC values (≥8 mg/L). Because non-mecA-mediated oxacillin resistant S. aureus phenotypes can also cause hard-to-treat diseases in humans, their misidentification as methicillin-susceptible S. aureus strains (MSSA) can compromise the efficiency of the antimicrobial therapy. These strains have been refereed as Borderline Oxacillin-Resistant S. aureus (BORSA) but their characterization and role in clinical microbiology have been neglected. Considering the increasing importance of livestock-associated methicillin-resistant S. aureus ST398 (LA-MRSA) as an emerging zoonotic pathogen worldwide, this study aimed to report the genomic context of oxacillin resistance in porcine S. aureus ST398 strains. S. aureus isolates were recovered from asymptomatic pigs from three herds. Oxacillin MIC values ranged from 4 to 32 mg/L. MALDI-TOF-confirmed isolates were screened for mecA and mecC by PCR and genotyped by means of PFGE and Rep-PCR. Seven isolates were whole genome sequenced. None of the isolates harbored the mecA gene or its variants. Although all seven sequenced isolates belonged to one sequence type (ST398), two different spa types (t571 and t1471) were identified. All isolates harbored conserved blaZ gene operon and no mutations on genes encoding for penicillin-binding-proteins were detected. Genes conferring resistance against other drugs such as aminoglycosides, chloramphenicol, macrolide, lincosamide and streptogramin (MLS), tetracycline and trimethoprim were also detected. Isolates also harbored virulence genes encoding for adhesins (icaA; icaB; icaC; icaD; icaR), toxins (hlgA; hlgB; hlgC; luk-PV) and protease (aur). Pigs can serve as reservoirs of non-mecA-mediated oxacillin-resistant ST398 strains potentially pathogenic to humans. Considering that mecA has been the main target to screen methicillin-resistant staphylococci, the occurrence of BORSA phenotypes is probably underestimated in livestock.     

Detection of methicillin‐resistant Staphylococcus aureus (MRSA) in the microbial flora from the conjunctiva of healthy donkeys from Sicily (Italy)

Objective To describe the bacterial flora present in the normal conjunctiva of donkeys from Sicily (Italy). Animals studied A total of 46 healthy donkeys housed in 3 locations within the territory of Palermo (Sicily, Italy) were studied. Donkeys ranged from 2 to 13 years of age, with a median age of 6 years. Procedures Forty‐six conjunctival swabs were obtained from both eyes of each animal, and specimens were cultured for aerobic bacteria. Furthermore, the antimicrobial activity of methicillin (1 μg) and oxacillin (5 μg) on Staphylococcus spp. isolates was evaluated, and a specific PCR assay, which allows the detection of mecA gene specific for methicillin‐resistant Staphylococcus aureus (MRSA) strains, was performed. Results Forty of 46 (86.9%) donkeys were positive for bacteria. Eighty bacterial isolates, representing 9 bacteria genera, were successfully cultured. The most frequently recovered bacterial genus was Staphylococcus (52/80 isolates; 65%). Several strains (20/80 isolates; 25%) belonging to the Enterobacteriaceae family were also isolated, among which the most frequently isolated genus was Enterobacter (eight isolates). Of the 52 Staphylococcus spp. isolates, 14 (26.9%) strains were oxacillin/methicillin resistant. The mecA gene was detected in 6/52 (11.5%) strains. Conclusions This study contributes to the knowledge about normal ocular flora and MRSA occurrence in donkey farms in Sicily.     

Antimicrobial susceptibilities and population structure of Staphylococcus epidermidis associated with ovine mastitis

Intramammary infections are a serious problem for dairy sheep farms, and Staphylococcus epidermidis is one of the main etiological agents of ovine mastitis. In this work, 131 S. epidermidis isolates, collected from 2201 dairy Sarda sheep belonging to 14 flocks with high somatic cell count scores, were studied. The flocks were located in diverse geographical areas of Sardinia, Italy. The aim of study was to assess the susceptibility of isolates to 13 antimicrobial agents, many of which are frequently used in mastitis therapy. Oxacillin was used for detecting methicillin-resistant S. epidermidis (MRSE) by disk diffusion test. Thirty-eight percent of the isolates (n = 50) were resistant to penicillin, 7.6% (n = 10) were resistant to tetracycline, and 2.3% (n = 3) were resistant to both penicillin and tetracycline (PTRSE). Two isolates were resistant to five antimicrobials including methicillin. Analysis of staphylococcal cassette chromosome mec (SCCmec) elements showed that both MRSE isolates harbored SCCmec type IVa. Based on pulsed-field gel electrophoresis (PFGE) typing by SmaI macrorestriction, S. epidermidis isolates were grouped into four clusters at 75% similarity level. The two multi-drug resistant MRSE isolates displayed distinct PFGE patters. This study indicates that S. epidermidis isolates from sheep milk samples may accumulate resistance markers for different antimicrobial agents. Furthermore, the occurrence of PTRSE and MRSE suggests to adopt adequate hygienic measures when handling animals with intramammary infections, in order to prevent spreading PTRSE and MRSE strains to humans through direct contact and/or consumption of contaminated food.     

Genotypic and phenotypic detection of capsular polysaccharide and biofilm formation in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolated from bovine milk collected from Brazilian dairy farms

Staphylococcus aureus is a pathogen that frequently causes mastitis in bovine herds worldwide. This pathogen produces several virulence factors, including cell-associated adhesins, toxic and cytolytic exoproteins, and capsular polysaccharides. The aim of the present study was to test for the presence of genes involved in capsular polysaccharide production and biofilm formation in S. aureus isolated from bovine mastitis samples collected from 119 dairy herds located in three different Brazilian regions, as well as to assay the production of capsular polysaccharides and biofilm, in vitro. The detection of the cap, icaAD, and bap genes was performed using PCR. The detection and quantification of capsular polysaccharide production was performed using ELISA assays. The ability of the isolates to form a biofilm was examined using the polystyrene surface of microtiter plates. All 159 S. aureus isolates investigated harboured the cap gene: 80 % carried the cap5 gene and 20 % carried the cap8 gene. Sixty-nine percent of the isolates expressed capsular polysaccharide (CP) in vitro, 58 % expressed CP5 and 11 % expressed CP8. All of the isolates harboured the icaA and icaD genes, and 95.6 % of the isolates carried the bap gene. Of the 159 isolates analysed, 97.5 % were biofilm producers. A significant association between the capsular genotype and phenotype and the amount of biofilm formation was detected: cap5/CP5 isolates tended to form more biofilm and to produce a thinner CP layer than cap8/CP8 isolates. The results indicate a high potential for pathogenicity among S. aureus isolated from bovine milk collected from three different regions in Brazil.     

Antibiotic resistance and phylogenetic comparison of human,pet animals and raw milk Staphylococcus aureus isolates

The present study was conducted to compare the S. aureus isolates from different sources in the basis of resistance phenotypic and genotypic features and phylogenetic differences. Total of 70 S. aureus isolates (including 25 human, 25 raw milk and 20 pet animal isolates) were subjected to the antimicrobial susceptibility testing, polymerase chain reaction (PCR) detection of the resistance genes and DNA fingerprinting using random amplification of polymorphic DNA–PCR (RAPD-PCR) to survey the variability of the isolates. Among 70 S. aureus, 55 (78.5%) isolates were MRSA. The isolates showed the highest antibiotic resistance to methicillin, ampicillin and penicillin (78.5%) and showed the lowest resistance to ciprofloxacin (12.8%). ErmB and tetM resistance genes were present in all isolates and the vanA gene was not detected in any of the isolates. Thirteen distinct clusters were identified in RAPD-PCR fingerprinting. Statistical analysis showed that the isolates without resistance to antibiotics were significantly in associated with raw milk origin (P < 0.05). According to the results of the study, S. aureus strains with pets and raw milk origin are significant sources of antibiotic-resistant isolates such as MRSA. They are also carriers of resistance genes that can be transmit to human isolates and cause drug resistance in human infections. Identifying the source of these infections is possible with a reliable genotyping method such as RAPD-PCR.     

Panton–Valentine leukocidin and some exotoxins of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and antimicrobial susceptibility profiles of staphylococci isolated from milks of small ruminants

The aims of this study were to determine the existence of pvl gene, some toxin genes, and mecA gene in Staphylococcus aureus strains isolated from sheep milk and to examine antimicrobial resistance profiles in staphylococci from sheep and goats' milk. The milk samples were collected from 13 different small ruminant farms in Kirikkale province from February to August 2009. A total of 1,604 half-udder milk samples from 857 ewes and 66 half-udder milk samples from 33 goats were collected. Staphylococcus spp. were isolated and identified from the samples. Toxin genes and mecA gene among S. aureus strains were determined by PCR. Antimicrobial susceptibility of staphylococci was examined by the disk diffusion method on Mueller-Hinton agar, and interpreted according to the Clinical Laboratory Standards Institute (CLSI) guidelines. The prevalence of subclinical intramammary infection in both ewes and goats was 5.2%. The most prevalent subclinical mastitis agents were coagulase-negative staphylococci and S. aureus with prevalences 2.8% (n:46) and 1.3% (n = 21), respectively. The prevalence of resistances in isolated Staphylococcus spp. to penicilin G, tetracycline, erythromycin, gentamicin, and enrofloxacin were found as 26.9% (18), 7.5% (5), 6.0% (4), 3.0% (2), and 1.5% (1), respectively. Only 3 of the 21 S. aureus ewe isolates (13.4%) were shown to harbor enterotoxin genes being either seh, sej or sec. However, fourteen (66.6%) of the 21 S. aureus isolates had pvl gene while none of the isolates harbored mecA gene. In conclusion, Staphylococci were shown to be the most prevalent bacteria isolated from subclinical mastitis of ewes and goats and these isolates were susceptible to most of the antibiotics. In addition, S. aureus strains isolated from ewes were harboring few staphylococcal enterotoxin genes. However, Panton–Valentine leukocidin produced by S. aureus could be an important virulence factor and contribute to subclinical mastitis pathogenicity.     

Genome analysis of methicillin‐resistant Staphylococcus aureus isolated from pigs: Detection of the clonal lineage ST398 in Cameroon and South Africa

Food animals are considered reservoirs of methicillin‐resistant Staphylococcus aureus (MRSA) and are implicated in their zoonotic transmission in the farm‐to‐plate continuum. LA‐MRSA has been reported as a zoonotic agent that has the potential to spread to humans and may cause infections in at‐risk groups. In this study, whole genome sequencing was used to describe the genetic environment (resistance mechanisms, virulence factors and mobile genetic elements) and investigate the genetic lineages of MRSA isolates from pigs in Cameroonian and South African abattoirs. During March–October 2016, 288 nasal and rectal pooled samples from 432 pigs as well as nasal and hand swabs from 82 humans were collected. Genomic DNA was sequenced using an Illumina MiSeq platform. Generated reads were de novo‐assembled using the Qiagen CLC Genomics Workbench and SPAdes. The assembled contigs were annotated, and antibiotic resistance genes, virulence factors, plasmids, SCCmec and phage elements were identified with ResFinder, Virulence Finder, PlasmidFinder, SCCmec Finder and PHAST, respectively. Core genome single nucleotide analysis was undertaken to assess clonal relatedness among isolates. A lower MRSA prevalence was observed in pigs in Cameroon (n = 1/13; 0.07%) compared with South Africa (n = 4/22; 18.18%), and none of the workers were colonized by MRSA. Genome analysis identified various antibiotic resistance genes along with six virulence factors in all isolates. All MRSA isolates belonged to the clonal lineage ST398 (spa‐type t011) and harboured the type Vc SCCmec and several plasmids. Our study shows that the livestock‐associated MRSA clonal lineage ST398 is already present in both Cameroon and South Africa and is probably underestimated in the absence of molecular epidemiological studies. It reveals the serious food safety and public health threat associated with this animal strain and underscores the need for interventions to contain this resistant clone.     

Antimicrobial susceptibility of Staphylococcus pseudintermedius colonizing healthy dogs in Saskatoon,Canada

This study reports antimicrobial susceptibility of Staphylococcus pseudintermedius carried by healthy dogs in Saskatoon, and describes changes in antimicrobial resistance since a 2008 study. One hundred healthy dogs presenting to the wellness service at the Western College of Veterinary Medicine were screened for S. pseudintermedius by culturing rectal and pharyngeal swabs. Staphylococcus pseudintermedius was identified biochemically and antimicrobial minimum inhibitory concentrations were determined by broth microdilution. Methicillin resistance was confirmed by polymerase chain reaction (PCR) and sequencing of the mecA gene. Of 221 S. pseudintermedius isolates from 78 dogs, 7 were methicillin resistant. No resistance to the fluoroquinolones, nitrofurantoin, tigecycline, vancomycin, quinupristin-dalfopristin, linezolid, or daptomycin was identified. Of the 78 positive dogs, isolates resistant to penicillin were found in 78%, to ampicillin in 61% and to tetracycline in 26%; resistance to oxacillin, erythromycin, clindamycin, trimethoprim + sulfamethoxazole, chloramphenicol, and gentamicin was found in < 10% of dogs. Compared to the 2008 study, the frequency of resistance to all drugs increased, and the frequency of colonization with pan-susceptible isolates decreased from 46% to 30%.     

Methicillin‐Resistant Staphylococcus aureus (MRSA) in Three Dairy Herds in Southwest Germany

The objective of this study was to analyse the occurrence of methicillin‐resistant Staphylococcus aureus (MRSA) in three dairy herds in the southwest of Germany that had experienced individual cases of clinical and subclinical mastitis associated with MRSA. The herds were identified by the detection of MRSA during routine resistance testing of mastitis pathogens. All quarters of all cows in the herds that were positive on California Mastitis Test were sampled for bacteriological analysis on two occasions. Bulk tank milk samples were also tested. Furthermore, nasal swabs were collected from people working on the farms and from cattle. Environmental samples were collected from associated pig holdings. Isolates were characterized using spa‐typing and testing for antimicrobial resistance. Our results revealed a substantial spread of MRSA in the three dairy herds. In the first of the two investigations carried out on all cows in the three herds, milk samples of 5.1–16.7% of dairy cows were found positive for MRSA. The respective proportions in the second herd level investigation were 1.4–10.0%. Quarters harbouring MRSA had higher somatic cell counts than quarters that were negative on culture. Methicillin‐resistant Staphylococcus aureus were also detected in nasal swabs of staff (7/9), cows (7/15) and calves (4/7), bulk tank milk samples (3/3) and environmental samples from pig premises (4/5) on the farm. Herds B and C had no contact to herd A. However, in all three herds MRSA of spa‐type t011 were detected in milk samples. Results show that MRSA of spa‐type t011 is a problem in dairy farms that needs urgent attention.     

Characterization of Methicillin‐Resistant Staphylococcus aureus Isolates from Pig Carcasses in Hong Kong

This study describes the isolation and characterization of methicillin‐resistant Staphylococcus aureus (MRSA) from slaughtered pigs sampled from local markets in Hong Kong. The nares of 400 slaughtered pigs were cultured and MRSA isolates characterized for the presence of antibiotic‐resistance determinants, toxins and SCCmec and spa types using PCR. Clonality was investigated using PFGE and MLST. The prevalence of MRSA colonization of slaughter pigs was 39.3%, the majority (92%) harbouring SCCmec type IVb. Of the 157 samples yielding MRSA, 13 had two distinct MRSA strains present. Spa type t899 was predominant, with only 5/170 isolates displaying closely related types (t4474, t1939, t2922 and t5390). PFGE with sma1 and MLST confirmed the strains as ST9. Most isolates were multidrug resistant. Tetracycline resistance (97%) was mainly attributable to tet(K) with only 3% of isolates additionally harbouring tet(M). Resistance to erythromycin (89%) and chloramphenicol (71%) was associated with the presence of erm(C), and fex( A), respectively. No strains carried cfr and there was no resistance to linezolid, although minimum inhibitory concentration (MICs) were close to the resistance break point. Resistance to clindamycin (99%), ciprofloxacin(78%), quinopristin–dalfopristin (44%) and cotrimoxazole (32%) was common, but remained low for fusidic acid (4%) and rifampicin (2%). All strains were negative for PVL, exfoliative, and enterotoxins. This survey confirmed the uniformity of MRSA isolates in pigs from several regions of China, in contrast to more diversified characteristics reported in European studies. Colonization rates were higher than previously reported. Isolates were resistant to a wide range of antibiotics, but resistance was not detected to linezolid, nitrofurantoin, vancomycin or tigecycline. Although the clinical importance of ST9 in humans is uncertain, continued surveillance, in particular of those occupationally‐exposed, is recommended.     

Whole genome sequencing of methicillin-resistant and methicillin-sensitive Staphylococcus aureus isolated from 4 horses in a veterinary teaching hospital and its ambulatory service

Genomic characterization was conducted on 2 methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from 2 horses hospitalized during an overlapping period of time and 2 methicillin-sensitive S. aureus (MSSA) strains isolated from 2 distinct horses. Phylogenetic proximity was traced and the genotypic and phenotypic characteristics of the antimicrobial resistance of the strains were compared.Whole genome sequencing of MRSA strains for this report was similar but differed from whole genome sequencing of MSSA strains. The MRSA strains were closely related, belonging to sequence type (ST) 612, spa type t1257, and SCCmec type IVd2B. The MSSA strains were also closely related, belonging to ST1660, spa type t3043, and having no detectable staphylococcal cassette chromosome mec elements. All MSRA and MSSA strains were Panton-Valentine leukocidin negative. There were discrepancies in the genotypic analysis and the antimicrobial susceptibility testing (phenotypic analysis) of MRSA strains for rifampin, trimethoprim-sulfamethoxazole, gentamicin, amikacin, and enrofloxacin.