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

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

Methicillin‐Resistant Staphylococcus aureus from Brazilian Dairy Farms and Identification of Novel Sequence Types

The aim of this study was to investigate the phenotypic and genotypic diversity and anti‐microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north‐eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti‐microbial susceptibility by Kirby–Bauer disc diffusion method. Confirmation of methicillin‐resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP‐2a latex agglutination. Clonal relatedness of isolates was determined by pulsed‐field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase‐negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622‐ST1625). The findings show that MRSA is prevalent in milk from semi‐extensive dairy cows in north‐eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm‐to‐table continuum is warranted.     

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

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

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.     

Risk factors for methicillin-resistant Staphylococcus aureus colonization in horses admitted to a veterinary teaching hospital

Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging problem in horses; however, the epidemiology of infection and colonization is poorly understood. This study evaluated factors associated with MRSA colonization at the time of admission to a veterinary teaching hospital. A case-control study evaluating historical factors was performed. Previous colonization of the horse, previous identification of colonized horses on the farm, antimicrobial administration within 30 days, admission to the neonatal intensive care unit, and admission to a service other than the surgical service were risk factors for community-associated colonization. A better understanding of risk factors for MRSA colonization is important to elucidate the epidemiology of this emerging veterinary and zoonotic pathogen, and to design evidence-based infection control programs.     

Epizootiology of endoparasitic infections in pet dogs and cats presented to a veterinary teaching hospital

Prevalences of, and risk factors associated with, protozoan and helminth parasite infections were determined for pet dogs and cats presented to the teaching hospital of the University of Pennsylvania School of Veterinary Medicine. Of 2294 canine fecal specimens, 34.8% were found to contain one or more kinds of parasite, consisting of: hookworms (14.4%); Trichuris vulpis (12.3%); Giardia (7.2%); ascarids (5.5%); coccidia (2.7%); cestodes (1.6%). The following risk factors were identified with infection in dogs: age less than 2 years; urban locality; male sex. Gonadectomy was associated with decreased parasite prevalences in both male and female dogs. Significant seasonal variations in prevalences of hookworm, ascarid and Giardia infections were found in dogs. Of 452 feline fecal specimens tested, 24.6% were positive for parasites, including: ascarids (16.4%); Giardia (3.5%); coccidia (2.9%); cestodes (2.4%); hookworm (1.1%). Host age of less than 2 years was a significant risk factor associated with parasitism in cats. Of 1571 dogs tested for Dirofilaria immitis microfilaremia, 3.7% were positive. Non-urban locality was a significant risk factor for D. immitis infection in dogs.     

Antibiotic sensitivity profiles do not reliably distinguish relapsing or persisting infections from reinfections in cats with chronic renal failure and multiple diagnoses of Escherichia coli urinary tract infection

Older cats with chronic renal failure (CRF) commonly develop urinary tract infections (UTI). Uropathogenic Escherichia coli (UPEC) is identified as the causal agent of UTI in most affected cats. Infections are often complicated, and UPEC infections may persist or recur in these cats. Antibiotic sensitivity profiles have been used to distinguish relapsing or persisting UTI from reinfection by different clones of the same species. However, the accuracy with which antibiograms discriminate different urinary E coli clones in cats is uncertain. We studied 17 cystocentesis-derived UPEC isolates collected from 5 cats with stable CRF and multiple diagnoses of UTI. UTIs were classified as relapses versus persistent infections or reinfections using antibiograms determined by Kirby-Bauer discs and Etests. Subsequently, clonality of UPEC isolates was determined by pulsed-field gel electrophoresis (PFGE). A comparison of PFGE results with antibiograms indicated that antibiotic resistance patterns varied considerably within several individual E coli clones. Both antibiotic susceptibility tests differentiated between relapsing or persistent infections and reinfections with only 58% overall efficiency. Thus, antibiotic sensitivity profiles cannot be relied upon to distinguish between persisting or relapsing infections as compared to reinfections in cats with CRF and multiple diagnoses of E coli UTI.     

Methicillin‐resistant Staphylococcus aureus in Resident Animals of a Long‐term Care Facility

Animals provide benefits to elderly and chronically ill people by decreasing loneliness, increasing social interactions, and improving mental health. As a result, many hospitals and long‐term care facilities allow family pets to visit ill or convalescing patients or support animal‐assisted therapy programs. These include programs that have resident animals in long‐term care facilities. Despite the benefits, there are concerns about disease transmission between pets and patients. Antibiotic‐resistant bacteria, such as methicillin‐resistant Staphylococcus aureus (MRSA), are a recognized problem in healthcare settings leading to refractory infections and potentially life‐threatening illnesses. MRSA has been isolated from numerous animal species, yet few studies are available on the carriage of this pathogen in animals residing in long‐term care facilities. Our objective was to characterize MRSA carriage among resident animals in a long‐term care facility. Methods: To document MRSA colonization, nasal swabs from 12 resident animals (one dogs and 11 cats) of a long‐term care facility were collected weekly for 8 weeks. Staphylococcus isolates were characterized by antimicrobial susceptibility and MRSA isolates were further characterized by pulsed‐field gel electrophoresis (PFGE). PFGE isolate patterns were compared with an existing database of MRSA isolate patterns at the Minnesota Department of Health. Results: Two of 11 cats were colonized with MRSA. MRSA was recovered from five of eight weekly samples in one cat and two of eight weekly samples in the other cat. All isolates were classified as USA100 (healthcare‐associated strains). Discussion: Long‐term care resident animals may acquire MRSA. Clonally related strains were identified over the 8‐week sampling period. It is unclear if pets serve as an on‐going source of infection to their human companions in long‐term care facilities.     

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.     

2012年-2015年我国部分地区禽偏肺病毒的分子流行病学分析

为了调查鸡群中禽偏肺病毒（aMPV）的流行情况,本研究从江苏、辽宁、河南、山东、河北等地有肿头症状的鸡群中取其鼻甲骨和鼻黏液进行 RT-PCR 检测,随机选取9株 PCR 阳性产物对其 F 基因进行序列测定与遗传进化分析。结果表明,280份病料中检出阳性样品142份,阳性检出率达50．7％;9个试验毒株之间 F 基因同源性为99．4％～100％,与 B 型 aMPV 代表株的同源性为97．7％～99．9％,而与 A 型、C型和 D 型 aMPV 代表株的同源性为71．9％～79．4％;由遗传进化树可见,这9株 aMPV 均属于 B 型分支。说明我国鸡群中目前存在 aMPV 感染,B 型毒株是优势流行基因型,从而为禽偏肺病毒疫苗的开发提供了流行病学资料。     

Genetic relatedness of methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolated from a dog and the dog owner

In the present study four methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains isolated from a dog (n = 3) and the anterior nares of the dog owner (n = 1) were investigated by conventional and molecular methods. The species identity of the four S. pseudintermedius strains was confirmed by conventional methods, by PCR mediated amplification of S. intermedius/S. pseudintermedius specific segments of thermonuclease encoding gene nuc and by restriction fragment length polymorphism analysis of phosphoacetyltransferase encoding gene pta. Investigation of the four S. pseudintermedius for toxinogenic potential revealed that all four strains were positive for the exfoliative toxin encoding gene siet and the leukotoxin encoding genes lukS, lukF. The oxacillin and penicillin resistance of the four S. pseudintermedius strains could be determined by cultivation of the strains on oxacillin resistant screening agar base, ChromID MRSA Agar and Brilliance MRSA Agar and by multiplex PCR detecting the resistance genes mecA and blaZ. The genetic relatedness of the strains was studied by macrorestriction analysis of their chromosomal DNA using pulsed field gel electrophoresis (PFGE). According to PFGE all four S. pseudintermedius strains represent an identical bacterial clone indicating a cross transmission between the dog and the dog owner.     

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.     

Risk factors for fecal shedding of Salmonella from horses in a veterinary teaching hospital

Identification of risk factors for horses shedding Salmonella in their feces helps identify patients at-risk of infection and protect the overall population through heightened biosecurity. Fecal samples from 230 hospitalized horses were cultured for Salmonella spp. Historical data were collected on 21 putative risk factors and assessed for association with the risk of a horse being culture positive using forwards stepwise logistic regression. Salmonella was isolated from 13 horses—most commonly from either the first (n=5) or second (n=4) sample collected. Only presenting complaint (confounded by age, breed and gender) was significantly (P≤0.05) associated with positive Salmonella-culture results. Analysis of residuals showed that the model was robust, but individual risk-factor estimates were changed by removal of outliers. Overall, presenting complaint (for example, lower-respiratory-tract disease) was the most important indicator of culture status.     

Prevalence of canine methicillin resistant Staphylococcus pseudintermedius in a veterinary diagnostic laboratory in Italy

The overall prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) was 2% (10/590) among 590 canine specimens submitted to an Italian veterinary diagnostic laboratory during a two-month period, and 21% (10/48) among Staphylococcus intermedius group (SIG) isolates. All methicillin-resistant strains exhibited additional resistance to fluoroquinolones, gentamicin, lincosamides, tetracyclines, and potentiated sulfonamides, belonged predominantly to spa type t02 and harboured SCCmec type II–III cassette.     

Methicillin-resistant Staphylococcus aureus in horses at a veterinary teaching hospital: frequency, characterization, and association with clinical disease

Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging equine pathogen. To attempt to control nosocomial and zoonotic transmission, an MRSA screening program was established for all horses admitted to the Ontario Veterinary College Veterinary Teaching Hospital, whereby nasal screening swabs were collected at admission, weekly during hospitalization, and at discharge. MRSA was isolated from 120 (5.3%) of 2,283 horses: 61 (50.8%) at the time of admission, 53 (44.2%) during hospitalization, and 6 from which the origin was unclear because an admission swab had not been collected. Clinical infections attributable to MRSA were present or developed in 14 (11.7%) of 120 horses. The overall rate of community-associated colonization was 27 per 1,000 admissions. Horses colonized at admission were more likely to develop clinical MRSA infection than those not colonized at admission (OR 38.9, 95% CI 9.49 160, P < 0.0001). The overall nosocomial MRSA colonization incidence rate was 23 per 1,000 admissions. The incidence rate of nosocomial MRSA infection was at the rate of 1.8 per 1,000 admissions, with an incidence density of 0.88 per 1,000 patient days. Administration of ceftiofur or aminoglycosides during hospitalization was the only risk factor associated with nosocomial MRSA colonization. MRSA screening of horses admitted to a veterinary hospital was useful for identification of community-associated and nosocomial colonization and infection, and for monitoring of infection control practices.     

The effect of Virkon S fogging on survival of Salmonella enterica and Staphylococcus aureus on surfaces in a veterinary teaching hospital

The objective of the study was to determine the disinfection efficacy of aerosolizing (cold fogging) Virkon S on survival of Stahpylococcus aureus and Salmonella enterica on different surfaces. Two experiments were conducted in different locations. Salmonella enterica and S. aureus were grown in broth culture and then seeded into pre-marked areas in each location and allowed to dry. Virkon S (1%) was aerosolized into the rooms (approximately 1L of per 30 m(3)). Samples were collected pre- and post-fogging for quantitative cultures to evaluate the efficacy of aerial disinfection. The reduction of S. enterica or S. aureus counts ranged from 3.40 to 0.95 log(10) (Salmonella) or 4.92 to 0.02 log(10) (Staphylococcus). The greatest reduction was evident in samples collected from non-porous horizontal surfaces, which were not obstructed from the air flow. These results indicate that fogging with Virkon S could be beneficial in routine disinfection of pre-cleaned surfaces. The benefits of routine use of cold fogging with Virkon S in veterinary hospital settings would include its wide-range antimicrobial action and minimal working-men power required to disinfect large areas. Also, fogging would potentially minimize microbial contamination in the hard to reach areas.