Kathleen O'Shea

The prevalence of carriage of meticillin-resistant staphylococci by veterinary dermatology practice staff and their respective pets

It has been shown that people and pets can harbour identical strains of meticillin-resistant (MR) staphylococci when they share an environment. Veterinary dermatology practitioners are a professional group with a high incidence of exposure to animals infected by Staphylococcus spp. The objective of this study was to assess the prevalence of carriage of MR Staphylococcus aureus (MRSA), MR S. pseudintermedius (MRSP) and MR S. schleiferi (MRSS) by veterinary dermatology practice staff and their personal pets. A swab technique and selective media were used to screen 171 veterinary dermatology practice staff and their respective pets (258 dogs and 160 cats). Samples were shipped by over-night carrier. Human subjects completed a 22-question survey of demographic and epidemiologic data relevant to staphylococcal transmission. The 171 human-source samples yielded six MRSA (3.5%), nine MRSP (5.3%) and four MRSS (2.3%) isolates, while 418 animal-source samples yielded eight MRSA (1.9%) 21 MRSP (5%), and two MRSS (0.5%) isolates. Concordant strains (genetically identical by pulsed-field gel electrophoresis) were isolated from human subjects and their respective pets in four of 171 (2.9%) households: MRSA from one person/two pets and MRSP from three people/three pets. In seven additional households (4.1%), concordant strains were isolated from only the pets: MRSA in two households and MRSP in five households. There were no demographic or epidemiologic factors statistically associated with either human or animal carriage of MR staphylococci, or with concordant carriage by person-pet or pet-pet pairs. Lack of statistical associations may reflect an underpowered study.

Detection of a blaSHV Extended-Spectrum β-Lactamase in Salmonella enterica Serovar Newport MDR-AmpC

ABSTRACT Salmonella enterica serovar Newport MDR-AmpC expressing TEM-1b and extended-spectrum β-lactamase SHV-12 was isolated from affected animals during an outbreak of salmonellosis that led to a 3-month closure of one of the largest equine hospitals in the United States.

Genotypic relatedness and phenotypic characterization of Staphylococcus schleiferi subspecies in clinical samples from dogs

OBJECTIVE To assess the degree of biological similarity (on the basis of genotype determined via pulsed-field gel electrophoresis [PFGE]) between isolates of 2 Staphylococcus schleiferi subspecies (S schleiferi subsp coagulans and S schleiferi subsp schleiferi) in clinical samples obtained from dogs. SAMPLE POPULATION 161 S schleiferi isolates from 160 canine patients. PROCEDURES A commercial microbiology identification system was used to identify each isolate as S schleiferi. Isolates underwent slide and tube coagulase testing and antimicrobial susceptibility testing. A mecA PCR assay and a latex agglutination test for penicillin-binding protein 2a (PBP2a) were also performed on each isolate. Clonal clusters with a similarity cutoff value of 80% were identified via PFGE. RESULTS Of the 161 isolates, 61 (38%), 79 (49%), and 21 (13%) were obtained from cutaneous sites, ears, and other sites, respectively; 110 (68%) were coagulase negative, and 51 (32%) were coagulase positive. Among the coagulase-negative and coagulase-positive isolates, 65% (71/110) and 39% (20/51) were oxacillin resistant, respectively. All oxacillin-resistant isolates yielded positive results via mecA PCR assay and PBP2a latex agglutination testing. Via PFGE, 15 major clusters and 108 individual pulsed-field profiles were identified. Oxacillin-resistant and oxacillin-susceptible isolates clustered separately. Clonal clusters were heterogeneous and contained representatives of both subspecies. CONCLUSIONS AND CLINICAL RELEVANCE Coagulase-positive and coagulase-negative isolates were not genotypically distinct and may represent a single S schleiferi sp with variable coagulase production, rather than 2 biologically distinct subspecies. Further studies are needed to characterize clinical or epidemiological differences associated with infections with coagulase-positive and coagulase-negative S schleiferi in dogs.

Optimization of an in vitro assay to detect Streptococcus equi subsp. equi

Streptococcus equi is the etiologic agent of a highly infectious upper respiratory disease of horses known as strangles. Bacterial culture methods and polymerase chain reaction (PCR) of nasopharyngeal washes and guttural pouch lavages are used routinely to test clinical and carrier animals for the presence of S. equi but no definitive or gold standard test method has been shown to be optimal. We hypothesized that (i) a flocked swab submerged in ten-fold serial dilution suspensions of S. equi prepared in 0.9% NaCl would detect more colony forming units (CFU) than a rayon swab when used to inoculate a blood agar plate, (ii) centrifugation of a 1 ml aliquot of each suspension would improve the limit of detection (LOD) by bacterial culture and PCR compared to the culture or PCR of submerged swab samples, (iii) PCR of the centrifuged samples from each suspension would be more sensitive than aerobic culture alone, and (iv) PCR of a 1 ml aliquot directly from a sample would be more sensitive than PCR of a sample following submersion of a flocked swab in 1 ml saline. Using 7 ten-fold serial dilutions of S. equi in 0.9% NaCl, the LOD for 4 bacterial culture methods and 3 PCR methods were compared. The LOD of direct PCR and flocked swab culture was determined at 1 cfu/ml. All PCR methods were equivalent to each other and were more sensitive than any of the culture methods at the lower dilutions. At higher cell densities (>100 cfu/ml) flocked swab culture was not statistically better than rayon swab culture, but it was superior to all other methods tested.

Pseudomonas fluorescens contamination of a feline packed red blood cell unit and studies of canine units

Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems.

Genotypic relatedness and antimicrobial resistance of Staphylococcus schleiferi in clinical samples from dogs in different geographic regions of the United States.

BACKGROUND Staphylococcus schleiferi is a known pathogen that can cause canine skin and ear infections. The aim of this study was to determine the molecular epidemiology and antimicrobial susceptibility of clinical veterinary isolates from different geographic regions in the United States. HYPOTHESIS It was hypothesized that S. schleiferi would maintain genotypic homogeneity across the different geographic regions and that meticillin-resistant (MR) isolates of S. schleiferi would predominate. METHODS Isolates were identified as S. schleiferi by a commercial microbiology identification system and confirmed by nuc gene PCR. Antibiotic susceptibility data were collected and PBP2a latex agglutination testing was performed on MR isolates. Pulsed-field gel electrophoresis (PFGE) was performed and clonal clusters were identified with a Dice coefficient similarity of >80%. RESULTS There were 116 isolates from the Mid-Atlantic region and 101 from across the United States. Of these 217 isolates, 209 (96%) were obtained from cutaneous sites. Of the Mid-Atlantic isolates, 62% (72 of 116) were MR and 16% (18 of 116) were multidrug-resistant (MDR). Of the isolates from the other geographic regions, 73% (74 of 101) were MR and 24% (24 of 101) were MDR. All MR isolates were positive by PBP2a latex agglutination. PFGE identified 155 individual pulsed-field profiles and three major pulsed-field types (PFT) that contained 61% (133 of 217) of the isolates. These pulsed-field types were geographically heterogeneous. CONCLUSIONS This study demonstrates the dissemination of successful MR pulsed-field types of S. schleiferi across the United States.

Clinical, microbiological, and molecular characterization of methicillin-resistantStaphylococcus aureusinfections 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.