David A. Bemis

Evaluation of susceptibility test breakpoints used to predict mecA-mediated resistance in Staphylococcus pseudintermedius isolated from dogs.

Clinical and Laboratory Standards Institute interpretive breakpoints for in vitro susceptibility tests that predict mecA-mediated oxacillin resistance in Staphylococcus pseudintermedius isolates from animals have been changed twice in the past decade. Moreover, there are no counterpart recommendations for human isolates of S. pseudintermedius. Individual medical and veterinary laboratories variably use interpretive breakpoints identical to those recommended for use with Staphylococcus aureus or identical to those recommended for use with coagulase-negative staphylococci. The purpose of the current study was to examine correlations between oxacillin disk diffusion, oxacillin gradient diffusion, oxacillin microbroth dilution, and cefoxitin disk diffusion tests used to predict mecA-mediated resistance in S. pseudintermedius and to retrospectively estimate, from disk diffusion zone diameter measurements, the prevalence and rate of increase of oxacillin resistance among canine S. pseudintermedius isolates submitted to a veterinary teaching hospital laboratory. Oxacillin disk diffusion zone diameters of ≤17 mm and oxacillin minimum inhibitory concentrations of ≥0.5 μg/ml were highly correlated with detection of mecA in canine S. pseudintermedius isolates by polymerase chain reaction. MecA-mediated resistance among S. pseudintermedius isolates from dogs increased from less than 5% in 2001 to near 30% in 2007. More than 90% of the methicillin-resistant S. pseudintermedius isolates in 2006 and 2007 were also resistant to representatives of ≥4 additional antimicrobial drug classes. Cefoxitin disk diffusion with the resistance breakpoint set at ≤24 mm significantly underestimated the presence of mecA in S. pseudintermedius.

Identification of a predominant multilocus sequence type, pulsed-field gel electrophoresis cluster, and novel staphylococcal chromosomal cassette in clinical isolates of mecA-containing, methicillin-resistant Staphylococcus pseudintermedius

Methicillin resistance encoded by the mecA gene is increasingly observed in Staphylococcus pseudintermedius. Little is known about the population genetics of veterinary staphylococci bearing methicillin resistance. The aim of this study was to determine the relatedness of resistant bacteria and to compare them with methicillin-susceptible isolates. Multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) fragment profiling were performed on methicillin-resistant S. pseudintermedius (MRSP) and methicillin-susceptible S. pseudintermedius (MSSP) isolates obtained from canine samples submitted to the veterinary teaching hospital bacteriology service between 2006 and 2008. Multilocus sequence typing detected 20 different sequence types, 16 of which were not previously described. Methicillin-resistant isolates were predominantly ST 68, possessed the Staphylococcus aureus-associated staphylococcal chromosomal cassette mec (SCCmec) type V(T) and fell within the largest PFGE cluster; whereas methicillin-susceptible strains were more genetically diverse. This suggests that most methicillin resistance within the population of isolates tested originated from a single source which has persisted and expanded for several years.

Antigen and antibody testing for the diagnosis of blastomycosis in dogs.

BACKGROUND Early diagnosis and treatment are associated with an improved prognosis in blastomycosis. The diagnosis of blastomycosis may be missed by cytology, histopathology, culture, or serology. An enzyme immunoassay (EIA) for detection of Blastomyces dermatitidis galactomannan antigen in body fluids has been used for rapid diagnosis of blastomycosis in humans. HYPOTHESIS Measurement of Blastomyces antigen in urine or serum by the MVista Blastomyces antigen EIA is more sensitive than measurement of anti-Blastomyces antibodies for diagnosis of blastomycosis in dogs. METHODS Serum and urine samples from 46 dogs with confirmed blastomycosis were tested for Blastomyces antigen and serum was tested for anti-Blastomyces antibodies. RESULTS The sensitivity for the detection of antigen in urine was 93.5% and it was 87.0% in serum. The sensitivity of antibody detection by agar gel immunodiffusion (AGID) was 17.4% and it was 76.1% by EIA. Antigen and antibody decreased during itraconazole treatment. CONCLUSIONS AND CLINICAL IMPORTANCE Antigen detection is a more sensitive test for diagnosis of blastomycosis than antibody testing by AGID, the only commercially available method. Antigen concentrations decreased with treatment.

Multilocus Sequence Typing for Characterization of Staphylococcus pseudintermedius

ABSTRACT Staphylococcus pseudintermedius is an opportunistic pathogen in dogs. Four housekeeping genes with allelic polymorphisms were identified and used to develop an expanded multilocus sequence typing (MLST) scheme. The new seven-locus technique shows S. pseudintermedius to have greater genetic diversity than previous methods and discriminates more isolates based upon host origin.

Risk of colonization or gene transfer to owners of dogs with meticillin‐resistant Staphylococcus pseudintermedius

To determine the zoonotic risk from meticillin-resistant staphylococcal species or transfer of resistance genes between dogs with pyoderma and their owners, 25 dog-owner pairs were studied. Cultures were obtained from the dogs lesions and the owners nasal cavity on the initial visit. Staphylococcus isolates were identified and antimicrobial susceptibility tests were performed by the Kirby-Bauer disk diffusion method. Presence of the mecA gene was determined by PCR. Staphylococcal Cassette Chromosome (SCCmec) typing was performed by multiplex PCR. Eighteen dogs had a meticillin-resistant staphylococcal species, with meticillin-resistant S. pseudintermedius (MRSP) isolated from 15 dogs. MRSP was isolated from two owners of dogs with MRSP skin infections. Both organisms had the same susceptibility pattern and SCCmec type. MRSP was not isolated from the owners after treating both dogs for 1 month. At least one coagulase-negative Staphylococcus spp. (CoNS) was isolated from each owner, with meticillin resistance found in 16 (64%) of the isolates. The mecA gene was identified in all but two of the meticillin-resistant Staphylococcus spp. Multiplex PCR identified SCCmec type V in all MRSP. The mecA gene-possessing CoNS isolates from owners contained either SCCmec type IVa or IVc. In conclusion, MRSP colonization of owners appeared to be uncommon and transient. Human nasal carriage of meticillin-resistant CoNS was common, but the SCCmec types were different from those in the canine MRSP isolates. Owners do not appear to be at great risk of zoonotic transfer of organisms or antimicrobial resistance genes from dogs with MRSP infections, but the findings should be confirmed with a much larger cohort.

Comparison of Tests To Detect Oxacillin Resistance in Staphylococcus intermedius, Staphylococcus schleiferi, and Staphylococcus aureus Isolates from Canine Hosts

ABSTRACT Multiple tests were compared to the reference standard PBP2a latex agglutination test for detection of mecA-mediated oxacillin resistance in canine staphylococci. Cefoxitin disk diffusion, using breakpoints for human isolates of coagulase-negative Staphylococcus spp., had low sensitivity for detection of oxacillin resistance in members of the Staphylococcus intermedius group.

Bordetella and Mycoplasma Respiratory Infections in Dogs and Cats

The consequences of B. bronchiseptica and mycoplasma infections in dogs and cats vary greatly. Only careful clinical judgment can dictate when to institute antimicrobial and other supportive treatments. Approaches to controlling diseases caused by these organisms should be tailored to meet individual needs. Management strategies that reduce natural exposure levels in the animals environment and maintain active immunity to contagious components of disease have the highest likelihood of success.

Mycobacterium tuberculosis transmission from human to canine.

A 71-year-old woman from Tennessee, USA with a 3-week history of a productive, nonbloody cough was evaluated. Chest radiograph showed infiltrates and atelectasis in the upper lobe of the right lung. A tuberculosis (TB) skin test resulted in a 14-mm area of induration. Sputum stained positive for acid-fast bacilli (AFB) and was positive for Mycobacterium tuberculosis by DNA probe and culture. Treatment was initiated with isoniazid, rifampicin, and pyrazinamide. After 14 days of daily, directly …

Treatment outcome of dogs with meticillin-resistant and meticillin-susceptible Staphylococcus pseudintermedius pyoderma.

BACKGROUND The prevalence of meticillin- and multidrug-resistant Staphylococcus pseudintermedius (MRSP) in canine pyoderma has been increasing in recent years; thus, treatment of these cases has become challenging. HYPOTHESIS/OBJECTIVES To compare treatment outcome (clinical resolution and treatment duration), adverse effects of medication, and concurrent diseases and medications in dogs with meticillin-susceptible S. pseudintermedius (MSSP) and MRSP pyoderma. ANIMALS/METHODS: Medical records were reviewed retrospectively, and 123 MSSP and 93 MRSP clinical cases between January 2008 and April 2010 were included. RESULTS In MSSP infections, cefalexin and cefpodoxime were the most commonly prescribed antimicrobials, accounting for 43.2 and 34.4% of cases, respectively. In MRSP infections, chloramphenicol and doxycycline were most commonly prescribed, accounting for 52.6 and 14.4% of cases, respectively. Adverse effects were reported in seven MSSP and 31 MRSP cases. The most commonly reported adverse effects were gastrointestinal, prompting antibiotic discontinuation in three MSSP and 20 MRSP cases. Chloramphenicol was associated with the highest incidence of adverse reactions (27 of 51 cases). Of 164 cases with follow up, 43 of 88 MSSP infections and 29 of 76 MRSP infections achieved complete clinical resolution at the first recheck examination. Three MSSP and seven MRSP cases failed to improve or resolve at subsequent visits assessed at 3-4 week intervals. CONCLUSIONS AND CLINICAL IMPORTANCE Results from this study showed that the majority of pyodermas resolved regardless of meticillin susceptibility. Although some cases of MRSP pyoderma took longer to treat, this is likely to be because of chronicity and not the organism. In addition, adverse effects were frequently associated with chloramphenicol administration.

OSTEOMYELITIS ASSOCIATED WITH SALMONELLA ENTERICA SS ARIZONAE IN A COLONY OF RIDGENOSE RATTLESNAKES (CROTALUS WILLARDI)

Abstract The identification of three Arizona ridgenose rattlesnakes (Crotalus willardi) with Salmonella arizonae–associated osteomyelitis led to a 5-yr prospective study of radiographic signs and Salmonella intestinal carriage rates in a 19-member colony of this rattlesnake species. Ventrodorsal radiographs were performed and cloacal swabs were cultured for Salmonella spp. annually. Ten snakes survived the 5-yr period, with six of them remaining free of bony lesions. Three snakes that had no bony lesions in 1995 developed radiographic signs of osteomyelitis during the study. Six snakes with bony lesions at the beginning of the study died or were euthanatized due to osteomyelitis during the study. The radiographic signs of osteomyelitis were progressive for five snakes that were serially radiographed. Only one snake with radiographic signs of osteomyelitis at the beginning of the study was still alive at the end of the study, and this animals bony lesions were more extensive at the end. Thirty-nine intestinal S. arizonae isolates, representing 13 serotypes, were obtained from the 19 snakes. Salmonella arizonae serotype 56:Z4,Z23 was isolated only once from a cloacal culture, from a snake that had no radiographic bone lesions. Twelve extraintestinal Salmonella isolates, representing two serotypes, were isolated from six snakes. All extraintestinal isolates except one were of S. arizonae serotype 56:Z4,Z23, and all isolates from bone were of this serotype. One snake with characteristic bone lesions died, and Providencia rettgeri was cultured from each of the tissues cultured, whereas no Salmonella spp. were isolated from this snake. Salmonella arizonae serotype 56:Z4,Z24 appears to have a tropism for bone and other extraintestinal sites in C. willardi and may cause a progressive, ultimately fatal disease in this species.