Richard Reid-Smith

Detection and quantification of methicillin-resistant Staphylococcus aureus (MRSA) clones in retail meat products

Aims:  The objective of the study was to determine the prevalence of methicillin‐resistant Staphylococcus aureus (MRSA) contamination of retail meat and to determine the level of contamination.

Detection and characterization of Clostridium difficile in retail chicken.

Aims:  This study was designed to evaluate the prevalence of Clostridium difficile contamination of retail chicken.

Acquisition of Resistance to Extended-Spectrum Cephalosporins by Salmonella enterica subsp. enterica Serovar Newport and Escherichia coli in the Turkey Poult Intestinal Tract

ABSTRACT Salmonella enterica subsp. enterica serovar Newport resistant to the extended-spectrum cephalosporins (ESCs) and other antimicrobials causes septicemic salmonellosis in humans and animals and is increasingly isolated from humans, animals, foods, and environmental sources. Mechanisms whereby serovar Newport bacteria become resistant to ESCs and other classes of antimicrobials while inhabiting the intestinal tract are not well understood. The present study shows that 25.3% of serovar Newport strains isolated from the turkey poult intestinal tract after the animals were dosed with Escherichia coli harboring a large conjugative plasmid encoding the CMY-2 β-lactamase and other drug resistance determinants acquired the plasmid and its associated drug resistance genes. The conjugative plasmid containing the cmy-2 gene was transferred not only from the donor E. coli to Salmonella serovar Newport but also to another E. coli serotype present in the intestinal tract. Laboratory studies showed that the plasmid could be readily transferred between serovar Newport and E. coli intestinal isolates. Administration of a single dose of ceftiofur, used to prevent septicemic colibacillosis, to 1-day-old turkeys did not result in the isolation of ceftiofur-resistant E. coli or Salmonella serovar Newport. There was a remarkable association between serotype, drug resistance, and plasmid profile among the E. coli strains isolated from the poults. This study shows that Salmonella serovar Newport can become resistant to ESCs and other antibiotics by acquiring a conjugative drug resistance plasmid from E. coli in the intestines.

Evaluation of the risks of shedding Salmonellae and other potential pathogens by therapy dogs fed raw diets in Ontario and Alberta.

Dogs that participate in animal‐assisted interventions (AAIs), often called ‘therapy dogs’, commonly interact with humans whose immune systems are not functioning optimally. The advisability of feeding raw meat (including poultry) to these animals remains a highly contentious issue, in spite of increasing evidence that raw meat is frequently contaminated with Salmonella. We set out to determine if consuming raw meat influences the risk of therapy dogs shedding Salmonella and other pathogens. Two hundred healthy therapy dogs from Ontario and Alberta were enrolled. Between May 2005 and November 2006, fecal specimens were collected from each dog every 2 months for 1 year, along with a log of places visited, antimicrobial use within the home, dog health status and diet. Specimens were cultured for Salmonella, methicillin‐resistant Staphylococcus aureus (MRSA), vancomycin‐resistant enterococci (VRE), extended‐spectrum cephalosporinase (ESC) Escherichia coli and Clostridium difficile. Forty (20%) of the dogs were reported to have been fed raw meat at some point during the year. The incidence rate of Salmonella shedding in the raw meat‐fed dogs was 0.61 cases/dog‐year, compared with 0.08 cases/dog‐year in dogs that were not fed raw meat (P < 0.001). Controlling for therapy dog group, the repeated measures, and pig ear consumption and diarrhoea in the 2 months prior to specimen submission, dogs that consumed raw meat were significantly more likely to test positive for Salmonellaat least once during the year than dogs that did not eat raw meat [odds ratio (OR) 22.7; 95% confidence interval (CI) 3.1–58.8; P < 0.001]. Specific Salmonella serovars were more common among dogs that consumed raw meat versus those that did not include S. Typhimurium, S. Heidelberg and S. Kentucky. Raw meat consumption was also significantly associated with shedding ESC E. coli (OR 17.2; 95% CI 9.4–32.3). No associations between C. difficile, MRSA or VRE and consumption of raw meat were detected. We recommend that dogs fed raw meat should be excluded from AAI programmes, particularly when the programmes involve interaction with humans at high risk of infection or adverse sequelae attributable to infection. Furthermore, although AAI dogs may not be representative of the general population of dogs, we also recommend that feeding of raw meat to dogs is to be avoided in homes where immunocompromised people live.

Antimicrobial Resistance in Generic Escherichia coli Isolates from Wild Small Mammals Living in Swine Farm, Residential, Landfill, and Natural Environments in Southern Ontario, Canada

ABSTRACT To assess the impacts of different types of human activity on the development of resistant bacteria in the feces of wild small mammals, we compared the prevalences and patterns of antimicrobial resistance and resistance genes in generic Escherichia coli and Salmonella enterica isolates from fecal samples collected from wild small mammals living in four environments: swine farms, residential areas, landfills, and natural habitats. Resistance to antimicrobials was observed in E. coli isolates from animals in all environments: 25/52 (48%) animals trapped at swine farms, 6/69 (9%) animals trapped in residential areas, 3/20 (15%) animals trapped at landfills, and 1/22 (5%) animals trapped in natural habitats. Animals trapped on farms were significantly more likely to carry E. coli isolates with resistance to tetracycline, ampicillin, sulfisoxazole, and streptomycin than animals trapped in residential areas. The resistance genes sul2, aadA, and tet(A) were significantly more likely to be detected in E. coli isolates from animals trapped on farms than from those trapped in residential areas. Three S. enterica serotypes (Give, Typhimurium, and Newport) were recovered from the feces of 4/302 (1%) wild small mammals. All Salmonella isolates were pansusceptible. Our results show that swine farm origin is significantly associated with the presence of resistant bacteria and resistance genes in wild small mammals in southern Ontario, Canada. However, resistant fecal bacteria were found in small mammals living in all environments studied, indicating that environmental exposure to antimicrobials, antimicrobial residues, resistant bacteria, or resistance genes is widespread.

Longitudinal Investigation of Methicillin‐Resistant Staphylococcus aureus in Piglets

Methicillin‐resistant Staphylococcus aureus (MRSA) has emerged as an important public health concern and pigs have been implicated in human infections. Cross‐sectional studies have demonstrated that MRSA can be commonly found in pigs internationally, but little is known about age‐related changes in MRSA colonization. This study evaluated MRSA colonization in piglets in a longitudinal manner. Serial nasal swabs were collected from piglets born to 10 healthy sows. The prevalence of MRSA colonization on days 1, 3, 7, 14 and 21 was 1% (1/100), 6.2% (3/97), 8.5% (8/94), 4.4% (4/91) and 20% (18/91) respectively, with an overall pre‐weaning prevalence of 34.5%. The prevalence on days 28, 42, 56 and 70 was 34% (31/91), 65% (57/88), 50% (44/88) and 42% (36/87) respectively, with an overall post‐weaning prevalence of 85%. Eighty‐four percent of piglets from negative sows and 100% of piglets from positive sows that survived at least until the time of weaning were colonized with MRSA at one or more times during the study. There was a significant association between sow and piglet colonization. The age of the piglet was significantly associated with the probability of colonization. No piglets or sows received antimicrobials during the study period. These results indicate that age must be considered when designing surveillance programmes and interpreting results of different studies on MRSA.

The Occurrence and Antimicrobial Susceptibility of Salmonellae Isolated from Commercially Available Canine Raw Food Diets in Three Canadian Cities

The purpose of the present study was to evaluate the prevalence and antimicrobial resistance patterns of Salmonella isolated from commercially available canine raw food diets in Canada. A total of 166 commercial frozen raw food diet samples were purchased from randomly selected local pet stores in three Canadian cities for a period of 8 months. All samples were evaluated for the presence of Salmonella, serotyped and tested for antimicrobial susceptibility. There was an overall Salmonella prevalence of 21%; chicken was an ingredient for 67% of the Salmonella‐positive diets. Eighteen different Salmonella serotypes were recovered, and resistance was observed to 12 of the 16 antimicrobials tested, with the majority of Ontario isolates exhibiting resistance to ampicillin and Calgary isolates to tetracycline. This study demonstrates the potential risk of raw food diets, especially for immunocompromised individuals, and stresses the need for implementing regulatory guidelines for the production of these diets in order to help control and ideally eliminate the bacterial risks associated with their use and consumption.

Clostridium difficile and methicillin-resistant Staphylococcus aureus shedding by slaughter-age pigs.

BackgroundClostridium difficile and methicillin-resistant Staphylococcus aureus are critical human pathogens and of increasing concern in food animals. Because of the apparent impact of age on prevalence of these organisms, studies of slaughter age pigs are important when considering the potential for contamination of food. This study evaluated C. difficile and MRSA shedding by slaughter age pigs from farms across Canada.ResultsClostridium difficile was isolated from 30/436 (6.9%) samples from 15/45 (33%) farms. After adjusting for clustering at the herd level, the prevalence was 3.4%. Ribotype 078 (toxinotype V, North American Pulsotype 7) was the most common strain, accounting for 67% of isolates. MRSA was isolated from 21/460 (4.6%) pigs from 5/46 (11%) farms. The prevalence in pigs after adjusting for clustering at the herd level was 0.2%. Seven different spa types were identified, with 3 related spa types (t011, t034, new) accounting for 16 (76%) consistent with ST398 predominating.Both MRSA and C. difficile samples were collected from 45 farms. Both MRSA and C. difficile were detected on 2 (4.4%), with C. difficile only on 13 (29%), MRSA only on 3 (6.7%) and neither on 27 (60%).ConclusionsThe prevalence of C. difficile and MRSA in slaughter age pigs was relatively low, particularly in comparison with studies involving younger pigs. The predominance of C. difficile ribotype 078 and MRSA ST398 was not surprising, but there was diversity in strain types and the majority of isolates of both organisms were strains that can be found in humans. While the prevalence of C. difficile and MRSA in slaughter age pigs was relatively low, there is clearly potential for contamination of meat from healthy pigs carrying this pathogen into slaughterhouses.

Phenotypic and genetic characterization of antimicrobial resistance in Salmonella serovars isolated from retail meats in Alberta, Canada

This study determined the prevalence of Salmonella serovars, antimicrobial resistance (AMR) and resistance genes in Salmonella isolated from retail meats purchased in Alberta, Canada. Samples were collected during one year period (May 2007-April 2008) on weekly basis from 19 census divisions in Alberta. A total of 564 samples including chicken (n = 206), turkey (n = 91), beef (n = 134) and pork (n = 133) were purchased. Salmonella were recovered from chicken (40%), turkey (27%) and pork (2%) samples and was not found in ground beef. A total of 21, 8, and 3 different serovars were recovered from chicken, turkey and pork meats, respectively. Salmonella Hadar was most common in chicken whereas S. Heidelberg was common in turkey meat. Overall 29% (32/110) of isolates were susceptible to tested antimicrobials and resistance to ciprofloxacin, amikacin and nalidixic acid was not found in any isolate. Multiresistance (≥2 antimicrobials) was found in 56% of isolates. Resistance to amoxicillin-clavulanic acid (AMC), ceftiofur (TIO), and ceftriaxone (CRO) was found in about 21% of chicken and 25% of turkey isolates. Resistance to either of tetracycline (TET), streptomycin (STR) or ampicillin (AMP) was unconditionally associated with S. Hadar but resistance to either of TET, AMP, AMC, TIO, CRO or cefoxitin was associated with S. Heidelberg. The strA/B (42% isolates), tet(A) (28% isolates), bla(CMY-2) (21% isolates) and bla(TEM) (17% isolates) were the most common resistance genes found. The bla(CMY-2) and bla(TEM) genes were unconditionally associated with S. Heidelberg; tet(A) and strA/B with S. Hadar and tet(B) gene with S. Kentucky. The strA/B genes were not associated with S. Heidelberg. Our data suggests that the prevalence of Salmonella serovars varied by the meat type and that AMR and resistance genes varied by the Salmonella serovars.

Disease transmission model for community-associated Clostridium difficile infection

Participating researchers and public health personnel at a Canadian workshop in 2007, noted considerable gaps in current understanding of community-associated Clostridium difficile infection (CA-CDI), specifically infection sources and risk factors. A disease transmission model for CA-CDI was requested as an initial step towards a risk assessment, to analyse infection sources and risk factors, addressing priority research areas. The developed model contains eight infection states (susceptible, gastrointestinal exposure, colonized, diseased, deceased, clinically resolved colonized, relapse diseased, and cleared) and notes directional transfers between the states. Most published research used focused on hospital-associated C. difficile infection (HA-CDI) and further studies are needed to substantiate the use of HA-CDI knowledge in the transmission of CA-CDI. The aim was to provide a consistent framework for researchers, and provide a theoretical basis for future quantitative risk assessment of CA-CDI.