Anne E. Deckert

Prevalence of Hepatitis E Virus Antibodies in Canadian Swine Herds and Identification of a Novel Variant of Swine Hepatitis E Virus

ABSTRACT Swine hepatitis E virus is a newly identified potentially zoonotic virus from pigs of particular concern for possible direct transmission to a human xenotransplant recipient by organ transplantation. In the present study, prevalence of serum antibodies to hepatitis E virus was examined in Canadian swine herds. A total of 998 serum samples collected from 6-month-old healthy slaughter hogs were examined by enzyme immunoassay and Western blot analysis for antibodies to the recombinant open reading frame 3 (ORF3) protein of hepatitis E virus expressed in Escherichia coli. These samples represented more than 80 different swine production units from five major swine-producing provinces across Canada. From this study, 594 samples (59.4%) were found to be positive for hepatitis E virus antibody. The seroprevalence was higher in Quebec (88.8%) and Ontario (80.1%) than in Alberta and Saskatchewan (38.3%). By PCR using a pair of oligonucleotide primers deduced from the ORF2 sequence of human hepatitis E virus, a specific hepatitis E virus sequence was recovered from feces of pigs. The nucleotide sequence identity between the U.S. swine hepatitis E virus and the Canadian isolate (SK3) was only 85.8%, suggesting that genotypic variations may exist in swine hepatitis E virus in North America. Among 165 serum samples collected from humans in Saskatchewan, 2.4% were found to be positive for antibodies to the hepatitis E virus ORF3 protein. Our data indicate that hepatitis E virus is highly prevalent in commercial swine populations in Canada and support the suggestion that the swine hepatitis E virus may be an important zoonotic agent for humans.

Associations between reported on-farm antimicrobial use practices and observed antimicrobial resistance in generic fecal Escherichia coli isolated from Alberta finishing swine farms.

Multilevel random intercept logistic and Poisson regression were used to model associations between various antimicrobial use practices and resistance to individual and multiple antimicrobials among generic fecal Escherichia coli isolated from Alberta finishing swine. In-feed antimicrobial use in finishers was significantly associated with increased risk of resistance to ampicillin, chloramphenicol, streptomycin, and sulfisoxazole in generic E. coli isolates. Chlortetracycline use in grower rations was associated with ampicillin and tetracycline resistance. Use of in-feed antimicrobials in finishers was significantly associated with increased risk of resistance of generic E. coli to multiple antimicrobials. The study findings suggest that certain farm-level interventions related to antimicrobial use might be beneficial in reducing development and emergence of antimicrobial resistance in swine populations.

Distribution of Sulfonamide Resistance Genes in Escherichia coli and Salmonella Isolates from Swine and Chickens at Abattoirs in Ontario and Québec, Canada

ABSTRACT Sulfonamide-resistant Escherichia coli and Salmonella isolates from pigs and chickens in Ontario and Québec were screened for sul1, sul2, and sul3 by PCR. Each sul gene was distributed differently across populations, with a significant difference between distribution in commensal E. coli and Salmonella isolates and sul3 restricted mainly to porcine E. coli isolates.

Prevalence and antimicrobial resistance in Campylobacter spp. isolated from retail chicken in two health units in Ontario.

Campylobacter is an important enteric pathogen of humans and can cause diarrhea, fever, and abdominal pain. Campylobacter infections have frequently been associated with the handling and consumption of raw and undercooked poultry. Antimicrobial resistance among Campylobacter strains is of concern in the treatment of campylobacteriosis in vulnerable populations. A 2-year multidisciplinary study was conducted in the Perth and Wellington-Dufferin-Guelph public health units in Ontario, Canada, to investigate the prevalence and antimicrobial resistance of Campylobacter spp. in retail chicken. Retail chicken samples were collected from randomly selected stores in these health units. Resulting Campylobacter isolates were tested for susceptibility to amoxicillin-clavulanic acid (AMC), ampicillin (AMP), chloramphenicol (CHL), ciprofloxacin (CIP), clindamycin (CLI), erythromycin (ERY), gentamicin (GEN), nalidixic acid (NAL), tetracycline (TCY), and trimethoprim-sulfamethoxazole (SXT) using the E test. The prevalence of Campylobacter in 1,256 retail chicken samples was 59.6%. Of these positive samples, 9% contained Campylobacter coli, 1% contained Campylobacter lari, and 90% contained Campylobacter jejuni. Of the chicken isolates that were resistant to one or more antimicrobial agents, 301 isolates (40%) were resistant to one agent, 374 (50%) were resistant to two, 39 (5%) were resistant to three, 20 (3%) were resistant to four, and 6 (1%) were resistant to five. Nine isolates (1%) were susceptible to all antimicrobial agents tested. All isolates were susceptible to AMC, CHL, and GEN. Less than 10% of isolates were resistant to NAL, CIP, CLI, ERY, and AMP. Resistance to TCY was common (56%). No isolates had a resistance pattern that included all three antimicrobials important in the treatment of human campylobacteriosis (CIP, ERY, and TCY); however, 24 isolates (3.2%) were resistant to at least two of these antimicrobials.

Ciprofloxacin-Resistant Campylobacter spp. in Retail Chicken, Western Canada

During 2005–2010, the Canadian Integrated Program for Antimicrobial Resistance Surveillance identified increased prevalence of ciprofloxacin (a fluororquinolone) resistance among Campylobacter isolates from retail chicken in British Columbia (4%–17%) and Saskatchewan (6%–11%), Canada. Fluoroquinolones are critically important to human medicine and are not labeled for use in poultry in Canada.

Novel Campylobacter isolation method using hydrophobic grid membrane filter and semisolid medium.

Culture procedures for isolation of thermophilic campylobacters from food matrices are complex, labor intensive, and time-consuming. Most available methods include the use of antibiotics as selective agents to prevent the growth of competing microflora. A simple procedure for isolation of thermophilic campylobacters after enrichment in Rosefs enrichment broth was developed using a hydrophobic grid membrane filter (HGMF) on semisolid medium (SSM). SSM contains no antibiotics, and the HGMF physically separates Campylobacter from the enrichment broth, allowing isolation based on differential motility. The HGMF-SSM method was compared to the Agriculture and Agri-Food Canada Food Safety Procedures Manual (FSPM-10) method (Isolation of Thermophilic Campylobacters from Fresh Pork, Beef Veal, Poultry and Ready-to-Eat Meat Products), which includes the use of selective antibiotics. During the initial study, after enrichment the HGMF-SSM method yielded pure cultures of campylobacters after 16 to 18 h (overnight) compared with 48 h for the FSPM-10 method. Ninety-four turkey samples collected at local retail stores and 38 frozen pig fecal samples were processed by both methods. Thirty-five samples (26.5%) were positive by the HGMF-SSM method; 24 (18.2%) of these positive samples contained Campylobacter jejuni and 11 (8.3%) contained Campylobacter coli. With the FSPM-10 method, 25 samples (18.9%) were positive: 21 (15.9%) with C. jejuni and 4 (3%) with C. coli. For a subsequent field study, only the HGMF-SSM method was used to isolate Campylobacter from 1,200 chicken samples and 454 turkey samples sold at retail. Analysis of five subisolates from various samples indicated that only one type of Campylobacter was recovered by the HGMF-SSM method, as ascertained by MICs for 10 antimicrobials, sequencing of the short variable region of the flaA gene, and fingerprinting based on amplified fragment length polymorphism. The absence of antibiotics in the SSM may explain the higher recovery of thermophilic campylobacters. The HGMF-SSM method resulted in improved isolation of campylobacters and is simpler, faster, cheaper, and less labor intensive than the FSPM-10 method. The recovery of one type of Campylobacter from the chicken samples may have important implications, particularly in epidemiological studies.

Antimicrobial resistance and antimicrobial use associated with laboratory-confirmed cases of Campylobacter infection in two health units in Ontario

AIM A population-based study was conducted over a two-year period in the Perth District (PD) and Wellington-Dufferin-Guelph (WDG) health units in Ontario to document antimicrobial resistance and antimicrobial use associated with clinical cases of laboratory-confirmed campylobacteriosis. METHODS Etest (bioMérieux SA, France) was used to determine the minimum inhibitory concentration of amoxicillin/clavulanic acid, ampicillin, chloramphenicol, ciprofloxacin (CIP), clindamycin, erythromycin (ERY), gentamicin, nalidixic acid and tetracycline. Data regarding antimicrobial use were collected from 250 cases. RESULTS Of the 250 cases, 165 (65.7%) reported staying home or being hospitalized due to campylobacteriosis. Fifty-four per cent of cases (135 of 249) reported taking antimicrobials to treat campylobacteriosis. In 115 cases (51.1%), fecal culture results were not used for treatment decisions because they were not available before the initiation of antimicrobial treatment and/or they were not available before the cessation of symptoms. Of the 250 cases, 124 (49.6%) had available Campylobacter isolates, of which 66 (53.2%) were resistant to at least one of the antimicrobials tested. No resistance to ampicillin, chloramphenicol or gentamicin was found in these isolates. Six isolates (4.8%) were resistant to CIP. Two isolates (1.6%) were resistant to ERY; however, no isolates were resistant to both CIP and ERY. CONCLUSION Prudent use practices should be promoted among physicians to reduce the use of antimicrobials for the treatment of gastroenteritis in general and campylobacteriosis in particular, as well as to minimize the future development of resistance to these antimicrobials in Campylobacter species.