Chunu Mainali

Enterococci, commonly found in the normal intestinal flora of humans and animals, have emerged as an important human pathogen. A total of 184 isolates (88 isolates in 2015 and 96 isolates in 2016) were collected from 46 flocks. Two predominant enterococcus species were identified: Enterococcus faecalis (59%) and Enterococcus faecium (~39%). Resistance to penicillin was significantly decreased in the overall enterococci community, while it remained unchanged in the multi-class drug resistant (MDR) community. We identified the emeA and efrAB genes, which encode efflux pump systems, in 93% (26/28) of the MDR isolates with (intermediate) resistance to levofloxacin. The ermB gene was present in all MDR strains with resistance to erythromycin. The lsa gene was detected in 87% (84/97) of the MDR isolates with resistance to quinupristin/dalfopristin. About 82.2% of MDR strains in 2015 and 100% of MDR strains in 2016 carried the insertion sequence IS256, which is known to be associated with AMR genes, conferring resistance to erythromycin, gentamicin and vancomycin in enterococci. These results support the need for monitoring AMR in Gram-positive bacteria in poultry production, specifically in broiler chicken farms, to complement current AMR data, and develop a timely intervention framework.

Background/Objectives: Campylobacteriosis in human populations is an ongoing issue in both developed and developing countries. Poultry production is recognized as a reservoir for antimicrobial resistance and main source of human Campylobacter infection. Methods: In this study, sixty-five Campylobacter isolates were cultured from fecal samples collected from 17 flocks of broiler chickens in Alberta, Canada over two years (2015–2016). Susceptibility assays and PCR assays were performed to characterize resistance phenotypes and resistance genes. Conjugation assays were used to examine the mobility of AMR phenotypes. Results: Campylobacter jejuni was the predominant species recovered during both years of sampling. There were no Campylobacter coli isolates found in 2015; however, approximately 33% (8/24) of isolates collected in 2016 were Campylobacter coli. The two most frequent antimicrobial resistance patterns in C. jejuni collected in 2015 were tetracycline (39%) and azithromycin/clindamycin/erythromycin/telithromycin resistance (29%). One isolate collected in 2015 has resistance pattern ciprofloxacin/nalidixic acid/tetracycline. The tetO gene was detected in all tetracycline resistant isolates from 2015. The cmeB gene was detected in all species isolates with resistance to azithromycin/clindamycin/erythromycin/telithromycin, and from two isolates with tetracycline resistance. Alignment of the nucleotide sequences of the cmeB gene from C. jejuni isolates with different resistance patterns revealed several single nucleotide polymorphisms. A variety of multi-drug resistance patterns were observed through conjugation experiments. Conclusions: These data suggest that poultry production may serve as a potential reservoir for and source of transmission of multi-drug resistant Campylobacter jejuni and supports the need for continued surveillance.

Campylobacteriosis in human populations has been an ongoing issue in both developed and developing countries over the last 10 years. There is a grave concern whether poultry production plays an intermediate role as a reservoir and source of transmission. In this study, sixty-five broiler chicken Campylobacter isolates were isolated from fecal samples collected from 17 flocks in Alberta, Canada over two years (2015-2016). Campylobacter jejuni was the predominantly recovered Campylobacter species in both years. Approximately 33% (8/24) Campylobacter coli was identified in year 2016. The two most common antimicrobial resistance patterns in Campylobacter in year 2015 were tetracycline resistance (39%) and azithromycin/clindamycin/erythromycin/telithromycin resistance (29%). Only one isolate in 2015 displayed resistance to ciprofloxacin/nalidixic acid/tetracycline. The tetO gene was detected in all tetracycline resistant isolates in 2015. The cmeB gene was detected in all isolates with resistance to azithromycin/clindamycin/erythromycin/ telithromycin, and two isolates with resistance to tetracycline. Comparing the sequence of the cmeB genes from isolates with different resistance patterns revealed several single nucleotide polymorphisms. Transconjugants with three distinctly combined resistance patterns were obtained via in vitro conjugation. In conclusion, our study suggested that poultry can be a potential reservoir for and source of transmission of Campylobacter for human infection and requires an ongoing monitoring.

Enterococci, commonly found in the normal intestinal flora of humans and animals, have emerged as an important human pathogen. A total of 184 isolates (88 isolates in 2015 and 96 isolates in 2016) were collected from 46 flocks. Two predominant enterococcus species were identified: Enterococcus faecalis (59%) and Enterococcus faecium (~39%). Resistance to penicillin was significantly decreased in the overall enterococci community, while it remained unchanged in the multi-class resistant (MCR) community. We identified emeA and efrAB genes, which encode efflux pump systems, in 93% (26/28) of the multi-class resistant (MCR) isolates with (intermediate) resistance to levofloxacin. The ermB gene was present in all MCR strains with resistance to erythromycin. The lsa gene was detected in 87% (84/97) of the MCR isolates with resistance to quinupristin/dalfopristin. About 82.2% of MCR strains in 2015 and 100% of MCR strains in 2016 carried insertion sequence IS256, which is known to be associated with AMR genes conferring resistance to erythromycin, gentamicin and vancomycin in enteroccocci. These results support the need for monitoring AMR in Gram positive bacteria in poultry production, specifically in broiler chicken farms, to complement current AMR data, and developing a timely intervention framework.

Horizontal gene transfer is an important mechanism which facilitates bacterial populations in overcoming antimicrobial treatment. In this study, a total of 120 Escherichia coli and 62 Salmonella enterica subsp. enterica isolates were isolated from poultry farms in Alberta. Fourteen serovars were identified among Salmonella isolates. Thirty one percent of E. coli isolates were multiclass drug resistant (resistant to ≥ 3 drug classes), while only about 16% of Salmonella isolates were multiclass drug resistant. Among those, eight E. coli isolates had an AmpC-type phenotype, and one Salmonella isolate had an extended-spectrum beta-lactamase (ESBL)-type β-lactamase phenotype. We identified both AmpC-type ( bla _CMY-2 ) and ESBL-type ( bla _TEM ) genes in both E. coli and Salmonella isolates. Plasmids from eight of nine E. coli and Salmonella isolates were transferred to recipient strain E. coli J53 through conjugation. Transferable plasmids in above total eight E. coli and Salmonella isolates were also transferred into a lab-made sodium azide-resistant Salmonella recipient through conjugation. The class 1 integrase gene, int1 , was detected on plasmids from two E. coli isolates. Further investigation of class 1 integron cassette regions revealed the presence of an aadA gene encoding streptomycin 3”-adenylyltransferase, an aadA1a/aadA2 gene encoding aminoglycoside 3”-O-adenyltransferase, and a putative adenylyltransferase gene. This study provides some insight into potential horizontal gene transfer events of antimicrobial resistance genes between E. coli and Salmonella in poultry production.

SUMMARY The distribution, composition, and management characteristics of small “backyard” poultry flocks may have important implications in the spread of both avian diseases and zoonoses of public health concern. Although the prevalence of small poultry flocks has increased in Alberta, Canada, in recent years, there is minimal demographic information available for these populations. To gain initial epidemiologic insight into this growing population and potential areas of risk, a survey was conducted to characterize the sector. Information on flock demographics and bird health, as well as production and biosecurity practices, were gathered and analyzed from 206 surveys, representing respondents from 43 counties. These results revealed great diversity of both owners and flocks, characterized by wide variations in flock sizes and composition. Laying hens were the most commonly reported type of bird (93.4%), followed by ducks and geese (35.3%), turkeys, (33.8%), and broiler chickens (33.1%). Notably, 58.1% of... RESUMEN Parvadas avícolas pequeñas en Alberta: Demografía y prácticas. La distribución, composición y características de manejo de las parvadas pequeñas de traspatio pueden tener importantes implicaciones en la propagación de las enfermedades aviares y de zoonosis de interés para la salud pública. Aunque en los últimos años ha aumentado la prevalencia de pequeñas parvadas de aves de corral en Alberta, Canadá, existe poca información demográfica disponible sobre estas poblaciones. Para obtener una visión epidemiológica inicial de esta población creciente y áreas potenciales de riesgo, se realizó una encuesta para caracterizar el sector. Se recopiló y analizó la información sobre la demografía de las parvadas, así como la información sobre las prácticas de producción y de bioseguridad, mediante 206 encuestas, que representaron a encuestados de 43 condados. Estos resultados revelaron gran diversidad tanto con relación a los propietarios como de las parvadas, caracterizados por amplias variaciones en el tamañ...

Antimicrobial-resistant Salmonella species are threatening to become a serious public health problem. Therefore, surveillance and prudent use of antimicrobials is needed in both the agricultural and human health sectors. The aim of this study was to describe the antimicrobial susceptibility profiles of Salmonella isolates recovered from healthy broiler chickens at slaughter from November 2004 to April 2005. Salmonella isolates recovered from 36 broiler flocks in Alberta, Canada, were serotyped and tested for antimicrobial susceptibility against 15 antimicrobials. Of 272 Salmonella isolates tested, 64.0% were resistant to one or more antimicrobials, 10.0% were resistant to three or more antimicrobials, and 1.8% were resistant to five antimicrobials. All isolates were susceptible to amikacin, amoxicillin-clavulanic acid, ceftiofur, cefoxitin, ceftriaxone, ciprofloxacin, and nalidixic acid. The highest prevalence of resistance was to tetracycline (54.8%), followed by streptomycin (24.2%) and sulfisoxazole (8.4%). The most common multiantimicrobial resistance patterns were to streptomycin-tetracycline (24.3%), streptomycin-sulfisoxazole-tetracycline (6.6%), and ampicillin-streptomycin-sulfisoxazole-tetracycline (3.7%). The strongest associations were observed between resistance to kanamycin and tetracycline (odds ratio = 65.7, P = 0.001) and to ampicillin and sulfisoxazole (odds ratio = 62.9, P = 0.001). Salmonella Hadar and Salmonella Heidelberg were the two most common serovars accounting for 40.4 and 13.6% of the total isolates, respectively. Eighty-one percent and 12.7% of Salmonella Hadar isolates and 62.0 and 8.1% of Salmonella Heidelberg isolates were resistant to 1 or more and three or more antimicrobials, respectively. The flock level prevalence of resistance ranged from 5.6% for trimethoprim-sulfamethoxazole to 83.3% for tetracycline. This study provides baseline information on antimicrobial susceptibility of Salmonella isolates of broiler chickens at slaughter in Alberta that can serve as a benchmark for future research.

This study was conducted to investigate antimicrobial resistance profiles of Escherichia coli isolates from broiler chickens in Alberta, Canada. Cecal contents of broiler chickens from 24 flocks were collected at slaughter between January and March 2005 for culture and antimicrobial susceptibility testing against a panel of 15 antimicrobials using a broth microdilution technique. Of 600 E. coli isolates tested, 475 (79.2%) were resistant to one or more antimicrobials, 326 (54.3%) were resistant to three or more antimicrobials, 65 (10.8%) were resistant to five or more antimicrobials, and 15 (2.5%) were resistant to seven or more antimicrobials. The most common resistance was to tetracycline (69.2%), followed by streptomycin (48.2%), kanamycin (40.3%), and sulfisoxazole (38.0%). None of the E. coli isolates were resistant to amikacin, ceftriaxone, or ciprofloxacin. Of the isolates that were resistant to two or more antimicrobials, the most common multidrug resistance patterns were streptomycinte-tracycline (44.0%), streptomycin-sulfisoxazole-tetracycline (30.7%), and kanamycin-streptomycin-sulfisoxazole-tetracycline (23.5%). Resistance to tetracycline and kanamycin (odds ratio = 46.7, P = 0.0001) was highly associated, followed by resistance to streptomycin and sulfisoxazole (odds ratio = 12.0, P = 0.0001), and streptomycin and tetracycline (odds ratio = 10.3, P = 0.0001). The flock level prevalence of resistance varied from 16.7% for chloramphenicol to 100.0% for ampicillin, streptomycin, sulfisoxazole, and tetracycline. The results of this study provided baseline information on antimicrobial susceptibility of E. coli isolates of broiler chickens at slaughter in Alberta, which can serve as a bench mark for future research.

Salmonellosis is one of the most common bacterial foodborne diseases of public health concern in industrialized countries. Poultry products are considered an important source of Salmonella-related ...