DNA identification and characterization of Campylobacter jejuni and Campylobacter coli isolated from caecal samples of chickens in Grenada

Department of Poultry Science, Auburn University, AL 36849, USA.
Journal of Applied Microbiology (Impact Factor: 2.48). 09/2009; 108(3):1041-9. DOI: 10.1111/j.1365-2672.2009.04507.x
Source: PubMed


To speciate Campylobacter strains from the caeca of chickens in Grenada using PCR and to evaluate DNA-based typing methods for the characterization of these isolates.
Isolates were speciated with two multiplex PCR assays and were typed with flaA-RFLP, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results confirmed that Campylobacter coli strains were more predominant than Campylobacter jejuni strains. From 56 isolates, 18 were misidentified using biochemical tests. PFGE typing gave the highest discriminatory power among the methods used (Simpson's index of diversity, D=0.9061). However, the combination of flaA-RFLP, PFGE and MLST results gave the highest discrimination for subtyping of these isolates (D=0.9857). A band position tolerance of 4% in BioNumerics was the most appropriate for the analysis of this database. MLST profiles were generally concordant with PFGE and/or flaA-RFLP types. Several isolates exhibited new MLST sequence types (STs), and 43 of the 49 Camp. coli strains belonged to the ST-828 clonal complex.
Campylobacter coli was the most prevalent species isolated from broilers and layers in Grenada, and a combination of restriction and sequence methods was most appropriate for the typing of Camp. coli isolates. Campylobacter coli STs clustered with described poultry-associated Camp. coli STs by phylogenetic analysis.
Further studies to understand the predominance of Camp. coli within Campylobacter spp. from chickens in Grenada may help elucidate the epidemiology of these pathogens in chickens.

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    • "High prevalence rates of C. coli in pigs have also been reported by many other researchers worldwide, ranging from 50% to 100% (Aarestrup et al., 1997; Ghimire et al., 2014; Guevremont et al., 2004; Mdegela et al., 2011; Munroe et al., 1983; Payot et al., 2004; Saenz et al., 2000; Steinhauserova et al., 2005; Thakur and Gebreyes, 2005b; Van Looveren et al., 2001; Varela et al., 2007). Although many studies have shown that C. coli is the most frequently isolated Campylobacter species from pigs, the predominance of C. jejuni (53.5%) observed in the present study may be due to the exposure of the tested pigs to C. jejuni from sources in Grenada other than pigs which include poultry (Hariharan et al., 2009; Miller et al., 2010; Sharma et al., 2015; Stone et al., 2013), sheep and goats (Stone et al., 2014), mongooses (Miller et al., 2014) and/or the environment (soil and water). "
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    ABSTRACT: This study determined the prevalence of C. jejuni and C. coli in feces from 180 randomly selected young, healthy, domestic pigs in Grenada, by culture and their identification and antimicrobial susceptibility profiles by phenotypic characteristics and Epsilometer test, respectively. Fecal samples from 172 of the 180 pigs (95.6%) were culture positive for Campylobacter species of which 53.5% were identified as C. jejuni and 46.5% as C. coli. Out of the 172 Campylobacter isolates, 119 viable isolates (65 C. jejuni and 54 C. coli) were tested for susceptibility against seven antibiotics. Low resistance rate (0 to 3.1%) of all the C. jejuni and C. coli isolates to four out of the seven antibiotics tested was observed. The highest resistance rates observed was against tetracycline with the resistance rates of 58.5% for C. jejuni and 61.1% for C. coli; followed by ampicillin, 18.5% for C. jejuni and 14.8% for C. coli; and metronidazole, 15.4% for C. jejuni and 13% for C. coli. The ampicillin/tetracycline resistance pattern was the most common pattern for both multidrug-resistant C. jejuni (12.3%) and C. coli (9.3%). This is the first study to report on multidrug-resistant patterns of C. jejuni and C. coli isolated from pigs in Grenada.
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    • "Multi-locus sequence typing (MLST) has become one of the most extensively used molecular typing methods for Campylobacter spp. (Miller et al., 2012; Müllner et al., 2010a; Sopwith et al., 2010). The first Campylobacter MLST scheme was developed for C. jejuni and C. coli. "
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    ABSTRACT: Campylobacter remains one of the most common bacterial causes of gastroenteritis worldwide. Tracking sources of this organism is challenging due to the large numbers of human cases, and the prevalence of this organism throughout the environment due to growth in a wide range of animal species. Many molecular subtyping methods have been developed to characterize Campylobacter species, but only a few are commonly used in molecular epidemiology studies. This review examines the applicability of these methods, as well as the role that emerging whole genome sequencing technologies will play in tracking sources of Campylobacter spp. infection.
    Journal of microbiological methods 07/2013; 95(1). DOI:10.1016/j.mimet.2013.07.007 · 2.03 Impact Factor
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    • "The major lineages identified in this study correspond to clonal complexes that are primarily associated with poultry and human disease (Table 2) and, except for ST-607, are geographically widely distributed (Table 3). Four of the six clonal complexes have been previously reported from Grenada [16, 24]. In general, results from this study support the previous observation that host association of Campylobacter genotypes transcends geographic variation [34]. "
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    ABSTRACT: This study determined whether multilocus sequence types (MLST) of Campylobacter from poultry in 2 farms in Grenada, West Indies, differed by farm, antimicrobial resistance and farm antibiotic use. Farm A used fluoroquinolones in the water and Farm B used tetracyclines. The E-test was used to determine resistance of isolates to seven antibiotics. PCR of the IpxA gene confirmed species and MLST was used to characterize 38 isolates. All isolates were either C. jejuni or C. coli. Farm antibiotic use directly correlated with antimicrobial resistance of Campylobacter isolates. Almost 80% of the isolates from Farm A were fluoroquinolone resistant and 17.9% of the isolates from Farm B were fluoroquinolone resistant. All Campylobacter isolates from Farm A were tetracycline sensitive, whereas 35.7% of isolates from Farm B were tetracycline resistant. Six previously recognized sequence types (STs) and 2 novel STs were identified. Previously recognized STs were those overwhelmingly reported from poultry and humans globally. Isolates with the same ST did not always have the same antibiotic resistance profile. There was little ST overlap between the farms suggesting that within-farm transmission of Campylobacter genotypes may dominate. MLST typing was useful for tracking Campylobacter spp. among poultry units and can help elucidate Campylobacter host-species population structure and its relevance to human health.
    02/2013; 2013:794643. DOI:10.1155/2013/794643
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