Arcobacter: Comparison of Isolation Methods, Diversity, and Potential Pathogenic Factors in Commercially Retailed Chicken Breast Meat from Costa Rica

Abstract

Arcobacter species have been recognized as potential food- and waterborne pathogens. The lack of standardized isolation methods and the relatively scarce knowledge about their prevalence and distribution as emerging pathogens are due to the limitations in their detection and identification. This study aimed to determine the presence and the identification of Arcobacter in chicken breast samples commercially retailed in San José, Costa Rica, as well as to describe the adherence and invasive potential of the strains to human cells (HEp-2). Fifty chicken breast samples were collected from retail markets in the metropolitan area of the country. Six different isolation methodologies were applied for the isolation of Arcobacter. Isolation strategies consisted of combinations of enrichments in de Boer or Houf selective broths and subsequent isolation in blood agar (directly or with a previous passive membrane filtration step) or Arcobacter selective agar. Suspicious colonies were identified with a genus-specific PCR, whereas species-level identification was achieved with a multiplex PCR. The overall isolation frequency of Arcobacter was 56%. From the isolation strategies, the combination of enrichment in Houf selective broth followed by filtration on blood agar showed the best performance, with a sensitivity of 89% and a specificity of 84%. A total of 46 isolates were confirmed as Arcobacter with the genus-specific PCR, from which 27 (59%) corresponded to Arcobacter butzleri, 9 (19%) to Arcobacter cryaerophilus, and 10 (22%) were not identified with this multiplex PCR. Regarding the potential pathogenicity, 75% of the isolates presented adherence to HEp-2 cells, while only 22% were invasive to that cell line. All invasive strains were A. butzleri or nonidentified strains. The results show the presence of potentially pathogenic Arcobacter in poultry and recognize the importance it should receive as a potential foodborne pathogen from public health authorities.