Evaluation of potassium clavulanate supplementation of Bolton broth for enrichment and detection of Campylobacter from chicken

Abstract

Culture-based detection of Campylobacter can be affected by competing flora, temperature, incubation time, and presence of blood. The presence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in poultry has become one of the most common factors interfering with the detection of Campylobacter. In the present study, we evaluated potassium clavulanate (ESBL inhibitor) as a supplement in Bolton broth (C-Bolton broth) for enrichment and detection of Campylobacter. First, we determined growth kinetics of Campylobacter in the presence of different concentrations of ESBL E. coli in C-Bolton broth during enrichment. The effects of other factors such as incubation time, incubation temperature, and presence of blood on Campylobacter detection in C-Bolton broth were also investigated. The growth of Campylobacter co-cultured at a low concentration (2 and 4 log10 CFU/mL) of ESBL E. coli was similar to that of Campylobacter alone in C-Bolton broth, and Campylobacter co-cultured at a high concentration (6 and 8 log10 CFU/mL) of ESBL E. coli showed slower growth than the pure Campylobacter culture. The Campylobacter detection limit was 1 log10 CFU/mL when mixed with 2, 4, or 6 log10 CFU/mL of E. coli and 3 log10 CFU/mL when mixed with 8 log10 CFU/mL of E. coli after 48 h enrichment in the broth. Campylobacter detection from chicken feces and litter samples was not affected by incubation time, or presence of blood in the broth. A modified procedure of enrichment in C-Bolton broth at 37°C for 24 h without blood showed a significantly (P ≤ 0.05) higher detection rate and a lower false-negative rate than the ISO 10272–1:2006 method for Campylobacter detection from chicken feces and litter samples. In summary, the present study demonstrates the efficacy of Bolton broth supplemented with potassium clavulanate in the detection of Campylobacter mixed with ESBL E. coli, and an improved procedure to detect Campylobacter from chicken feces and litter samples.

Human campylobacteriosis: A public health concern of global importance

“…Owing to this problem, Bolton and Robertson in 1977 developed a selective Preston medium suitable for Campylobacter isolation from environmental and food samples (Bolton and Robertson, 1982). Several other selective broths and media latter developed for Campylobacter isolation includes Bolton broth, Preston broth and Campylobacter enrichment broth (Baylis et al., 2000), modified charcoal cefoperazone deoxycholate agar (mCCDA) (Wei et al., 2018), CampyFood agar (CFA) and broth, RAPID’ Campylobacter agar (Seliwiorstow et al., 2014), Campylobacter agar base (CAB) and Campylobacter Cefex agar (Kashappanavar et al., 2018). Campylobacter species are microaerobic, fastidious bacteria capable of growing in a temperature between 37 °C and 42 °C (Davis and DiRita, 2017).…”

Campylobacteriosis, Shigellosis and Salmonellosis in Hospitalized Children with Acute Inflammatory Diarrhea in Georgia

“…Incubation was performed at 42 • C in a microaerobic gas mixture consisting of 85% nitrogen, 10% carbon dioxide and 5% oxygen [30]. Modified charcoal, cefoperazone, deoxycholate agar (mCCDA) (Thermo Fisher Specialty Diagnostics Ltd., Hampshire, UK) [31] and Chromogenic agar Campylobacter (CHROMagar, France) [32] served as two selective media for the isolation of Campylobacter species. Confirmation of suspected colonies grown on the media was performed applying the Biomerieux system ApiCampy (Biomerieux Inc, Marcy-l'Etoile, Lyon, France).…”