A New and Convenient Method (TEMPO® “CAM”) for Enumeration of Campylobacter spp. from Poultry-associated Matrices

Introduction: Campylobacter jejuni and C. coli are the leading causes of human foodborne bacterial gastroenteritis worldwide, with poultry meat almost exclusively implicated as the vehicle of transmission. Conventional procedures for the enumeration of Campylobacter spp., such as direct plating according to ISO 10272-2:2006, are time-consuming and resource-intensive. The TEMPO® system, which has been developed for other bacterial populations, is an easy-to-use automated enumeration technique based on most probable number (MPN) principles.

Purpose: To develop a prototype assay and compare to standard methods for the enumeration of Campylobacter in poultry matrices using the TEMPO® platform.

Methods: A proprietary medium was formulated, consisting of CampyFood Broth®, two additional selective antibiotics and a fluorescent indicator. Ninety-six samples were used for evaluation of the assay against ISO/TS 10272-2:2006 (mCCDA), including broiler chicken feces, ceca, neck skin and carcasses collected at different points along the processing chain. All cards and plates were incubated at 41.5°C for 48h under microaerobic conditions. Counts were converted to log cells/ml and analyzed statistically.

Results: A linear association (R² = 0.96; t-test P ≥ 0.05) was observed between TEMPO® “CAM” and ISO/TS 10272-2:2006. Quantitative analysis showed 97% of enumerations were less than ± 1 log cells/ml different between the two methods. A sensitivity and specificity of 88% each, and good agreement (KAPPA = 0.62) were also determined.

Significance: In light of recent performance standards and objectives, which have set quantitative limits for campylobacters on processed dressed poultry carcasses, this study presents preliminary data for a sensitive, automated method for the enumeration of Campylobacter. Optimization and commercialization of TEMPO® “CAM” is in progress.