Detection of Campylobacter colonies using hyperspectral imaging

Abstract

The presence of Campylobacter in foods of animal origin is the leading cause of bacterially induced human gastroenteritis. Isolation and detection of Campylobacter in foods via direct plating involves lengthy laboratory procedures including enrichments and microaerobic incubations, which take several days to a week. The incubation time for growing Campylobacter colonies in agar media usually takes 24–48 h. Oftentimes the problem is the difficulty of visually differentiating Campylobacter colonies from non-Campylobacter contaminants that frequently grow together with Campylobacter on many existing agars. In this study, a new screening technique using non-destructive and non-contact hyperspectral imaging was developed to detect Campylobacter colonies in Petri dishes. A reflectance spectral library of Campylobacter and non-Campylobacter contaminants was constructed for characterization of absorption features in wavelengths from 400 to 900 nm and for developing classification methods. Blood agar and Campy-Cefex agar were used as culture media. The study found that blood agar was the better culture medium than Campy-Cefex agar in terms of Campylobacter detection accuracy. Classification algorithms including single-band thresholding, band-ratio thresholding and spectral feature fitting were developed for detection of Campylobacter colonies as early as 24 h of incubation time. A band ratio algorithm using two bands at 426 and 458 nm chosen from continuum-removed spectra of the blood agar bacterial cultures achieved 97–99% of detection accuracy. This research has profound implications for early detection of Campylobacter colonies with high accuracy. Also, the developed hyperspectral reflectance imaging protocol is applicable to other pathogen detection studies.