Identification of Escherichia coli Pathotypes in Natural Cocoa Mass Produced in Côte d’Ivoire from March to May 2025: Indicators of Recent Fecal Contamination

Abstract

Natural cocoa mass is an intermediate product widely used in the chocolate industry and constitutes a major economic resource in Africa, particularly in Côte d’Ivoire. However, production and handling conditions may promote faecal microbiological contamination, thereby compromising its sanitary quality. Objective: This study aimed to assess the microbial risk associated with natural cocoa masses produced in Côte d’Ivoire during the cocoa harvesting season from March to May 2025. Study Design: This study was a descriptive, experimental, and cross-sectional study focusing on the assessment of microbiological hygiene indicators of natural cocoa mass. Place and duration of Study: The study was conducted in the San-Pedro area, Côte d’Ivoire, from March to May 2025, over a period of 03 months. Methodology: Six representative samples (E1–E6), collected from the industrial processing units SACO, OLAM, and ICP located in San Pedro, were analysed. Five Escherichia coli pathotypes, recognised as indicators of recent faecal contamination and a potential public health risk, were investigated: Shiga toxin-producing E. coli (STEC), enterohaemorrhagic E. coli (EHEC), enterotoxigenic E. coli (ETEC), E. coli O157, and lactose-positive E. coli. Analyses were performed according to standardised ISO methods, combining observation of typical colony morphological characteristics (colour, size, shape, margin, elevation, surface, and opacity), confirmatory biochemical tests, and molecular analyses (PCR) targeting the virulence genes (elt, est, rfbO157, fliCH7, stx1, stx2, and eae). Results: The identification results revealed the complete qualitative and quantitative absence of all characteristic colonies and targeted virulence genes, confirming the absence of all investigated E. coli pathotypes in every analysed sample (0 ± 0 CFU/g). Conclusion: These findings indicate that the natural cocoa masses produced by SACO, OLAM, and ICP during the study period present no microbiological risk associated with pathogenic E. coli. They therefore comply with microbiological safety requirements and may be considered suitable for human consumption and for subsequent industrial processing under satisfactory sanitary conditions.