Agradecimentos: A.P.M. Pereira, A.S. Sant'Ana, and D.P. Amorim-Neto are thankful for the support of "Coordenação de Aperfeiçoamento de Pessoal de Nível Superior" (CAPES) (Finance Code 001) and "Fundação de Amparo à Pesquisa do Estado de São Paulo" (FAPESP) (#21/15224–8). The authors are also thankful to Stéphanie Oriol for expert technical assistance in Bacillus cereus genotyping and to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the financial support (Grant #CNPq #400806/2013–4; #302763/2014–7, #305804/2017–0 and #306644/2021–5) Ver menos

Abstract: Sporeforming bacteria are a concern in some food raw materials, such as cocoa powder. Samples (n = 618) were collected on two farms and at several stages during cocoa powder manufacture in three commercial processing lines to determine the impact of each stage on bacterial spore populations. Mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic, and Bacillus cereus spore populations were enumerated in all the samples. Genetic diversity in B. cereus strains (n = 110) isolated from the samples was examined by M13 sequence-based PCR typing, partial sequencing of the panC gene, and the presence/absence of ces and cspA genes. The counts of different groups of sporeforming bacteria varied amongst farms and processing lines. For example, the counts of mesophilic aerobic spore-forming (MAS) populations of cocoa bean fermentation were lower than 1 log spore/g in Farm 1 but higher than 4 log spore/g in Farm 2. B. cereus isolated from cocoa powder was also recovered from cocoa beans, nibs, and samples after roasting, refining, and pressing, which indicated that B. cereus spores persist throughout cocoa processing. Phylogenetic group IV was the most frequent (73%), along with processing. Strains from phylogenetic group III (14 %) did not show the ces gene's presence Ver menos