Effet de la digestion gastrique sur la viabilité et l'expression de gènes de stress de bactéries probiotiques dans une matrice de sève d'érable

Les bactéries probiotiques doivent être en mesure de survivre pendant la digestion gastrique afin d’atteindre le côlon et ainsi conférer des bienfaits sur la santé humaine. Le potentiel des matrices d’origine laitière pour soutenir les probiotiques a souvent été décrit, mais celui des produits végétaux tels que la sève d'érable pour cette application reste peu connue, alors que l’engouement pour ces produits est en croissance. L'objectif de ce mémoire était d'évaluer la viabilité cellulaire par l’utilisation du propidium monoazide (PMA) couplé à la qPCR, ainsi que d'évaluer l'expression de cinq gènes reliés au stress atpA, dnaK, galK, groEL et hrcA par RT-qPCR lors de la digestion gastrique in vitro (TIM-1)de trois souches commerciales (Bifidobacterium animalis ssp. lactis A026, Lacticaseibacillus paracaseiA234, Lacticaseibacillus rhamnosus A119) ainsi que de la souche de référence Lacticaseibacillus rhamnosus GG (ATCC 53103) contenues dans de la sève d'érable. Les résultats ont indiqué que toutes les souches ont maintenu un taux de survie élevé suite à la digestion gastrique; B. animalis ssp. lactisA026 (7.14 log de cellules viables/mL), L. paracasei A234 (8.12 log de cellules viables/mL), L.rhamnosus A 119 (8.48 log de cellules viables/mL) et L. rhamnosus GG (8.36 log de cellules viables/mL)et que l’expression de certains gènes de stress ciblés avait été modulée pendant la digestion gastrique. Seule la souche L. rhamnosus GG a montré une modulation au niveau de l’expression de tous les gènes ciblés. Dans l'ensemble, l’expression d’au moins un gène associé à la réponse au stress a été modulée pour toutes les souches probiotiques pendant la digestion gastrique dans la matrice de sève d’érable. Face aux stress de la digestion gastrique, les différentes réponses du métabolisme des bactéries probiotiques permettent une meilleure survie cellulaire. En définitive, ces quatre souches ont le potentiel d'être utilisées comme probiotiques dans une matrice de sève d'érable, ce qui pourrait mener au développement d'une boisson fonctionnelle.The probiotic bacteria must be able to survive during gastric digestion in order to reach the colon and thus provide human health benefits. The potential of dairy matrices to support probiotics hasoften been described, but plant products such as maple sap for this application remains little known,while the interest for these products is growing. The objective of this thesis was to evaluate cell viability by the use of propidium monoazide (PMA) coupled to qPCR, as well as to evaluate the expression offive genes linked to stress atpA, dnaK, galK, groEL and hrcA by RT-qPCR during in vitro gastricdigestion (TIM-1) of three commercial strains (Bifidobacterium animalis ssp. lactis A026,Lacticaseibacillus paracasei A234, Lacticaseibacillus rhamnosus A119) as well as the reference strain Lacticaseibacillus rhamnosus GG (ATCC 53103) delivered in maple sap. The results indicated that all strains maintained a high survival rate after gastric digestion; B. animalis subsp. lactis A026 (7.14 log ofviable cells/mL), L. paracasei A234 (8.12 log of viable cells/mL), L. rhamnosus A119 (8.48 log of viablecells/mL), and L. rhamnosus GG (8.36 log of viable cells/mL) and that the expression of certain target stress genes was modulated during gastric digestion. Only L. rhamnosus GG strain showed modulationin the expression of all target genes. Overall, the expression of at least one gene associated with the stress response was modulated for all probiotic strains during gastric digestion in the maple sap matrix. Faced with the stress of gastric digestion, the different responses of the metabolism of probiotic bacteria allow better cell survival. Ultimately, these four strains have the potential to be used asprobiotics in a maple sap matrix, which could lead to the development of a functional drink

Effect of recycled manure solids as bedding on bulk tank milk and implications for cheese microbiological quality

The dairy farm environment influences the raw milk microbiota and consequently affects milk processing. Therefore, it is crucial to investigate farm management practices such as the bedding materials. The aim of this study was to evaluate the effect of recycled manure sol-ids (RMS) as bedding material on bulk tank milk and microbiological implications for cheese quality. Bulk tank samples were collected from 84 dairy farms using RMS or straw bedding. The use of RMS did not influ-ence thermophilic and mesophilic aerobic viable counts from spores. However, straw-milk samples gave higher values for mesophilic anaerobic spore-forming bacteria (0.44 log cfu/mL) than RMS-milk samples (0.17 log cfu/mL). The presence of thermoresistant lactic acid bacteria was not increased in milk from farms using RMS. Nevertheless, taxonomic profiles of thermoresis-tant bacteria isolated were different between the 2 types of milk. More Enterococcus faecalis and Streptococcusspp. were identified in RMS-milk samples. Thermore-sistant enterococci and streptococci could easily end up in cheese. Therefore, milk proteolytic activities of these isolates were tested. Neither Streptococcus spp. nor Enterococcus faecium isolates exhibited proteolytic ac-tivities, whereas 53% of E. faecalis showed some. Also, only 1 vancomycin-resistant enterococcus was detected. Survival of selected RMS-milk samples isolates (3 E. faecalis and 1 Streptococcus thermophilus) was evalu-ated during a model Cheddar cheese manufacture. Al-though those strains survived well, they did not modify the acidification curve of milk. However, they might cause organoleptic defects during cheese maturing