Bacterial diversity and its evolution during storage of fresh beef from different origins under different atmosphere and temperature conditions

The purpose of this study was to evaluate the bacterial diversity and its evolution during storage of fresh beef, depending on its origin, packaging and storage temperature. Two batches of three vacuum packed striploins from United Kingdom and Belgium were obtained from a food wholesaler located in the Walloon Region. Fifteen days after slaughter, the striploins were sliced and individually kept under vacuum for 30 days: i) at −1 °C; ii) at +4 °C and iii) at −1 °C for 15 days and then at +4 °C for 15 days. The bacterial diversity was evaluated by metagenomic approach 15, 30 and 45 days after slaughter. Furthermore, each 15 days part of the vacuum packed striploin slices were repacked under modified atmosphere (70 % O2/30 % CO2), stored at +4 °C for 2 days and at +8 °C for 5 days, and then analyzed. Metagenomic analysis revealed a selection of the initial flora depending on atmosphere and temperature conditions. The development of Lactobacillus algidus was favored in samples preserved under vacuum at −1 °C, while a predominance of Lactococcus piscium was observed for samples stored at +4 °C. Moreover, storage under modified atmosphere favored the development of Leuconostoc gasicomitatum. These microorganisms have already been isolated from beef, but no study has evaluated their role in food conservation. The next step of this study will be to isolate and characterize strains of Lactobacillus algidus from meat and to assess their bioprotective potential.Conservation longue durée de la viande fraîche de bovins Blanc Bleu Belge : contraintes, évaluation et recommandations, Projet D31-127

Effects of protein source and cooking procedure on intestinal microbiota and on fermentation end-products in rats

Animal and plant proteins are major proteins sources in the human diet. After their enzymatic degradation in the upper gastro-intestinal tract, the undigested fraction of these proteins is available for fermentation by the microbiota of the large intestine leading to the production of short-chain fatty acids (SCFA), branched-chain fatty acids (BCFA), ammonia, biogenic amines, sulphur metabolites, phenols and indoles. As some of these compounds have genotoxic and cytotoxic effects, protein fermentation is considered as detrimental to the host’s epithelial health. BCFA are usually used as a marker of intestinal protein fermentation. We studied in vivo the impact of proteins from animal and plant origin, raw or after a cooking procedure, on the composition of gut microbiota and on fermentation end-products. Weanling rats were used as models of the human gut microbiota. Eight experimental diets were formulated with beef meat (Longissimus dorsi), chicken meat (Pectoralis major), white pea beans (Phaseolus vulgaris), soybeans (Glycine max), used raw and cooked, as sole source of protein in the diet. One casein diet was used as control. All diets, formulated to contain 15% of raw protein, were given to seven rats for four weeks. After euthanasia, caecal contents were collected. Pyrosequencing analyses (Roche 454 GS Junior Genome Sequencer) were performed to study the microbial composition. SCFA and BCFA were measured using HPLC (Waters 2690). Microbial composition in the caecum is associated to the type of dietary protein and to the cooking procedure applied. The proportion of BCFA in the caecal content is mainly affected by the type of protein. So BCFA represent respectively 04-06% and 35-44% of total SCFA with diets based on plant and on animal proteins. In conclusion, both the type of protein and the cooking procedure could impact the gut microbiota in terms of composition and of fermentative capacity.Crédit aux Chercheurs 1.5180.1

Metagenomic analysis targeted on the 16S ribosomal DNA to study the quality of meat : a example with raw minced beef meat

Introduction: Steak tartare is a popular meat dish in Belgium and some other European countries. This meat preparations due to their raw nature, is highly sensitive to bacterial spoilage. A better understanding of the bacterial content of this product will thus be insightful to control the risk of spoilage. Metagenomics targeted on the 16S ribosomal DNA has appeared as a powerful tool to study bacterial composition of food samples. The aim of this study is to identify the bacterial population sof steak tartare from different origin along their shelf life. Material and methods: A total of 59 samples were analysed from seven butcheries, six restaurants, six sandwich bars, 8 supermarkets without intern butcheries and 8 supermarkets with intern butcheries. Samples where directly analysed the day of receipt (day 0) and at the end their shelf life after storage at 4°C (day 2), except for six restaurants and sandwich bars who were analysed only at day 0. Classical microbiological analyses were performed in order to determine psychotrophic aerobic colony counts using modified ISO 4833 method. Metagenomic analysis targeting the 16S rDNA was performed using the Roche GS junior. Raw sequences were treated by bioinformatics in order to obtain identification and proportion of bacteria in food sample. Results: Remarkable differences appear between the origins of steaks tartare. The bacterial concentration is between 3 and 7 log CFU/g depending of the origin and the day of analysis. The samples from the butcheries are mainly composed of Lactobacillus populations and to a lesser extend of environmental contaminants like Xanthomonas campestris. On the opposite, the samples from some of the restaurants are contaminated with an estimated level of 6 to 7 log CFU/g of Brochotrix thersmosphacta, Leuconostocaceae like Leuconostoc carnosum or an uncultured Weissella sp., or, with a lesser extend, with some contaminants like Pseudomonas sp. or Psychrobacter sp. These last samples were characterized with some spoilage characteristics (slime, off odor) that can thus be put in relation with the identified bacterial populations. The samples from sandwich bars were characterized by a lower level of bacterial population (3-4 log CFU/g), but with a greater diversity in the microflora along with a higher number of environmental contaminants that are not usually found in meat products. The products at the end of the shelf life have a higher bacterial concentration but with a lower diversity with spoiled bacteria as Brochotrix thermosphacta. Significance: Compared to culture based methods on selective media and previous independent culture techniques, metagenomic analysis combined with the enumeration of psychrotrophic flora gives more valuable information, and its use should be considered as a technique for quality control or for accurately determining the shelf life and the quality of the meat.Metagenomic on meat product