Régine Talon

Bacterial role in flavour development

The role of bacteria in the production of non volatile and volatile compounds involved in the fermented meat flavour is discussed. Lactic acid bacteria produce D-lactic and acetic acids which may give a sour note. By reducing the pH, they also modulate the other bio-chemical bacterial activities. In muscle tissue proteins are degraded into peptides and lipids into fatty acids mainly by endogenous enzymes. In fermented meat products with a high pH lipases from very lipolytic species of Staphylococcus could increase lipolysis. Bacteria could also play a role in the production and degradation of free amino acids. Staphylococcus and to a lesser extent, lactic acid bacteria could participate in the production of methyl-branched aldehydes and their corresponding alcohols and acids from branchedchain amino acids. By their nitrate reductase and catalase Staphylococcus species limit fatty acid oxidation and aldehyde production. Staphylococcus could contribute to the ester content as they can produce or hydrolyse esters in vitro.

Effects of starter cultures on the formation of flavour compounds in dry sausage

The purpose of this work was to study the impact of starter cultures on the production of flavour compounds in dry sausages. The effect of six starter cultures corresponding to different combinations of lactic acid bacteria (Lactobacillus sake L110, Pediococcus acidilactici 725, P. pentosaceus 716) and different Staphylococcus species (S. carnosus 833, S. warneri 863, S. saprophyticus M31) strains were tested in a total of 30 dry sausages without spices. The analysis of flavour compounds using a dynamic headspace apparatus coupled to a gas chromatograph and a mass spectrometer enabled us to identify about 80 volatile compounds. They were of various origins-lipids oxidation, fermentations, amino acid catabolism and animal feedstuffs. The influence of the starters and especially the flavouring strains proved to have a major effect on the level of volatile compounds in dry sausages. The flavour tests led to more accurate determination of the sensory characteristics of important molecules in the flavour of dry sausages. The sensory analyses showed that the butter odour of dry sausages largely depends on the catabolism of carbohydrates and that curing and rancid odours were correlated with some typical compounds of lipid oxidation.

Identification of a Variety of Staphylococcus Species by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

ABSTRACT Whole-cell fingerprinting by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) in combination with a dedicated bioinformatic software tool (MALDI Biotyper 2.0) was used to identify 152 staphylococcal strains corresponding to 22 staphylococcal species. Spectra of the 152 isolates, previously identified at the species level using a sodA gene-based oligonucleotide array, were analyzed against the main spectra of 3,030 microorganisms. A total of 151 strains out of 152 (99.3%) were correctly identified at the species level; only one strain was identified at the genus level. The MALDI-TOF MS method revealed different clonal lineages of Staphylococcus epidermidis that were of either human or environmental origin, which suggests that the MALDI-TOF MS method could be useful in the profiling of staphylococcal strains. The topology of the dendrogram generated by the MALDI Biotyper 2.0 software from the spectra of 120 Staphylococcus reference strains (representing 36 species) was in general agreement with that inferred from the 16S rRNA gene-based analysis. Our findings indicate that the MALDI-TOF MS technology, associated with a broad-spectrum reference database, is an effective tool for the swift and reliable identification of Staphylococci.

Identification of a variety of Staphylococcus species by MALDI-TOF mass spectrometry

ABSTRACT Whole-cell fingerprinting by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) in combination with a dedicated bioinformatic software tool (MALDI Biotyper 2.0) was used to identify 152 staphylococcal strains corresponding to 22 staphylococcal species. Spectra of the 152 isolates, previously identified at the species level using a sodA gene-based oligonucleotide array, were analyzed against the main spectra of 3,030 microorganisms. A total of 151 strains out of 152 (99.3%) were correctly identified at the species level; only one strain was identified at the genus level. The MALDI-TOF MS method revealed different clonal lineages of Staphylococcus epidermidis that were of either human or environmental origin, which suggests that the MALDI-TOF MS method could be useful in the profiling of staphylococcal strains. The topology of the dendrogram generated by the MALDI Biotyper 2.0 software from the spectra of 120 Staphylococcus reference strains (representing 36 species) was in general agreement with that inferred from the 16S rRNA gene-based analysis. Our findings indicate that the MALDI-TOF MS technology, associated with a broad-spectrum reference database, is an effective tool for the swift and reliable identification of Staphylococci.

Microbial ecosystems of traditional fermented meat products: The importance of indigenous starters

This paper reviews the diversity of microbiota, both in the environment and in traditional fermented European sausages. The environments of processing units were colonised at variable levels by resident spoilage and technological microbiota, with sporadic contamination by pathogenic microbiota. Several critical points were identified such as the machines, the tables and the knives - knowledge crucial for the improvement of cleaning and disinfecting practices. Traditionally fermented sausages generally did not present a sanitary risk. The great diversity of lactic acid bacteria and staphylococci was linked to manufacturing practices. Development of indigenous starters is very promising because it enables sausages to be produced with both high sanitary and sensory qualities. Our increasing knowledge of the genomes of technological bacteria will allow a better understanding of their physiology in sausages.

Analysis of biogenic amines in northern and southern European sausages and role of flora in amine production

The biogenic amine contents, microbial counts and flora producing amines were investigated in four types of fermented sausages. Southern type European sausages (Italian and Belgian) showed higher tyramine and phenylethylamine values than northern type ones (Norwegian and Belgian). The spontaneous non-starter lactic acid bacteria could be responsible for the production of these amines in the Italian products, and the cocci Gram positive in the Belgian South ones. The Norwegian sausages showed the lowest total amine content of those studied. The two Belgian types were characterised by the highest putrescine contents, associated with high counts of Enterococcus. The production of amines in vitro by the starter cultures used in the manufacture of the sausages revealed that none of the Lactobacillus species produced any amines and only Kocuria varians and Staphylococcus carnosus showed phenylethylamine and tryptamine production. High correlations were found between the content of putrescine, histamine and cadaverine.

Origin and ecological selection of core and food-specific bacterial communities associated with meat and seafood spoilage

The microbial spoilage of meat and seafood products with short shelf lives is responsible for a significant amount of food waste. Food spoilage is a very heterogeneous process, involving the growth of various, poorly characterized bacterial communities. In this study, we conducted 16S ribosomal RNA gene pyrosequencing on 160 samples of fresh and spoiled foods to comparatively explore the bacterial communities associated with four meat products and four seafood products that are among the most consumed food items in Europe. We show that fresh products are contaminated in part by a microbiota similar to that found on the skin and in the gut of animals. However, this animal-derived microbiota was less prevalent and less abundant than a core microbiota, psychrotrophic in nature, mainly originated from the environment (water reservoirs). We clearly show that this core community found on meat and seafood products is the main reservoir of spoilage bacteria. We also show that storage conditions exert strong selective pressure on the initial microbiota: alpha diversity in fresh samples was 189±58 operational taxonomic units (OTUs) but dropped to 27±12 OTUs in spoiled samples. The OTU assemblage associated with spoilage was shaped by low storage temperatures, packaging and the nutritional value of the food matrix itself. These factors presumably act in tandem without any hierarchical pattern. Most notably, we were also able to identify putative new clades of dominant, previously undescribed bacteria occurring on spoiled seafood, a finding that emphasizes the importance of using culture-independent methods when studying food microbiota.

Effect of nitrate and incubation conditions on the production of catalase and nitrate reductase by staphylococci

The objective of this work was to study the production of catalase and nitrate reductase by staphylococci in order to understand their role in lipid oxidation during sausage manufacturing. Catalase and nitrate reductase were measured in resting cells and supernatants of staphylococci grown in different conditions. All staphylococci (except S. warneri) synthetized nitrate reductase. In static condition, the synthesis was maximal during exponential growth phase, whereas in shaking condition, the synthesis was maximal at the beginning of stationary phase. The production of nitrate reductase was increased in presence of nitrate, this effect was particularly important for the two S. carnosus strains which exhibited the highest activity. For all staphylococci, the production of catalase was maximal at the end of the exponential growth phase. The lowest amount of catalase was produced by S. warneri and the highest by S. carnosus. Only S. xylosus 873 and S. saprophyticus 852 released high amounts of catalase in the supernatant growth. Staphylococci produced higher amounts of catalase in shaking conditions. Addition of nitrate in the growth media favoured the synthesis of catalase, with a pronounced effect for S. carnosus. Nitrate also favoured the release of catalase.

Histamine and tyramine production by bacteria from meat products

A series of 94 strains of lactic acid bacteria and Micrococcaceae were tested for their ability to decarboxylate histidine and tyrosine in a laboratory medium. Histamine and tyramine were quantified by using a fluorimetric and a HPLC method. There was no significant difference between the results obtained with either method. Among the strains tested, only three released histamine. On the other hand, all the strains of Carnobacterium produced high concentrations of tyramine (2193 micrograms/ml). Some strains of Lactobacillus curvatus and also Lactobacillus plantarum showed tyramine production. Micrococcaceae and Lactobacillus sake did not produce tyramine.

Traditional dry fermented sausages produced in small-scale processing units in Mediterranean countries and Slovakia. 1: Microbial ecosystems of processing environments

Microbial ecosystems were surveyed in 314 environmental samples from 54 Southern and Eastern European small-scale processing units (PUs) manufacturing traditional dry fermented sausages. The residual microflora contaminating the surfaces and the equipment were analysed after cleaning and disinfection procedures. All the PU environments were colonised at various levels by spoilage and technological microflora with excessive contamination levels in some of the PUs. Sporadic contamination by pathogenic microflora was recorded. Salmonella and Listeria monocytogenes were detected in 4.8% and 6.7% of the samples, respectively, and Staphylococcus aureus was enumerated in 6.1% of the samples. Several critical points were identified, such as the machines for S. aureus and the tables and the knives for L. monocytogenes; this knowledge is crucial for the improvement of hygiene control systems in small and traditional meat processing industries. The variability of the residual contamination emphasized the different cleaning, disinfecting and manufacturing practices routinely followed by these small-scale processing units.