Phenotypic Traits and Immunomodulatory Properties of Leuconostoc carnosum Isolated From Meat Products

Twelve strains of Leuconostoc carnosum from meat products were investigated in terms of biochemical, physiological, and functional properties. The spectrum of sugars fermented by L. carnosum strains was limited to few mono- and disaccharides, consistently with the natural habitats of the species, including meat and fermented vegetables. The strains were able to grow from 4 to 37°C with an optimum of approximately 32.5°C. The ability to grow at temperatures compatible with refrigeration and in presence of up to 60 g/L NaCl explains the high loads of L. carnosum frequently described in many meat-based products. Six strains produced exopolysaccharides, causing a ropy phenotype of colonies, according to the potential involvement on L. carnosum in the appearance of slime in packed meat products. On the other side, the study provides evidence of a potential protective role of L. carnosum WC0321 and L. carnosum WC0323 against Listeria monocytogenes, consistently with the presence in these strains of the genes encoding leucocin B. Some meat-based products intended to be consumed without cooking may harbor up to 108 CFU/g of L. carnosum; therefore, we investigated the potential impact of this load on health. No strains survived the treatment with simulated gastric juice. Three selected strains were challenged for the capability to colonize a mouse model and their immunomodulatory properties were investigated. The strains did not colonize the intestine of mice during 10 days of daily dietary administration. Intriguingly, despite the loss of viability during the gastrointestinal transit, the strains exhibited different immunomodulatory effect on the maturation of dendritic cells in vivo, the extent of which correlated to the production of exopolysaccharides. The ability to stimulate the mucosal associated immune system in such probiotic-like manner, the general absence of antibiotic resistance genes, and the lack of the biosynthetic pathways for biogenic amines should reassure on the safety of this species, with potential for exploitation of selected starters.

Comparative Genomics of Leuconostoc carnosum

Leuconostoc carnosum is a known colonizer of meat-related food matrices. It reaches remarkably high loads during the shelf life in packaged meat products and plays a role in spoilage, although preservative effects have been proposed for some strains. In this study, the draft genomes of 17 strains of L. carnosum (i.e., all the strains that have been sequenced so far) were compared to decipher their metabolic and functional potential and to determine their role in food transformations. Genome comparison and pathway reconstruction indicated that L. carnosum is a compact group of closely related heterofermentative bacteria sharing most of the metabolic features. Adaptation to a nitrogen-rich environment, such as meat, is evidenced by 23 peptidase genes identified in the core genome and by the autotrophy for nitrogen compounds including several amino acids, vitamins, and cofactors. Genes encoding the decarboxylases yielding biogenic amines were not present. All the strains harbored 1–4 of 32 different plasmids, bearing functions associated to proteins hydrolysis, transport of amino acids and oligopeptides, exopolysaccharides, and various resistances (e.g., to environmental stresses, bacteriophages, and heavy metals). Functions associated to bacteriocin synthesis, secretion, and immunity were also found in plasmids. While genes for lactococcin were found in most plasmids, only three harbored the genes for leucocin B, a class IIa antilisterial bacteriocin. Determinants of antibiotic resistances were absent in both plasmids and chromosomes.

Growth Inhibitory and Selective Pressure Effects of Sodium Diacetate on the Spoilage Microbiota of Frankfurters Stored at 4 °C and 12 °C in Vacuum

This study evaluated microbial growth in commercial frankfurters formulated with 1.8% sodium lactate (SL) singly or combined with 0.25% sodium diacetate (SDA), vacuum-packaged (VP) and stored at 4 °C and 12 °C. Standard frankfurters without SDA, containing 0.15% SL, served as controls (CN). Lactic acid bacteria (LAB) were the exclusive spoilers in all treatments at both storage temperatures. However, compared to the CN and SL treatments, SL + SDA delayed growth of LAB by an average of 5.1 and 3.1 log units, and 3.0 and 2.0 log units, respectively, after 30 and 60 days at 4 °C. On day 90, the SL + SDA frankfurters were unspoiled whereas the SL and CN frankfurters had spoiled on day 60 and day 30 to 60, respectively. At 12 °C, LAB growth was similar in all treatments after day 15, but strong defects developed in the CN and SL frankfurters only. Differential spoilage patterns were associated with a major reversal of the LAB biota from gas- and slime-producing Leuconostoc mesenteroides and Leuconostoc carnosum in the CN and SL frankfurters to Lactobacillus sakei/curvatus in the SL + SDA frankfurters. Thus, SL + SDA extends the retail shelf life of VP frankfurters by delaying total LAB growth and selecting for lactobacilli with a milder cured meat spoilage potential than leuconostocs, particularly under refrigeration.

Draft Genome Sequences of 12 Leuconostoc carnosum Strains Isolated from Cooked Ham Packaged in a Modified Atmosphere and from Fresh Sausages

Leuconostoc carnosum is a lactic acid bacterium that preferentially colonizes meat. In this work, we present the draft genome sequences of 12 Leuconostoc carnosum strains isolated from modified-atmosphere-packaged cooked ham and fresh sausages. Three strains harbor bacteriocin genes.

Effect of taxifolin on physicochemical and microbiological parameters of dry-cured pork sausage

The effect of taxifolin (TXF) with starter cultures (SC), such as Leuconostoc carnosum, or a mixture of strains Pediococcus pentosaceus and Staphylococcus xylosus, on the TXF stability was evaluated. UPLC analysis demonstrated that after 181 days of storage total TXF content was the highest in samples with TXF and L. carnosum (60%), compared to the 1st day of storage. The sausages with TXF and the mixture of P. pentosaceus and S. xylosus (56%) followed next. The samples treated only with TXF retained 40% of TXF, compared to the 1st day of storage. TXF had no significant effect on the growth of lactic acid bacteria. The accumulation of biogenic amines (BA), including histamine and putrescine, was more effectively reduced in sausages inoculated with the TXF plus P. pentosaceus and S. xylosus mixture. Using this mixture, the rate of lipolysis and processes of lipid oxidation were effectively slowed down. Fatty acid (FA) composition was stable in all cases.

Effect of Biofermentation with Taxifolin on Physicochemical and Microbiological Properties of Cold-Smoked Pork Sausages

The aim of this work is to evaluate the effect of taxifolin on the physicochemical and microbiological properties of cold-smoked pork sausages produced using different commercial starter cultures with Leuconostoc carnosum and with a mixture of Pediococcus pentosaceus and Staphylococcus xylosus. Ultra performance liquid chromatography analysis demonstrated that after 181 days of storage total taxifolin content was the highest in samples with taxifolin and L. carnosum (60 %), compared to the first day of storage. The sausages with taxifolin and the mixture of P. pentosaceus and S. xylosus (56 %) followed next. Taxifolin improved the hygienic quality of sausages without significant effect on the growth of lactic acid bacteria. The accumulation of biogenic amines, including histamine and putrescine, was more effectively reduced in sausages inoculated with the taxifolin and P. pentosaceus and S. xylosus mixture. Using this mixture, the rate of lipolysis and lipid oxidation were effectively slowed down. Samples with taxifolin and L. carnosum showed the highest free radical scavenging activity on the first day of the study ((77.4±1.3) %) (p<0.05 in all samples). Mixtures containing taxifolin and starter cultures bound free radicals better than taxifolin alone. The colour parameters (L*, a* and b*) of preparations and final products were significantly influenced by taxifolin and starter cultures and storage time (p<0.05 in all samples).

Impact of Nisin-Activated Packaging on Microbiota of Beef Burgers during Storage

ABSTRACTBeef burgers were stored at 4°C in a vacuum in nisin-activated antimicrobial packaging. Microbial ecology analyses were performed on samples collected between days 0 and 21 of storage to discover the population diversity. Two batches were analyzed using RNA-based denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. The active packaging retarded the growth of the total viable bacteria and lactic acid bacteria. Culture-independent analysis by pyrosequencing of RNA extracted directly from meat showed thatPhotobacterium phosphoreum,Lactococcus piscium,Lactobacillus sakei, andLeuconostoc carnosumwere the major operational taxonomic units (OTUs) shared between control and treated samples. Beta diversity analysis of the 16S rRNA sequence data and RNA-DGGE showed a clear separation between two batches based on the microbiota. Control samples from batch B showed a significant high abundance of some taxa sensitive to nisin, such asKocuria rhizophila,Staphylococcus xylosus,Leuconostoc carnosum, andCarnobacterium divergens, compared to control samples from batch A. However, only from batch B was it possible to find a significant difference between controls and treated samples during storage due to the active packaging. Predicted metagenomes confirmed differences between the two batches and indicated that the use of nisin-based antimicrobial packaging can determine a reduction in the abundance of specific metabolic pathways related to spoilage. The present study aimed to assess the viable bacterial communities in beef burgers stored in nisin-based antimicrobial packaging, and it highlights the efficacy of this strategy to prolong beef burger shelf life.