Elina J. Vihavainen

Characterization of psychrotrophic bacterial communities in modified atmosphere-packed meat with terminal restriction fragment length polymorphism.

Characterization of psychrotrophic lactic acid bacteria (LAB) and Brochothrix thermosphacta communities is needed to understand the microbial ecology of spoilage of modified atmosphere-packed (MAP) meats. To overcome the limitations of the currently used methods for the characterization of psychrotrophic bacterial communities in meat, we developed a culture-independent, 16S rRNA gene-targeted terminal restriction fragment length polymorphism (T-RFLP) method. An identification library consisting of 100 Gram-positive and 30 Gram-negative meat-associated bacterial strains was set up to identify the terminal restriction fragments derived from the communities. The taxonomic resolution level of the T-RFLP method was in between genus and species within the investigated LAB strains and within family and genus within the investigated Gram-negative strains. The established library was applied to identify the members of bacterial communities in MAP minced meat at the end of the shelf life. The T-RFLP results and plate counts on Man-Rogosa-Sharpe, Violet Red Bile Glucose, and Streptomycin sulfate thallium acetate actidione agars indicated that LAB and B. thermosphacta predominated in meat. The bacterial taxa associated with the T-RFLP results were compared to those identified among plate-grown LAB isolates by numerical ribopattern analysis. Both methods agreed that Leuconostoc spp. and Carnobacterium spp. prevailed in the LAB community in minced meat followed by Lactobacillus algidus, Lactococcus spp. and Weissella spp. Colony identification revealed that Leuconostoc gasicomitatum, L. gelidum, Carnobacterium divergens and C. maltaromaticum were the predominant LAB species. The T-RFLP results were shown to correlate with viable counts of Leuconostoc spp. and B. thermosphacta. The T-RFLP method was found to be a useful tool enabling rapid and high-throughput characterization of psychrotrophic bacteria prevailing in MAP meat.

Diversity of Leuconostoc gasicomitatum associated with meat spoilage.

Leuconostoc gasicomitatum isolates (n=384) associated with spoilage of meat and vegetable-based foods were characterised by pulsed-field gel electrophoresis (PFGE) typing. Our aim was to evaluate the diversity and distribution of spoilage-associated L. gasicomitatum isolates from meat products, and to determine whether the PFGE genotypes are specific to product, producer, or isolation year (1997-2007). PFGE typing differentiated the isolates into 68 genotypes, and revealed that none one of the 54 genotypes associated with meat products was recovered from vegetable-based foods. Generally, the meat-derived genotypes were not specific to meat animal species, and many genotypes included isolates from products of different types or processors, as well as isolates collected in different years. Furthermore, certain genotypes were repeatedly identified from products of the same processing plant suggesting that the processing environment may have an impact on L. gasicomitatum contamination of meat products.

Leuconostoc spoilage of vacuum-packaged vegetable sausages.

The present study was conducted to assess the role of lactic acid bacteria (LAB) in spoilage of a vacuum-packaged vegetable sausage product. This spoilage problem was characterized by formation of gas and slime, and was limiting the shelf life of the product. To investigate the LAB populations, LAB were enumerated in vegetable sausages graded as either spoiled or acceptable. From these vegetable sausages, 110 prevailing LAB isolates were recovered and identified using an LAB ribotyping database, which uses HindIII restriction fragment length polymorphism patterns of the 16S and 23S rRNA genes as operational taxonomic units. Finally, to determine the effects of the prevailing LAB on the sensory properties of the product, fresh vegetable sausages were inoculated with six LAB strains. The results revealed that Leuconostoc gelidum, Leuconostoc gasicomitatum, and Leuconostoc mesenteroides were the predominant LAB in the commercial vegetable sausages. The inoculation of these LAB onto vegetable sausages resulted in the formation of gas, slime, and a sour off-odor. Based on these findings, L. gelidum, L gasicomitatum, and L. mesenteroides were responsible for spoilage of the vegetable sausage product.

Chapter 11:Culture Media for Leuconostoc gasicomitatum

Leuconostoc gasicomitatum and the closely related species L. gelidum cause spoilage of various foods stored chilled in modified atmosphere. This chapter describes methods for their detection and identification. There is no selective medium available, although these bacteria grow on media suitable for lactic acid bacteria. Phenotypic methods of identification are not suitable for distinguishing between different Leuconostoc species, and several alternative methods are described.