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.

Use of results of microbiological analyses for risk-based control of Listeria monocytogenes in marinated broiler legs.

Microbial risk assessment provides a means of estimating consumer risks associated with food products. The methods can also be applied at the plant level. In this study results of microbiological analyses were used to develop a robust single plant level risk assessment. Furthermore, the prevalence and numbers of Listeria monocytogenes in marinated broiler legs in Finland were estimated. These estimates were based on information on the prevalence, numbers and genotypes of L. monocytogenes in 186 marinated broiler legs from 41 retail stores. The products were from three main Finnish producers, which produce 90% of all marinated broiler legs sold in Finland. The prevalence and numbers of L. monocytogenes were estimated by Monte Carlo simulation using WinBUGS, but the model is applicable to any software featuring standard probability distributions. The estimated mean annual number of L. monocytogenes-positive broiler legs sold in Finland was 7.2x10(6) with a 95% credible interval (CI) 6.7x10(6)-7.7x10(6). That would be 34%+/-1% of the marinated broiler legs sold in Finland. The mean number of L. monocytogenes in marinated broiler legs estimated at the sell-by-date was 2 CFU/g, with a 95% CI of 0-14 CFU/g. Producer-specific L. monocytogenes strains were recovered from the products throughout the year, which emphasizes the importance of characterizing the isolates and identifying strains that may cause problems as part of risk assessment studies. As the levels of L. monocytogenes were low, the risk of acquiring listeriosis from these products proved to be insignificant. Consequently there was no need for a thorough national level risk assessment. However, an approach using worst-case and average point estimates was applied to produce an example of single producer level risk assessment based on limited data. This assessment also indicated that the risk from these products was low. The risk-based approach presented in this work can provide estimation of public health risk on which control measures at the plant level can be based.

Spoilage of value-added, high-oxygen modified-atmosphere packaged raw beef steaks by Leuconostoc gasicomitatum and Leuconostoc gelidum.

Moisture-enhancing and marinating of meats are commonly used by the meat industry to add value to raw, retail products. Recently in Finland, certain value-added beef steak products have proven to be unusually susceptible to microbial spoilage leading to untoward quality deteriorations during producer-defined shelf-life. This study was conducted to evaluate the role of lactic acid bacteria (LAB) in the premature spoilage of value-added beef packaged under high-oxygen modified atmospheres. Spoilage was characterised by green discolouration and a buttery off-odour. The predominant LAB in eight packages of spoiled, marinated or moisture-enhanced beef steaks were identified by reference to a 16 and 23S rRNA gene restriction fragment length polymorphism pattern (ribotype) database. Leuconostoc gasicomitatum, Leuconostoc gelidum, Lactobacillus algidus, Lactobacillus sakei and Carnobacterium divergens were found to predominate in the LAB populations at numbers above 10(8) CFU/g. Inoculation of moisture-enhanced steaks with LAB strains and strain mixtures originating from the spoiled products demonstrated the spoilage potential of L. gasicomitatum and L. gelidum isolates. These two species produced green surface discolouration and buttery off-odours similar to these found in the spoiled, commercial products.

Role of broiler carcasses and processing plant air in contamination of modified-atmosphere-packaged broiler products with psychrotrophic lactic acid bacteria.

Some psychrotrophic lactic acid bacteria (LAB) are specific meat spoilage organisms in modified-atmosphere-packaged (MAP), cold-stored meat products. To determine if incoming broilers or the production plant environment is a source of spoilage LAB, a total of 86, 122, and 447 LAB isolates from broiler carcasses, production plant air, and MAP broiler products, respectively, were characterized using a library of HindIII restriction fragment length polymorphism (RFLP) patterns of the 16 and 23S rRNA genes as operational taxonomic units in numerical analyses. Six hundred thirteen LAB isolates from the total of 655 clustered in 29 groups considered to be species specific. Sixty-four percent of product isolates clustered either with Carnobacterium divergens or with Carnobacterium maltaromaticum type strains. The third major product-associated cluster (17% of isolates) was formed by unknown LAB. Representative strains from these three clusters were analyzed for the phylogeny of their 16S rRNA genes. This analysis verified that the two largest RFLP clusters consisted of carnobacteria and showed that the unknown LAB group consisted of Lactococcus spp. No product-associated LAB were detected in broiler carcasses sampled at the beginning of slaughter, whereas carnobacteria and lactococci, along with some other specific meat spoilage LAB, were recovered from processing plant air at many sites. This study reveals that incoming broiler chickens are not major sources of psychrotrophic spoilage LAB, whereas the detection of these organisms from the air of the processing environment highlights the role of processing facilities as sources of LAB contamination.

Enterococcus devriesei sp. nov., associated with animal sources.

The taxonomic position of two bovine strains, LMG 13603 and LMG 14595, assigned to the species Enterococcus raffinosus on the basis of biochemical features, was reinvestigated. Both reference strains and two other isolates, 6/1 (=LMG 22829) originating from a charcoal-broiled river lamprey and IE38.4 (=LMG 22830) from the air of a poultry slaughter by-product processing plant, occupied a clearly separate position, on the basis of sequence analysis of the housekeeping gene pheS (encoding the phenylalanyl-tRNA synthase alpha-subunit), relative to the type strain of E. raffinosus and all other enterococcal species with validly published names. 16S rRNA gene sequencing of strains LMG 13603, LMG 14595, 6/1 and IE38.4 confirmed their phylogenetic position in the Enterococcus avium species group, there being more than 99 % 16S rRNA gene sequence similarity to most members of the group, including E. raffinosus, and revealed Enterococcus pseudoavium as the closest phylogenetic relative (99.8-99.9 %). Further phenotypic and genotypic analyses using whole-cell-protein electrophoresis, (GTG)(5)-PCR fingerprinting, ribotyping and DNA-DNA hybridization experiments demonstrated that all four strains represent a novel enterococcal species, for which the name Enterococcus devriesei sp. nov. is proposed. The type strain is LMG 14595T (=CCM 7299T).