The population structure and diversity of Lactococcus lactis subsp. lactis, a major industrial bacterium involved in milk fermentation, was determined at both gene and genome level. Seventy-six lactococcal isolates of various origins were studied by different genotyping methods and thirty-six strains displaying unique macrorestriction fingerprints were analyzed by a new multilocus sequence typing (MLST) scheme. This gene-based analysis was compared to genomic characteristics determined by pulsed-field gel electrophoresis (PFGE). The MLST analysis revealed that L. lactis subsp. lactis is essentially clonal with infrequent intra- and intergenic recombination; also, despite its taxonomical classification as a subspecies, it displays a genetic diversity as substantial as that within several other bacterial species. Genome-based analysis revealed a genome size variability of 20%, a value typical of bacteria inhabiting different ecological niches, and that suggests a large pan-genome for this subspecies. However, the genomic characteristics (macrorestriction pattern, genome or chromosome size, plasmid content) did not correlate to the MLST-based phylogeny, with strains from the same sequence type (ST) differing by up to 230 kb in genome size. The gene-based phylogeny was not fully consistent with the traditional classification into dairy and non-dairy strains but supported a new classification based on ecological separation between “environmental” strains, the main contributors to the genetic diversity within the subspecies, and “domesticated” strains, subject to recent genetic bottlenecks. Comparison between gene- and genome-based analyses revealed little relationship between core and dispensable genome phylogenies, indicating that clonal diversification and phenotypic variability of the “domesticated” strains essentially arose through substantial genomic flux within the dispensable genome.

Leuconostoc citreum NRRL B‐1299 has long been known to produce α‐glucans containing both α‐(1→6) and α‐(1→2) linkages, which are synthesized by α‐transglucosylases of the GH70 family. We sequenced the genome of Leuconostoc citreum NRRL B‐1299 to identify the full inventory of GH70 enzymes in this strain. Three new genes (brsA, dsrM and dsrDP) putatively encoding GH70 enzymes were identified. The corresponding recombinant enzymes were characterized. Branching sucrase A (BRS‐A) grafts linear α‐(1→6) dextran with α‐(1→2)‐linked glucosyl units, and is probably involved in the α‐(1→2) branching of L. citreum NRRL B‐1299 dextran. This is the first report of a naturally occurring α‐(1→2) branching sucrase. DSR‐M and DSR‐DP are dextransucrases that are specific for α‐(1→6) linkage synthesis and mainly produce oligomers or short dextrans with molar masses between 580 and 27 000 g·mol⁻¹. In addition, DSR‐DP contains sequences that diverge from the consensus sequences that are typically present in enzymes that synthesize linear dextran. Comparison of the genome with five other L. citreum genomes further revealed that dsrDP is unique to L. citreum NRRL B‐1299. The presence of this gene in a prophage represents the first evidence of phage‐mediated horizontal transfer of genes encoding such enzymes in lactic acid bacteria. Finally, brsA and dsrM are located in a chromosomal region in which genes encoding strain‐specific GH70 enzymes are consistently located. This region may be a good target on which to focus in order to rapidly evaluate the diversity of GH70 enzymes in L. citreum strains.

Lactococcus lactis subsp. lactis strain A12 was isolated from sourdough. Combined genomic, transcriptomic, and phenotypic analyses were performed to understand its survival capacity in the complex sourdough ecosystem and its role in the microbial community. The genome sequence comparison of strain A12 with strain IL1403 (a derivative of an industrial dairy strain) revealed 78 strain-specific regions representing 23% of the total genome size. Most of the strain-specific genes were involved in carbohydrate metabolism and are potentially required for its persistence in sourdough. Phenotype microarray, growth tests, and analysis of glycoside hydrolase content showed that strain A12 fermented plant-derived carbohydrates, such as arabinose and α-galactosides. Strain A12 exhibited specific growth rates on raffinose that were as high as they were on glucose and was able to release sucrose and galactose outside the cell, providing soluble carbohydrates for sourdough microflora. Transcriptomic analysis identified genes specifically induced during growth on raffinose and arabinose and reveals an alternative pathway for raffinose assimilation to that used by other lactococci.

As fragile food commodities, microbial and organoleptic qualities of fishery and seafood can quickly deteriorate. In this context, microbial quality and security improvement during the whole food processing chain (from catch to plate), using hurdle technology, a combination of mild preserving technologies such as biopreservation, modified atmosphere packaging and superchilling, are of great interest. Thirty-five lactic acid bacteria (LAB) belonging to the species Carnobacterium maltaromaticum, Carnobacterium inhibens, Lactococcus piscium, Leuconostoc gelidum, Vagococcus fluvialis, Vagococcus penaei and Aerococcus viridans, known to possess interesting antimicrobial activity, were chosen for their potential application as bioprotective agents as a part of hurdle technology applied to fishery products. The selection approach was based on 7 criteria including antimicrobial activity, alteration potential, resistance to chitosan coating and superchilling process, cross inhibition, biogenic amines production (histamine, tyramine) and antibiotics resistance. Antimicrobial activity was assessed against 6 common spoiling bacteria in fishery products (Shewanella baltica, Photobacterium phosphoreum, Brochothrix thermosphacta, Lactobacillus sakei, Hafnia alvei, Serratia proteamaculans) and 1 pathogenic bacteria (Listeria monocytogenes) in co-culture inhibitory assays miniaturized in 96-well microtiter plates. Antimicrobial activity and spoilage evaluation, both performed in cod and salmon juice, highlighted the existence of sensory signatures and inhibition profiles, which seem to be species related. Finally 6 LAB with no unusual antibiotics resistance profile nor histamine production ability were selected as bioprotective agents for further in situ inhibitory assays in cod and salmon based products, alone or in combination with other hurdles (chitosan, modified atmosphere packing and superchilling).