This mini review deals with some controversial non-starter lactic acid bacteria (NSLAB) species known to be both human and animal pathogens but also health-promoting and probiotic. The focus is on Lactococcus garvieae, two Streptococcus species (S. uberis and S. parauberis), four Weissella species (W. hellenica, W. confusa, W. paramesenteroides, and W. cibaria), and Mammalicoccus sciuri, which worldwide, are often found within the microbiotas of different kinds of cheese, mainly traditional artisanal cheeses made from raw milk and/or relying on environmental bacteria for their ripening. Based on literature data, the virulence and health-promoting effects of these bacteria are examined, and some of the mechanisms of these actions are reviewed. Additionally, their possible roles in cheese ripening are also discussed. The analysis of the literature data available so far showed that, in general, the pathogenic and the beneficial strains, despite belonging to the same species, show somewhat different genetic constitutions. Yet, when the safety of a given strain is assessed, genomic analysis on its own is not enough, and a polyphasic approach including additional physiological and functional tests is needed.

Weissella strains are often detected in spontaneously fermented foods. Because of their abilities to produce lactic acid and functional exopolysaccharides as well as their probiotic traits, Weissella spp. improve not only the sensorial properties but also nutritional values of the fermented food products. However, some Weissella species have been associated with human and animal diseases. In the era of vast genomic sequencing, new genomic/genome data are becoming available to the public on daily pace. Detailed genomic analyses are due to provide a full understanding of individual Weissella species. In this study, the genomes of six Weissella paramesenteroides strains were de novo sequenced. The genomes of 42 W. paramesenteroides strains were compared to discover their metabolic and functional potentials in food fermentation. Comparative genomics and metabolic pathway reconstructions revealed that W. paramesenteroides is a compact group of heterofermentative bacteria with good capacity of producing secondary metabolites and vitamin Bs. Since the strains rarely harbored plasmid DNA, they did not commonly possess the genes associated with bacteriocin production. All 42 strains were shown to bear vanT gene from the glycopeptide resistance gene cluster vanG. Yet none of the strains carried virulence genes.

In this study, the lactic fermentation of immature tomatoes as a tool for food ingredient production was evaluated as a circular economy-oriented alternative for valorising industrial tomatoes that are unsuitable for processing and which have wasted away in large quantities in the field. Two lactic acid bacteria (LAB) were assessed as starter cultures in an immature tomato pulp fermentation to produce functional food ingredients with probiotic potential. The first trial evaluated the probiotic character of Lactiplantibacillus plantarum (LAB97, isolated from immature tomato microbiota) and Weissella paramesenteroides (C1090, from the INIAV collection) through in vitro gastrointestinal digestion simulation. The results showed that LAB97 and C1090 met the probiotic potential viability criterion by maintaining 6 log10 CFU/mL counts after in vitro simulation. The second trial assessed the LAB starters\' fermentative ability. Partially decontaminated (110 °C/2 min) immature tomato pulp was used to prepare the individually inoculated samples (Id: LAB97 and C1090). Non-inoculated samples, both with and without thermal treatment (Id: CTR-TT and CTR-NTT, respectively), were prepared as the controls. Fermentation was undertaken (25 °C, 100 rpm) for 14 days. Throughout storage (0, 24, 48, 72 h, 7, and 14 days), all the samples were tested for LAB and Y&M counts, titratable acidity (TA), solid soluble content (SSC), total phenolic content (TPC), antioxidant capacity (AOx), as well as for organic acids and phenolic profiles, and CIELab colour and sensory evaluation (14th day). The LAB growth reached ca. 9 log10 CFU/mL for all samples after 72 h. The LAB97 samples had an earlier and higher acidification rate than the remaining ones, and they were highly correlated to lactic acid increments. The inoculated samples showed a faster and higher decrease rate in their SSC levels when compared to the controls. A nearly two-fold increase (p < 0.05) during the fermentation, over time, was observed in all samples\' AOx and TPC (p < 0.05, r = 0.93; similar pattern). The LAB97 samples obtained the best sensory acceptance for flavour and overall appreciation scores when compared to the others. In conclusion, the L. plantarum LAB97 starter culture was selected as a novel probiotic candidate to obtain a potential probiotic ingredient from immature tomato fruits.

Artisanal cheesemaking is still performed using practices and conditions derived from tradition. Feta and Kefalograviera cheeses are very popular in Greece and have met worldwide commercial success. However, there is a lack of knowledge regarding their lactic acid microecosystem composition and species dynamics during ripening. Thus, the aim of the present study was to assess the microecosystem as well as the autochthonous lactic acid microbiota during the ripening of artisanal Feta and Kefalograviera cheeses. For that purpose, raw sheep\'s milk intended for cheesemaking, as well as Feta and Kefalograviera cheeses during early and late ripening were analyzed, and the lactic acid microbiota was identified using the classical phenotypic approach, clustering with PCR-RAPD and identification with sequencing of the 16S-rRNA gene, as well as with the Biolog GEN III microplates. In addition, the functional properties of the bacterial community were evaluated using the Biolog EcoPlates, which consists of 31 different carbon sources. In general, concordance between the techniques used was achieved. The most frequently isolated species from raw sheep\'s milk were Enteroroccus faecium, Lactiplantibacillus plantarum and Pediococcus pentosaceus. The microecosystem of Feta cheese in the early ripening stage was dominated by Lp. plantarum and E. faecium, whereas, in late ripening, the microecosystem was dominated by Weissella paramesenteroides. The microecosystem of Kefalograviera cheese in the early ripening stage was dominated by Levilactobacillus brevis and E. faecium, and in late ripening by W. paramesenteroides and E. faecium. Finally, Carbohydrates was the main carbon source category that metabolized by all microbial communities, but the extent of their utilization was varied. Kefalograviera samples, especially at early ripening, demonstrated higher metabolic activity compared to Feta cheese. However, dominating species within microbial communities of the cheese samples were not significantly different.

Probiotics are viable microorganisms that confer general health benefits to the host when consumed in an adequate concentration. Probiotics may also possess strain-specific therapeutic properties and therefore finding novel strains with probiotic properties is becoming increasingly important. The present study has focused on the isolation of probiotic bacteria from dairy products which possessed potential therapeutic properties. Of the 79 strains isolated, eight were selected for further studies based on a number of traits including biofilm formation, deoxyribonuclease (DNase) activity, agglutination activity, auto-aggregation activity, antibiotic resistance, and antagonistic activity. Strain MYPS5.1 was selected from the eight isolates as the best potential probiotic candidate strain and was subsequently identified as Weissella paramesenteroides by 16S rDNA gene sequencing and sequence analysis. W. paramesenteroides strain MYPS5.1 was resistant to a number of antibiotics and the strain produced a high concentration of exopolysaccharide (EPS) (380.42 mg/L). The functional groups C-H, C = C, N = N, N-H, and C-O in the EPS were identified by using Fourier transform infrared (FTIR) spectroscopy. Computational studies showed that it interacted with cyclin-dependent kinase (CDK), a molecule which is thought to play a role in cancer pathogenesis (REF). Collectively, these results suggest that Weissella paramesenteroides MYPS5.1 is a potential probiotic strain with potential therapeutic properties.
BACKGROUND: The online version contains supplementary material available at 10.1007/s13205-021-03074-2.

Lactic fermentation of unripe green tomatoes as a tool to produce food ingredients is a viable alternative for adding value to industrial tomatoes unsuitable for processing and left in large quantities in the fields. Fermentation using starter cultures isolated from the fruit (plant-matrix adapted) can have advantages over allochthonous strains in obtaining fermented products with sensory acceptability and potentially probiotic characteristics. This paper details the characterisation of the unripe green tomato lactic microbiota to screen LAB strains for use as starter cultures in fermentation processes, along with LAB strains available from INIAV\'s collection. Morphological, biochemical (API system), and genomic (16S rDNA gene sequencing) identification showed that the dominant LAB genera in unripe green tomato are Lactiplantibacillus, Leuconostoc, and Weissella. Among nine tested strains, autochthonous Lactiplantibacillus plantarum and allochthonous Weissella paramesenteroides showed tolerance to added solanine (200 ppm) and the best in vitro probiotic potential. The results indicate that the two LAB strains are promising candidates for manufacturing probiotic fermented foods from unripe green tomatoes.

Weissella paramesenteroides has gained a considerable attention as bacteriocin and exopolysaccharide producers. However, potential of W. paramesenteroides to utilize different prebiotics is unexplored area of research. Fruits being vectors of various probiotics, five W. paramesenteroides strains, namely, FX1, FX2, FX5, FX9, and FX12, were isolated from different fruits. They were screened and selected based on their ability to survive at pH 2.5 and in 1.0% sodium taurocholate, high cell surface hydrophobicity, mucin adhesion, bile-induced biofilm formation, antimicrobial activity (AMA) against selected enteropathogens, and prebiotic utilization ability, implicating the functional properties of these strains. In vitro safety evaluation showed that strains were susceptible to antibiotics except vancomycin and did not harbor any virulent traits such as biogenic amine production, hemolysis, and DNase production. Based on their functionality, two strains FX5 and FX9 were selected for prebiotic utilization studies by thin layer chromatography (TLC) and short-chain fatty acids (SCFAs) production by high performance liquid chromatography. TLC profile evinced the ability of these two strains to utilize low molecular weight galactooligosaccharides (GOS) and fructooligosaccharides (FOS), as only the upper low molecular weight fractions were disappeared from cell-free-supernatants (CFS). Enhanced β-galactosidase activity correlated with galactose accumulation in residual CFS of GOS displayed GOS utilization ability. Both the strains exhibited AMA against E. coli and Staph. aureus and high SCFAs production in the presence of prebiotic, suggesting their synbiotic potential. Thus, W. paramesenteroides strains FX5 and FX9 exhibit potential probiotic properties with prebiotic utilization and can be taken forward to evaluate synergistic synbiotic potential in detail.

Lactic acid bacteria are observed during early stages of almost all food waste composting. Among them, 2 types of lactic acid bacteria, Pediococcus (homofermentative lactic acid bacterium) and Weissella (heterofermentative lactic acid bacterium) have been often reported. In this study, the roles of these 2 types of lactic acid bacteria in the composting were tried to elucidate. It has been pointed out that Pediococcus accelerates the composting process by producing lactic acid which prevented acetic acid generation, thus activating indigenous composting microorganisms. On the other hand, this study elucidated that Weissella produced acetic acid of 20 mg g-1 DS, which is harmful to composting microorganisms, resulting in the inhibition of vigorous organic matter degradation. When these 2 coexist in the starting material, whether the composting succceeds or not depends on the ratio of these 2 lactic acid bacteria. If Pediococcus and Weissella ratio was higher than 101.5, acetic acid level was almost 3 times lower than that observed in the composting with their lower ratios of 1 and 10-1, probably because of the interaction of Pediococcus and Weissella resulting in the suppression of Weissella activity, and thus composting was accelerated.

This case report describes the isolation and differentiation of Weissella (W.) spp. from the milk of two cows (A and B) with clinical mastitis (milk changes, asymmetry of the udder and increased somatic cell counts). Quarter milk samples obtained from two dairy cows of different farms had been submitted to the diagnostic laboratory of the Clinic for Ruminants in Vienna for bacteriological examination. Alpha-hemolytic catalase-negative gram-positive cocci in pure culture on Columbia blood agar were isolated and could not be assigned to a Lancefield group. The isolates were biochemically characterized as Leuconostoc spp. (API® 20 Strep, bioMérieux). A control examination of cow B within 7 weeks confirmed these findings. 16S rDNA sequencing indicated W. paramesenteroides (cow A) and W. cibaria (cow B). The analysis by pulsed-field gel electrophoresis (PFGE) showed identical SmaI/ApaI profiles for both W. cibaria isolates (cow B), which differed from the W. paramesenteroides fingerprint of cow A (67% similarity). This study indicates a possible relationship between the detection of Weissella spp. and the occurrence of bovine intramammary infections.

The present investigation sought to provide a reliable and highly efficient electrotransformation method for the lactic acid bacterium Weissella paramesenteroides DX. Experiments were carried out with the shuttle vectors pVS44 (2910 bps), pTRKH3 (7766 bps) and its derivative pTRKH3-1 (4855 bps). Several parameters, including the concentration of transforming plasmid DNA, plasmid size, electric field strength, age of the culture, cell density, and the pretreatment of cells with dl-threonine, lysozyme, and combined treatment with lithium acetate and dithiothreitol, were investigated and proved to influence the efficiency of transformation. Electrocompetence was found to peak in the early stationary phase (OD600 1.2). Other optimized conditions included: the concentration of 10 μg/ml transforming DNA, the cell density of 10(10) cells/ml, a high-density electric field pulse of 2.5 kV, 25 μF and 200 Ω, pretreatment of cells with 40 mM dl-threonine and 2000 U/ml lysozyme, and yielded 3.5×10(4) transformants/μg DNA for pVS44 while 1.2×10(4) transformants/μg DNA for the large plasmid TRKH3. Compared to previously reported data, the obtained transformation efficiencies provided an 8.75-fold increase for pVS44 and ensured plasmid stability for 120 generations in non-selective medium.