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The aim of this research was to find out the effect of different combinations of starter and non-starter cultures on the proteolysis of Castellano cheese during ripening. Four cheese batches were prepared, each containing autochthonous lactobacilli and or Leuconostoc, and were compared with each other and with a control batch, that used only a commercial starter. To achieve this, nitrogen fractions (pH 4.4-soluble nitrogen and 12 % trichloroacetic acid soluble nitrogen, polypeptide nitrogen and casein nitrogen), levels of free amino acids and biogenic amines were assessed. Texture and microstructure of cheeses were also evaluated. Significant differences in nitrogen fractions were observed between batches at different stages of ripening. The free amino acid content increased throughout the cheese ripening process, with a more significant increase occurring after the first 30 days. Cheeses containing non-starter lactic acid bacteria exhibited the highest values at the end of the ripening period. Among the main amino acids, GABA was particularly abundant, especially in three of the cheese batches at the end of ripening. The autochthonous lactic acid bacteria were previously selected as non-producers of biogenic amines and this resulted in the absence of these compounds in the cheeses. Analysis of the microstructure of the cheese reflected the impact of proteolysis. Additionally, the texture profile analysis demonstrated that the cheese's hardness intensified as the ripening period progressed. The inclusion of autochthonous non-starter lactic acid bacteria in Castellano cheese production accelerated the proteolysis process, increasing significantly the free amino acids levels and improving the sensory quality of the cheeses., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Lactic acid bacteria can synthesize dextran and oligosaccharides with different functionality, depending on the strain and fermentation conditions. As natural structure-forming agent, dextran has proven useful as food additive, improving the properties of several raw materials with poor technological quality, such as cereal by-products, fiber-and protein-rich matrices, enabling their use in food applications. In this study, we assessed dextran biosynthesis in situ during fermentation of brewers´ spent grain (BSG), the main by-product of beer brewing industry, with Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16. The starters performance and the primary metabolites formed during 24 h of fermentation with and without 4% sucrose (w/w) were followed., Results: The starters showed similar growth and acidification kinetics, but different sugar utilization, especially in presence of sucrose. Viscosity increase in fermented BSG containing sucrose occurred first after 10 h, and it kept increasing until 24 h concomitantly with dextran formation. Dextran content after 24 h was approximately 1% on the total weight of the BSG. Oligosaccharides with different degree of polymerization were formed together with dextran from 10 to 24 h. Three dextransucrase genes were identified in L. pseudomesenteroides DSM20193, one of which was significantly upregulated and remained active throughout the fermentation time. One dextransucrase gene was identified in W. confusa A16 also showing a typical induction profile, with highest upregulation at 10 h., Conclusions: Selected lactic acid bacteria starters produced significant amount of dextran in brewers' spent grain while forming oligosaccharides with different degree of polymerization. Putative dextransucrase genes identified in the starters showed a typical induction profile. Formation of dextran and oligosaccharides in BSG during lactic acid bacteria fermentation can be tailored to achieve specific technological properties of this raw material, contributing to its reintegration into the food chain.

Achieving a high monosaccharide composition in malt wort is instrumental to achieve successful lactic acid bacteria fermentation of malt based beverages. The conversion of monosaccharides to alternative metabolites such as the sweet polyol, mannitol with heterofermentative strains presents a novel approach for sugar reduction and to compensate for the loss of sweetness. This work outlines the application of an adopted mashing regimen with the addition of exogenous enzymes to produce wort with high fructose content which can be applied to different malted grain types with consistently efficacious monosaccharide production for bacterial fermentation. The so produced worts are then fermented with Leuconostoc citreum TR116 a mannitol hyper-producer. Malted barley, oat and wheat were mashed to stimulate protein degradation and release of free amino acids along with the enzymatic conversion of starch to fermentable sugars. Amyloglucosidase and glucose isomerase treatment converted di- and oligo-saccharides to glucose and provided a moderate fructose concentration in malt worts which was consistent across the three cereals. Fructose was completely depleted during fermentation with Lc. Citreum TR116 and converted to mannitol with high efficiency (>90%) while overall sugar reduction was >25% in all malt worts. Differences in amino acid composition of malt worts did not significantly affect growth of Lc. Citreum TR116 but did affect the formation of the aroma compounds diacetyl and isoamyl alcohol. Organic acid production and acidification of wort was similar across cereal substrates and acetic acid formation was linked to yield of mannitol. The results suggest that differences in amino acid and fructose content of malt worts considerably change metabolite formation during fermentation with Lc. Citreum TR116, a mannitol hyper-producer. This work gives new insight into the development of consumer acceptable malt based beverages which will provide further options for the health conscious and diabetic consumer, an important step in the age of sugar overconsumption., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Control of Listeria monocytogenes in queso fresco and other fresh cheeses continues to be a challenge in the United States. These cheese types are particularly challenging due to their high moisture and high pH, which provide favorable conditions for the growth of L. monocytogenes. Protective cultures (i.e., viable strains of lactic acid bacteria that inhibit other microorganisms) have been investigated in foods such as meat as an alternative, clean-label control strategy for L. monocytogenes. However, the efficacy of protective cultures can vary by food matrix. In this study, we were interested in whether protective cultures used to control L. monocytogenes in meats could be applied to control the pathogen in queso fresco. We selected 4 commercially available bacterial cultures used for the control of L. monocytogenes in meat: Lactobacillus curvatus, Lactobacillus sakei, Pediococcus acidilactici, and Leuconostoc carnosum. We incorporated these cultures into batches of queso fresco during manufacturing and evaluated them for their ability to inhibit the growth of surface-applied L. monocytogenes at levels of 1 × 10 2 and 1 × 10 4 cfu/g. We stored the queso fresco at 6 and 21°C for up to 21 d. After 14 d, Listeria was able to grow to 1 × 10 7 cfu/g on the cheese. Our data show that the bacterial cultures did not significantly inhibit the growth of L. monocytogenes in queso fresco. The results from this study highlight the complexity of antagonistic bacterial interactions and their potential variability across food matrices. Protective cultures represent an important, clean-label tool for the control of L. monocytogenes in foods, but each strain must be evaluated in the food environment it is intended for to ensure its efficacy., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The fermentation conditions for exopolysaccharides (EPS) with a highly branched structure extracted from Leuconostoc citreum B-2 were optimized by response surface methodology (RSM). The results showed that under the optimal fermentation conditions of sucrose, yeast extract and pH of 99.80 g/L, 5.94 g/L and 6.12, respectively, the B-2 EPS yield was 59.33 ± 1.34 g/L, which was 5.93 times greater than that from the initial nonoptimized conditions. Chemical composition analysis showed that the carbon, hydrogen, nitrogen and sulfur contents of the B-2 pure EPS (P-EPS) were 39.05 ± 0.25%, 7.19 ± 0.08%, 0.72 ± 0.03% and 0%, respectively. The content of uronic acid was high, reaching 20.94 ± 6.30%. B-2 EPS had certain 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl (·OH), superoxide anion (O 2- ), hydrogen peroxide (H 2 O 2 ), nitroso radical (NO 2- ) scavenging abilities and reducing power. The degradation temperature of the B-2 EPS was 313 °C, indicating that B-2 EPS have high thermal stability. The emulsification effect on vegetable oils was better than that of hydrocarbons. The EPS produced by Leu. citreum B-2 was activated in different concentrations of sucrose-supplemented skimmed milk, which caused the skimmed milk to have different degrees of solidification. Moreover, B-2 EPS promoted the growth of probiotic bacteria, especially Lactobacillus delbrueckii., (Copyright © 2019 Elsevier B.V. All rights reserved.)

To develop a starter culture system for improving the shelf-life and quality of kimchi, we prepared a mixed starter composed of Lactococcus lactis and Leuconostoc citreum. Two strains, L. lactis WiKim0098 and Leu. citreum WiKim0096, showed high antimicrobial activity and mannitol productivity, respectively. These lactic acid bacteria (LAB) were introduced as a starter into kimchi following cultivation in foodgrade liquid medium. Two kimchi samples, with and without starter, were fermented for 12 days at 10°C. Compared to the control kimchi without starter, a lower initial pH and higher number of LAB were observed in kimchi with starter at 0 day. However, the starter in kimchi prolonged the period taken by kimchi to reach to pH 4.2 by approximately 1.5-fold compared to that in the control kimchi. To estimate the effect of the starter on the flavor of kimchi, metabolite changes were evaluated by gas chromatography/mass spectrometry. In starter fermented kimchi, the levels of mannitol and amino acid, which are associated with the flavor of kimchi, were increased following fermentation. The amount of mannitol was confirmed by high-performance liquid chromatography analysis, showing concentrations of 3.4 and 5.1 mg/ml for the control and starter fermented kimchi, respectively. Thus, mixed starter inoculated with L. lactis WiKim0098 and Leu. citreum WiKim0096 may extend the shelf-life of kimchi and improve its sensory characteristics.

This study aimed to understand the survival and growth patterns of bacteriophage-sensitive Weissella and Leuconostoc strains involved in kimchi fermentation. Dongchimi kimchi was prepared, and Weissella and Leuconostoc were co-cultivated in the dongchimi broth. Weissella cibaria KCTC 3807 growth was accompanied by rapid lysis with an increase in the bacteriophage quantity. Leuconostoc citreum KCCM 12030 followed the same pattern. The bacteriophage-insensitive strains W. cibaria KCTC 3499 and Leuconostoc mesenteroides KCCM 11325 survived longer under low pH as their growth was not accompanied by bacteriophages. The bacteriophage lysate of W . cibaria KCTC 3807 accelerated and promoted the growth of Leuconostoc . Overall, our results show that bacteriophages might affect the viability and population dynamics of lactic acid bacteria during kimchi fermentation.

This study aimed to gain deeper insights into the microbiota composition and population dynamics, monitor the dominant bacterial populations and identify the specific spoilage microorganisms (SSOs) of vacuum-packed bacon during refrigerated storage using both culture-independent and dependent methods. High-throughout sequencing (HTS) showed that the microbial composition changed greatly with the prolongation of storage time. The diversity of microbiota was abundant at the initial stage then experienced a continuous decrease. Lactic acid bacteria (LAB) mainly Leuconostoc and Lactobacillus dominated the microbial population after seven days of storage. A total of 26 isolates were identified from different growth media using traditional cultivation isolation and identification method. Leuconostoc mesenteroides and Leuconostoc carnosum were the most prevalent species since day 15, while Lactobacillus sakei and Lactobacillus curvatus were only found on day 45, suggesting that they could be responsible for the spoilage of bacon. Serratia, Rahnella, Fusobacterium and Lactococcus underwent a dramatic increase at some point in individual batchs which may be considered as potential contributors to the spoilage., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The ability of twelve strains belonging to three Leuconostoc species (Leuconostoc mesenteroides, Leuconostoc lactis and Leuconostoc pseudomesenteroides) to grow under diverse sub-lethal technological stress conditions (cold, acidic, alkaline and osmotic) was evaluated in MRS broth. Two strains, Leuconostoc lactis Ln N6 and Leuconostoc mesenteroides Ln MB7, were selected based on their growth under sub-lethal conditions, and volatile profiles in RSM (reconstituted skim milk) at optimal and under stress conditions were analyzed. Growth rates under sub-lethal conditions were strain- and not species-dependent. Volatilomes obtained from the two strains studied were rather diverse. Particularly, Ln N6 (Ln. lactis) produced more ethanol and acetic acid than Ln MB7 (Ln. mesenteroides) and higher amounts and diversity of the rest of volatile compounds as well, at all times of incubation. For the two strains studied, most of stress conditions applied diminished the amounts of ethanol and acetic acid produced and the diversity and levels of the rest of volatile compounds. These results were consequence of the different capacity of the strains to grow under each stress condition tested., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A water-soluble exopolysaccharide (EPS)-producing strain YF32 was isolated from soybean paste, which was then identified as Leuconostoc pseudomesenteroides. After culturing the strain in Man-Rogosa-Sharpe (MRS) medium containing 5% sucrose at 30°C for 48h, the EPS was purified, and a yield of 12.5g/L was achieved. The weight-average molecular weight (Mw) was 5.54×10 6 Da by high-performance size-exclusion chromatography (HPSEC). The structural characterization of the purified EPS was determined by gas chromatography (GC), Fourier transform infrared (FT-IR), 1 H, 13 C nuclear magnetic resonance (NMR) spectroscopy. The results demonstrated that the exopolysaccharide was glucan with a peak, a linear backbone composed of consecutive α-(1→6)-linked d-glucopyranose units. No branching was observed in the dextran structure. The degradation temperature (Td) of EPS was 307.62°C, which suggested that dextran exhibited high thermal stability. YF32 dextran also showed high water solubility and emulsibility. All results suggested that dextran has the potential to be applied in food fields as a food additive., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Psychrotrophic lactic acid bacteria (LAB) are the prevailing spoilage organisms in packaged cold-stored meat products. Species composition and metabolic activities of such LAB spoilage communities are determined by the nature of the meat product, storage conditions, and interspecies interactions. Our knowledge of system level responses of LAB during such interactions is very limited. To expand it, we studied interactions between three common psychrotrophic spoilage LAB ( Leuconostoc gelidum , Lactococcus piscium , and Lactobacillus oligofermentans ) by comparing their time course transcriptome profiles obtained during their growth in individual, pairwise, and triple cultures. The study revealed how these LAB employed different strategies to cope with the consequences of interspecies competition. The fastest-growing bacterium, Le. gelidum , attempted to enhance its nutrient-scavenging and growth capabilities in the presence of other LAB through upregulation of carbohydrate catabolic pathways, pyruvate fermentation enzymes, and ribosomal proteins, whereas the slower-growing Lc. piscium and Lb. oligofermentans downregulated these functions. These findings may explain the competitive success and predominance of Le. gelidum in a variety of spoiled foods. Peculiarly, interspecies interactions induced overexpression of prophage genes and restriction modification systems (mechanisms of DNA exchange and protection against it) in Lc. piscium and Lb. oligofermentans but not in Le. gelidum Cocultivation induced also overexpression of the numerous putative adhesins in Lb. oligofermentans These adhesins might contribute to the survival of this slowly growing bacterium in actively growing meat spoilage communities. IMPORTANCE Despite the apparent relevance of LAB for biotechnology and human health, interactions between members of LAB communities are not well known. Knowledge of such interactions is crucial for understanding how these communities function and, consequently, whether there is any possibility to develop new strategies to interfere with their growth and to postpone spoilage of packaged and refrigerated foods. With the help of controlled experiments, detailed regulation events can be observed. This study gives an insight into the system level interactions and the different competition-induced survival strategies related to enhanced uptake and catabolism of carbon sources, overexpression of adhesins and putative bacteriocins, and the induction of exchange of genetic material. Even though this experiment dealt with only three LAB strains in vitro , these findings agreed well with the relative abundance patterns typically reported for these species in natural food microbial communities., (Copyright © 2018 American Society for Microbiology.)

Serpa cheese is one of the traditional regional Portuguese cheeses having the Protected Denomination of Origin (PDO) designation. This study investigated the bacterial community in the traditional Portuguese Serpa cheese. The microorganisms identified at the end of ripening (30 days) mainly were lactic acid bacteria (LAB). Lactobacillus paracasei/Lactobacillus casei was the main species in cheese from PDO registered industries, whereas in non-PDO registered industries Lactobacillus brevis was highlighted, among other LAB. Enterobacteriaceae species were detected at 20% to 40% of the total isolates. The results obtained by high-throughput sequencing analysis confirmed that LAB was the main microbial group, with Lactococcus genus contributing to approximately 40% to 60% of the population, followed by Leuconostoc and Lactobacillus. The Enterobacteriaceae family was also important. The differences between bacterial communities from PDO and non-PDO registered industries suggest that the lack of regulation of the cheese-making practices may influence unfavorably. The new knowledge about bacterial diversity in Serpa cheese could be useful to set up new ripening conditions, which favor the development of desirable microorganisms., Practical Application: The control of the manufacturing process of traditional cheeses can be improved through the knowledge of the bacterial diversity that develops. Thus, the growth of desirable microorganisms can be promoted to homogenize the final product., (© 2018 Institute of Food Technologists®.)

In artisanal tanneries, the skins are immersed in cereals fermented by natural microbial flora in order to reduce the pH of the skin, an essential condition for carrying out the final step. The environmental thermal variation alters the fermentation process and affects the quality of the final product. The aim of this work was to isolate lactic acid bacteria from cereals mixture fermented in an artisanal tannery and to evaluate in vitro the acidifying activity of the strains as a first step for the formulation of a starter culture. In most samples, a prevalence of cocci (95%) was observed with respect to bacilli. The best acidifying strains were identified by phenotypic and genotypic techniques as Enterococcus faecium CRL 1943 (rapid acidification at 37 °C) and Leuconostoc citreum CRL 1945 (high acidifying activity at 18 °C). In addition, the biomass production of the selected strains was analyzed at free and controlled pH (bioreactors 1.5 L). The production of biomass was optimal at controlled pH, with a higher growth (0.5-1.1 log units). Both strains were compatible, allowing their inclusion in a mixed culture. These lactic strains could contribute to the systematization of the tanning process., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.

Lactic acid bacteria produce diverse functional metabolites in fermented foods. However, little is known regarding the metabolites and the fermentation process in kimchi. In this study, the culture broth from Leuconostoc lactis, a lactic acid bacterium isolated from kimchi, was analysed by liquid chromatography-tandem mass spectrometry and identified by the MS-DIAL program. The MassBank database was used to analyse the metabolites produced during fermentation. A mass spectrum corresponding to 2-hydroxyisocaproic acid (HICA) was validated based on a collision-induced dissociation (CID) fragmentation pattern with an identified m/z value of 131.07. HICA production by lactic acid bacteria was monitored and showed a positive correlation with hydroxyisocaproate dehydrogenases (HicDs), which play a key role in the production of HICA from leucine and ketoisocaproic acid. Interestingly, the HICA contents of kimchi varied with Leuconostoc and Lactobacillus content during the early stage of fermentation, and the addition of lactic acid bacteria enhanced the HICA content of kimchi. Our results suggest that HICA production in kimchi is dependent on the lactic acid bacterial composition.

A systematic study about the intrinsic resistance of 29 strains (26 autochthonous and 3 commercial ones), belonging to Leuconostoc genus, against diverse stress factors (thermal, acidic, alkaline, osmotic and oxidative) commonly present at industrial or conservation processes were evaluated. Exhaustive result processing was made by applying one-way ANOVA, Student's test (t), multivariate analysis by Principal Component Analysis (PCA) and Matrix Hierarchical Cluster Analysis. In addition, heat adaptation on 4 strains carefully selected based on previous data analysis was assayed. The strains revealed wide diversity of resistance to stress factors and, in general, a clear relationship between resistance and Leuconostoc species was established. In this sense, the highest resistance was shown by Leuconostoc lactis followed by Leuconostoc mesenteroides strains, while Leuconostoc pseudomesenteroides and Leuconostoc citreum strains revealed the lowest resistance to the stress factors applied. Heat adaptation improved thermal cell survival and resulted in a cross-resistance against the acidic factor. However, all adapted cells showed diminished their oxidative resistance. According to our knowledge, this is the first study regarding response of Leuconostoc strains against technological stress factors and could establish the basis for the selection of "more robust" strains and propose the possibility of improving their performance during industrial processes., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The effects of purified salt (PS) and mineral-rich sea salt (MRS), both with different mineral profiles, on kimchi fermentation were studied using a culture-dependent 16S rRNA sequencing technique and mass-based metabolomic analysis. The different mineral profiles in the fermentation medium caused changes in the bacterial profiles of the 2 kimchi products. An increase of Leuconostoc species in MRS-kimchi decreased the Lactobacillus/Leuconostoc ratio, which led to changes in metabolites (including sugars, amino acids, organic acids, lipids, sulfur compounds, and terpenoids) associated with kimchi quality. Although further studies on the relationship between these salt types and kimchi fermentation are needed, these results suggested that the MRS treatment had positively affected the changes of the kimchi mineral contents, bacterial growth, and metabolite profiles, which are linked to kimchi quality., (© 2017 Institute of Food Technologists®.)

Four obligate heterofermentative lactic acid bacteria (LAB) strains (Weissella cibaria PON10030 and PON10032 and Leuconostoc citreum PON 10079 and PON10080) were tested as single strain starters, mono-species dual strain starters, and multiple strain starter for the preparation and propagation of sourdoughs for the production of a typical bread at industrial level. The kinetics of pH and TTA during the daily sourdough refreshments indicated a correct acidification process for all trials. The concentration of lactic and acetic acid increased consistently during fermentation. The resulting molar ratios between these two organic acids in the experimental trials were lower than those observed in the control trial. The microbiological investigation showed levels of approximately 10(9) CFU/mL in almost all sourdoughs and the comparison of the genetic polymorphisms of the dominating LAB with those of the pure cultures evidenced the persistence of the added strains over time. The resulting breads were evaluated for several quality parameters. The breads with the greatest height were obtained with the quadruple combination of leuconostocs and weissellas. The highest softness was registered for the breads obtained from fermentations performed by W. cibaria PON10032 alone and in combination. The different inocula influenced also the color, the void fraction, the cell density and the mean cell area of the breads. Different levels of acids, alcohols, aldehydes, esters, hydrocarbons, ketones, terpenes, furans and phenol were emitted by the breads. The sensory tests indicated the breads from the sourdoughs fermented with the seven LAB inocula as sweeter and less acidic than control breads and the breads from the trials with the highest complexity of LAB inoculums were those more appreciated by tasters. A multivariate approach found strong differences among the trials. In particular, control breads and the breads obtained with different starter LAB were quite distant and a more strict relation was found among the productions carried out by W. cibaria strains. This study proved the suitability of the selected strains of L. citreum and W. cibaria for industrial-scale level applications in sourdough bread production., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Staphylococcus aureus is presently regarded as an emerging zoonotic agent due to the spread of specific methicillin-resistant S. aureus (MRSA) clones in pig farms. Studying the microbiota can be useful for the identification of bacteria that antagonize such opportunistic veterinary and zoonotic pathogen in animal carriers. The aim of this study was to determine whether the nasal microbiome of pig S. aureus carriers differs from that of non-carriers. The V3-V5 region of the 16S rRNA gene was sequenced from nasal swabs of 44 S. aureus carriers and 56 non-carriers using the 454 GS FLX titanium system. Carriers and non-carriers were selected on the basis of quantitative longitudinal data on S. aureus carriage in 600 pigs sampled at 20 Danish herds included in two previous studies in Denmark. Raw sequences were analysed with the BION meta package and the resulting abundance matrix was analysed using the DESeq2 package in R to identify operational taxonomic units (OTUs) with differential abundance between S. aureus carriers and non-carriers. Twenty OTUs were significantly associated to non-carriers, including species with known probiotic potential and antimicrobial effect such as lactic acid-producing isolates described among Leuconostoc spp. and some members of the Lachnospiraceae family, which is known for butyrate production. Further 5 OTUs were significantly associated to carriage, including known pathogenic bacteria such as Pasteurella multocida and Klebsiella spp. Our results show that the nasal microbiome of pigs that are not colonized with S. aureus harbours several species/taxa that are significantly less abundant in pig carriers, suggesting that the nasal microbiota may play a role in the individual predisposition to S. aureus nasal carriage in pigs. Further research is warranted to isolate these bacteria and assess their possible antagonistic effect on S. aureus for the pursuit of new strategies to control MRSA in pig farming.

The D-form of lactate, which causes metabolic stress upon excessive dietary intake, is mainly produced by Leuconostoc sp., the predominant species in sauerkraut. To shift the metabolic flux of d-lactate from pyruvate to l-lactate, we expressed the l-lactate dehydrogenase (ldhL) gene in Leuconostoc mesenteroides ATCC 8293. The ldhL gene from Lactobacillus plantarum was introduced into L. mesenteroides using the shuttle vectors pLeuCM and pLeuCM42. To elevate the expression level of ldhL in L. mesenteroides, the nucleotides for pyruvate kinase promoter were fused to ldhL and cloned into above vectors to construct pLC18pkL and pLC42pkL. As results, introduction of pLC42pkL in L. mesenteroides significantly improved both l-LDH activity and l-lactate productivity during fermentation, decreasing the d-/l-lactate ratio. When used as a starter culture for sauerkraut fermentation, recombinant L. mesenteroides harboring pLC42pkL increased l-lactate concentration and decreased d-lactate concentration compared to the wild type strain. We newly developed a recombinant L. mesenteroides which has high l-lactate dehydrogenase activity and applied this strain to minimize the harmful effect of d-lactate during the sauerkraut fermentation. To the best of our knowledge, we demonstrate for the first time the effective use of recombinant Leuconostoc sp. for quality improvement of fermented foods., (Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

New strains are desirable to diversify flavour of fermented dairy products. The objective of this study was to evaluate the potential of Leuconostoc spp. and Lactobacillus spp. in the production of aroma compounds by metabolic fingerprints of volatiles. Eighteen strains, including five Lactobacillus species (Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus sakei) and three Leuconostoc species (Leuconostoc citreum, Leuconostoc lactis, and Leuconostoc mesenteroides) were incubated for 5 weeks in a curd-based slurry medium under conditions mimicking cheese ripening. Populations were enumerated and volatile compounds were analysed by headspace trap gas chromatography-mass spectrometry (GC-MS). A metabolomics approach followed by multivariate statistical analysis was applied for data processing and analysis. In total, 12 alcohols, 10 aldehydes, 7 esters, 11 ketones, 5 acids and 2 sulphur compounds were identified. Very large differences in concentration of volatile compounds between the highest producing strains and the control medium were observed in particular for diacetyl, 2-butanol, ethyl acetate, 3-methylbutanol, 3-methylbutanoic acid and 2-methylbutanoic acid. Some of the characterized strains demonstrated an interesting aromatizing potential to be used as adjunct culture.

Interest in natural/organic meat products has resulted in the need to validate the effectiveness of clean label antimicrobials to increase safety and shelf life of these products. A Response Surface Methodology (RSM) was used to investigate the effects of varying levels of moisture, pH, and a commercial "clean-label" antimicrobial (cultured sugar-vinegar blend; CSVB) on the growth rate of Listeria monocytogenes and Leuconostoc mesenteroides in uncured turkey stored at 4 °C for 16 wk. Twenty treatment combinations of moisture (60% to 80%), pH (5.8 to 6.4), and CSVB (2.5% to 5.0%) were evaluated during phase I to develop growth curves for both microbe types, whereas the interactive effects of pH (5.8 to 6.4) and CSVB (0.0 to 4.75) were tested in 16 treatment combinations during Phase II at a single moisture level using L. monocytogenes only. CSVB inhibited L. monocytogenes growth in 14 of the 20 treatments tested in Phase I and in 12 of the 16 treatments in Phase II through 16 and 8 wk, respectively. In contrast, CSVB had little effect on L. mesenteroides, with growth inhibited in only 4 of 20 treatments in Phase I and was therefore not tested further in Phase II. Significant interactions of the RSM design coefficients yielded a predictive model for L. mesenteroides growth rate, but due to lack of growth, no growth rate model was developed for L. monocytogenes. CSVB was found to be an effective antilisteral antimicrobial, while having little effect on a spoilage microorganism., (© 2016 Institute of Food Technologists®)

This study evaluated the effect of residual O2 level (0% to 5%) on microbial growth and volatile metabolite production on par-fried French fries packaged in a modified atmosphere with 60% CO2 (rest N2 ) at 4 °C. The results obtained showed that the initial headspace (IH) O2 level had an effect on growth of Leuconostoc mesenteroides on French fry simulation agar, whereby growth was slightly faster under 5% O2 . In terms of quantity, ethanol, 2-methyl-1-propanol, and dimethyl disulphide were the most significant volatile metabolites produced by L. mesenteroides. The production of ethanol by L. mesenteroides was highest on simulation agar packaged under low IH O2 levels (0% to 1%), indicating that the fermentative metabolism was induced under these conditions. In agreement with the results observed on the simulation medium, growth of native lactic acid bacteria was faster under an IH O2 level of 5%. In addition, ethanol, 2-methyl-1-propanol, and dimethyl disulphide were also quantitatively the most important volatile metabolites. However, in contrast, greater quantities of ethanol and dimethyl disulphide were produced on par-fried French fries packaged under 5% O2 . This was attributed to the limited growth of the native flora on the par-fried French fries under residual O2 levels of 0% and 1%. Although some significant differences (P < 0.05) occurred between the French fries packaged in 0%, 1%, and 5 % residual O2 during storage, all products were considered to be acceptable for consumption. The results of this study can be used to optimize the shelf-life of packaged chill stored potato products., (© 2016 Institute of Food Technologists®)

In spite of a global concern on the transfer of antibiotic resistances (AR) via the food chain, limited information exists on this issue in species of Leuconostoc and Weissella, adjunct cultures used as aroma producers in fermented foods. In this work, the minimum inhibitory concentration was determined for 16 antibiotics in 34 strains of dairy origin, belonging to Leuconostoc mesenteroides (18), Leuconostoc citreum (11), Leuconostoc lactis (2), Weissella hellenica (2), and Leuconostoc carnosum (1). Atypical resistances were found for kanamycin (17 strains), tetracycline and chloramphenicol (two strains each), and erythromycin, clindamycin, virginiamycin, ciprofloxacin, and rifampicin (one strain each). Surprisingly, L. mesenteroides subsp. mesenteroides LbE16, showed resistance to four antibiotics, kanamycin, streptomycin, tetracycline and virginiamycin. PCR analysis identified tet(S) as responsible for tetracycline resistance in LbE16, but no gene was detected in a second tetracycline-resistant strain, L. mesenteroides subsp. cremoris LbT16. In Leuconostoc mesenteroides subsp. dextranicum LbE15, erythromycin and clindamycin resistant, an erm(B) gene was amplified. Hybridization experiments proved erm(B) and tet(S) to be associated to a plasmid of ≈35 kbp and to the chromosome of LbE15 and LbE16, respectively. The complete genome sequence of LbE15 and LbE16 was used to get further insights on the makeup and genetic organization of AR genes. Genome analysis confirmed the presence and location of erm(B) and tet(S), but genes providing tetracycline resistance in LbT16 were again not identified. In the genome of the multi-resistant strain LbE16, genes that might be involved in aminoglycoside (aadE, aphA-3, sat4) and virginiamycin [vat(E)] resistance were further found. The erm(B) gene but not tet(S) was transferred from Leuconostoc to Enterococcus faecalis both under laboratory conditions and in cheese. This study contributes to the characterization of AR in the Leuconostoc-Weissella group, provides evidence of the genetic basis of atypical resistances, and demonstrates the inter-species transfer of erythromycin resistance.

Background: In the process of enrichment of dairy products a priority element is the proper selection of compounds that are a mineral carrier. Calcium bisglycinate is better absorbed by the body than inorganic forms of calcium. Moreover, the lactic acid which is produced in kefir fermentation and the presence of lactose have also a positive effect on the improvement of absorption of calcium. The aim of the present study was to determine the influence of the applied dose of calcium in the form of calcium bisglycinate on the physicochemical and sensory properties and texture of kefirs during 21-day period of cold storage., Methods: Processed cow milk was enriched with 0, 5, 10, 15, 20, 25 and 30 mg of calcium (for 100 g of milk), repasteurized (72°C, 1 min), cooled down (26°C), inoculated with Commercial VITAL kefir culture (Danisco, Poland) and fermented for 16 hours (26°C). The assessment of the influence of addition   of calcium bisglycinate on acidity, syneresis, texture and sensory characteristics (1-9 points) of kefirs was conducted at four fixed dates (after 1 day, 7 days, 14 days and 21 days of storage)., Results: During successive weeks of cold storage in all experimental groups there was observed a tendency to decrease general acidity. On the 1st and 7th days of cold storage reduced whey leakage was observed in kefirs enriched with 25 mg and 30 mg Ca/100 g of milk. With increasing doses of enrichment with calcium both the hardness, adhesiveness and gumminess of kefirs decreased. The applied doses of calcium did not cause changes in the sensory characteristics such as colour and consistency of the fermented beverages., Conclusions: Calcium bisglycinate may be used to enrich kefirs with calcium even with 30 mg of calcium in 100 g of milk without the modification of the product's parameters.

Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile saltresistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

A levan-producing strain, BD1707, was isolated from Tibetan kefir and identified as Leuconostoc citreum. The effects of carbon sources on the growth of L. citreum BD1707 and levan production in tomato juice were measured. The changes in pH, viable cell count, sugar content, and levan yield in the cultured tomato juice supplemented with 15% (wt/vol) sucrose were also assayed. L. citreum BD1707 could synthesize more than 28 g/liter of levan in the tomato juice-sucrose medium when cultured at 30°C for 96 h. Based on the monosaccharide composition, molecular mass distribution, Fourier transform infrared (FTIR) spectra, and nuclear magnetic resonance (NMR) spectra, the levan synthesized by L. citreum BD1707 was composed of a linear backbone consisting of consecutive β-(2→6) linked d-fructofuranosyl units, with an estimated average molecular mass of 4.3 × 10(6) Da., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Some Leuconostoc spp. have the ability to produce slime and undesirable compounds in cooked sausage. The objectives of this research were to identify Leuconostoc sources in a Vienna-type sausage processing plant and to evaluate the genetic diversity of the isolated strains. Three hundred and two samples of sausage batter, sausages during processing, spoiled sausage, equipment surfaces, chilling brine, workers' gloves and aprons, and used casings were collected (March to November 2008 and February to April 2010) from a sausage processing plant. Lactic acid bacteria (LAB) were quantified, and Leuconostoc were detected using PCR. Strains were isolated and identified in Leuconostoc-positive samples. Leuconostoc strains were genotyped using randomly amplified polymorphic DNA and pulsed-field gel electrophoresis. LAB content of nonspoiled and spoiled sausage ranged from <0.8 to 4.4 log CFU/g and from 4.9 to 8.3 log CFU/g, respectively. LAB levels on equipment surfaces ranged from <1.3 to 4.8 log CFU/100 cm(2). Leuconostoc was detected in 35% of the samples, and 88 Leuconostoc spp. strains were isolated and genotyped. The main Leuconostoc spp. isolated were L. mesenteroides (37 genotypes), L. fallax (29 genotypes), and L. lactis (6 genotypes). Some strains of Leuconostoc isolated from equipment surfaces and sausages showed the same genotype. One L. lactis genotype included strains isolated from spoiled sausages analyzed in April 2008 and March to April 2010. Equipment and conveyor belts constitute Leuconostoc contamination sources. Leuconostoc persistence in the sausage processing environment and in the final product suggests the existence of microbial reservoirs, possibly on equipment surfaces.

Objective: Our aim was to compare the composition and diversity of Lactobacillus microbiota in infants living in Malawi and Southwestern Finland., Methods: The composition and diversity of the Lactobacillus group was analyzed in the feces of healthy 6-month-old infants living in rural Malawi (n = 44) and Southwestern Finland (n = 31), using the quantitative polymerase chain reaction method and PCR-denaturing gradient gel electrophoresis fingerprinting., Results: Malawian infants had higher counts of lactobacilli than their Finnish counterparts (7.45 log cells/g vs 6.86 log cells/g, P < 0.001, respectively) and the Lactobacillus community was richer and more diverse in the Malawian infants. Leuconostoc citreum and Weissella confusa were the predominant species in both study groups, but Malawian infants were more often colonized by these species (100% vs 74.2%, P < 0.001; 95.5% vs 41.9%, P < 0.001, respectively). Moreover, Lactobacillus ruminis, Lactobacillus gasseri, Lactobacillus acidophilus, and Lactobacillus mucosae were detected more often in the Malawian infants (59.1% vs 0.0%, P < 0.001; 38.6% vs 9.7%, P = 0.004; 29.5% vs 0.0%, P < 0.001; 22.7% vs 3.2%, P = 0.017, respectively). Lactobacillus casei group species, however, were only detected in the Finnish infants., Conclusions: Malawian infants have a more abundant Lactobacillus microbiota with a distinct composition compared with Finnish infants. The environment, including diet and hygiene, may be among the factors influencing these differences.

This study aimed at evaluating raw materials as potential lactic acid bacteria (LAB) sources for kimchi fermentation and investigating LAB successions during fermentation. The bacterial abundances and communities of five different sets of raw materials were investigated using plate-counting and pyrosequencing. LAB were found to be highly abundant in all garlic samples, suggesting that garlic may be a major LAB source for kimchi fermentation. LAB were observed in three and two out of five ginger and leek samples, respectively, indicating that they can also be potential important LAB sources. LAB were identified in only one cabbage sample with low abundance, suggesting that cabbage may not be an important LAB source. Bacterial successions during fermentation in the five kimchi samples were investigated by community analysis using pyrosequencing. LAB communities in initial kimchi were similar to the combined LAB communities of individual raw materials, suggesting that kimchi LAB were derived from their raw materials. LAB community analyses showed that species in the genera Leuconostoc, Lactobacillus, and Weissella were key players in kimchi fermentation, but their successions during fermentation varied with the species, indicating that members of the key genera may have different acid tolerance or growth competitiveness depending on their respective species., (© 2015 Institute of Food Technologists®)

Microbial community structure and population dynamics during spontaneous bamboo shoot fermentation for production of 'soidon' (indigenous fermented food) in North-east India were studied using cultivation-dependent and cultivation-independent molecular approaches. Cultivation-dependent analyses (PCR-amplified ribosomal DNA restriction analysis and rRNA gene sequencing) and cultivation-independent analyses (PCR-DGGE, qPCR and Illumina amplicon sequencing) were conducted on the time series samples collected from three independent indigenous soidon fermentation batches. The current findings revealed three-phase succession of autochthonous lactic acid bacteria to attain a stable ecosystem within 7 days natural fermentation of bamboo shoots. Weissella spp. (Weissella cibaria, uncultured Weissella ghanensis) and Lactococcus lactis subsp. cremoris predominated the early phase (1-2 days) which was joined by Leuconostoc citreum during the mid-phase (3 days), while Lactobacillus brevis and Lactobacillus plantarum emerged and became dominant in the late phase (5-7 days) with concurrent disappearance of W. cibaria and L. lactis subsp. cremoris. Lactococcus lactis subsp. lactis and uncultured Lactobacillus acetotolerans were predominantly present throughout the fermentation with no visible dynamics. The above identified dominant bacterial species along with their dynamics can be effectively utilized for designing a starter culture for industrialization of soidon production. Our results showed that a more realistic view on the microbial ecology of soidon fermentation could be obtained by cultivation-dependent studies complemented with cultivation-independent molecular approaches. Moreover, the critical issues to be considered for reducing methodological biases while studying the microbial ecology of traditional food fermentation were also highlighted with this soidon fermentation model., (© 2015 John Wiley & Sons Ltd.)

Phages infecting Leuconostoc mesenteroides strains can be overlooked during milk fermentation because they do not slowdown the acidification process. However, they can negatively impact the flavor profile of the final product. Yet, the information about these phages is still scarce. In this work, we investigated diverse factors influencing the adsorption of seven virulent Ln. mesenteroides phages, isolated from blue cheese manufacture in Argentina, to their host cells. The addition of calcium ions was generally necessary to observe complete cell lysis and plaque formation for four of the seven phages, but adsorption was very high even in the absence of this cation for all phages. The temperature barely influenced the adsorption process as it was high within the temperature range tested (0 to 50 °C). Moreover, the kinetics of adsorption were similar on viable and non-viable cells, revealing that phage adsorption does not depend on physiological state of the bacterial cells. The adsorption rates were also high at pH values from 4 to 9 for all Ln. mesenteroides phages. We also analyzed the complete genome sequences of two of these phages. Complete nucleotide analysis of phages Ln-8 and Ln-9 showed dsDNA genomes with sizes of 28.5 and 28.9 kb, and the presence of 45 and 48 open reading frames (ORFs), respectively. These genomes were highly similar to those of previously characterized Φ1-A4 (USA, sauerkraut, fermentation) and ΦLN25 (England, whey), both virulent Ln. mesenteroides phages. A detailed understanding of these phages will lead to better control strategies., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Growing expectations of consumers of fermented dairy products urge the search for novel solutions that would improve their organoleptic properties and in the case of rennet cheeses-that would also accelerate their ripening process. The aim of this study was to determine the peptidolytic activities and growth of heat-treated commercial culture of lactic acid bacteria. The analyzed culture was characterized by a relatively high peptidolytic activity. The growth of bacterial culture subjected to heat treatment at 50-80 °C for 15 s, 10 and 3 min was delayed by a few or 10-20 h compared to the control culture. Based on the results achieved, it may be concluded that in the production of rennet cheeses, the application of additional, fermentation-impaired starter cultures (via heating for ten or so minutes) may serve to accelerate their ripening and to improve their sensory attributes.

Leuconostoc gelidum subsp. gasicomitatum is a common spoilage bacterium in meat products packaged under oxygen-containing modified atmospheres. Buttery off-odors related to diacetyl/acetoin formation are frequently associated with the spoilage of these products. A whole-genome microarray study, together with gas chromatography (GC)-mass spectrometry (MS) analyses of the pathway end products, was performed to investigate the transcriptome response of L. gelidum subsp. gasicomitatum LMG18811(T) growing on semidefined media containing glucose, ribose, or inosine, which are essential carbon sources in meat. Generally, the gene expression patterns with ribose and inosine were quite similar, indicating that catabolism of ribose and nucleosides is closely linked. Diacetyl/acetoin concentrations as high as 110 or 470 μM were measured when growth was based on inosine or ribose, respectively. The gene expression results for pyruvate metabolism (upregulation of α-acetolactate synthase, downregulation of l-lactate dehydrogenase and pyruvate dehydrogenase) were as expected when diacetyl and acetoin were the end products. No diacetyl production (<7.5 μM) was detected with the glucose-containing medium, even though the cell counts of LMG18811(T) was 6 or 10 times higher than that on inosine or ribose, respectively. Although glucose was the most effective carbon source for the growth of L. gelidum subsp. gasicomitatum, utilization of inosine and ribose resulted in the production of the unwanted buttery-odor compounds. These results increase our understanding of which compounds are likely to enhance the formation of buttery odors during meat spoilage caused by L. gelidum subsp. gasicomitatum., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The aim of this study was to develop a competitive quantitative-PCR (CQ-PCR) method for rapid analysis of the population dynamics of lactic acid bacteria (LAB) in kimchi. For this, whole chromosome sequences of Leuconostoc mesenteroides, Lactobacillus plantarum, and Lb. brevis were compared and species-specific PCR primers targeting dextransucrase, 16S rRNA, and surface layer protein D (SlpD) genes, respectively, were constructed. The tested strains were quantified both in medium and kimchi by CQ-PCR and the results were compared with the data obtained using a conventional plate-counting method. As a result, the three species were successfully detected and quantified by the indicated primer sets. Our results show that the CQ-PCR method targeting species-specific genes is suitable for rapid estimation of LAB population to be used in the food fermentation industry.

The characteristics of the growth of Leuconostoc mesenteroides BD1710 and the synthesis of dextran in tomato juice supplemented with 15% sucrose were assayed. L. mesenteroides BD1710 could synthesize approximately 32 g L(-1) dextran in the tomato-juice-sucrose medium when cultured at 28 °C for 48 h, which was on the same level as the dextran yield in a chemically defined medium. The viscosity of the cultured tomato-juice-sucrose medium with various dextran contents was also measured. The results of the monosaccharide composition, molecular-weight distribution, Fourier transform infrared spectra (FTIR) and nuclear magnetic resonance spectra (NMR) showed that the polysaccharide synthesized by L. mesenteroides BD1710 in the tomato-juice-sucrose medium was dextran with a peak molecular weight of 6.35 × 10(5)Da, a linear backbone composed of consecutive α-(1 → 6)-linked d-glucopyranosyl units and approximately 6% α-(1 → 3) branches., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Aims: Leuconostoc citreum is an important lactic acid bacterium in fermented foods, but dextran production often causes undesired ropiness. To prevent this side effect, a dextran-free mutant needs to be created., Methods and Results: Homologous recombination of the dextransucrase gene (dsrC) was conducted using a segregationally unstable plasmid, pCBM32-DSUDs. A mutant was obtained on sucrose agar medium, and a site-specific insertional inactivation in the gene was confirmed. When cultured in sucrose medium, the mutant strain produced no dextransucrase or dextran. Additionally, it showed a longer lag phase (9 h) than the wild-type strain (3 h), providing new insights into the role of dextransucrase in carbohydrate metabolism of Leuconostoc., Conclusions: In this study, a dextransucrase knockout mutant was constructed. It was found that Leuc. citreum dextransucrase not only synthesizes dextran for cell protection but also provides fructose as an important carbon source for cell growth., Significance and Impact of the Study: This knockout mutation was generated for the first time in Leuc. citreum. The dextran-free mutant has the potential to be used for various industrial purposes, including as a starter culture for production of nonviscous fermented foods and as a dextran-free host for production of recombinant proteins., (© 2014 The Society for Applied Microbiology.)

Furfural is an inhibitor of growth and ethanol production by Zymomonas mobilis. This study used a naturally occurring (not GMO) biological pre-treatment to reduce that amount of furfural in a model fermentation broth. Pre-treatment involved inoculating and incubating the fermentation broth with strains of Leuconostoc mesenteroides or Leuconostoc pseudomesenteroides. The Leuconostoc strains converted furfural to furfuryl alcohol without consuming large amounts of dextrose in the process. Coupling this pre-treatment to ethanolic fermentation reduced furfural in the broth and improved growth, dextrose uptake and ethanol formation. Pre-treatment permitted ethanol formation in the presence of 5.2 g L(-1) furfural, which was otherwise inhibitive. The pre-treatment and presence of the Leuconostoc strains in the fermentation broth did not interfere with Z. mobilis ethanolic fermentation or the amounts of ethanol produced. The method suggests a possible technique for reducing the effect that furfural has on the production of ethanol for use as a biofuel., (Published by Elsevier Ltd.)

The microbiota associated with spontaneous fermentation of vegetables in a saline substrate may represent an important group of bacteria in the food industry. In this work, the lactic acid bacteria (LAB) Weissella cibaria, Lactobacillus plantarum, Lactobacillus paraplantarum, and Leuconostoc citreum were identified by partial 16S rRNA gene sequence analysis. In addition, entophytic bacteria such as Pantoea eucalypti, Pantoea anthophila, Enterobacter cowanii, and Enterobacter asburiae were detected, but they were irrelevant for the fermentation process and were inhibited after 12 h of fermentation when the pH decreased from 6.5 to 4.9. Moreover, 2 species of yeast were isolated and identified as Hanseniaspora pseudoguilliermondii and Kodamaea ohmeri by their partial 26S rRNA gene sequence. The growth of LAB was evaluated at different sodium chloride contents. L. citreum was the most halotolerant species followed by L. plantarum and W. cibaria with a concentration index to obtain a 50% population reduction (IC(50)) of 7.2%, 6.6%, and 5.2%, respectively. Furthermore, the growth of LAB and Escherichia coli O157:H7 was evaluated in the presence of the main phenylpropanoids from chilli peppers such as p-coumaric and ferulic acid. It was determined that LAB can grow in both acids at 4 mM, unlike E. coli O157:H7, whose growth is inhibited in the presence of these acids., (© 2014 Institute of Food Technologists®)

Two Bacillus subtilis strains isolated from the fruiting body of a basidiomycete fungus Pholiota squarrosa exhibited a broad range of antibacterial activity, including those against methicillin-resistant Staphylococcus aureus INA 00761 (MRSA) and Leuconostoc mes6nteroides VKPM B-4177 resistant to glycopep-> tide antibiotics, as well as antifungal activity. The strains were identified as belonging to the "B. subtilis" com- plex based on their morphological and physiological characteristics, as well as by sequencing of the 16S rRNA gene fragments. Both strains (INA 01085 and INA 01086) produced insignificant amounts of polyene antibiotics (hexaen and pentaen, respectively). Strain INA 01086 produced also a cyclic polypeptide antibiotic containing Asp, Gly, Leu, Pro, Tyr, Thr, Trp, and Phe, while the antibiotic of strain INA 01085 contained, apart from these, two unidentified nonproteinaceous amino acids. Both polypeptide antibiotics were new compounds efficient against gram-positive bacteria and able to override the natural bacterial antibiotic resistance.

Pyrosequencing of the 16S rRNA gene, community-level physiological profiles determined by the use of Biolog EcoPlates, and proteolysis analyses were used to characterize Canestrato Pugliese Protected Designation of Origin (PDO) cheese. The number of presumptive mesophilic lactococci in raw ewes' milk was higher than that of presumptive mesophilic lactobacilli. The numbers of these microbial groups increased during ripening, showing temporal and numerical differences. Urea-PAGE showed limited primary proteolysis, whereas the analysis of the pH 4.6-soluble fraction of the cheese revealed that secondary proteolysis increased mainly from 45 to 75 days of ripening. This agreed with the concentration of free amino acids. Raw ewes' milk was contaminated by several bacterial phyla: Proteobacteria (68%; mainly Pseudomonas), Firmicutes (30%; mainly Carnobacterium and Lactococcus), Bacteroidetes (0.05%), and Actinobacteria (0.02%). Almost the same microbial composition persisted in the curd after molding. From day 1 of ripening onwards, the phylum Firmicutes dominated. Lactococcus dominated throughout ripening, and most of the Lactobacillus species appeared only at 7 or 15 days. At 90 days, Lactococcus (87.2%), Lactobacillus (4.8%; mainly Lactobacillus plantarum and Lactobacillus sakei), and Leuconostoc (3.9%) dominated. The relative utilization of carbon sources by the bacterial community reflected the succession. This study identified strategic phases that characterized the manufacture and ripening of Canestrato Pugliese cheese and established a causal relationship between mesophilic lactobacilli and proteolysis., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Background: Spontaneous fermentation of plant-derived material is mainly performed on a small scale, with the exception of fermented olives, cucumbers, sauerkraut and kimchi, which have met worldwide commercial significance., Results: This study of spontaneous fermentation of green tomatoes at different stages of ripening revealed a significant effect on the growth kinetics of lactic acid bacteria and the final pH value. Leuconostoc mesenteroides dominated spontaneous fermentation when the initial pH value ranged from 3.8 to 4.8 whereas at higher pH values (4.9-5.4) it co-dominated with Leu. citreum and Lactobacillus casei. Application of RAPD-PCR and rep-PCR allowed differentiation at sub-species level, suggesting a microbial succession at that level accompanying the respective at species level., Conclusion: Ripening stage affected the development of the micro-ecosystem through the growth of lactic acid bacteria and concomitant pH value reduction; however, the outcome of the fermentation was only marginally different., (© 2013 Society of Chemical Industry.)

Sweet bell peppers are a significant constituent of retail, chilled-stored and packaged food products like fresh salads, marinades and ready-to-eat (RTE) meals. Previously, through general screening of the Belgian market and by means of source tracking analysis in a plant manufacturing minimally processed, vegetable salads the susceptibility of fresh-cut sweet bell peppers to lactic acid bacterium (LAB) contamination was substantiated. The determination of the metabolic profiles of Leuconostoc gelidum subsp. gasicomitatum and Lactococcus piscium, two major psychrotrophic, spoilage-related LAB species, on sweet bell pepper (SBP) simulation medium under different packaging conditions - 1.) vacuum: 100% N2, 2.) air: 21% O2, 79% N2, 3.) MAP1: 30% CO2, 70% N2 and 4.) MAP2: 50% O2, 50% CO2 - facilitated a better understanding of the spoilage potential of these microbes as well as the presumptive contribution of O2 in the spectrum of produced volatile organic compounds (VOCs) associated with poor organoleptic properties of food products. Generally, none of the applied gas compositions inhibited the growth of the 4 L. gelidum subsp. gasicomitatum isolates, however the presence of O2 resulted in buttery off-odors by inducing primarily the accumulation of diacetyl and pungent "vinegar" smell due to acetic acid. The 3 tested isolates of L. piscium varied greatly among their growth dynamics and inhibition at MAP2. They exhibited either weak spoilage profile or very offensive metabolism confirming significant intraspecies diversity., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Previously, a considerable underestimation (+0.5-3.2 log CFU/g) on the contamination levels of psychrotrophic lactic acid bacteria (LAB) was observed for 33 retail, packaged food products stored at chilling temperature when the mesophilic enumeration technique was implemented as reference shelf-life parameter. In the present study, the microbial diversity of the dominant psychrotrophic LAB recovered after incubation of plates at 22 °C for 5 days was determined using a polyphasic taxonomic approach. A total of 212 LAB isolates were identified using a combination of rep-PCR fingerprinting, amplified fragment length polymorphism (AFLP) analysis and pheS gene sequencing. Leuconostoc gasicomitatum, Leuconostoc gelidum, Leuconostoc spp., Lactococcus piscium and Lactobacillus algidus proved to be the most competent and predominant species that may go undetected by the widely applied mesophilic enumeration protocols (ISO 4833:2003 and ISO 15214:1998). This study has assessed the interspecific variation among potential spoilage LAB, and highlights the significance of implementing a reference shelf-life parameter based on the enumeration of the total psychrotrophic bacterial load for industrial microbiological routine analyses., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Kimchi, a traditional Korean food made by the fermentation of vegetables, has become popular globally because of its organoleptic, beneficial, and nutritional properties. Spontaneous kimchi fermentation in unsterilized raw materials leads to the growth of various lactic acid bacteria (LAB), which results in variations in the taste and sensory qualities of kimchi products and difficulties in the standardized industrial production of kimchi. Raw materials, kimchi varieties, ingredients, and fermentation conditions have significant effects on the microbial communities and fermentative characteristics of kimchi during fermentation. Heterofermentative LAB belonging to the genera Leuconostoc, Lactobacillus, and Weissella are likely to be key players in kimchi fermentation and have been subjected to genomic and functional studies to gain a better understanding of the fermentation process and beneficial effects of kimchi. The use of starter cultures has been considered for the industrial production of high quality, standardized kimchi. Here, we review the composition and biochemistry of kimchi microflora communities, functional and genomic studies of kimchi LAB, and perspectives for industrial kimchi production.

The purpose of this study was to identify the major bacteria in Chinese traditional sourdough (CTS). Five CTS samples (Hn-87, Sx-91, Gs-107, Hf-112, and Hr-122) were collected from different Chinese steamed breads shops or private households. The total bacterial DNA was extracted from sourdough samples and sequenced using Illumina Hiseq 2000 system. Illumina tags were assigned to BLASTN server based on 16S rRNA libraries to reveal a genetic profile. Phylogenetic analysis revealed that the bacteria in traditional sourdough samples were dominated by the genera Leuconostoc and Lactobacillus. Beta diversity analysis, principal component analysis, and cluster analysis compared the bacterial differences in traditional sourdough samples. The results showed that Leuconostoc, Lactobacillus, and Weissella were the predominant genera among the 5 samples. This differentiated the sourdoughs into 3 typologies, namely, 1) Gs-107 and Sx-91, 2) Hr-122 and Hn-87, and 3) Hf-112. This study identified 3 unique major bacteria genus in CTS bread ecosystems., (© 2013 Institute of Food Technologists®)

The individual and combined influences of dimethyl dicarbonate (DMDC) and nisin (200 IU/mL) at mild heat on the inactivation of indigenous microorganisms in litchi juice, including bacteria, molds and yeasts (M&Y), were investigated. The fresh litchi juice with or without nisin were exposed to 250 mg/L DMDC at 30, 40, or 45 °C for 0.5, 1, 2, 3, 4, or 6 h. A complete inactivation of M&Y in the litchi juice with or without nisin was achieved as exposed to 250 mg/L DMDC at 30, 40, or 45 °C for 0.5 h. The bacteria, especially Bacillus sp. and Leuconstoc mesenteroides showed higher resistance than M&Y in the litchi juice. Bacillus sp. and Leuconstoc mesenteroides in the litchi juice was not completely inactivated by 250 mg/L DMDC at 30, 40, or 45 °C. However, nisin addition can enhanced the inactivation of these bacteria by DMDC, and nisin and DMDC also showed a synergistic effect on the inactivation of bacteria. M&Y and bacteria were not detected in the litchi juice added with 200 IU/mL nisin as exposed to 250 mg/L DMDC at 45 °C for 3 h. In addition, microbial shelf life of the litchi juice during storage at 4 °C also was evaluated as treated by 250 mg/L DMDC or combination with nisin at 45 °C for 3 h., (© 2013 Institute of Food Technologists®)

Leuconostoc are known to produce dextran, which have great commercial importance in chemical, medical and food industry. The present study is an attempt to select the best medium for the isolation of indigenous dextran producing Leuconostoc, measuring their enzyme activities for dextransucrase, production of dextran and identification of dextran producing Leuconostoc CMG706, CMG707, CMG710 and CMG713. Since, dextran producing Leuconostoc produce slimy colonies, twenty-four slime producing bacterial strains were isolated from different food sources, fruits and vegetables. Three different isolation medium were evaluated for the isolation of Leuconostoc only and the best one was found to be one containing sucrose and sodium azide. Further, all slime producing bacterial strains were screened for enzyme activity of dextransucrase, which is responsible for dextran production. Four bacterial strains CMG706, CMG707, CMG710 and CMG713 giving high enzyme activities were selected for dextran production and identified.

Dongchimi, one of the most common types of watery kimchi in Korea, was prepared using radish and its pH values, microbial cell numbers, bacterial communities, and metabolites were monitored periodically to investigate the fermentation process of watery kimchi. The bacterial abundance increased quickly during the early fermentation period and the pH values concurrently decreased rapidly without any initial pH increase. After 15 days of fermentation, the bacterial abundance decreased rapidly with the increase of Saccharomyces abundance and then increased again with a decrease of Saccharomyces abundance after 40 days of fermentation, suggesting that bacteria and Saccharomyces have a direct antagonistic relationship. Finally, after 60 days of fermentation, a decrease in bacterial abundance and the growth of Candida were concurrently observed. Community analysis using pyrosequencing revealed that diverse genera such as Leuconostoc, Lactobacillus, Pseudomonas, Pantoea, and Weissella were present at initial fermentation (day 0), but Leuconostoc became predominant within only three days of fermentation and remained predominant until the end of fermentation (day 100). Metabolite analysis using (1)H NMR showed that the concentrations of free sugars (fructose and glucose) were very low during the early fermentation period, but their concentrations increased rapidly although lactate, mannitol, and acetate were produced. After 30 days of fermentation, quick consumption of free sugars and production of glycerol and ethanol were observed concurrently with the growth of Saccharomyces, levels of which might be considered for use as a potential indicator of dongchimi quality and fermentation time., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: An adaptive-network-based fuzzy inference system (ANFIS) was compared with an artificial neural network (ANN) in terms of accuracy in predicting the combined effects of temperature (10.5 to 24.5°C), pH level (5.5 to 7.5), sodium chloride level (0.25% to 6.25%) and sodium nitrite level (0 to 200 ppm) on the growth rate of Leuconostoc mesenteroides under aerobic and anaerobic conditions., Methods: THE ANFIS AND ANN MODELS WERE COMPARED IN TERMS OF SIX STATISTICAL INDICES CALCULATED BY COMPARING THEIR PREDICTION RESULTS WITH ACTUAL DATA: mean absolute percentage error (MAPE), root mean square error (RMSE), standard error of prediction percentage (SEP), bias factor (Bf), accuracy factor (Af), and absolute fraction of variance (R (2)). Graphical plots were also used for model comparison., Conclusions: The learning-based systems obtained encouraging prediction results. Sensitivity analyses of the four environmental factors showed that temperature and, to a lesser extent, NaCl had the most influence on accuracy in predicting the growth rate of Leuconostoc mesenteroides under aerobic and anaerobic conditions. The observed effectiveness of ANFIS for modeling microbial kinetic parameters confirms its potential use as a supplemental tool in predictive mycology. Comparisons between growth rates predicted by ANFIS and actual experimental data also confirmed the high accuracy of the Gaussian membership function in ANFIS. Comparisons of the six statistical indices under both aerobic and anaerobic conditions also showed that the ANFIS model was better than all ANN models in predicting the four kinetic parameters. Therefore, the ANFIS model is a valuable tool for quickly predicting the growth rate of Leuconostoc mesenteroides under aerobic and anaerobic conditions.