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The effect of lactocin AL705, bacteriocin produced by Latilactobacillus (Lat.) curvatus CRL1579 against Listeria biofilms on stainless steel (SS) and polytetrafluoroethylene (PTFE) coupons at 10 °C was investigated. L. monocytogenes FBUNT showed the greatest adhesion on both surfaces associated to the hydrophobicity of cell surface. Partially purified bacteriocin (800 UA/mL) effectively inhibited L. monocytogenes preformed biofilm through displacement strategy, reducing the pathogen by 5.54 ± 0.26 and 4.74 ± 0.05 log cycles at 3 and 6 days, respectively. The bacteriocin-producer decreased the pathogen biofilm by ∼2.84 log cycles. Control and Bac- treated samples reached cell counts of 7.05 ± 0.18 and 6.79 ± 0.06 log CFU/cm2 after 6 days of incubation. Confocal scanning laser microscopy (CLSM) allowed visualizing the inhibitory effect of lactocin AL705 on L. monocytogenes preformed biofilms under static and hydrodynamic flow conditions. A greater effect of the bacteriocin was found at 3 days independently of the surface matrix and pathogen growth conditions at 10 °C. As a more realistic approach, biofilm displacement strategy under continuous flow conditions showed a significant loss of biomass, mean thickness and substratum coverage of pathogen biofilm. These findings highlight the anti-biofilm capacity of lactocin AL705 and their potential application in food industries.

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Lactic acid bacteria (LAB) were isolated, identified, and characterized from pig feces at various growth stages and feed rations in order to be used as probiotic feed additives. Lactic acid bacteria numbers ranged from 7.10 ± 1.50 to 9.40 log CFUs/g for growing and lactating pigs, respectively. Isolates (n = 230) were identified by (GTG)5-polymerase chain reaction and partial sequence analysis of 16S rRNA. Major LAB populations were Limosilactobacillus reuteri (49.2%), Pediococcus pentosaceus (20%), Lactobacillus amylovorus (11.4%), and L. johnsonii (8.7%). In-vitro assays were performed, including surface characterization and tolerance to acid and bile salts. Several lactobacilli exhibited hydrophobic and aggregative characteristics and were able to withstand gastrointestinal tract conditions. In addition, lactobacilli showed starch- and phytate-degrading ability, as well as antagonistic activity against Gram-negative pathogens and the production of bacteriocin-like inhibitory substances. When resistance or susceptibility to antibiotics was evaluated, high phenotypic resistance to ampicillin, gentamicin, kanamycin, streptomycin, and tetracycline and susceptibility towards clindamycin and chloramphenicol was observed in the assayed LAB. Genotypic characterization showed that 5 out of 15 resistance genes were identified in lactobacilli; their presence did not correlate with phenotypic traits. Genes erm(B), strA, strB, and aadE conferring resistance to erythromycin and streptomycin were reported among all lactobacilli, whereas tet(M) gene was harbored by L. reuteri and L. amylovorus strains. Based on these results, 6 probiotic LAB strains (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T, and L. johnsonii S92R) can be selected to explore their potential as direct feed additives to promote swine health and replace antibiotics.

Des bactéries lactiques (LAB) ont été isolées, identifiées et caractérisées à partir de matières fécales de porc à différents stades de croissance et de rations alimentaires afin d'être utilisées comme additifs alimentaires probiotiques. Le nombre de bactéries lactiques variait de 7,10 ± 1,50 à 9,40 log UFC/g pour les porcs en croissance et en lactation, respectivement. Les isolats (n = 230) ont été identifiés par réaction d'amplification en chaîne par la (GTG)5-polymérase et analyse partielle de la séquence de l'ARNr 16S. Les principales populations de LAB étaient Limosilactobacillus reuteri (49,2 %), Pediococcus pentosaceus (20 %), Lactobacillus amylovorus (11,4 %) et L. johnsonii (8,7 %). Des tests in vitro ont été effectués, y compris la caractérisation de surface et la tolérance aux acides et aux sels biliaires. Plusieurs lactobacilles présentaient des caractéristiques hydrophobes et agrégatives et étaient capables de résister aux conditions du tractus gastro-intestinal. De plus, les lactobacilles ont montré une capacité de dégradation de l'amidon et des phytates, ainsi qu'une activité antagoniste contre les agents pathogènes à Gram négatif et la production de substances inhibitrices de type bactériocine. Lorsque la résistance ou la sensibilité aux antibiotiques a été évaluée, une résistance phénotypique élevée à l'ampicilline, à la gentamicine, à la kanamycine, à la streptomycine et à la tétracycline et une sensibilité à la clindamycine et au chloramphénicol ont été observées dans les LAB testés. La caractérisation génotypique a montré que cinq gènes de résistance sur 15 ont été identifiés dans les lactobacilles; leur présence n'était pas corrélée aux traits phénotypiques. Les gènes erm(B), strA, strB et aadE conférant une résistance à l'érythromycine et à la streptomycine ont été signalés parmi tous les lactobacilles, tandis que le gène tet(M) était hébergé par les souches L. reuteri et L. amylovorus. Sur la base de ces résultats, six souches probiotiques LAB (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T et L. johnsonii S92R) peuvent être sélectionnées pour explorer leur potentiel en tant qu'additifs alimentaires directs pour promouvoir la santé des porcs et remplacer les antibiotiques.(Traduit par Docteur Serge Messier).

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Human infection by Enterohemorrhagic Escherichia coli (EHEC) constitutes a serious threat to public health and a major concern for the meat industry. Presently, consumers require safer/healthier foods with minimal chemical additives, highlighting the need for sustainable solutions to limit and prevent risks. This work evaluated the ability of two antagonistic lactic acid bacteria (LAB) strains, Lactiplantibacillus plantarum CRL681 and Enterococcus mundtii CRL35, and their combination in order to inhibit EHEC in beef (ground and vacuum sealed meat discs) at 8 °C during 72 h. The effect of lower lactic acid (LA) concentrations was evaluated. Meat color was studied along with how LAB strains interfere with the adhesion of Escherichia coli to meat. The results indicated a bacteriostatic effect on EHEC cells when mixed LAB strains were inoculated. However, a bactericidal action due to a synergism between 0.6% LA and LAB occurred, producing undetectable pathogenic cells at 72 h. Color parameters (a*, b* and L*) did not vary in bioprotected meat discs, but they were significantly modified in ground meat after 24 h. In addition, LAB strains hindered EHEC adhesion to meat. The use of both LAB strains plus 0.6% LA, represents a novel, effective and ecofriendly strategy to inactivate EHEC in meat.

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Lactic acid bacteria (LAB) selected on the basis of probiotic characteristics were administered to beef feedlot catlle and the effect on body condition/growth and nutritional-metabolic status as well as on E. coli O157:H7 fecal shedding, were investigated. A feeding trials involving 126 steers were used to evaluate the effects of Lactobacillus acidophilus CRL2074, Limosilactobacillus fermentum CRL2085 and Limosilactobacillus mucosae CRL2069 and their combinations (5 different probiotic groups and control) when 107-108 CFU/animal of each probiotic group were in-feed supplemented. Cattle were fed a high energy corn-based diet (16 to 88%) and samples from each animal were taken at 0, 40, 104 and 163 days. In general, animals body condition and sensorium state showed optimal muscle-skeletal development and behavioral adaption to confinement; no nasal/eye discharges and diarrheic feces were observed. The nutritional performance of the steers revealed a steady increase of biometric parameters and weight. Animals supplied with L. mucosae CRL2069 for 104 days reached the maximum mean live weight (343.2 kg), whereas the greatest weight daily gain (1.27 ± 0.16 Kg/day) was obtained when CRL2069 and its combination with L. fermentum CRL2085 (1.26 ± 0.11 kg/day) were administered during the complete fattening cycle. With several exceptions, bovine cattle blood and serum parameters showed values within referential ranges. As a preharvest strategy to reduce Escherichia coli O157:H7 in cattle feces, CRL2085 administered during 40 days decreased pathogen shedding with a reduction of 43% during the feeding period. L. fermentum CRL2085 and L. mucosae CRL2069 show promise for feedlot cattle feeding supplementation to improve metabolic-nutritional status, overall productive performance and to reduce E. coli O157:H7 shedding, thus decreasing contamination chances of meat food products.

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Despite the recent approval of stingless bee honey to the Argentine Food Code, there are still many gaps in information. Likely, the main reason for this is that multiple ecological and chemical factors influence their production and antimicrobial properties. This work combined metabolomic, microbiological, and physicochemical analyses to characterize the honey ofTetragonisca fiebrigifrom Northeastern Argentina. The antimicrobial activity tests showed that honey samples (n = 24) inhibited some Gram-positive and Gram-negative bacteria at different sensitivity levels. Furthermore, samples selected for their high bioactivity revealed crystallizations, a positive correlation with fungal growth, and the presence of flavonoids. The major polyphenols annotated by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis and supported by metabolomic tools were quercetin 3,4'-dimethyl ether, pachypodol, jaceoside, irigenin trimethyl ether, corymboside, chrysoeriol 7-neohesperidoside, and corymboside. In contrast, samples missing antimicrobial activity did not crystallize, lacked flavonoids, and were enriched in phenylethylamides. Based on these findings, we discuss the significance of flavonoids and phenylethylamides on honey's antimicrobial activity and food quality and how they may indeed reflect essential parameters of the hive, such as microbial balance and eubiosis.

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Modulation of animal gut microbiota is a prominent function of probiotics to improve the health and performance of livestock. In this study, a large-scale survey to evaluate the effect of lactic acid bacteria probiotics on shaping the fecal bacterial community structure of feedlot cattle during three experimental periods of the fattening cycle (163 days) was performed. A commercial feedlot located in northwestern Argentina was enrolled with cattle fed mixed rations (forage and increasing grain diet) and a convenience-experimental design was conducted. A pen (n = 21 animals) was assigned to each experimental group that received probiotics during three different periods. Groups of n = 7 animals were sampled at 40, 104 and 163 days and these samples were then pooled to one, thus giving a total of 34 samples that were subjected to high-throughput sequencing. The microbial diversity of fecal samples was significantly affected (p < 0.05) by the administration period compared with probiotic group supplementation. Even though, the three experimental periods of probiotic administration induced changes in the relative abundance of the most representative bacterial communities, the fecal microbiome of samples was dominated by the Firmicutes (72-98%) and Actinobacteria (0.8-27%) phyla, while a lower abundance of Bacteroidetes (0.08-4.2%) was present. Probiotics were able to modulate the fecal microbiota with a convergence of Clostridiaceae, Lachnospiraceae, Ruminococcaceae and Bifidobacteriaceae associated with health and growth benefits as core microbiome members. Metabolic functional prediction comparing three experimental administration periods (40, 104 and 163 days) showed an enrichment of metabolic pathways related to complex plant-derived polysaccharide digestion as well as amino acids and derivatives during the first 40 days of probiotic supplementation. Genomic-based knowledge on the benefits of autochthonous probiotics on cattle gastrointestinal tract (GIT) microbiota composition and functions will contribute to their selection as antibiotic alternatives for commercial feedlot.

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The effect of bioprotective extracts (BEs) from Lactobacillus acidophilus CRL641 (BE-1) and Latilactobacillus curvatus CRL705 (BE-2) against the exopolysaccharide producer Latilactobacillus sakei CRL1407 in vacuum-packaged meat discs at 4 °C was evaluated. Lat. sakei CRL1407 was able to grow in control samples from 2.80 to 7.77 log CFU/g after 38 days. BE-1 and BE-2 reduced bacterial growth by 2.11 and 1.35 log CFU/g, respectively, but their combination led to a greater growth reduction (3.31 log CFU/g). The antimicrobial activity was detected in treated samples with BE-1 and BE-1 + BE-2 until day 16, while with BE-2 only at the initial time. The pH values remained constant in the discs treated with the BEs combination, whereas the greatest drop in pH was observed in control samples. The minor lipid oxidation without perceptible color changes was detected in the presence of BE-1 and BE-1 + BE-2. The combination of BEs as biocontrol agent plus conventional preservation barriers could extend the fresh meat shelf-life without quality loss.

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The effect of bioprotective extracts (BEs) from Latilactobacillus curvatus CRL705 and Lactobacillus acidophilus CRL641 against Latilactobacillus sakei CRL1407 was evaluated in a refrigerated meat model system under vacuum and aerobic conditions at 4 and 10 °C. As shown by culturing, the BE-1 from L. acidophilus completely inhibited the spoilage strain, while that from Lat. Curvatus CRL705 (BE-2) and its combination with BE-1 exerted a bacteriostatic effect. The antimicrobial activity and exopolysaccharide production correlated with the efficacy of inhibitory treatment while final pH decrease was higher in control samples. When flow cytometry was applied, a lack of correlation with plate counting was found; counts under the detection limit for BE-1 at 21 and 28 days at 4 and 10 °C represented between 64.15 and 73.70% of dead cells. Thus, the concurrence of lactic acid bacteria as biocontrol agents and the use of more accurate tools to prevent the growth of deteriorating species will contribute to the extension of fresh meat shelf-life without quality loss.

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Listeria monocytogenes is one of the major food-related pathogens and is able to survive and multiply under different stress conditions. Its persistence in industrial premises and foods is partially due to its ability to form biofilm. Thus, as a natural strategy to overcome L. monocytogenes biofilm formation, the treatment with lactocin AL705 using a sublethal dose (20AU/ml) was explored. The effect of the presence of the bacteriocin on the biofilm formation at 10°C of L. monocytogenes FBUNT was evaluated for its proteome and compared to the proteomes of planktonic and sessile cells grown at 10°C in the absence of lactocin. Compared to planktonic cells, adaptation of sessile cells during cold stress involved protein abundance shifts associated with ribosomes function and biogenesis, cell membrane functionality, carbohydrate and amino acid metabolism, and transport. When sessile cells were treated with lactocin AL705, proteins' up-regulation were mostly related to carbohydrate metabolism and nutrient transport in an attempt to compensate for impaired energy generation caused by bacteriocin interacting with the cytoplasmic membrane. Notably, transport systems such as ß-glucosidase IIABC (lmo0027), cellobiose (lmo2763), and trehalose (lmo1255) specific PTS proteins were highly overexpressed. In addition, mannose (lmo0098), a specific PTS protein indicating the adaptive response of sessile cells to the bacteriocin, was downregulated as this PTS system acts as a class IIa bacteriocin receptor. A sublethal dose of lactocin AL705 was able to reduce the biofilm formation in L. monocytogenes FBUNT and this bacteriocin induced adaptation mechanisms in treated sessile cells. These results constitute valuable data related to specific proteins targeting the control of L. monocytogenes biofilm upon bacteriocin treatment.