Selected Lactic Acid Bacteria Strains Research Articles

The Influence of the Degree of Thermal Inactivation of Probiotic Lactic Acid Bacteria and Their Postbiotics on Aggregation and Adhesion Inhibition of Selected Pathogens.

The study aimed to evaluate the effect of thermal inactivation of potentially probiotic lactic acid bacteria (LAB) strains isolated from food on their ability to compete with pathogenic microorganisms. Five strains of LAB, previously isolated from food and characterized, one commercial reference strain of Lactiplantibacillus plantarum 299v, and two indicator strains of Staphylococcus aureus 25923 and Listeriamonocytogenes 15313 were used in the study. The experiment consisted in applying a stress factor (high temperature: 80 °C, at a different time: 5, 15, and 30 min) to the tested LAB cells to investigate the in vitro properties such as hydrophobicity abilities (against p-xylene and n-hexadecane), auto-aggregation, co-aggregation with pathogens, and inhibition of pathogens adhesion to the porcine gastric mucin. The bacterial strains showed various hydrophobicity to p-xylene (36-73%) and n-hexadecane (11-25%). The affinity for solvents expanded with increasing thermal inactivation time. All LAB isolates were able to auto-aggregate (ranging from 17 to 49%). Bacterial strains subjected to 5 and 15 min of thermal inactivation had the highest auto-aggregation ability in comparison to viable and heat-killed cells for 30 min. The LAB strains co-aggregated with pathogens to different degrees; among them, the highest scores of co-aggregation were observed for L. monocytogenes, reaching 27% (with 15 min of heat-killed LAB cells). All LAB strains reduced the adherence of pathogenic bacteria in the competition test, moreover, heat-killed cells (especially 15 min inactivated) were more efficient than viable cells. The properties of selected LAB strains as moderately heat-stressed forms analyzed in the study increased the prevention of colonization and elimination of pathogenic bacteria in the in vitro model of gastrointestinal tract. The thermal inactivation process may therefore preserve and modifies some characteristics of bacterial cells.

Bioaccessibility of Phenolic Acids and Flavonoids from Buckwheat Biscuits Prepared from Flours Fermented by Lactic Acid Bacteria.

The literature reports that the consumption of common buckwheat (Fagopyrum esculentum Moench), exactly the polyphenols it contains, is associated with a wide spectrum of health benefits. Therefore, the determination of the bioaccessibility of phenolic acids and flavonoids from buckwheat biscuits formulated from liquid-state fermented flours (BBF) by selected lactic acid bacteria (LAB) after gastrointestinal digestion was addressed in this study. Bioaccessibility could be defined as the fraction of a compound that is released from the food matrix in the gastrointestinal lumen and used for intestinal absorption. The bioaccessibility of eight phenolic acids (protocatechuic, vanillic, syringic ferulic, caffeic, sinapic, p-coumaric, and t-cinnamic) and six flavonoids (epicatechin, vitexin, orientin, apigenin, kaempferol, and luteolin) were provided for BBF and BBC (buckwheat biscuits prepared from fermented and unfermented flours, respectively). The bioaccessibility indexes (BI) indicated the high bioaccessibility of phenolic acids and improved bioaccessibility of flavonoids from BBF. Moreover, the data provide evidence for the suitability of selected LAB strains to be used as natural sour agents for further bakery product development rich in phenolic acids and flavonoids with LAB-dependent bioaccessibility.

Antimicrobial activity against Staphylococcus aureus and genome features of Lactiplantibacillus plantarum LR-14 from Sichuan pickles.

The persistence of Staphylococcus aureus within biofilm can lead to contamination of medical devices and life-threatening infections. Luckily, lactic acid bacteria (LAB) have an inhibitory effect on the growth of these bacteria. This study aims to select LAB strains from fermented vegetables, and analyze their potential inhibition activities against S. aureus. In total, 45 isolates of LAB were successfully isolated from Sichuan pickles, and the CFS of Lactiplantibacillus plantarum LR-14 exerted the strongest inhibitory effect against S. aureus. Moreover, S. aureus cells in planktonic and biofilm states both wrinkled and damaged when treated with the CFS of L. plantarum LR-14. In addition, whole genome sequencing analysis indicates that L. plantarum LR-14 contains various functional genes, including predicted extracellular polysaccharides (EPS) biosynthesis genes, and genes participating in the synthesis and metabolism of fatty acid, implying that L. plantarum LR-14 has the potential to be used as a probiotic with multiple functions.

Selection of a Potential Synbiotic against Cronobacter sakazakii

Cronobacter sakazakii is an opportunistic foodborne pathogen that can be fatal to infants; it is commonly associated with powdered infant formula due to contamination during manufacturing processes or during preparation in hospitals or homes. This project aimed to select a potential synbiotic, a combination of probiotic strains with a prebiotic product, to inhibit the growth of C. sakazakii in an in vitro dynamic infant gut model (Simulator of the Human Intestinal Microbial Ecosystem). A total of 16 lactic acid bacteria (LAB) were tested for their inhibitory properties against four different C. sakazakii strains by a zone of inhibition test. Lactobacillus and Pediococcus species were able to inhibit the growth (>15-mm inhibition zones) of all C. sakazakii strains tested, and only one strain from the two genera exhibited atypical resistance to tetracycline. All C. sakazakii strains and the selected LAB strains, which inhibited C. sakazakii and did not exhibit atypical antibiotic resistance, were grown in Luria-Bertani or de Man Rogosa Sharpe broth, respectively, containing 1% dextrose or 1% commercial prebiotic (w/v) to compare their ability to metabolize the prebiotic product. Overall, based on the growth inhibition of C. sakazakii, antibiotic susceptibility, and prebiotic metabolism, 6 of the 16 LAB were chosen to be part of a potential synbiotic. This study has provided valuable information that will help with the development of a synbiotic that can be used in powdered infant formula to reduce the potential for C. sakazakii-related illnesses in infants.

Screening and Molecular Identification of Lactic Acid Bacteria Producing β-Glucan in Boza and Cider

The goal of this study was screening and molecular identification of Lactic Acid Bacteria (LAB) producing β-glucan from different species isolated from boza and cider compared to a standard strain for Lactobacillus rhamnosus NRRL 1937 (LGG). From 48 unknown isolates, four LAB strains were selected. Based on the NCBI database, their nomenclature was A3, B6, and C9 for Limosilactobacillus fermentum SH1, SH2, and SH3 along with D6 for Leuconostoc mesenteroides SH4. Also, their similarity values were 100%, 99.8%, 100%, and 100%, respectively. The potential of Exopolysaccharide (EPS) (as β-glucan) production for selected LAB strains by gtf gene, conventional PCR and gene expression using both LGG as a control and LAB 16S rRNA gene as a house-keeping gene was investigated. In addition, EPS (mg/100 mL), cell mass (mg/100 mL), pH, total carbohydrate%, total protein% and β-glucan% by the HPLC for all selected LAB isolates were studied. All results of genetic and chemical tests proved the superiority of B6 treatment for L. fermentum SH2. The results showed the superiority of B6 treatment in gtf gene expression (14.7230 ± 0.070-fold) followed by C9 and A3 treatments, which were 10.1730 ± 0.231-fold and 8.6139 ± 0.320-fold, respectively. while D6 treatment recorded the lowest value of gene expression (0.8566 ± 0.040-fold) compared to the control LGG (one-fold). The results also demonstrated that B6 treatment was superior to the other treatments in terms of EPS formation, with a value of 481 ± 1.00 mg/100 mL, followed by the C9 treatment at 440 ± 2.00 mg/100 mL, compared to the LGG (control) reaching 199.7 ± 3.51 mg/100 mL. Also, the highest % of quantitative and qualitative β-glucan in EPS was observed in B6 followed by C9, D6 and A3 which were 5.56 ± 0.01%, 4.46 ± 0.01%, 0.25 ± 0.008% and 0.12 ± 0.008%, respectively compared to control (0.31 ± 0.01%). Finally, the presented results indicate the importance of screening the local LAB isolates to obtain a superior strain for β-glucan production which will be introduced in a subsequent study under optimum conditions.

The Cultivation, Growth, and Viability of Lactic Acid Bacteria: A Quality Control Perspective

Lactic acid bacteria (LAB) are essential dairy starter cultures that are significantly employed for the manufacture of fermented dairy products such as yogurt and cheese. LAB predominantly produce lactic acid as a major end product of fermentation, and they synthesize important metabolites that impart the organoleptic characteristics of fermented food products. LAB are fastidious bacteria that thrive in many environments when adequate nutritional requirements are fulfilled. The demand for superior LAB dairy starter cultures for fermentation applications in the food and dairy industry, has resulted in the need to provide viable and active cultures for all bioprocessing operations. The development of a standard protocol for ensuring the viability and enhanced functionality of LAB cultures in the laboratory as well as dairy processing environments is thus very critical. In addressing concerns linked to resuscitating weak, stressed, and injured LAB culture cells, a protocol that vividly outlines salient steps to recover, enhance cell regeneration, and improve metabolic functionality of LAB strains is of the utmost importance. The maintenance of culture purity, functionality, and viability for LAB starter cultures is likewise critical. Therefore, adherence to a unique protocol guideline will result in the promotion of fermentation performance for many LAB strains dedicated to fermentation and biotechnology processes. As a result, the Food Microbiology and Biotechnology Laboratory at North Carolina Agriculture and Technical State University has developed a standard protocol for the activation and quality control of selected LAB strains that has resulted in highly functional and viable LAB culture strains employed for fermentation research. The adaptation and recommendation of a protocol such as this for use in the dairy and food industry will help to ensure LAB viability and functionality for many applications.

The Cultivation, Growth, and Viability of Lactic Acid Bacteria: A Quality Control Perspective.

Lactic acid bacteria (LAB) are essential dairy starter cultures that are significantly employed for the manufacture of fermented dairy products such as yogurt and cheese. LAB predominantly produce lactic acid as a major end product of fermentation, and they synthesize important metabolites that impart the organoleptic characteristics of fermented food products. LAB are fastidious bacteria that thrive in many environments when adequate nutritional requirements are fulfilled. The demand for superior LAB dairy starter cultures for fermentation applications in the food and dairy industry, has resulted in the need to provide viable and active cultures for all bioprocessing operations. The development of a standard protocol for ensuring the viability and enhanced functionality of LAB cultures in the laboratory as well as dairy processing environments is thus very critical. In addressing concerns linked to resuscitating weak, stressed, and injured LAB culture cells, a protocol that vividly outlines salient steps to recover, enhance cell regeneration, and improve metabolic functionality of LAB strains is of the utmost importance. The maintenance of culture purity, functionality, and viability for LAB starter cultures is likewise critical. Therefore, adherence to a unique protocol guideline will result in the promotion of fermentation performance for many LAB strains dedicated to fermentation and biotechnology processes. As a result, the Food Microbiology and Biotechnology Laboratory at North Carolina Agriculture and Technical State University has developed a standard protocol for the activation and quality control of selected LAB strains that has resulted in highly functional and viable LAB culture strains employed for fermentation research. The adaptation and recommendation of a protocol such as this for use in the dairy and food industrywill help to ensure LAB viability and functionality for many applications.

Lactobacillus fermentum as a new inhibitor to control advanced glycation end-product formation during vinegar fermentation

The inhibitory activity of lactic acid bacteria (LAB) toward advanced glycation end-products (AGEs) during vinegar fermentation was studied, and its relationships with the substrate consumption, antioxidant capacity, total phenolic content, total flavonoid compounds, α-glucosidase, and α-amylase activity inhibition were evaluated. The vinegar was made from rice powder flour by liquid-state fermentation (LSF). The selected LAB strains were separately co-cultivated with Saccharomyces cerevisiae and Acetobacter pasteurianus 1.41 in alcoholic and acetic acid fermentation, respectively. Among 3 strains, Lactobacillus fermentum showed the strongest inhibitory effect on the formation of total fluorescent AGEs and carboxymethyl lysine (CML)/carboxyethyl lysine (CEL) in the fermentation process. The corresponding mechanisms included the acceleration of substrate consumption, improvement of antioxidant activities, and inhibition of α-glucosidase and α-amylase. In addition, the fluorescent AGEs and the CML/CEL were negatively correlated with the antioxidant activities, while the α-glucosidase and α-amylase activities were positively correlated with the total phenols and total flavonoids. Moreover, the variety of main flavor compounds increased, including esters, alcohols, phenols and acids. The results of the study support the potential use of screened LAB strains to inhibit the formation of fluorescent AGEs, CML and CEL on fermented products and in the food processing industry, without associated risks to consumers.

Isolation and identification of lactic acid bacteria in fruit processing residues from the Brazilian Cerrado and its probiotic potential

Fruit residues may contain lactic acid bacteria (LAB) with probiotic properties. This study aimed to isolate, characterize and identify LAB isolated from Brazilian Cerrado fruit residues, as well as to evaluate the safety and functionality properties in vitro. Twenty colonies were isolated and characterized. Biochemical and morphological analyses showed 14 LAB isolates, which were identified by sequencing the 16S rRNA gene in Lactiplantibacillus plantarum/pentosus, Lacticaseibacillus casei/paracasei, Pediococcus adicilactici and Weissella cibaria/confusa. Most of the isolates showed susceptibility to antibiotics and none showed hemolytic, gelatinase, coagulase, and DNAse activity. At pH 2, the survival of most strains examined was reduced after 3h exposure -. Four showed a survival rate similar to control, while L. casei/paracasei from guapeva fruit showed a higher rate. 11 isolates inhibited the growth of Esherichia coli, Staphylococcus aureus and Salmonella sp. Cluster analysis grouped the isolates according to similarity. The results highlight the specificity of the food matrix, genus, species and strain, however selected LAB strains can be submitted for further investigation of new probiotic strains.

Strategies for Biocontrol of Listeria monocytogenes Using Lactic Acid Bacteria and Their Metabolites in Ready-to-Eat Meat- and Dairy-Ripened Products.

Listeria monocytogenes is one of the most important foodborne pathogens. This microorganism is a serious concern in the ready-to-eat (RTE) meat and dairy-ripened products industries. The use of lactic acid bacteria (LAB)-producing anti-L. monocytogenes peptides (bacteriocins) and/or lactic acid and/or other antimicrobial system could be a promising tool to control this pathogen in RTE meat and dairy products. This review provides an up to date about the strategies of use of LAB and their metabolites in RTE meat products and dairy foods by selecting the most appropriate strains, by analysing the mechanism by which they inhibit L. monocytogenes and methods of effective application of LAB, and their metabolites in these kinds of products to control this pathogen throughout the processing and storage. The selection of LAB with anti-L. monocytogenes activity allows to dispose of effective strains in meat and dairy-ripened products, achieving reductions form 2–5 logarithmic cycles of this pathogen throughout the ripening process. The combination of selected LAB strains with antimicrobial compounds, such as acid/sodium lactate and other strategies, as the active packaging could be the next future innovation for eliminating risk of L. monocytogenes in meat and dairy-ripened products.

Fermentation and Storage Characteristics of "Fuji" Apple Juice Using Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus plantarum: Microbial Growth, Metabolism of Bioactives and in vitro Bioactivities.

Fruit juices have been widely used as the substrates for probiotic delivery in non-dairy products. In this study, three lactic acid bacteria (LAB) strains, including Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus plantarum, were selected to ferment apple juice. During 72-h of fermentation, these LAB strains grew well in the apple juice with significant increases in viable cell counts (from 7.5 log CFU/mL to 8.3 log CFU/mL) and lactic acid content (from 0 to 4.2 g/L), and a reduction of pH value (from 5.5 to around 3.8). In addition, the antioxidant and antibacterial capacities of fermented apple juice in vitro were significantly improved through the phenolic and organic acid metabolisms. After storage at 4°C for 30 days, the total amino acid content of fermented apple juice was significantly increased, although the viable cell counts and total phenolic content were decreased (p < 0.05). Furthermore, the stored fermented apple juices still possessed antibacterial and in vitro antioxidant activities. Overall, all the selected LAB strains could be suitable for apple juice fermentation and can effectively improve their biological activities.

Screening of Lactic Acid Bacteria with Anti- Helicobacter pylori Activity and Evaluation of Their Probiotic Characteristics

Objective: To screen lactic acid bacteria (LAB) with anti-Helicobacter pylori activity and great probiotic properties from fermented food, dairy products and infant feces. Method: A total of 39 LAB-like strains with stable curd state were screened from 175 samples. 9 Strains with anti-H. pylori activity were initially isolated by agar diffusion assay and H. pylori urease activity test and identified by 16S rDNA sequence analysis. Subsequently, the acid and bile tolerance, antibiotic resistance, antibacterial activity, cell surface hydrophobicity, and cell auto-aggregation of the selected LAB strains were determined. Results: Two probiotic strains with anti-Helicobacter pylori activity were screened: Lactobacillus helveticus GQ1702 and Lactobacillus plantarum D1, and their cell culture supernatants exhibited anti-H. pylori activity equivalent to the anti-H. pylori activity of 0.038 and 0.034 mg/mL metronidazole, respectively. The survival rates of GQ1702 and D1 were above 70% after incubation at pH3 for 3 h, the survival rates of GQ1702 and D1 after incubation at 0.3% bile salt were 0.47% and 1.08%, respectively. Cell surface hydrophobicities of GQ1702 and D1 were 25.10% and 11.79%, respectively. The self-agglutination rate reached 71.18% and 50.78% respectively after 24 hours. The auto-aggregation percentages of GQ1702 and D1 were 61.18% and 57.25%, respectively, after 24 hours. The two LAB strains were not resistant to chloramphenicol, penicillin, erythromycin, ampicillin, or cefazolin, and exhibited antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, E. coli O157:H7, and Salmonella typhimurium. Conclusion: Lactobacillus helveticus GQ1702 and Lactobacillus plantarum D1 with anti-Helicobacter pylori activity were obtained in this experiment, which could provide a source of strains for development and research of functional dairy products.

In-Vitro Characterization of Growth Inhibition against the Gut Pathogen of Potentially Probiotic Lactic Acid Bacteria Strains Isolated from Fermented Products.

Lactic acid bacteria (LAB) are probiotic candidates that may restore the balance of microbiota populations in intestinal microbial ecosystems by controlling pathogens and thereby promoting host health. The goal of this study was to isolate potential probiotic LAB strains and characterize their antimicrobial abilities against pathogens in intestinal microbiota. Among 54 LAB strains isolated from fermented products, five LAB strains (NSMJ15, NSMJ16, NSMJ23, NSMJ42, and NFFJ04) were selected as potential probiotic candidates based on in vitro assays of acid and bile salt tolerance, cell surface hydrophobicity, adhesion to the intestinal epithelium, and antagonistic activity. Phylogenetic analysis based on 16S rRNA genes showed that they have high similarities of 99.58–100% to Lacticaseibacillus paracasei strains NSMJ15 and NFFJ04, Lentilactobacillus parabuchneri NSMJ16, Levilactobacillus brevis NSMJ23, and Schleiferilactobacillus harbinensis NSMJ42. To characterize their antimicrobial abilities against pathogens in intestinal microbiota, the impact of cell-free supernatant (CFS) treatment in 10% (v/v) fecal suspensions prepared using pooled cattle feces was investigated using in vitro batch cultures. Bacterial community analysis using rRNA amplicon sequencing for control and CFS-treated fecal samples at 8 and 16 h incubation showed the compositional change after CFS treatment for all five LAB strains. The changed compositions were similar among them, but there were few variable increases or decreases in some bacterial groups. Interestingly, as major genera that could exhibit pathogenicity and antibiotic resistance, the members of Bacillus, Escherichia, Leclercia, Morganella, and Vagococcus were decreased at 16 h in all CFS-treated samples. Species-level classification suggested that the five LAB strains are antagonistic to gut pathogens. This study showed the probiotic potential of the five selected LAB strains; in particular, their antimicrobial properties against pathogens present in the intestinal microbiota. These strains would therefore seem to play an important role in modulating the intestinal microbiome of the host.

Selective Thermotolerant Lactic Acid Bacteria Isolated From Fermented Juice of Epiphytic Lactic Acid Bacteria and Their Effects on Fermentation Quality of Stylo Silages.

The aim of the present study was to isolate and identify lactic acid bacteria (LAB) from fermented juice of tropical crops such as Napier grass, Ruzi grass, Purple guinea grass, Stylo legume, and Leucaena and their application to improve the quality of tropical crop silage. Fifteen strains of LAB were isolated. The LAB strains were Gram-positive and catalase-negative bacteria and could be divided into three groups, i.e., Pediococcus pentosaceus, Lactiplantibacillus (para)plantarum, and Limosilactobacillus fermentum according to the biochemical API 50CH test. Based on the analysis of 16S rRNA sequence, the strains isolated in the group L. (para)plantarum were distinguished. Two isolates (N3 and G4) were identified as Lactiplantibacillus plantarum. Three isolates (St1, St2, and St3) were identified as L. paraplantarum. In addition, the identification of other isolates was confirmed in the group P. pentosaceus (R1, R4, R5, R8, R11, and L1) and the group L. fermentum (N4, G6, G7, and N4). All selected strains were able to grow at 50°C. All LAB strains showed antimicrobial activity against Escherichia coli ATCC 25922, Shigella sonnei ATCC 25931, Pseudomonas aeruginosa ATCC 27853, and Bacillus cereus ATCC 11778. Four selected LAB strains (St1, St3, N4, and R4) were tested for their capacity to successfully ensile Stylo legume (Stylosanthes guianensis CIAT184). Stylo silages treated with LAB were well preserved, the NH3–N and butyric acid contents were lower, and the lactic acid content was higher than those in the control (p < 0.05). The acetic acid content was the highest in R4-treated silage among the treatments (p < 0.05). The crude protein (CP) content of St1-silage was significantly (p < 0.05) higher than the others. The inoculation of thermotolerant LAB selected from fermented juice of epiphytic lactic acid bacteria (FJLB) was found to be highly instrumental to obtain well-preserved silage from the Stylo legume.

In vitro assessment of safety and functional probiotic properties of Lactobacillus mucosae strains isolated from Iranian native ruminants intestine

The search for host isolated probiotic bacteria from animal intestine may discover new probiotic candidates with promising health properties. This study evaluated the safety and functional probiotic potential of the lactic acid bacteria (LAB) isolated from the Iranian native ruminants intestine under in vitro assays. The isolates‌ were selected according to criteria including survivability in low pH, bile salts, pancreatic enzymes, different temperatures, NaCl concentrations, antibacterial activities, presence of adhesion genes and safety characteristics. The selected LAB were then identified to species level using 16S rRNA gene sequencing. The results showed that out of one hundred and eighty-seven LAB isolates, only six strains (NABRII50, NABRII51, NABRII52, NABRII53, NABRII54 and NABRII55) were tolerant to low pH, bile salt, pancreatin enzyme, 45 °C temperature and 2% sodium chloride. The six selected isolates belonged to Lactobacillus mucosae. Two of the adhesion genes (mub and map) were detected in all strains except NABRII53. The virulence factors were observed in NABRII50, NABRII53 and NABRII55. The tetracycline resistance gene (tet (S)) was detected in NABRII55. This study was the first effort to select Lb. mucosae strains with the probiotic potential from the Iranian ruminants intestine. These results revealed that the ruminant intestinal ecosystem could be considered as a valuable origin of probiotic candidates and all the selected LAB strains except NABRII50, NABRII53 and NABRII55 could be considered as promising probiotics. HIGHLIGHTS Lactobacillus mucosae strains isolated from Iranian native ruminants intestine including NABRII51, NABRII52 and NABRII54 showed probiotic potential under in vitro assays. The Lb. mucosae strains including NABRII51, NABRII52 and NABRII54 were able to survive in intestinal physiological conditions, and carried the adhesion genes, such as mub and map. In vitro biosafety assays confirmed that Lb. mucosae strains (NABRII51, NABRII52 and NABRII54) were safe to further studies.

Growth inhibition of Fusarium graminearum and deoxynivalenol detoxification by lactic acid bacteria and their application in sourdough bread

SummaryEight lactic acid bacteria (LAB) strains were screened for their ability to inhibit Fusarium graminearum (F. graminearum) growth and remove deoxynivalenol (DON). Furthermore, the selected LAB strains were applied as starter cultures to evaluate their performance during the breadmaking process. Lactobacillus plantarum (L. plantarum) AR524 exhibited strong inhibitory activity against F. graminearum growth in a pH‐dependent manner and removed up to 40.9% of DON, mainly through cell wall binding. Compared with the control, L. plantarum AR524 inhibited 60.19% of F. graminearum growth and removed 50.53% of DON during breadmaking. The texture properties of the bread were also remarkably improved by L. plantarum AR524, and its shelf life was extended from 3 days to 6 days without the application of any chemical preservatives. Therefore, L. plantarum AR524 shows excellent potential for application as a bioprotective starter culture in bakery products.

Functional probiotics of lactic acid bacteria from Hu sheep milk

BackgroundProbiotics are being considered as valuable microorganisms related to human health. Hu sheep is referred as one of the important sheep breeds in China. Goat milk produced by Hu sheep is characterized with high nutritional value and hypoallergenic in nature. Particularly, this milk contains plenty of milk prebiotic and probiotic bacteria. This study was aimed to scrutinize more bacterial strains from Hu sheep milk with potential probiotic activity.ResultsBased on 16S rRNA sequence analysis, pool of forty bacterial strains were identified and evaluated their antimicrobial activities against Staphylococcus aureus, enterohemorrhagic Escherichia coli (EHEC), Salmonella typhimurium, Listeria monocytogenes enterotoxigenic E. coli (ETEC) and Aeromonas caviae. Four out of these isolated strains demonstrated their efficient bacteriostatic ability and potential healthy properties. We also examined the safety aspects of these bacterial candidates including three Lactococcus lactis strains (named as HSM-1, HSM-10, and HSM-18) and one Leuconostoc lactis strain (HSM-14), and were further evaluated via in vitro tests, including antimicrobial activity, cell surface characteristics (hydrophobicity, co-aggregation, and self-aggregation), heat treatment, antibiotic susceptibility, simulated transport tolerance in the gastrointestinal tract, and acid/bile tolerance. The obtained results revealed that HSM-1, HSM-10, HSM-14, and HSM-18 showed high survival rate at different conditions for example low pH, presence of bovine bile and demonstrated high hydrophobicity. Moreover, HSM-14 had an advantage over other strains in terms of gastrointestinal tract tolerance, antimicrobial activities against pathogens, and these results were significantly better than other bacterial candidates.ConclusionHu sheep milk as a source of exploration of potential lactic acid bacteria (LAB) probiotics open the new horizon of probiotics usage from unconventional milk sources. The selected LAB strains are excellent probiotic candidates which can be used for animal husbandry in the future. Rationale of the study was to utilize Hu sheep milk as a source of potential probiotic LABs. The study has contributed to the establishment of a complete bacterial resource pool by exploring the Hu sheep milk microflora.

Behavior of Listeria monocytogenes in the presence or not of intentionally-added lactic acid bacteria during ripening of artisanal Minas semi-hard cheese

The fate of Listeria monocytogenes during ripening of artisanal Minas semi-hard cheese, as influenced by cheese intrinsic properties and by autochthonous (naturally present) or intentionally-added anti-listerial lactic acid bacteria (LAB) was modeled. Selected LAB strains with anti-listerial capacity were added or not to raw or pasteurized milk to prepare 4 cheese treatments. Counts of LAB and L. monocytogenes, pH, temperature and water activity were determined throughout cheese ripening (22 days, 22±1ᵒC). Different approaches were adopted to model the effect of LAB on L. monocytogenes: an independent approach using the Huang primary model to describe LAB growth and the linear decay model to describe pathogen inactivation; the Huang-Cardinal [pH] model using the effect of pH variation in a dynamic tertiary approach; and the Jameson-effect with Nmax tot model which simultaneously describes L. monocytogenes and LAB fate. L. monocytogenes inactivation occurred in both treatments with added LAB and inactivation was faster in raw milk cheese (−0.0260 h−1) vs. pasteurized milk cheese (−0.0182 h−1), as estimated by the linear decay model. Better goodness-of-fit was achieved for the cheeses without added LAB when the Huang primary model was used. A faster and great pH decline was detected for cheeses with added LAB, and the Huang-Cardinal [pH] model predicted higher pathogen growth rate in cheese produced with raw milk, but greater L. monocytogenes final concentration in pasteurized milk cheese. The Jameson-effect model with Nmax tot predicted that LAB suppressed pathogen growth in all treatments, except in the treatment with pasteurized milk and no LAB addition. The Huang-Cardinal [pH] model was more accurate in modeling L. monocytogenes kinetics as a function of pH changes than was the Jameson-effect model with Nmax tot as a function of LAB inhibitory effect based on the goodness-of-fit measures. The Jameson-effect model may however be a better competition model since it can more easily represent L. monocytogenes growth and death. This study presents crucial kinetic data on L. monocytogenes behavior in the presence of competing microbiota in Minas semi-hard cheese under dynamic conditions.

Protection of human intestinal epithelial cells from oxidative stress caused by mercury using lactic acid bacteria.

Heavy metals are harmful to human health. Therefore, we investigated the biosorption of heavy metals by lactic acid bacteria (LAB). Of all the tested heavy metals, biosorption by LAB was highest for mercury, followed by lead, cadmium, and finally arsenic. The viability of HCT-116 cells was reduced by half in the presence of 7.5 µg/mL mercury but recovered after the addition of selected LAB strains. HCT-116 cells showed increased superoxide dismutase and catalase activities, whereas glutathione peroxidase activities decreased significantly. Addition of Lactobacillus sakei TOKAI 57m recovered all antioxidant enzyme activities. Our results suggest that this strain can be used for cellular detoxification.

Unveiling the potentials of bacteriocin (Pediocin L50) from Pediococcus acidilactici with antagonist spectrum in a Caenorhabditis elegans model

Human-milk-based probiotics play a major role in the early colonization and protection of infants against gastrointestinal infection. We investigated potential probiotics in human milk. Among 41 Lactic acid bacteria (LAB) strains, four strains showed high antimicrobial activity against Escherichia coli 0157:H7, Listeria monocytogenes ATCC 15313, Bacillus cereus ATCC 14576, Staphylococcus aureus ATCC 19095, and Helicobacter pylori. The selected LAB strains were tested in simulated gastrointestinal conditions for their survival. Four LAB strains showed high resistance to pepsin (82%–99%), bile with pancreatine stability (96%–100%), and low pH (80%–94%). They showed moderate cell surface hydrophobicity (22%–46%), auto-aggregation abilities (12%–34%), and 70%–80% co-aggregation abilities against L. monocytogenes ATCC 15313, S. aureus ATCC 19095, B. cereus ATCC 14576, and E. coli 0157:H7. All four selected isolates were resistant to gentamicin, imipenem, novobiocin, tetracycline, clindamycin, meropenem, ampicillin, and penicillin. The results show that Pediococcus acidilatici is likely an efficient probiotic strain to produce < 3 Kda pediocin-based antimicrobial peptides, confirmed by applying amino acid sequences), using liquid chromatography mass spectrometry and HPLC with the corresponding sequences from class 2 bacteriocin, and based on the molecular docking, the mode of action of pediocin was determined on LipoX complex, further the 13C nuclear magnetic resonance structural analysis, which confirmed the antimicrobial peptide as pediocin.