Selected Lactic Acid Bacteria Strains Research Articles

Application of essential oils as natural antimicrobials in lactic acid bacteria contaminating fermentation for the production of organic cachaça

Sugarcane-based fermentation is an essential process for different sectors of economic importance, such as the food industry with fermented and distilled beverages. However, this process can suffer from high contamination by wild yeasts and bacteria, especially lactic acid bacteria (LAB). This makes it necessary to use decontamination strategies and search for new methods that have a low environmental impact and contribute to the production of organic products. Among the options, oregano and thyme essential oils stand out for their antibacterial compounds. The aim of this study was to use oregano and thyme essential oils as natural antimicrobials in the alcoholic fermentation of sugar cane juice. Initially, the minimum inhibitory concentration of the essential oils in the fermentation was assessed through turbidity in the sensitivity test, which allowed us to determine which concentrations of essential oils would inhibit the contaminants, 3 morphologically selected LAB strains, as well as assessing the viability of CA-11. For LAB, 3 concentrations of each essential oil were tested, ranging from 0.1 to 0.4 μL/mL, while for CA-11 it was 0.06, to 0.1 μL/mL. The results indicated a maximum total value of essential oils per mL of 0.06 μL. Based on this result, a 24-1 fractional factorial was established, with 8 conditions +3 central points, with 4 variables, oregano essential oil (0, 0.03 and 0.06 μL/mL), thyme (0, 0.03 and 0.06 μL/mL), initial soluble solids (14, 16 and 18°Brix) and initial yeast concentration (2.5, 3 and 3.5 g/L), with the temperature set at 32 °C for a period of 12 h. The results showed that the center point condition with 0.03 μL/mL of oregano EO, 0.03 μL/mL of thyme controlled the proliferation of contaminating bacteria compared to the control condition. In the experimental validation, the treatment with essential oils had a lower final population of LAB (5.95 log) than the final population of the control treatment (6.53 log), and it was also observed that the treatment with EOs had an alcohol production around 3 % higher than the treatment without antimicrobials. The experimental validation phase confirmed the synergistic action of oregano and thyme essential oils in controlling the proliferation of contaminating bacteria. In conclusion, it was possible to determine the synergistic antimicrobial action of essential oils against LAB during alcoholic fermentation based on organic sugar cane.

Characterization and Assessment of Native Lactic Acid Bacteria from Broiler Intestines for Potential Probiotic Properties.

Probiotics are the most promising alternative to antibiotics for improving animal production and controlling pathogenic infections, while strains derived from natural hosts are considered highly desirable due to their good adaptation to the gastrointestinal tract. The aim of this study was to screen Lactobacillus with broad-spectrum antibacterial activity from broilers fed an antibiotic-free diet and evaluate their potential as poultry probiotics. A total of 44 lactic acid bacteria (LAB) strains were isolated from the intestines of healthy broilers, among which 3 strains exhibited outstanding antimicrobial activity and were subsequently identified through 16S rRNA sequencing as Enterococcus faecium L8, Lactiplantibacillus plantarum L10, and Limosilactobacillus reuteri H11. These three isolates demonstrated potent bacteriostatic activity against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, and Salmonella cholerae, with inhibition zones ranging from 15.67 ± 1.53 to 21.33 ± 0.58 mm. The selected LAB strains exhibited high tolerance to acid and bile salts, with L. reuteri H11 displaying the highest survival rate (ranging from 34.68% to 110.28%) after exposure to 0.3% (w/v) bile salts for 6 h or a low pH environment (pH 2, 2.5, and 3) for 3 h. Notably, L. reuteri H11 outperformed other strains in terms of hydrophobicity (84.31%), auto-aggregation (53.12%), and co-aggregation with E. coli ATCC 25922 (36.81%) and S. aureus ATCC 6538 (40.20%). In addition, the three LAB isolates were either fully or moderately susceptible to the tested antibiotics, except for strain L8, which resisted gentamycin and vancomycin. Consequently, these three LAB strains, especially L. reuteri H11, isolated from the intestines of broiler chickens, represent promising probiotic candidates that can be employed as feed additives to enhance production performance and control poultry pathogens.

Isolation of 60 strains from fermented milk of mares and donkeys in Algeria and identification by 16S rRNA sequencing of lactobacilli: Assessment of probiotic skills of important strains and aromatic productivity power

Background and Aim: Donkey and mare milk have high nutritional and functional values, but their lactic acid bacteria (LAB) content remains poorly studied and undervalued in the Algerian dairy industry. This study aimed to isolate and select LAB strains that produce antimicrobial substances during fermentation and to characterize the probiotic profiles of each extracted strain to indicate their potential for antioxidant and proteolytic activity. Materials and Methods: This study focuses on isolating and identifying lactic acid bacterial strains from 10 Equid-fermented milk samples collected in two regions of El Bayed Wilaya (Algeria). Identification of LAB strains was obtained by 16S rRNA sequencing. The probiotic properties of important strains and their aromatic productivity power are assessed. To evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, Chryseobacterium joostei, Pseudomonas aeruginosa, and Escherichia coli, we selected 21 strains. Different induction methods have been used to amplify the antibacterial effects against these pathogenic strains. Results: Among a total of 60 identified strains, 31 had a probiotic profile, and most were catalase-negative. Aromatic productivity power was observed in eight strains: Lactiplantibacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Weissella confusa, Weissella cibaria, Leuconostoc mesenteroides, Leuconostoc lactis, and Lactobacillus sp1. Conclusion: Our results provide insight into the considerable diversity of LAB present in fermented donkey and mare milk. To meet the expectations of the Algerian dairy industry, it is important that the probiotic skills of the nine selected strains are met. In addition, a significant number of these strains may have important probiotic activity and biotechnological potential. Keywords: Algeria, aromatic productivity, lactic acid bacteria, mare and donkey milk, probiotic skills.

Characterization of the broad-spectrum antibacterial activity of bacteriocin-like inhibitory substance-producing probiotics isolated from fermented foods

Antimicrobial peptides, such as bacteriocin, produced by probiotics have become a promising novel class of therapeutic agents for treating infectious diseases. Selected lactic acid bacteria (LAB) isolated from fermented foods with probiotic potential were evaluated for various tests, including exopolysaccharide production, antibiotic susceptibility, acid and bile tolerance, antibacterial activity, and cell adhesion and cytotoxicity to gastric cell lines. Six selected LAB strains maintained their high viability under gastrointestinal conditions, produced high exopolysaccharides, showed no or less cytotoxicity, and adhered successfully to gastric cells. Furthermore, three strains, Weissella confusa CYLB30, Lactiplantibacillus plantarum CYLB47, and Limosilactobacillus fermentum CYLB55, demonstrated a strong antibacterial effect against drug-resistant Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica serovar Choleraesuis, Enterococcus faecium, and Staphylococcus aureus. Whole genome sequencing was performed on these three strains using the Nanopore platform; then, the results showed that all three strains did not harbor genes related to toxins, superantigens, and acquired antimicrobial resistance, in their genome. The bacteriocin gene cluster was found in CYLB47 genome, but not in CYLB30 and CYLB55 genomes. In SDS-PAGE, the extract of CYLB30 and CYLB47 bacteriocin-like inhibitory substance (BLIS) yielded a single band with a size of less than 10 kDa. These BLIS inhibited the growth and biofilm formation of drug-resistant P. aeruginosa and methicillin-resistant S. aureus (MRSA), causing membrane disruption and inhibiting adhesion ability to human skin HaCaT cells. Moreover, CYLB30 and CYLB47 BLIS rescued the larvae after being infected with P. aeruginosa and MRSA infections. In conclusion, CYLB30 and CYLB47 BLIS may be potential alternative treatment for multidrug-resistant bacteria infections.

Lactic Acid Bacteria isolated from traditional and innovative alheiras as potential biocontrol agents

From a selection of seven traditional and 14 innovative alheiras, 491 lactic acid bacteria (LAB) were isolated and tested for their antimicrobial activity against several food-borne pathogens. Among these, six strains revealed antimicrobial activity through potential bacteriocin production against 14 Listeria monocytogenes strains, Enterococcus faecalis ATCC 29212, Clostridium sporogenes ESB050, and Clostridium perfringens ESB054.Through whole genome sequencing (WGS), these strains were identified as Lactiplantibacillus plantarum (2), Leuconostoc mesenteroides (1), and Pediococcus acidilactici (3). Furthermore, several orthologues of class II bacteriocins genes were identified, including Plantaricin E, Plantaricin F, Pediocin PA, Enterocin X, Leucocin A, and Coagulin A. No virulence or antibiotic resistance genes’ orthologues were detected by WGS analysis. However, the selected LAB strains showed variable phenotypic patterns related to virulence genes and antibiotic resistance when assessed through classical methodologies. None of these strains demonstrated the production of biogenic amines, gelatinase or DNase. Additionally, no hemolytic activity or lipase enzyme production was observed. However, only Lpb. plantarum 9A3 was sensitive to all tested antibiotics and was thus chosen for further examination. The bacteriocins produced by Lpb. plantarum (9A3) exhibited stability across a broad range of conditions, including temperatures from 4 to 100 °C, pH values ranging from 2 to 8, exposure to surfactants and detergents (Tween 20 and 80, SDS, EDTA 0.1, 2 and 5 mM, urea and sodium deoxycholate), and enzymes (papain and catalase). Their maximum activity (AU/mL = 12,800) against four L. monocytogenes strains was observed between 21 and 36 h of growth of Lbp. plantarum 9A3, indicating a bacteriostatic mode of action. Therefore, this strain appears to be a robust candidate for potential application as a protective strain to be used in the food industry. Not only is it safe, but it also produces stable bacteriocins (harbouring genes encoding for the production of three) effectively inhibiting significant pathogens such as L. monocytogenes and C. perfringens.

Sustainable extraction of bioactive compound from apple pomace through lactic acid bacteria (LAB) fermentation

Apple pomace (AP), a by-product of the juice industry, is a rich and inexpensive source of natural bioactive substances, including phenolic compounds, that exhibit health–promoting effects. The recovery of these compounds from plant material using only classical extraction techniques and environmentally friendly solvents is often ineffective due to the entrapment of some compounds in the complex structures of plant cell walls. Lactic Acid Bacteria (LAB) fermentation can be a simple technology to increase the content of phenolic compounds, as well as the antioxidant activity of plant material. In this study, pomace from conventionally grown apples (Malus Domestica) of the Ligol cultivar were fermented with selected LAB strains (Lpb. plantarum KKP 3182, Lpb. plantarum KKP 1527, Lpb. plantarum ZFB 200), commercial starter cultures of Lpb. plantarum, and spontaneously. The fermented material was then subjected to ultrasound-assisted extraction, and the resulting extracts were analysed for their composition (phenolic compounds, triterpenoids, simple organic acids), and antioxidant activity. We found that: (1) the total phenolic content of AP extracts fermented with Lpb. plantarum KKP 1527 was about 30% higher than that of non-fermented AP extracts, (2) extracts of AP fermented with Lpb. plantarum KKP 1527 characterized a higher value of the antioxidant activity, (3) an increase in gallic acid procyanidin A2, protocatechuic acid, and procyanidin B2, while a decrease in rutin and quercetin was observed. The results indicated that AP fermented with Lpb. plantarum KKP 1527 may be a powerful and low–cost source of natural antioxidants which have applications in many industries.

Biodiversity and biotechnological properties of lactic acid bacteria isolated from traditional Moroccan sourdoughs.

The present study aimed at characterizing lactic acid bacteria (LAB) strains isolated from traditional sourdoughs collected in different regions of Morocco. Isolated strains were firstly identified using Gram staining and catalase reaction test. Presumptive LAB strains were then checked for various phenotypical properties including growth at 45°C, resistance to NaCl, enzyme production, acidification capacity, diacetyl and exopolysaccharide (EPS) production, and antifungal activity. Finally, selected LAB strains were identified using 16S rDNA sequencing. Results showed that 32.1% of the isolates were thermophilic (45°C) and 83.9% were resistant to NaCl (6.5%). Moreover, 51.7 and 37.5% were able to produce diacetyl and EPS, respectively. Regarding enzyme production, 55.3 and 7.1% of the isolates showed lipolytic and proteolytic activities, respectively. Low pH values (3.37-3.76) were obtained after 24h of incubation of LAB strains in de Man, Rogosa and Sharpe (MRS) broth. Antifungal activity test against Aspergillus flavus, Aspergillus niger and Penicillium spp. showed an inhibition rate up to 50%. Bacterial DNA sequencing showed that LAB isolates belong to seven species, chiefly Levilactobacillus brevis, Lentilactobacillus parabuchneri, Lactiplantibacillus plantarum, Pediococcus pentosaceus, Enterococcus hirae, Bifidobacterium pseudocatenulatum, and Companilactobacillus paralimentarius. These findings, for the first time in Moroccan sourdoughs, indicate that the isolated LAB strains have good multifunctional properties and could be suitable as good starters for sourdough bread production under controlled conditions.

Identification and Characterization of Lactic Acid Bacteria Isolated from Effluents Generated During Cassava Fermentation as Potential Candidates for Probiotics

ABSTRACT Fermented cassava is obtained through fermentation process where the main role is played by various lactic acid bacteria (LAB), while the by-products (effluents) are discarded. The objective of this study was to isolate, identify and analyze potential probiotics in fermented cassava by-products/effluents using molecular techniques. We quantified the LAB’s metabolites and tested their antimicrobial properties. We isolated 13 LAB strains which are all Gram-positive, and catalase, oxidase, indole and coagulase negative. Only 6 out of 13 selected isolates survived the low pH of 1.5 and 0.3% bile salt. These 6 isolates showed different levels of antimicrobial activity against the tested pathogens with zones of inhibition ranging from 5.10 mm to 22.80 mm. The isolated LAB were identified as Lactiplantibacillus plantarum and Limosilactobacillus fermentum. The results indicated that the selected LAB strains have potential as probiotics due to their ability to survive in acidic and bile-rich environments as well as their antimicrobial properties.

Screening and Metabolomic Analysis of Lactic Acid Bacteria-Antagonizing Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a conditional Gram-negative pathogen that produces extracellular virulence factors that can lead to bloodstream invasion, severely harm tissues, and disseminate bacteria, ultimately leading to various diseases. In this study, lactic acid bacteria (LAB) with strong antagonistic ability against P. aeruginosa were screened, and the regulatory mechanism of LAB against P. aeruginosa was evaluated. The results showed that the three selected LAB strains had strong inhibition ability on the growth, biofilm formation, and pyocyanin expression of P. aeruginosa and a promoting effect on the expression of autoinducer-2. Among them, Lactipantibacillus plantarum (Lp. plantarum) LPyang is capable of affecting the metabolic processes of P. aeruginosa by influencing metabolic substances, such as LysoPC, oxidized glutathione, betaine, etc. These results indicate that LPyang reduces the infectivity of P. aeruginosa through inhibition of its growth, biofilm formation, pyocyanin expression, and regulation of its metabolome. This study provides new insights into the antagonistic activity of Lp. plantarum LPyang against P. aeruginosa.

Potential application of lactic acid bacteria for controlling discoloration in tuna (Thunnus orientalis)

Pacific bluefin tuna (Thunnus orientalis) is a highly valued seafood consumed globally. The red color of tuna meat is considered a crucial indicator of its freshness and value, influencing consumer purchasing decisions. Therefore, the present study aimed to investigate the effect of lactic acid bacteria (LAB) isolated from traditional Korean fermented seafood (jeotgal) on preventing the discoloration of tuna meat during storage. Thirty-six strains of LAB (M1–M36) were isolated and examined for superior antioxidant and metmyoglobin conversion activities. Two strains, M1 (Lactococcus lactics) and M8 (Leuconostoc citreum), were selected. Subsequently, the tuna meat was treated with the cell-free supernatant (CFS) of the selected LAB strains and stored at 4 °C. Treatment with CFS maintained a significantly high level of redness (a*) in the tuna meat for up to 48 h; furthermore, this tendency was consistent with the relative quantification of myoglobin derivatives. The oxymyoglobin content in CFS-treated tuna meat remained above 60% for up to 60 h. Conversely, the metmyoglobin content in the untreated group reached over 60% only after 48 h, resulting in complete discoloration. Conclusively, this study demonstrated that LAB-CFS can prevent tuna meat discoloration during refrigerated storage, indicating its potential for maintaining the quality and color of tuna meat.

Changes in Lacto-Fermented Agaricus bisporus (White and Brown Varieties) Mushroom Characteristics, including Biogenic Amine and Volatile Compound Formation

This study aimed to evaluate the changes in Agaricus bisporus (white and brown) characteristics (colour and acidity parameters, lactic acid bacteria (LAB) and mould/yeast counts, biogenic amine content, fatty acid (FA) and volatile compound (VC) profiles, overall acceptability, and emotions induced for consumers) during a 48 h lactic acid fermentation with Lacticaseibacillus casei No. 210, Lactiplantibacillus plantarum No. 135, Lacticaseibacillus paracasei No. 244, and Pediococcus acidilactici No. 29 strains. Fermented white and brown A. bisporus showed higher LAB count and lower pH, lightness, redness, and yellowness than non-fermented ones. Yeast and fungi counts were similar between non-fermented and fermented samples. All samples contained spermidine (on average, 191.5 mg/kg) and some of the fermented samples had tyramine (on average, 80.7 mg/kg). Saturated FA was the highest in non-fermented brown A. bisporus. The highest monounsaturated and polyunsaturated FA contents were found in Lp. plantarum No. 135 fermented white and brown A. bisporus, respectively. For the first time, the VC profile of fermented A. bisporus was analysed. 1-Octen-3-ol content significantly decreased while benzyl alcohol, acetoin, and 2,3-butanediol increased in most fermented samples. Fermented A. bisporus received good acceptability scores. The emotional evaluation showed that the LAB strain and the interaction of the LAB strain and A. bisporus variety were significant on the intensity of emotions “happy” and “sad”, while all analysed factors and their interactions were significant on the intensity of “angry” and “disgusted” (p ≤ 0.05). The findings of this study show the potential of the selected LAB strains and contribute to the increasing body of research on fermented mushrooms.

Evaluation of antifungal activity of lactic acid bacteria against fungi in simulated cheese matrix

This study aimed to investigate the antagonist activity of lactic acid bacteria (LAB) isolated from Brazilian artisanal cheese and dairy environments against filamentous fungi commonly found in dairy products. In the in vitro test, a total of fifty-two LAB isolates were screened for their inhibitory effects against Aspergillus niger IOC 207 and Penicillium chrysogenum IOC 132. Among them, five strains (Weissella confusa W5, W8; Weissella paramesenteroides W9; Weissella cibaria W25; Lactiplantibacillus plantarum Q4C3) were selected for further in situ evaluation using a cheese matrix. The selected LAB strains exhibited excellent antifungal effect in simulated cheese matrix against both fungi targets; however, when combined with a commercial starter culture, their antifungal activity was reduced. Nevertheless, the results of this study suggest that these LAB strains may have potential as bioprotective cultures in food applications; they have the ability to produce antifungal compounds that could be characterized and potentially used as adjunct cultures in future food processing. Further research in this direction could shed light on their metabolic capabilities and pave the way for their potential use as biopreservatives in the food industry.

Influence of Biotreatment on Hordeum vulgare L. Cereal Wholemeal Contamination and Enzymatic Activities

Crop contamination with mycotoxins is a global problem with a negative impact on human and animal health as well as causing economical losses in food and feed chains. This study was focused on the evaluation of the effect of lactic acid bacteria (LAB) strain (Levilactobacillus brevis-LUHS173, Liquorilactobacillus uvarum-LUHS245, Lactiplantibacillus plantarum-LUHS135, Lacticaseibacillus paracasei-LUHS244 and Lacticaseibacillus casei-LUHS210) fermentation on the changes in the level of deoxynivalenol (DON) and its conjugates in Fusarium spp.-contaminated barley wholemeal (BWP). Samples, with different contamination of DON and its conjugates, were treated separately (for 48 h). In addition to mycotoxin content, enzymatic activities (amylolytic, xylanolytic, and proteolytic) of BWP (before and after fermentation) were evaluated. It was established that the effect of decontamination depends on the LAB strain used, and a significant reduction in DON and the concentration of its conjugates in Lc. casei fermented samples was achieved: the amount of DON decreased on average by 47%, and the amount of D3G, 15-ADON and 3-ADON decreased by 82.4, 46.1, and 55.0%, respectively. Lc. casei also showed viability in the contaminated fermentation medium and an effective production of organic acids was obtained. Additionally, it was found that enzymes are involved to the detoxification mechanism of DON and its conjugates in BWP. These findings indicate that fermentation with selected LAB strains could be applied for contaminated barley treatment in order to significantly reduce Fusarium spp. mycotoxin levels in BWP and improve the sustainability of grain production.

Efficacy of Lactic Acid Bacteria as a Biocontrol Agent against Anthracnose (Persea americana Miller) Decay in Avocado (Persea americana) cv Fuerte Fruit

Anthracnose disease is still a threat to avocado fruit quality, and the use of fungicide (Plochloraz®) for its control has generated safety concerns that necessitate the search for alternatives. Therefore, the efficiency of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against Colletotrichum gloeosporioides was investigated in this study. Weissella cibaria 21 (LAB 21), Leuconostoc pseudomesenteroides 56 (LAB 56), Weissella confusa 17 (LAB 17), Lactiplantibacillus plantarum 75 (LAB 75), and Lactiplantibacillus plantarum 171 (LAB 171) were evaluated in vitro as potential biocontrol agents to replace the Prochloraz® that is currently used in susceptible avocado (Persea americana Miller) Fuerte fruit. To confirm the biocontrol activity of the selected LAB strains, the antagonistic growth, spore germination, LAB recovery, nutrient competition, acid tolerance, and biofilm formation were assessed. In fruit treated with a LAB cell suspension, curatively inoculated with C. gloeosporioides, or naturally infected avocado cv Fuerte fruit, the epicatechin content and expression of defense-related genes (PAL, LOX, AVFADl 2–3, AVFAEL, and FLS) were compared with Prochloraz® and sterile water (control) treatments. With LAB 56, LAB 75, and LAB 21, significant inhibition of radial mycelial growth (MGI) (>90%) and spore germination (100%) was observed similar to those due to Prochloraz®. The MGI increased with a reduction in nutrient concentration. LAB strains reduced anthracnose disease incidence and severity compared with Prochloraz® and were the highest in LAB 21 and LAB 56. The LAB 21 and LAB 56 strains produced strong biofilms against C. gloeosporioides. In contrast to LAB 56, the control, and Prochloraz®, and LAB 21 had the highest epicatechin content (406 mg/g) and upregulated the PAL, AVFADl 2–3, AVFAEl, and FLS genes, thereby reducing the incidence of anthracnose in avocado fruit. As a result, LAB 21 suspensions can be used as an alternative to Prochloraz® in the control of anthracnose disease.

(Meta)genomics -assisted screening of novel antibacterial lactic acid bacteria strains from traditional fermented milk from Western China and their bioprotective effects on cheese

In this study, the bacterial composition of five traditional fermented milk samples from western China was evaluated by 16S metagenomic analysis. Forty lactic acid bacteria (LAB) strains were isolated from each sample and their antilisterial activity was tested. The inter-strain genetic diversity of the LAB isolated was investigated by randomly amplified polymorphic DNA (RAPD) analysis with reference to their antilisterial activity. The antimicrobial potential of selected LAB strains was studied by whole genome sequencing and mining, which resulted in the identification of several novel biosynthetic gene clusters (BGCs). Lactiplantibacillus plantarum TXZ2-35, Limosilactobacillus fermentum TZ-22 and Companilactobacillus crustorum QHS-4, carrying novel BGCs, were used as bio-protective adjunct cultures in Cheddar cheese. These three strains were able to control the proliferation of Listeria monocytogenes in cheese and some strains could improve cheese quality in terms of texture and sensory quality. In summary, we applied metagenomic and genomic approaches to quickly identify novel antilisterial LAB strains from traditional fermented food. Our results demonstrate the effectiveness of incorporating novel bioprotective adjunct cultures in improving the safety and quality of cheese.

Isolation and Characterization of Lactic Acid Bacteria from Fermented Milk Produced in Jimma Town, Southwest Ethiopia, and Evaluation of their Antimicrobial Activity against Selected Pathogenic Bacteria.

Raw milk is usually contaminated with pathogenic bacteria. Fermentation of milk is important to inhibit the growth of contaminants, spoilage, and pathogenic bacteria. The objective of this study was to isolate lactic acid bacteria from fermented milk and evaluate their antimicrobial activity against selected pathogenic bacteria. Laboratory-based experimental study design was conducted from May-July, 2021.Three samples of Ergo (each of 250 ml) were collected from Jimma town. Lactic acid bacteria (LAB) isolates were identified through integrated phenotypic techniques. Further identification was conducted through using API 50 CHL strips. Antimicrobial activities (AMAs) of LAB isolates were tested against clinical isolates of E. coli, S. aureus, and Salmonella spp. using agar well diffusion method. The data were analyzed by using SPSS software version 21 and Microsoft Excel spreadsheet. Tables and figures were applied to describe characteristics of data. Twelve LAB isolates were identified. Those LAB isolates include six Lactococcus lactis subsp. lactis, Lactobacillus acidophilus (2), Lactiplantibacillus plantarum (1), Limosilactobacillus fermentum (2), and Leuconostoc lactis (1). Based on primary screening of LAB, isolates/strains ESCIa, ESBIa, and ESCIc show strong AMA against S. aureus, E. coli, and Salmonella spp. The CFS of ESCIc showed the highest AMA against S. aureus and Salmonella spp. with a zone of inhibition of 14.12 ± 1.6 mm and 12.9 ± 3.6 mm, respectively, while ESBIa showed the highest AMA against E. coli with a zone of inhibition of 13.5 ± 2.1 mm. The CFSs of selected LAB strains were heat tolerant at varying temperatures up to 100°C. The CFSs of selected LAB strains were inactivated by proteinase enzymes, but they are not inactivated with amylase enzymes. Conclusions and Recommendation. All 12 LAB isolates exhibited antimicrobial activity against tested bacterial strains. Lactobacillus isolates showed the highest antagonistic activity on tested indicator strains. Thus, they are possible alternatives to antibiotics in the era of antimicrobial resistance. S. aureus was the most sensitive to antimicrobial effects/agents of selected LAB isolates. Consumption of fermented foods is advisable since they support the growth of healthy GIT microbiota. Fermentation serves as biopreservation of food. However, analysis of probiotic features and in vivo probiotic effects of those LAB isolates will be subject of future research/study.

Effect of stress factors on the production of biogenic amines by lactic acid bacteria isolated from fermented Mexican foods (cheese and beer)

The growth of lactic acid bacteria (LAB) in fermented foods, especially in ripened foods, could be associated with the production of biogenic amines (BAs). BAs are small, nitrogen-containing, thermostable, non-volatile organic compounds, which can induce toxic effects in food consumers. This work aimed to determine potential BAs-producer LAB strains isolated from ripened cheese and beer and to evaluate stress factors that induce the production of these metabolites. Determination of BAs-related genes in eighteen strains of LAB was done by polymerase chain reaction (PCR). The effect of pH, NaCl concentration and ethanol concentration on the LAB growth and BAs production was evaluated. The concentration of BAs (histamine, tyrosine, putrescine and cadaverine) was determined by high performance liquid chromatography (HPLC). Additionally, survival of selected LAB strains during beer making process was evaluated. All LAB strains could tolerate concentrations of 4 and 6 mL/100 mL of ethanol. More than one strain showed the ability to produce all the BAs studied and all beer strains were able to produce Tyramine. The pH level was the most influent factor in BAs production; pH 4 was optimal for the production of histamine (cheese and beer strains) and pH 5 for tyramine (beer strains). LAB population showed a 2-log reduction along the two initial days of fermentation. At the end of the beer making process 1.6 log of LAB were recovered. BAs production has been considered a mechanism that increases bacterial survival against acidic environment. Some LAB strains in fermented products may represent a health risk for consumers. Sanitary practices during food production and raw material quality should be complement measurements to a good control of fermentation process to minimize BAs presence in foods.

Immunostimulatory Activity of Lactic Acid Bacteria Cell-Free Supernatants through the Activation of NF-κB and MAPK Signaling Pathways in RAW 264.7 Cells.

Lactic acid bacteria (LAB) can improve host health and has strong potential for use as a health functional food. Specific strains of LAB have been reported to exert immunostimulatory effects. The primary goal of this study was to evaluate the immunostimulatory activities of novel LAB strains isolated from humans and foods and to investigate the probiotic properties of these strains. Cell-free supernatants (CFS) obtained from selected LAB strains significantly increased phagocytosis and level of nitric oxide (NO) and pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 in RAW264.7 macrophage cells. The protein expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2, which are immunomodulators, was also upregulated by CFS treatment. CFS markedly induced the phosphorylation of nuclear factor-κB (NF-κB) and MAPKs (ERK, JNK, and p38). In addition, the safety of the LAB strains used in this study was demonstrated by hemolysis and antibiotic resistance tests. Their stability was confirmed under simulated gastrointestinal conditions. Taken together, these results indicate that the LAB strains selected in this study could be useful as probiotic candidates with immune-stimulating activity.

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.