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This study focused on L. paracasei strains isolated from fermented palm sap in southern Thailand that exhibit potential probiotic characteristics, including antibiotic susceptibility, resistance to gastrointestinal stresses, and antimicrobial activity against various pathogens. However, a thorough investigation of the whole genome sequences of L. paracasei isolates is required to ensure their safety and probiotic properties for human applications. This study aimed to sequence the genome of L. paracasei isolated from fermented palm sap, to assess its safety profile, and to conduct a comprehensive comparative genomic analysis with other Lacticaseibacillus species. The genome sizes of the seven L. paracasei strains ranged from 3,070,747 bp to 3,131,129 bp, with a GC content between 46.11% and 46.17% supporting their classification as nomadic lactobacilli. In addition, the minimal presence of cloud genes and a significant number of core genes suggest a high degree of relatedness among the strains. Meanwhile, phylogenetic analysis of core genes revealed that the strains possessed distinct genes and were grouped into two distinct clades. Genomic analysis revealed key genes associated with probiotic functions, such as those involved in gastrointestinal, oxidative stress resistance, vitamin synthesis, and biofilm disruption. This study is consistent with previous studies that used whole-genome sequencing and bioinformatics to assess the safety and potential benefits of probiotics in various food fermentation processes. Our findings provide valuable insights into the potential use of seven L. paracasei strains isolated from fermented palm sap as probiotic and postbiotic candidates in functional foods and pharmaceuticals. [ABSTRACT FROM AUTHOR]

Insect farming is gaining attention as a promising area for exploring probiotic bacteria, which can benefit both insect health and various industries. Silkworm farming is a key industry in Thailand; however, challenges such as disease susceptibility and optimising growth require innovative solutions for sustainable practices. Our study addresses this by assessing lactic acid bacteria (LAB) in native Thai silkworm faeces, which accumulate as natural by-products during the rearing process. We conducted biochemical tests, including those for catalase, haemolytic activity, bile salt tolerance, antimicrobial activity, antibiotic susceptibility, and cell surface hydrophobicity, along with taxonomic classification. Out of 102 isolates, eight potential probiotics were selected, with five showing strong probiotic traits like acid and bile salt tolerance and cell surface hydrophobicity, enhancing gut survivability. These isolates also displayed antagonistic activity against pathogens like Staphylococcus aureus, Salmonella typhimurium, Escherichia coli, and Pseudomonas aeruginosa. Safety assessments confirmed their safety, with no haemolytic activity and sensitivity to antibiotics like chloramphenicol and amoxicillin. These LAB isolates (SP04, SP06, SP44, SP64, and SP67), identified as Enterococcus faecalis strain NBRC 100481, show promise as in vitro probiotics for silkworm rearing, calling for further in vivo evaluation. [ABSTRACT FROM AUTHOR]

Khao-Mak is a traditional dessert from Thailand with a unique flavor profile from low alcohol and lactic acid. Khao-Mak is made from cooked glutinous rice fermented with the starter culture (Look-Pang), Look-Pang including yeast, molds, lactic acid bacteria, and herbs. Khao-Mak is made from white glutinous rice, pigmented rice (Kam-Doi, Leum-Pua), and germinated pigmented rice (Kam-Doi, Leum-Pua). This investigation focused on the physico-chemical, metabolites and flavor profiles of Khao-Mak in the fermentation period to optimize the best time for fermentation and biomarkers. The best time for fermentation was 2 days, because after day 2 of fermentation the physico-chemical properties slightly changed. Leum-Phua is the best rice variety for producing Khao-Mak in this research, because Leum-Phua was found to have the highest total anthocyanin content (TAC), total phenolic content (TPC) and %DPPH. In addition, metabolomics and flavoromics analysis results discovered 37 and 48 compounds, respectively, produced during fermentation processes. The metabolites and flavor compounds found were used to differentiate all samples using principal component analysis (PCA). Khao-Mak had strong alcohol, wine-like, whiskey-like, solvent-like, sweet and fruity flavors. Heat plot of metabolites and flavor compounds showed that lactic acid, acetic acid, 3-methyl-1-butanol, 2-methyl-1-propanol, and 1-propanol were in greater proportions than other compounds and used as biomarkers for fermentation. [ABSTRACT FROM AUTHOR]

Background: Lactic acid bacteria (LAB) are widely used as probiotics in poultry production due to their resilience to low pH and high bile salt concentrations, as well as their beneficial effects on growth performance and antagonistic activity against enteric pathogens. However, the efficacy of probiotics depends on strain selection and their ability to colonize the host's intestine. This study aimed to select, identify, and evaluate LAB strains isolated from chicken feces in Thailand for potential use as probiotics in the chicken industry. Methods: LAB strains were isolated from 58 pooled fresh fecal samples collected from chicken farms in various regions of Thailand, including commercial and backyard farms. Gram-positive rods or cocci with catalase-negative characteristics from colonies showing a clear zone on MRS agar supplemented with 0.5% CaCO3 were identified using MALDI-TOF mass spectrometry. The LAB isolates were evaluated for acid (pH 2.5 and pH 4.5) and bile salt (0.3% and 0.7%) tolerance. Additionally, their cell surface properties, resistance to phenol, antimicrobial activity, hemolytic activity, and presence of antimicrobial resistance genes were determined. Results: A total of 91 LAB isolates belonging to the Pediococcus, Ligilactobacillus, Limosilactobacillus, and Lactobacillus genera were obtained from chicken feces samples. Backyard farm feces exhibited a greater LAB diversity compared to commercial chickens. Five strains, including Ligilactobacillus salivarius BF12 and Pediococcus acidilactici BF9, BF14, BYF20, and BYF26, were selected based on their high tolerance to acid, bile salts, and phenol. L. salivarius BF12 and P. acidilactici BF14 demonstrated strong adhesion ability. The five LAB isolates exhibited significant cell-cell interactions (auto-aggregation) and co-aggregation with Salmonella. All five LAB isolates showed varying degrees of antimicrobial activity against Salmonella strains, with P. acidilactici BYF20 displaying the highest activity. None of the LAB isolates exhibited beta-hemolytic activity. Whole genome analysis showed that L. salivarius BF12 contained ermC, tetL, and tetM, whereas P. acidilactici strains BF9 and BF14 carried ermB, lnuA, and tetM. Conclusion: The selected LAB isolates exhibited basic probiotic characteristics, although some limitations were observed in terms of adhesion ability and the presence of antibiotic resistance genes, requiring further investigation into their genetic location. Future studies will focus on developing a probiotic prototype encapsulation for application in the chicken industry, followed by in vivo evaluations of probiotic efficacy. [ABSTRACT FROM AUTHOR]

Edible coatings are pivotal in enhancing the quality of processed meat products, acting as barriers to environmental and microbial influences by adhering directly to the food surface. Arrowroot flour, a widely produced edible tuber in Thailand, is uncharted in terms of its capability and effectiveness as an edible coating on food materials. This study aims to elucidate the composition and spectral properties of arrowroot tuber flour (ATF) to discern its viability as an edible coating for pork sausages. ATF exhibited a composition predominantly featuring carbohydrates (74.78%), moisture (9.59%), and protein (8.89%), underlining its appropriateness as an edible coating. Rapid visco amylograph revealed ATF's significant pasting capability. This study incorporated kaffir lime leaves essential oil (KEO) into the ATF coating in diverse concentrations (0–3%). Fourier-transform Infrared spectroscopy illuminated characteristic peaks and bands, showing observable shifts with the integration of KEO, yet the majority of peak placements remained essentially unchanged. The microstructure of the coatings maintained its homogeneity at heightened KEO concentrations, reflecting compatibility with ATF. The efficacy of the ATF-KEO coatings was evaluated on pork sausages, using uncoated samples as controls. While color modifications were evident, coated sausages maintained consistent moisture content, water activity, and pH levels throughout the storage duration. The coated samples also manifested enhanced textural attributes and a decline in lipid oxidation, as evidenced by reduced TBARS levels compared to controls. A subsequent microbial examination corroborated the inhibitory capacity of the ATF-KEO coatings on the microbial proliferation in pork sausages, encapsulating Total Viable Count (TVC), psychrotrophic bacteria, and lactic acid bacteria. In conclusion, the findings substantiate the promising application of ATF, especially in synergy with KEO, as a proficient edible coating for meat products. This combination aids in preserving color and texture, impeding microbial advancement, and moderating lipid oxidation, thereby contributing to the overall quality and safety of the products. [ABSTRACT FROM AUTHOR]

Fourteen lactic acid bacteria isolated from fermented foods and healthy animal feces in Thailand were characterized for their potential as probiotics. The 16S rRNA gene sequence analyses identified them as Lactiplantibacillus, Levilactobacillus, Limosilactobacillus, Lacticaseibacillus, Campanilactobacillus, Pediococcus, and Enterococcus genera. All strains survived in simulated gastrointestinal fluid (pH 2) and bile salt solution (pH 8) at over 70% and 63%, compared with initial cell concentration, respectively. In vitro adhesion testing showed their adhesive property of over 70%, while the results of antibiotic susceptibility indicated that all strains were susceptible to amoxicillin, ampicillin, erythromycin, chloramphenicol, clindamycin, imipenem, kanamycin, norfloxacin, penicillin, tetracycline, and vancomycin. All strains exhibited antimicrobial ability against Staphylococcus aureus TISTR 1466, Listeria monocytogenes TISTR 2196, Escherichia coli TISTR 780, Salmonella Enteritidis TISTR 2202, and Salmonella Typhimurium TISTR 292. Moreover, Limosilactobacillus reuteri MF67.1 and Companilactobacillus farciminis R7-1 showed bile salt hydrolase activity. Cellfree culture supernatants of all 14 strains were screened for immunomodulating effects on Tumor Necrosis Factor alpha (TNF-α) production. Lactiplantibacillus paraplantarum R26-3 and Lacticaseibacillus zeae M2/5 showed high inhibition of TNF-α production at 34% and 29% reduction, respectively; while the other 12 strains decreased TNF-α production at various lower levels. Results suggested that all 14 strains met the general criteria of probiotics. Lac. zeae M2/5, Lac. paraplantarum R26-3, Lim. reuteri MF67.1, and Com. farciminis R7-1 were interesting candidates for further studies as anti-inflammatory (M2/5, R26-3) or cholesterol-reducing agents (MF67.1, R7-1) in in vivo animal models. [ABSTRACT FROM AUTHOR]

Background: Pathogenic Escherichia coli (E. coli) is an important causative agent for infectious diseases in pigs and causes significant economic loss. The global concern of antimicrobial resistance of bacteria raises awareness of the alternative ways of using antimicrobial peptides (AMPs). The study was aimed to identify and test the efficacy of AMPs from Lactobacillus spp. against the growth of pathogenic E. coli isolated from pigs in Thailand. Briefly, cell-free culture supernatants (CFCS) from 3 strains of lactic acid bacteria (LAB) consisting of Lactobacillus acidophilus (strain KMP), Lactobacillus plantarum (strain KMP), and Pediococcus pentosaceus (strain KMP) were tested against pathogenic E. coli via agar well diffusion assay in quadruplicates. The presence of a zone of inhibition (ZOI) around wells was evaluated at different incubation time. Acid and bile tolerance test was performed for bacterial viability in acid and bile salt conditions. In addition, LAB cross-streaking assay was evaluated for antagonist activity. Results: The study showed that CFCS from L. acidophilus KMP, L. plantarum KMP, and P. pentosaceus KMP could inhibit the growth of pathogenic E. coli isolated from pigs in a time-dependent manner. To exemplify, the ZOI of L. plantarum KMP against E. coli (ETEC) at 8, 10, 12, 14, and 16 h incubation, were 26.6 ± 1.1, 24.9 ± 1.9, 22.5 ± 2.4, 20.3 ± 2.9, and 17.9 ± 3.3 mm, respectively. The ZOI was significantly different between 8, 10, 12, 14 h incubation, and the ZOI of the CFCS from L. plantarum KMP was larger than others (P-value < 0.05). Furthermore, L. acidophilus KMP, L. plantarum KMP, and P. pentosaceus KMP showed viability in pH 3.0, 0.3, and 0.5% (w/v) bile salt concentration. They exhibited no antagonist activity among each other. Conclusions: According to the results, the CFCS from LAB including L. acidophilus KMP, L. plantarum KMP and P. pentosaceus KMP can inhibit the growth of pathogenic E. coli, isolated from pigs in Thailand. The antimicrobial activity observed was incubation time dependent. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB) originated from fermented foods, are generally known to possess the lipolytic activity which provides the desirable flavor of the products. However, the researches on their lipolytic activity are limited. Therefore, LAB with lipolytic activity derived from traditional fermented foods and plants in Thailand were isolated and identified based on the phenotypic characteristics and 16S rRNA gene sequences. The isolates were identified as Lactiplantibacillus pentosus (four isolates), Lactiplantibacillus plantarum (three isolates), Limosilactobacillus fermentum (four isolates), Enterococcus hirae (two isolates), and each isolate of Lactiplantibacillus paraplantarum, Companilactobacillus tucceti, Companilactobacillus farciminis, Lacticaseibacillus porcinae, Lactobacillus amylovorus, Pediococcus pentosaceus, and Enterococcus faecalis. Lipolytic activity of isolates was in the range of 0.16 ± 0.01-1.07 ± 0.02 U/mL. Enterococcus faecalis MP1-3 from mum displayed the highest lipolytic activity (1.07 ± 0.02 U/mL). Various inducers including tributyrin, Tween 20, Tween 40, Tween 60, Tween 80, and palm oil were used for lipolytic activity assay of isolate MP1-3 which cultivated in production medium at 30°C for 72 h. The greatest lipolytic activity (3.28 ± 0.22 U/mL) was found when supplemented the medium with Tween 20. The results suggest that Enterococcus faecalis MP1-3 could be chosen for further lipase purification and characterization. [ABSTRACT FROM AUTHOR]

Sixteen gamma-aminobutyric acid (GABA)-producing lactic acid bacteria (LAB) strains were isolated from plant-based Thai fermented foods, such as fermented bamboo shoots (naw-mai-dong), pickled green mustard (phakgard-dong), salted mango (ma-muang-dong), and sweetened radish (hua-chai-po), obtained in Central and Eastern Thailand. These strains were placed in a monophyletic cluster consisting of Lactobacillus, Pediococcus, and Leuconostoc species and divided into seven groups based on phenotypic characteristics and 16S rRNA gene sequences. Eleven rod-shape strains of Lactobacillus were identified as L. brevis (Group I, 6 strains), L. pentosus (Group II, 2 strains), L. fermentum (Group III, 2 strains), and L. plantarum (Group IV, 1 strain). Two tetrad-forming strains of Pediococcus were identified as P. acidilactici (Group V, 1 strain) and P. pentosaceus (Group VI, 1 strain). In addition, three coccus chain bacteria were identified as Leuconostoc mesenteroides (Group VII, 3 strains). The produced GABA from the 16 strains ranged from 0.76±0.02 to 13.42±0.28 g/l. Among them, L. brevis GPB7-4 (Group I) gave the highest level of GABA production at 13.42±0.28 g/l from 30 g/l monosodium glutamate (MSG) when cultivated at 30 °C for 72 h. Based on genome analysis, the GPB7-4 strain was closely related to L. brevis ATCC 367 with an ANIb value of 99.94%. The strain consisted of the complete genes of GABA producer gadA and gadB indicating its ability to produce GABA, thus it is a promising candidate for fermented food applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Naw Mai Dong is an ethnic fermented bamboo shoot product that is popular in the upper northern region of Thailand. Xylan is a component of hemicellulose that can be found in bamboo shoots. It is a raw material utilized in xylooligosaccharide production. To produce Naw Mai Dong, bamboo shoots are sliced and pickled in bottles or jars that contain water rinsed from rice crops with or without salt. This would differ from the simple brine comprised of water and salt that is associated with most other pickled products. Naw Mai Dong is sour due to its acidity. Importantly, fermented fruits and vegetables are a potential source of lactic acid bacteria, which have been well-documented to possess a number of probiotic characteristics. This would imply that some of the lactic acid bacteria present during bamboo shoot fermentation could serve as efficient xylooligosaccharides-fermenting probiotic lactic acid bacteria. Xylooligosaccharides (XOSs) are produced from xylan, which is a component of the hemicellulose that can be found in bamboo shoots. Naw Mai Dong, an ethnic pickled bamboo shoot product of northern Thailand, is generally characterized as acidic and has a sour taste. It can be considered a potential source of probiotic lactic acid bacteria (LAB). This study aimed to isolate efficient XOSs-fermenting probiotic LAB from ethnic pickled bamboo shoot products. A total of 51 XOSs-fermenting LAB were recovered from 24 samples of Naw Mai Dong, while 17 strains exhibited luxuriant growth in xylose and XOSs. Among these, seven strains belonging to Levicaseibacillus brevis and Pediococcus acidilactici exhibited similar growth in glucose, xylose, and XOSs, while the rest showed a weaker degree of growth in xylose and XOSs than glucose. Sixteen strains exhibited resistance under gastrointestinal tract conditions and displayed antimicrobial activity against foodborne pathogens. Notably, Lv. brevis FS2.1 possessed the greatest probiotic properties, with the highest %hydrophobicity index and %auto-aggregation. Effective degradation and utilization of XOSs by probiotic strains are dependent upon xylanase and β-xylosidase production, as well as xylose metabolism. It can be concluded that pickled bamboo shoot products can be a beneficial source of XOSs-fermenting probiotic LAB. [ABSTRACT FROM AUTHOR]

Miang, a traditional fermented tea from Northern Thailand, potentially hosts beneficial probiotic bacteria. A total of 133 isolates of lactic acid bacteria (LAB) isolated from Miang were evaluated for probiotic potential. Among them, 5 strains showed high tolerance to bile and acidic conditions and were selected for further evaluation. All selected strains showed inhibitory activity against human pathogens, including Bacillus cereus, Staphylococcus aureus, and Salmonella ser. Typhimurium. Nucleotide sequences analysis of the 16S rRNA gene revealed that 3 isolates were identified as Lactobacillus pentosus; the remaining were L. plantarum and Pediococcus pentosaceus, respectively. All 5 strains showed a high survival rate of more than 90% when exposed to simulated gastrointestinal conditions and were also susceptible to antibiotics such as erythromycin, tetracycline, and gentamycin, and resistant to vancomycin, streptomycin, and polymycin. In addition, the selected isolates exhibited different degrees of cell surface hydrophobicity (58.3–92.9%) and auto-aggregation (38.9–46.0%). The antioxidant activity reflected in DPPH scavenging activities of viable cells and their cell-free culture supernatants (CFCS) were also found in selected LAB isolates. Moreover, selected LAB isolates showed ability to grow on commercial prebiotics (GOS, FOS or XOS). The preliminary study of spray-drying using cyclodextrin as thermoprotectant suggested that all strains can be designed as a powdered formulation. L. pentosus A14-6 was the best strain, with high tolerance against simulated gastrointestinal conditions, high cell surface hydrophobicity, effective response to tested commercial oligosaccharides, especially XOS, and the highest cell antioxidant properties. L. pentosus A14-6 was therefore targeted for further applications in food and synbiotic applications. [ABSTRACT FROM AUTHOR]

Many indigenous fermented foods of Northern Thailand and neighbouring regions have traditionally been known for their health benefits. In this study, we explored the communities of bacteria in selected fermented foods which are commonly consumed among ethnic groups around Northern Thailand, for which information on their microbial compositions or their functional properties is still limited. The selected food groups included Thua Nao (alkaline fermented soybean product), Nham (fermented pork sausage/loaf), Nam phak (fermented Chinese cabbage) and Miang (fermented leaves from Miang Tea trees). Bacteria in these fermented foods were isolated and enumerated. Bacterial communities were determined using a culture-independent (pyrosequencing) approach. Lactic acid bacteria were recovered from all of these fermented food samples, with levels ranging from 3.1 to 7.5 log CFU/g throughout the fermentation processes. Analysis of the 16S rRNA gene from the fermented food samples using 454-pyrosequencing resulted in 113,844 sequences after quality evaluation. Lactic acid bacteria were found in high proportions in Nham, Nam phak and Miang. Bacillus was predominant in Thua nao, in which significant proportions of Lactic acid bacteria of the family Leuconostocaceae were also found. Groups of lactic acid bacteria found varied among different food samples, but three genera were predominant: Lactococcus, Lactobacillus and Leuconostoc, of which many members are recognised as probiotics. The results showed that these traditional Thai fermented food products are rich sources of beneficial bacteria and can potentially be functional/probiotic foods. [ABSTRACT FROM AUTHOR]

Miang , a traditional fermented tea leaf (Camellia sinensis var. assamica) consumed in northern Thailand, was simulated in laboratory conditions using non-filamentous fungi process (NFP) and microbial community was periodically investigated for over 6 months of fermentation by both culture-dependent and -independent techniques. The viable cell numbers of lactic acid bacteria (LAB), yeast, and Bacillus enumerated by the culture-dependent technique markedly surged over 3 days of initial fermentation and then smoothly declined by the end of fermentation. LAB were found as the main microbial population throughout the fermentation period followed by yeast and Bacillus. High-throughput sequencing of microbial community during fermentation revealed that Firmicutes (86.9–96.0%) and Proteobacteria (4.0–12.4%) were the dominant bacterial phyla, whereas Ascomycota was found to be the main fungal phylum with an abundance of over 99% in the fungal community. The dominant bacterial family was Lactobacillaceae (39.7–79.5%) followed by Acetobacteraceae , Enterobacteriaceae , Bacillaceae , Aeromonadaceae , Staphylococcaceae , Moraxellaceae , Clostridiaceae , Exiguobacteraceae , Streptococcaceae , and Halomonadaceae. Meanwhile, the main fungal family was incertae sedis Saccharomycetales (75.6–90.5%) followed by Pichiaceae , Pleosporaceae , Botryosphaeriaceae , Davidiellaceae , Mycosphaerellaceae , and Saccharomycodaceae. In addition, Lactobacillus (29.2–77.2%) and Acetobacter (3.8–22.8%), and the unicellular fungi, Candida (72.5–89.0%) and Pichia (8.1–14.9%), were the predominant genera during the fermentation process. The profiles of physical and chemical properties such as Miang texture, pH, organic acids, polysaccharide-degrading enzyme activities, and bioactive compounds have rationally indicated the microbial fermentation involvement. β-Mannanase and pectinase were assumed to be the key microbial enzymes involved in the Miang fermentation process. Total tannin and total polyphenol contents were relatively proportional to the antioxidant activity. Lactic acid and butyric acid reached maximum of 50.9 and 48.9 mg/g dry weight (dw) at 9 and 63 days of fermentation, respectively. This study provided essential information for deeper understanding of the Miang fermentation process based on the chemical and biological changes during production. [ABSTRACT FROM AUTHOR]

Salmonella is the leading cause of foodborne disease and a public health concern. In this study, food samples were collected from fresh markets within northeastern Thailand. Antibiotic-resistant Salmonella were detected in fresh vegetables and fermented meat products. Of 172 collected food samples, 25 (14.53%) were contaminated with Salmonella resistant to at least 1 of 9 antibiotics. Eight strains of 7 serovars, including S. Derby, S. Paratyphi B, S. Stanley, S. Dabou, S. Rissen, S. Hvittingfoss, and S. Brenderup, were resistant to S, TE, CN, SXT, and NA, accounting for 20%, 16%, 12%, 12%, and 12% of all antibiotics, respectively. To determine the anti-Salmonella activity of lactic acid bacteria, Pediococcus pentosaceus PS34 isolated from fermented fish was selected from 5 strains of food-derived lactic acid bacteria according to the antibacterial and acid-resistance properties. When added to 3 log CFU/ml cultures of Salmonella, the PS34 cell-free supernatant (CFS) reduced the numbers of Salmonella after 1, 15, 30, and 60 minutes of incubation; the reduction reached 99.9% within 30 minutes. The minimal arbitrary inhibition concentration of freeze-dried PS34 CFS was a 1:4 dilution. With the exception of neutralization to pH 6.5, treatment with proteolytic enzymes and heating at 100°C and 120°C did not reduce the freeze-dried PS34 CFS anti-Salmonella activity, indicating that the extracellular PS34 bioactive substances were heat stable and highly effective against Salmonella in vitro. Thus, bioactive substances produced by P. pentosaceus PS34 have the potential to reduce the risk of antibiotic-resistant Salmonella from contaminated food reaching consumers. [ABSTRACT FROM AUTHOR]

Background: Lactic acid bacteria (LAB) are widely used as probiotics in poultry production due to their resilience to low pH and high bile salt concentrations, as well as their beneficial effects on growth performance and antagonistic activity against enteric pathogens. However, the efficacy of probiotics depends on strain selection and their ability to colonize the host's intestine. This study aimed to select, identify, and evaluate LAB strains isolated from chicken feces in Thailand for potential use as probiotics in the chicken industry., Methods: LAB strains were isolated from 58 pooled fresh fecal samples collected from chicken farms in various regions of Thailand, including commercial and backyard farms. Gram-positive rods or cocci with catalase-negative characteristics from colonies showing a clear zone on MRS agar supplemented with 0.5% CaCO 3 were identified using MALDI-TOF mass spectrometry. The LAB isolates were evaluated for acid (pH 2.5 and pH 4.5) and bile salt (0.3% and 0.7%) tolerance. Additionally, their cell surface properties, resistance to phenol, antimicrobial activity, hemolytic activity, and presence of antimicrobial resistance genes were determined., Results: A total of 91 LAB isolates belonging to the Pediococcus, Ligilactobacillus, Limosilactobacillus , and Lactobacillus genera were obtained from chicken feces samples. Backyard farm feces exhibited a greater LAB diversity compared to commercial chickens. Five strains, including Ligilactobacillus salivarius BF12 and Pediococcus acidilactici BF9, BF14, BYF20, and BYF26, were selected based on their high tolerance to acid, bile salts, and phenol. L. salivarius BF12 and P. acidilactici BF14 demonstrated strong adhesion ability. The five LAB isolates exhibited significant cell-cell interactions (auto-aggregation) and co-aggregation with Salmonella . All five LAB isolates showed varying degrees of antimicrobial activity against Salmonella strains, with P. acidilactici BYF20 displaying the highest activity. None of the LAB isolates exhibited beta-hemolytic activity. Whole genome analysis showed that L. salivarius BF12 contained ermC, tetL , and tetM , whereas P. acidilactici strains BF9 and BF14 carried ermB, lnuA , and tetM ., Conclusion: The selected LAB isolates exhibited basic probiotic characteristics, although some limitations were observed in terms of adhesion ability and the presence of antibiotic resistance genes, requiring further investigation into their genetic location. Future studies will focus on developing a probiotic prototype encapsulation for application in the chicken industry, followed by in vivo evaluations of probiotic efficacy., Competing Interests: The authors declare that they have no competing interests., (© 2023 Khurajog et al.)

A Gram-stain-positive, facultatively anaerobic and endospore-forming rod-shaped bacterium, designed strain CPB3-1 T , was isolated from tree bark. This homofermentative strain produced dl-lactic acid from glucose. It grew at 20-45 °C, pH 4.0-9.5 and in 0-3.0 % (w/v) NaCl. It contained meso -diaminopimelic acid in cell-wall peptidoglycan and had menaquinone with seven isoprene units (MK-7) as the predominant component. The major fatty acid was anteiso-C 17 : 0 . The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, an unknown phospholipid and an unknown lipid. Based on the results of 16S rRNA gene sequence analysis, strain CPB3-1 T belonged to the genus Sporolactobacillus and was closely related to Sporolactobacillus kofuensis DSM 11701 T and Sporolactobacillus spathodeae BK117-1 T (both 96.7 % similarity), Sporolactobacillus inulinus NRIC 1133 T and Sporolactobacillus terrae DSM 11697 T (both 96.6 % similarity), and Sporolactobacillus shoreicorticis MK21-7 T , Sporolactobacillus laevolacticus DSM 442 T , Sporolactobacillus shoreae BK92 T and Sporolactobacillus pectinivorans GD201205 T (all 95.8-96.5 % similarity). The draft genome of strain CPB3-1 T contained 2 930 919 bps with 3117 coding genes. The DNA G+C content was 45.1 mol%. The digital DNA-DNA hybridization values between strain CPB3-1 T and closely related type strains were 19.2-24.0 %. The average nucleotide identity (84.0-87.6 %) and average amino acid identity (66.5-76.3 %) values were lower than the cut-off values for species delineation. Strain CPB3-1 T was clearly distinguished from related Sporolactobacillus species based on its phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence similarity and the results of draft genome analysis. Therefore, the strain represents a novel species of the genus Sporolactobacillus , for which the name Sporolactobacillus mangiferae sp. nov. is proposed. The type strain is CPB3-1 T (=JCM 35082 T =TISTR 10004 T ).

[Display omitted] • Three novel spore-forming bacteria, designated strains PP-18T, JC-4, and JC-7, isolated from soils in Thailand, were taxonomically characterized using polyphasic approach. • Weizmannia acidilactici sp. nov. is proposed for PP-18T, JC-4 and JC-7 based on 16S rRNA gene sequence similarity, ANI, and DNA-DNA hybridization (dDDH) values compared to those of W. coagulans LMG 6326T and W. acidiproducens KCTC 13078T. • Weizmannia strains PP-18T, JC-4 and JC-7 produced l -lactic acid from glucose in the range of 45.35–47.75 g/L with the yield of 0.63–0.76 g/g and productivity of 0.63–0.66 g/L·h. The strains designed PP-18T, JC-4 and JC-7 isolated from soils, were Gram-stain-positive rods, facultative anaerobe, endospore-forming bacteria. The strains produced l -lactic acid from glucose. They showed positive for catalase but negative for oxidase, nitrate reduction and arginine hydrolysis. Strains P-18T, JC-4 and JC-7 were closely related to Weizmannia coagulans LMG 6326T (97.27–97.64%) and W. acidiproducens KCTC 13078T (96.46–96.74%) based on 16S rRNA gene sequence similarity, respectively. They contained meso -diaminopimelic acid in cell wall peptidoglycan and had seven isoprene units (MK-7) as the predominant menaquinone. The major cellular fatty acids of strain PP-18T were iso -C 15:0 , anteiso-C 17:0 , iso-C 16:0 and anteiso-C 15:0. The ANIb and ANIm values among the genomes of strains PP-18T, JC-4 and JC-7 are above 99.4% while their ANIb and ANIm values among them and W. coagulans LMG 6326T and W. acidiproducens KCTC 13078T were ranged from 76.61 to 79.59%. These 3 strains showed the digital DNA-DNA hybridization (dDDH) values of 20.7–23.6% when compared with W. coagulans LMG 6326T and W. acidiproducens DSM 23148T. The DNA G + C contents of strains PP-18T, JC-4 and JC-7 were 45.82%, 45.86% and 45.86%, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphoglycolipids. The results of phenotypic and chemotaxonomic characteristics and whole-genome analysis indicated that the strains PP-18T, JC-4 and JC-7 should be represented as a novel species within the genus Weizmannia for which the name Weizmannia acidilactici sp. nov. is proposed. The type strain is PP-18T (=KCTC 33974T = NBRC 113028T = TISTR 2515T). [ABSTRACT FROM AUTHOR]

A total of 48 lactic acid bacteria (LAB) exhibited antagonistic activity against Lactobacillus sakei subsp. sakei JCM 1157 or Staphylococcus aureus DMST 8840. Only strain K2N7 was selected for characterization of bacteriocin activity. It was identified as Lactobacillus pentosus based on 16S rDNA analysis. The maximum bacteriocin production was detected in early stationary phase of growth. It was found to be sensitive to proteolytic enzymes (trypsin, proteinase K, pronase E and Ą-chymotrypsin). The bacteriocin K2N7 was heat stable (2 h at 100oC) and retained activity over a wide pH range (2.0-12.0). Bacteriocin K2N7 has a narrow inhibitory spectrum restricted to genus Lactobacillus including Lactobacillus plantarum D6SM3, a bacterial strain known to cause overfermentation in Kung-Som. The peptide was purified by 60% ammonium sulphate precipitation followed by sequential cation exchange chromatography and hydrophobic interaction characteristic. The molecular mass of bacteriocin K2N7 (2.017 kDa) was determined by matrix-assisted laser desorption/ionization time-offlight mass spectrometry analysis (MALDI-TOF MS). [ABSTRACT FROM AUTHOR]

The aim of this work was to purify and characterize the bacteriocin produced by Enterococcus faecalis KT2W2G isolated from the mangrove forest in southern Thailand, in order to evaluate its potential as a new food protective agent. The active peptide from the cell-free supernatant of Ent. faecalis KT2W2G was purified in 4 steps: (i) precipitation with 70% saturated ammonium sulfate, (ii) elution on a reversed phase cartridge (Sep-Pak C 8 ) using different concentrations of acetonitrile, (iii) cation-exchange chromatography and (iv) final purification by reversed phase-HPLC on a C 8 column. Each purification step increased the specific activity and reduced the amount of contaminating non-bacteriocin proteins. The specific activity of purified bacteriocin was 13,470.53 AU/mg of protein, which corresponded to a 48.10-fold increase. Tricine–SDS-PAGE of the purified bacteriocin gave molecular weight ranging between 3.5 and 6.5 kDa. The activity of the partially purified bacteriocin was unaffected by pH (2.0–12.0) and thermostable, but was sensitive to proteolytic enzymes. This bacteriocin maintained full stability after storage at −20, 4 and 37 °C for 2 months. It was stable when incubated for 1 month at 4 °C in 0–30% NaCl. Inhibitory spectrum of this bacteriocin showed a wide range of activities against other LAB, food-spoilage and food-borne pathogens. Ent. faecalis KT2W2G was sensitive to kanamycin, tetracycline and vancomycin but resistant to ampicillin, gentamicin and penicillin. PCR amplification demonstrated that Ent. faecalis KT2W2G does not harbor virulence genes cylA , cylB and esp but has virulence genes ace , asa1 and efaAfs. The bacteriocin and its producing strain may find application as bio-preservatives for reduction of food-spoilage and food-borne pathogens in food products. [ABSTRACT FROM AUTHOR]

Coffee is a significant agricultural commodity that is well-known and enjoyed all over the world for its distinct flavor and aroma. The quality of coffee is affected by various parameters. The existing coffee processes, which rely on natural microorganisms, are uncontrollable, resulting in product inconsistency. To overcome this problem, this study aimed to investigate the potential use of selected microbial inoculants in wet method processing and the effects of fermentation time and temperature on coffee quality. Selected yeasts, bacteria, and lactic acid bacteria, isolated from the coffee fermentation process, were used as the starter cultures. Twenty-four conditions were set. At the 0 and 24 h of fermentation, total bacterial and yeast counts, as well as pH, were measured and trend to change during fermentation. After fermentation, coffee beans were subjected to sensory evaluation by Quality-Arabica Graders. Results showed that specific microbial starter culture under controlled conditions successfully improved coffee quality. Ten of the twenty-four types coffee beans tested were classified as specialty coffee with distinct cupping characteristics. Scaling up experiment was done by a mixture of three strains of Pichia kluyveri and Wickerhamomyces anomalus (condition Y4-04) at Doi Saket District, Chiang Mai, Thailand. According to the flavor profile derived from Q-graders, the cupping results was related to the volatile and water-soluble compounds identified by GC-MS and 1H NMR analyses. It suggests that the coffee quality can be improved by using starter cultures and carefully managing fermentation settings. • This is the first report that demonstrates the improvement of the arabica coffee fermentation process in Thailand. • Novel combinations of microorganisms were used in this study. • Ten of the twenty-four coffee beans tested were classified as specialty coffee with distinct cupping characteristics. • Scaling up experiment was done at Chiang Mai, Thailand. The cupping score was higher than the control experiment. • GC-MS and NMR were used to determine the quality of fermented roasted coffee. [ABSTRACT FROM AUTHOR]

Twenty-two bacterial isolates from wild elephant faeces and 4 isolates from buffalo faeces in Thailand were investigated for probiotic properties and their lactic acid production. On the basis of their phenotypic characteristic clustering and 16S rRNA gene sequence analysis, the isolates were divided into seven groups. Group I (10 isolates) was identified as Enterococcus hirae, group II (4 isolates) as E. avium, group III (3 isolates) as Pediococcus pentosaceus, IV (3 isoaltes) as P. acidilactici, group V (3 isolates) as Lactobacillus pentosus, group VI (1 isolate) as Lactococcus garvieae and group VII (2 isolates) as Weissella paramesenteroides. All of the bacterial isolates were examined for their potentially probiotic properties in acid and bile tolerance as well as in inhibition of cancer cells proliferation by MTT assay. Although the isolates showed no significant effects on human monocytic U937 cells and colon carcinoma Caco-2 cells proliferation, P. acidilactici EL8-2 exhibited the ability to tolerate in the acidic conditions and bile salts. Therefore, P. acidilactici EL8-2 may be regarded as probiotic strains. For lactic acid fermentation with glucose, it was found that E. hirae BF14-1 produced 100% optically pure L(+)-lactic acid while W. paramesenteroides BF13-3 produced 99.79% D (-)-lactic acid. [ABSTRACT FROM AUTHOR]

Three strains of lactic acid bacteria (LAB), Pediococcus acidilactici (CP7-3) and Lactobacillus plantarum (CP1-15 and CP2-11) isolated from Thai fermented sausage "Sai-Krok-Prew" were tested for their bacteriocin-like activity against foodborne pathogen, Staphylococcus aureus. The strains CP7-3 and CP1-15 showed the inhibitory activity against the test organism, while CP2-11 had no inhibition. The three LAB strains were further investigated for use as single and dual starter culture in Sai-Krok-Prew. With a single inoculum of either CP2-11 or CP1-15, only the number of total lactic acid bacteria in the sausage with CP 2-11 was higher than that in the other samples. However, the colony count of fungi, Escherichia coli and S. aureus were not significantly different. With the dual starters of CP7-3 and CP2-11 and of CP7-3 and CP1-15, the numbers of S. aureus reduced almost 1 log CFU/g. Furthermore, the counts of Salmonella in both inoculated sausages were reduced to an undetectable level. The reduction was 3 log CFU/g, which was significantly different from that in the control (P = 0.05). Sensory assessment indicated that most panelists gave higher scores for characteristic, taste, texture and overall likeness on the sausage with the dual starter culture CP7-3 and CP1-15 than the uninoculated sausage. Therefore, uses of the mixed cultures of CP1-15 and CP7-3 may be beneficial to both safety and characteristics of Sai Krok Prew. [ABSTRACT FROM AUTHOR]

Abstract: Plaa-som is a Thai fermented fish product for which whole fish or fish fillets are fermented with either cooked rice or steamed sticky rice, salt, and garlic. A total of 762 lactic acid bacteria (LAB) were isolated during plaa-som fermentation by culture on CaCO3-MRS agar plates. They were screened and grouped by amplified ribosomal DNA restriction analysis (ARDRA), giving six groups that were identified by ribosomal DNA sequencing as Lactococcus garvieae, Streptococcus bovis, Weissella cibaria, Pediococcus pentosaceus, Lactobacillus plantarum, and Lactobacillus fermentum. Freshly mixed ingredients contained low populations of LAB (less than 10CFU/g) that subsequently grew during fermentation to final populations of approximately 107 CFU/g. Early stages of the process were dominated by the presence of Lc. garvieae, S. bovis, and W. cibaria. At 48h into fermentation, W. cibaria, P. pentosaceus, and Lb. plantarum were prevalent, and gave way to a dominance of Lb. plantarum that completed the fermentation. A mixture of these LAB species could be considered as species for development of a starter culture for plaa-som fermentation. [Copyright &y& Elsevier]

Since 1996, Vibrio parahaemolyticus serotype O3:K6 and its clones have been reported to cause human infections worldwide. In Thailand, infections caused by these pandemic strains are still high. In order to control these infections, an attempt has been made to use probiotic bacteria. In this study, 327 strains of lactic acid bacteria were isolated from 22 types of fermented Thai foods. They were screened for probiotic properties. One isolate designated as PSU-LAB 71 strongly inhibited the pandemic strains of V. parahaemolyticus and also Vibrio cholerae. PSU-LAB71 was isolated from naturally fermented pork and was identified as Lactobacillus plantarum. The mechanism of its ability to inhibit V. parahaemolyticus was most likely caused by acid production. PSU-LAB71 showed high adhesion to an enterocyte-like Caco-2 cell line. This study highlighted the possibility of using PSU-LAB71 as a probiotic strain to control infections by pandemic V. parahaemolyticus. PRACTICAL APPLICATIONS Asian countries use many fermented foods containing lactic acid bacteria. This study describes a method to isolate potential probiotic bacteria from fermented foods that have inhibitory properties against pathogenic Vibrio species, especially pandemic strains of Vibrio parahaemolyticus. This report may initiate other countries that face problems caused by this pathogen to isolate their local lactic acid bacteria that have the potential to act as probiotics to control infections. [ABSTRACT FROM AUTHOR]

Abstract: Isolation of lactic acid bacteria (LAB) and yeasts from the starter dough of Chinese steamed buns from four different commercial sources in Thailand was carried out. Thirty-one lactic acid bacteria and eight yeast strains were isolated. Total counts of LAB were from 1.8×104 to 108 colonies/g sample, whilst yeasts were very low from 10 to 2.3×102 colonies/g sample. The pH values of all starter doughs ranged from 3.36 to 3.52 and the Total Titratable Acidity (TTA) varied from 11.1 to 17.0ml of 0.1N NaOH/10g dough. All LAB isolates were identified as Lactobacillus. The phenotypic characteristics were used to cluster all the LAB isolates into two major groups (Group A and Group B), with the B group subdivided into four groups. Phylogenetic analysis, based upon partial 16S rRNA gene sequences, showed that isolates of Group A, which all contained meso-diaminopimelic acid in their cell wall and produced dl-lactic acid, were closely related to Lactobacillus plantarum, whilst the strains of Group B that produced l-lactic acid were closely related to Lactobacillus casei. For yeasts, eight isolates based on the D1/D2 domain sequences of 26S rRNA were identified as Candida tropicalis, Pichia stipitis, Candida parapsilosis, Issatchenkia orientalis and Saccharomyces cerevisiae. [Copyright &y& Elsevier]

Examines the lactic acid bacteria from healthy oral cavity of volunteers in Thailand. Inhibition of oral pathogens; Production of hydrogen peroxide and bacteriocins; Antimicrobial sensitivity of isolates.

From independent swab samples of the cloaca of indigenous gamecocks (CIG), anus of healthy baby goats (AHG), and vagina of goats (VG) originating from Phitsanulok, Thailand, a total of 263 isolates of lactic acid bacteria (LAB) were collected. Only three isolates, designated C707, G502, and V202, isolated from CIG, AHG, and VG, respectively, exhibited an excellent inhibitory zone diameter against foodborne pathogenic bacteria when evaluated by agar spot test. Isolates C707 and G502 were identified as Enterococcus faecium , whereas V202 was identified as Pediococcus acidilactici , based on 16S rRNA sequence analysis. When foodborne pathogenic bacteria were co-cultured with chosen LAB in mixed BHI-MRS broth at 39°C, their growth was suppressed. These LAB were found to be capable of surviving in simulated stomach conditions. Only the isolate G502 was able to survive in the conditions of simulated intestinal juice. This research suggests that selected LAB could be used as a food/feed supplement to reduce foodborne pathogenic bacteria and improve the safety of animal-based food or feed.

The microbial community in ethnic fermented tea, "Miang " prepared from Camellia sinensis var. assamica in northern Thailand was investigated during the 10 days of a filamentous fungi growth-based process (FFP) using both culture-dependent and culture-independent techniques. During the aerobic phase (days 1–3), filamentous fungi were visually observed growing on steamed tea leaves during which yeast and lactic acid bacteria (LAB) viable cell numbers substantially increased up to 108 and 109 CFU/g Miang, respectively, and became the main microbial population both in semi-submerged fermentation (SSF) and submerged fermentation (SMF) processes. Next generation sequencing indicated the high diversity of the filamentous fungal population in the initial stage of aerobic fermentation. Aspergillus niger was the predominant fungus on day 3 and was the most important microbe in association with yeast, LAB and Bacillus sp. in both the SSF and SMF processes. Pectinases, β-mannanase and cellulases were mainly found during the aerobic fermentation phase, suggesting they were important during fermentation. The organic acids found in Miang were acetic, gallic, glucuronic, lactic, oxalic and succinic acids, while FFP Miang showed similar bioactive compounds including total tannin, total flavonoids and total polyphenols with both SSF and SMF processes. However, SMF resulted in higher antioxidant activity than SSF, while the hardness of both was similar. This study showed an important product processing platform for assessing the relationship between microbes, biotransformation and also functional metabolites accumulated in Miang fermented using the FFP. [ABSTRACT FROM AUTHOR]

Bacteriophage infection is a serious problem in milk fermentation where it causes a delay in the fermentation process and reduces product quality. This study provides the first molecular characterisation of a lactic acid bacteriophage from spoiled fermented milk tanks in the dairy industry in Thailand. Bacteriophage ΦT25 is virulent, and shows strong lytic activity against Lacticaseibacillus paracasei LPC. Genome analysis demonstrated that phage ΦT25 possesses a linear, double-stranded DNA genome of 38,421 bp. To assay endolysin activity of phage ΦT25 (ORF35), Lysin vector pET21a was constructed by cloning the LysinT25 gene into plasmid pET21a. The size of LysinT25 endolysin was predicted to be ∼41 kDa LysinT25 was able to lyse a wide range of lactic acid bacteria and other Gram-positive bacteria, such as Staphylococcus aureus and Listeria monocytogenes. In addition, this endolysin has a synergistic inhibitory effect against Escherichia coli when combined with chloroform as an outer-membrane permeabiliser. [ABSTRACT FROM AUTHOR]

Gamma-aminobutyric acid (GABA), the inhibitory neurotransmitter, can be naturally synthesized by a group of lactic acid bacteria (LAB) which is commonly found in rich carbohydrate materials such as fruits and fermented foods. Thirty-six isolates of GABA-producing LAB were obtained from Thai fermented foods. Among these, Levilactobacillus brevis F064A isolated from Thai fermented sausage displayed high GABA content, 2.85 ± 0.10 mg/mL and could tolerate acidic pH and bile salts indicating a promising probiotic. Mulberry (Morus sp.) is widely grown in Thailand. Many mulberry fruits are left to deteriorate during the high season. To increase its value, mulberry juice was prepared and added to monosodium glutamate (MSG), 2% (w/v) prior to inoculation with 5% (v/v) of L. brevis F064A and incubated at 37 °C for 48 h to obtain the GABA-fermented mulberry juice (GABA-FMJ). The GABA-FMJ obtained had 3.31 ± 0.06 mg/mL of GABA content, 5.58 ± 0.52 mg gallic acid equivalent/mL of antioxidant activity, 234.68 ± 15.53 mg cyanidin-3-glucoside/mL of anthocyanin, an ability to inhibit growth of Bacillus cereus TISTR 687, Salmonella Typhi DMST 22842 and Shigella dysenteriae DMST 1511, and 10.54 ± 0.5 log10 colony-forming units (CFU)/mL of viable L. brevis F064A cell count. This GABA-FMJ was considered as a potential naturally functional food for human of all ages. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB), which are the most frequently used probiotics in foods, confer health benefits such as antimicrobial activity, immune stimulation, and anticancer activity. Fermented palm sap is a potential source of LAB. This study aimed to evaluate in vitro antimicrobial and probiotic properties of LAB isolated from traditional fermented palm sap in Thailand. Among 40 isolated LAB species, 10 were preliminarily selected for their antimicrobial activity. These 10 isolates were identified and confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA sequencing as Lactobacillus paracasei (8/10), Lactobacillus fermentum (1/10), and Lactobacillus brevis (1/10). They were evaluated for probiotic characteristics and antimicrobial activities against pathogens. These isolates were tolerant toward simulated gastrointestinal tract conditions, including low pH, pepsin, pancreatin, and bile salts. The 10 isolates retained strong auto-aggregation and cell surface hydrophobicity, and they adhered tightly to human intestinal epithelial cells. The isolates were susceptible to ampicillin, erythromycin, clindamycin, tetracycline, and chloramphenicol but resistant to vancomycin, kanamycin, and streptomycin. Moreover, all isolates exhibited no hemolytic activity. All isolates exhibited good antibacterial activity against nine pathogenic bacteria. Thus, these 10 Lactobacillus isolates from fermented palm sap are promising potential candidates for use as probiotics in functional fermented foods and pharmaceutical products., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)

A Gram-stain-positive, catalase-negative, rod-shaped, non-motile, non-spore-forming, and facultatively anaerobic strain CRM56-3 T , isolated from fermented tea leaves collected from Chiang Rai province, Thailand, was characterized based on a polyphasic approach. The strain produced dl-lactic acid heterofermentatively from glucose. It grew at 15-42 °C (optimum at 30 °C), pH 3.5-8.0 (optimum pH 6.0) and in 1-4 % (w/v) NaCl. Strain CRM56-3 T contained C 16:0 , C 19:0 cyclo ω 8 c , and C 18:1 ω 7 c , and/or C 18:1 ω 6c as major cellular fatty acids. Based on 16S rRNA gene sequence analysis, strain CRM56-3 T belongs to the genus Secundilactobacillus and was closely related to Secundilactobacillus odoratitofui DSM 19909 T (99.2 %), S. collinoides JCM 1123 T (98.9 %), and S. paracollinoides DSM 15502 T (98.7 %). The draft genome of strain CRM56-3 T contained 2681617 bp with 2413 coding sequences and DNA G+C content determined from genome sequence of 44.5 mol%. The digital DNA-DNA hybridization (dDDH) between strain CRM56-3 T and S. odoratitofui DSM 19909 T , S. collinoides JCM 1123 T , and S. paracollinoides DSM 15502 T were 19.5, 20.4, and 21.6 %, respectively. The average nucleotide identity (ANIm) and the average amino acid identity (AAI) between strain CRM56-3 T and closely related strains were lower than 85.0 and 80.0 %, respectively. The strain CRM56-3 T was clearly distinguished from related Secundilactobacillus species by its phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence similarity, and the draft genome analysis. Therefore, the strain represents a novel species of the genus Secundilactobacillus, for which the name of Secundilactobacillus folii sp. nov. is proposed. The type strain is CRM56-3 T (=JCM 34223 T =LMG 31663 T =TISTR 2851 T ).

A Gram-stain-positive, catalase-positive, facultatively anaerobic, terminal-spore-forming rod, designated strain BCM23-1 T , was isolated from bark of Tamarindus indica collected from Chiang Mai Province, Thailand. This strain produced d-lactic acid from glucose. It grew at 20-45 °C (optimum, 30 °C), pH 3.5-9 (optimum, pH 7.0) and in the presence of 1-4 % (w/v) NaCl. The cell-wall peptidoglycan contained meso -diaminopimelic acid (A1γ). The major isoprenoid quinone was menaquinone 7 (MK-7). Polar lipids analysis revealed the presence of diphosphatidylglycerol, phosphatidylglycerol, an unidentified aminophospholipid, an unidentified phospholipid and an unidentified lipid. The predominant cellular fatty acids were anteiso-C 17 : 0 , anteiso-C 15:0, and iso-C 16 : 0 when cultivated on GYP agar plates. The 16S rRNA gene sequence similarity between strain BCM23-1 T and Terrilactibacillus laevilacticus NK26-11 T was 98.3 %. The draft genome of BCM23-1 T was 3.24 Mb in size and contained 3088 coding sequences with an in silico DNA G+C content of 37.1 mol%. The values of ANIb, ANIm and digital DNA-DNA hybridization between strain BCM23-1 T and T. laevilacticus NK26-11 T were 89.9, 90.8 and 40.4 %, respectively. The results of phenotypic and chemotaxonomic, 16S rRNA gene sequence similarity, and whole genome analyses support strain BCM23-1 T as representing a novel species of Terrilactibacillus for which the name Terrilactibacillus tamarindi sp. nov. is proposed. The type strain is BCM23-1 T (=LMG 31662 T =JCM 33748 T =TISTR 2841 T ).

A Gram-stain-positive and catalase negative coccus, designated strain Gos25-1 T , isolated from a cotton flower (Gossypium hirsutum L.) collected from Khao Wong district, Kalasin province, Thailand. The taxonomic position of this strain was systematically studied based upon polyphasic taxonomic methods. The strain was facultatively anaerobic and produced l-lactic acid from glucose. The predominant cellular fatty acids were the straight-chain fatty acids C18 : 1ω9c and C16 : 0. According to 16S rRNA and phenylalanyl-tRNA synthase alpha subunit (pheS) gene sequence similarity, this strain was closely related to Enterococcus pallens NBRC 100697 T , E. hermanniensis CIP 108559 T , E. avium NBRC 100477 T and E. raffinosus NBRC 100492 T with 98.9-99.1 % and 77.0-82.0 % sequence similarities, respectively. Phylogenetic analysis indicated that strain Gos25-1 T was clearly distinguished from closely related species of the genus Enterococcus. Draft genome of Gos25-1 T had a size of 3.99 Mb which was contained 3788 coding sequences with in silico G+C content of 42.4 mol%. The ANIb and a digital DNA-DNA hybridisation (dDDH) values between strain Gos25-1 T and the closest related species, E. pallens NBRC 100697 T were 73.65 and 21.10 %, respectively. According to polyphasic characterisation, this strain represents a novel species of the genus Enterococcus, for which the name Enterococcus florum sp. nov. is proposed. The type strain is Gos25-1 T (=CIP 110956 T =LMG 29007 T =NBRC 111461 T =TISTR 2382 T ).

A total of 32 lactic acid bacteria (LAB) were isolated from Khanom-jeen, a Thai traditional fermented rice noodle. They belonged to the genus Leuconostoc (Ln), Lactobacillus (Lb), Enterococcus (E), Lactococcus (Lc), and Weissella (W), based on their phenotypic characteristics and 16S rRNA gene sequence analyses. The strains were identified as Ln. pseudomesenteroides (group 1, two strains), Ln. citreum (group 2, three strains), Ln. lactis (group 3, three strains), Lb. paracasei subsp. tolerans (group 4, two strains), E. faecium (group 5, three strains), Lc. lactis subsp. lactis (group 6, one strain), W. confusa (group 7, six strains), Lb. fermentum (group 8, seven strains), and Lb. plantarum subsp. plantarum and Lb. pentosus (group 9, five strains). Fifteen strains exhibited the inhibitory activity against Helicobacter pylori clinical isolates by spot-on-lawn method. Lb. fermentum P43-01 resisted to bile acids showed the broad spectrum of antimicrobial activity against H. pylori strains MS83 and BK364. These antagonistic effects were associated with proteinaceous compounds which are sensitive to α-chymotrypsin and pepsin. Results indicated that production of bacteriocin-like substances of selected strain might be the significant mechanism that exerted the inhibition on H. pylori. A potential strain could be used as probiotics in alternative or adjunctive therapy for a patient suffering from H. pylori infection.

Porcine epidemic diarrhea virus (PEDV) is a coronavirus which causes severe diarrhea and fatal dehydration in piglets. In general, probiotic supplements could enhance recovery and protect piglets against enteric pathogens. Seven local lactic acid bacteria (LAB), (Ent. faecium 79N and 40N, Lact. plantarum 22F, 25F and 31F, Ped. acidilactici 72N and Ped. pentosaceus 77F) from pig feces were well-characterized as high potential probiotics. Cell-free supernatants (CFS) and live LAB were evaluated for antiviral activities by co-incubation on Vero cells and challenged with a pandemic strain of PEDV isolated from pigs in Thailand. Cell survival and viral inhibition were determined by cytopathic effect (CPE) reduction assay and confirmed by immunofluorescence. At 1:16, CFS dilution (pH 6.3-6.8) showed no cytotoxicity in Vero cells and was therefore used as the dilution for antiviral assays. The diluted CFS of all Lact. plantarum showed the antiviral effect against PEDV; however, the same antiviral effect could not be observed in Ent. faecium and Pediococcus strains. In competitive experiment, only live Lact. plantarum 25F and Ped. pentosaceus 77F showed CPE reduction in the viral infected cells to <50% observed field area. This study concluded that the CFS of all tested lactobacilli, and live Lact. plantarum (22F and 25F) and Pediococcus strains 72N and 77F could reduce infectivity of the pandemic strain of PEDV from pigs in Thailand on the target Vero cells.

The microbiota of lactic acid bacteria (LAB) in thirty-five samples of Miang, a traditional fermented tea leaf product, collected from twenty-two different regions of eight provinces in upper northern Thailand was revealed through the culture-dependent technique. A total of 311 presumptive LAB strains were isolated and subjected to clustering analysis based on repetitive genomic element-PCR (rep-PCR) fingerprinting profiles. The majority of the strains belonged to the Lactobacillus genera with an overwhelming predominance of the Lb. plantarum group. Further studies of species-specific PCR showed that 201 of 252 isolates in the Lb. plantarum group were Lb. plantarum which were thus considered as the predominant LAB in Miang, while the other 51 isolates belonged to Lb. pentosus. In contrast to Lb. plantarum, there is a lack of information on the tannase gene and the tea tannin-tolerant ability of Lb. pentosus. Of the 51 Lb. pentosus isolates, 33 were found to harbor the genes encoding tannase and shared 93-99% amino acid identity with tannase obtained from Lb. pentosus ATCC 8041 T . Among 33 tannase gene-positive isolates, 23 isolates exhibited high tannin- tolerant capabilities when cultivated on de Man Rogosa and Sharpe agar-containing bromocresol purple (0.02 g/L, MRS-BCP) supplemented with 20% (v/v) crude tea extract, which corresponded to 2.5% (w/v) tannins. These Lb. pentosus isolates with high tannin-tolerant capacity are expected to be the high potential strains for functional tannase production involved in Miang fermentation as they will bring about certain benefits and could be used to improve the fermentation of tea products.

Lactic acid bacteria (LAB) are very important in converting of agricultural products into safe, delicious and shelf stable foods for human consumption. The preservative activity of LAB in foods is mainly attributed to the production of anti-microbial metabolites such as organic acids and bacteriocins which enables them to grow and control the growth of pathogens and spoilage microorganisms. Besides ensuring safety, bacteriocin-producing LAB with their probiotic potentials could also be emerging as a means to develop functional meat products with desirable health benefits. Nevertheless, to be qualified as a candidate probiotic culture, other prerequisite probiotic properties of bacteriocin-producing LAB have to be assessed according to regulatory guidelines for probiotics. Nham is an indigenous fermented sausage of Thailand that has gained popularity and acceptance among Thais. Since Nham is made from raw meat and is usually consumed without cooking, risks due to undesirable microorganisms such as Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes, are frequently observed. With an ultimate goal to produce safer and healthier product, our research attempts on the development of a variety of new Nham products are discussed., (Copyright © 2015 Elsevier Ltd. All rights reserved.)