The objective of this study was to isolate, identify, and assess the safety and functionality in vitro of putative probiotic bacterial strains. Isolation procedures were based on standard methods using elective and selective media. The isolates were identified by comparative 16S rRNA sequencing analysis while their safety was determined according to the safety tests recommended by the FAO/WHO such as antibiotic resistance, hemolysin, and biogenic amine production. Most of the isolates did not pass the in vitro safety tests; therefore, only Lactiplantibacillus plantarum (from ant intestine and cheese), Lacticaseibacillus paracasei (from goat milk and kimchi), Enterococcus faecium (from chili doenjang and vegetables with kimchi ingredients), Limosilactobacillus fermentum (from saliva), and Companilactobacillus alimentarius (from kimchi) were identified and selected for further studies. The isolates were further differentiated by rep-PCR and identified to the strain level by genotypic (16S rRNA) and phenotypic (Gen III) approaches. Subsequently, the strain tolerance to acid and bile was evaluated resulting in good viability after simulated gastrointestinal tract passage. Adhesion to mucin in vitro and the presence of mub, mapA, and ef-tu genes confirmed the adhesive potential of the strains and the results of features associated with adhesion such as hydrophobicity and zeta potential extended the insights. This study reflects the importance of fermented and non-fermented food products as a promising source of lactic acid bacteria with potential probiotic properties. Additionally, it aims to highlight the challenges associated with the selection of safe strains, which often fail in the in vitro tests, thus hindering the possibilities of "uncovering" novel and safe probiotic strains.

• Klíčová slova
• Adhesion, Lactic acid bacteria, Safety, Viability,
• MeSH
• bakteriální adheze MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• Lactobacillales genetika   izolace a purifikace   klasifikace   metabolismus   MeSH
• potravinářská mikrobiologie * MeSH
• probiotika * izolace a purifikace   MeSH
• RNA ribozomální 16S * genetika   MeSH
• sekvenční analýza DNA MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA bakterií MeSH
• RNA ribozomální 16S * MeSH

Lactic acid bacteria (LABs) are known to secrete species-specific secondary metabolites that could be utilized as novel therapeutics against multi-drug resistant pathogens. This study aimed to investigate the antagonistic and probiotic properties of LABs isolated from the vaginal ecosystem of healthy women and to assess the stability of their antagonistic metabolites. Among 43 strains isolated from healthy women, eight LAB strains exhibited detectable BLISs (bacteriocin-like substances) producing ability against E. faecalis (JH-86), S. aureus (JH-68), Streptococcus sp. (JH-80), and E. coli (JH-101), with zone of inhibition (ZI) ranging from 9.00 to 20.33 mm and minimum inhibitory concentrations (MICs) from 62.5 to 500 μL/mL, respectively. The partially purified compounds extracted from cell free supernatant (CFS) displayed an increase in antagonistic activity based on ZI, 9.67-30.17 mm and MICs, 3.91-15.63 mg/mL, respectively. In a time-kill study, both crude and partially purified compounds of Limosilactobacillus reuteri (MT180537), Pediococcus pentosaceus (MT176555), Limosilactobacillus pontis (MW362838), and Levilactobacillus brevis (MW362790) exhibited significant bactericidal action against E. faecalis (MW051601), the most frequent etiological agent of aerobic vaginitis (AV). The active secondary metabolites from L. reuteri (MT180537), P. pentosaceus (MT176555), and L. pontis (MW362838) were protein in nature and remained stable under different physicochemical conditions. Regarding probiotic properties, the strains presented probiotic characteristics, i.e., good acid, bile salt tolerance, aggregation properties, and biofilm formation. The strains were susceptible to most of the commonly used antibiotics and had no hemolytic activity. In conclusion, antagonistic compounds or BLIS produced by L. reuteri (MT180537) could be investigated further for preparation of ointments to treat AV.

• MeSH
• ekosystém MeSH
• Escherichia coli MeSH
• Lactobacillales * MeSH
• lidé MeSH
• Limosilactobacillus reuteri * MeSH
• probiotika * farmakologie   MeSH
• Staphylococcus aureus MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Before use in practice, it is necessary to precisely identify and characterize a new probiotic candidate. Eight animal lactobacilli and collection strain Lactobacillus reuteri CCM 3625 were studied from the point of saccharide fermentation profiles, bile salt resistance, antibiogram profiles, and influence of bile on sensitivity to antibiotics. Studied lactobacilli differed in their sugar fermentation ability determined by API 50CHL and their identification based on these profiles did not correspond with molecular-biological one in most cases. Survival of strains Lactobacillus murinus C and L. reuteri KO4b was not affected by presence of bile. The resistance of genus Lactobacillus to vancomycin and quinolones (ofloxacin, ciprofloxacin) was confirmed in all strains tested. This study provides the new information about oxgall (0.5 and 1 %) effect on the lactobacilli antibiotic susceptibility. Antibiotic profiles were not noticeably affected, and both bile concentrations tested had comparable impact on the lactobacilli antibiotic sensitivity. Interesting change was noticed in L. murinus C, where the resistance to cephalosporins was reverted to susceptibility. Similarly, susceptibility of L. reuteri E to ceftazidime arose after incubation in both concentration of bile. After influence of 1 % bile, Lactobacillus mucosae D lost its resistance to gentamicin. On the base of gained outcomes, the best probiotic properties manifested L. reuteri KO4b, Lactobacillus plantarum KG4, and L. reuteri E due to their survival in the presence of bile.

• MeSH
• antibakteriální látky farmakologie   MeSH
• kozy MeSH
• Lactobacillus účinky léků   izolace a purifikace   MeSH
• ovce MeSH
• probiotika analýza   MeSH
• tolerance léku MeSH
• žaludek mikrobiologie   MeSH
• žlučové kyseliny a soli farmakologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• žlučové kyseliny a soli MeSH

Twelve lactobacilli isolates from mucosa of 3-5-week-old weaned pigs were found to exert good antimicrobial activity against common porcine pathogens (S. aureus, B. cereus, E. coli, C. perfringens). Two of them produced in addition to lactic acid also considerable amounts of acetic acid, and 6 of them produced hydrogen peroxide and metabolites other than organic acids. Isolates 4/26 and 2/25 (identified as L. crispatus or L. amylovorus) were inhibitory against most strains of S. aureus, B. cereus and E. coli, and especially the strain 4/26 survived well in simulated gastric and intestinal juice. Diarrhea-causing E. coli O8K88H9 Ent(+) was successfully inhibited by the growing culture as well as by the catalase-treated and neutralized supernatant of L. reuteri 12/26. Mucin degradation and multiple resistance to antibiotics were not observed.

• MeSH
• antibiotická rezistence MeSH
• Bacillus cereus účinky léků   MeSH
• Clostridium perfringens účinky léků   MeSH
• Escherichia coli účinky léků   MeSH
• feces mikrobiologie   MeSH
• ileum mikrobiologie   MeSH
• kultivační média speciální farmakologie   MeSH
• kultivační média chemie   farmakologie   MeSH
• kyselina mléčná farmakologie   MeSH
• Lactobacillus účinky léků   izolace a purifikace   metabolismus   MeSH
• muciny metabolismus   MeSH
• nemoci prasat prevence a kontrola   MeSH
• odstavení MeSH
• peroxid vodíku metabolismus   MeSH
• přirozená imunita MeSH
• probiotika MeSH
• Staphylococcus aureus účinky léků   MeSH
• střevní sliznice mikrobiologie   MeSH
• Sus scrofa mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kultivační média speciální MeSH
• kultivační média MeSH
• kyselina mléčná MeSH
• muciny MeSH
• peroxid vodíku MeSH