In the present study, the evolution of the physicochemical and microbiological characteristics of lactic acid bacteria (LAB) in traditional Kırklareli white brined cheese collected from 14 different cheese manufacturing facilities were investigated on different days of the 90-day ripening period. The obtained LAB within the species Lactococcus (Lc.) lactis, Latilactobacillus (Lt.) curvatus, Lactobacillus (Lb.) casei and Lb. plantarum, Enterococcus (E.) durans, E. faecium, E. faecalis, Streptococcus macedonicus, and Weissella paramesenteroides were characterized in terms of their influence on technological properties and their potential as starter cultures for traditional white brined cheese production. The results of the microbiological and physicochemical investigations showed that a few selected isolates of Lc. lactis, Lb. casei, and Lb. plantarum had certain functions as starter germs. Moderate acidification capacity, antibacterial activity and proteolytic activity, which are characteristic of their use as starter lactic acid bacteria, were found. Importantly, antibiotic resistance among selected Lc. lactis, Lb. casei, and Lb. plantarum isolates was extremely low, whereas some of these isolates demonstrated antibacterial activity against major foodborne pathogenic bacteria. Based on the results obtained in this study, selected Lc. and Lb. isolates can also be considered as starter culture in traditional cheese production.
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.
- 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
The order Lactobacillales represents a morphologically, metabolically, and physiologically diverse group of bacteria. Lactic acid bacteria represent the core of this phylogenetic group. They are a part of epiphytic microflora, fermented dairy, meat, fruit and vegetable products, and the digestive tract of humans and animals. Despite the fact that these bacteria form a phenotypically and genotypically heterogeneous group, their phylogenetic relationship enables to propose a common genetic marker usable in classification, typing, and phylogeny. By creation of consensus sequence based on available genomic sequences of some representatives of order Lactobacillales, a specific primer-pair binding variable region of aspS gene (length of 615 nts) encoding the aspartyl-tRNA synthetase was designed. This gene has not yet been used in classification and phylogeny of the order Lactobacillales, although it meets the requirements of molecular markers (distribution and single copy in bacterial genomes, functional constancy and genetic stability, sequence variability among taxonomic units, irreplaceable role in proteosynthesis). Primers were applied on 54 type and wild Lactobacillales strains. Obtained sequences allowed to provide alignments for purpose of phylogenetic tree reconstructions that uncovered particular phylogenetic clusters of vagococci/enterococci, obligately homofermentative and heterofermentative lactobacilli. Although a relatively short fragment of the aspS gene (approximately 33% of the complete gene sequence) was evaluated, much higher sequence variability (61.8% of pairwise identity) among strains examined compared with 16S rRNA gene (90.7%, length of 1318 nt) provides a relatively simple and effective tool for classification and typing of selected representatives of the order Lactobacillales.
- MeSH
- aspartát-tRNA-ligasa genetika MeSH
- bakteriální geny MeSH
- bakteriální proteiny genetika MeSH
- DNA bakterií genetika MeSH
- esenciální geny MeSH
- fylogeneze * MeSH
- genetické markery MeSH
- Lactobacillales klasifikace enzymologie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Microorganisms develop biofilms on indwelling medical devices and are associated with biofilm-related infections, resulting in substantial morbidity and mortality. Therefore, to prevent and control biofilm-associated infections, the present study was designed to assess the anti-biofilm potential of postbiotics derived from probiotic organisms against most prevalent biofilm-forming Pseudomonas aeruginosa PAO1. Eighty lactic acid bacteria isolated from eight neonatal fecal samples possessed antibacterial activity against P. aeruginosa PAO1. Among these, only four lactic acid bacteria produced both bacteriocin and exopolysaccharides but only one isolate was found to maximally attenuate the P. aeruginosa PAO1 biofilm. More specifically, the phenotypic and probiotic characterization showed that the isolated lactic acid bacteria were gram positive, non-motile, and catalase and oxidase negative; tolerated acidic and alkaline pH; has bile salt concentration; showed 53% hydrophobicity; and was found to be non-hemolytic. Phylogenetically, the organism was found to be probiotic Lactobacillus fermentum with accession no. KT998657. Interestingly, pre-coating of a microtiter plate either with bacteriocin or with exopolysaccharides as well as their combination significantly (p < 0.05) reduced the number of viable cells forming biofilms to 41.7% compared with simultaneous coating of postbiotics that had 72.4% biofilm-forming viable cells as observed by flow cytometry and confocal laser scanning microscopy. Therefore, it can be anticipated that postbiotics as the natural biointerventions can be employed as the prophylactic agents for medical devices used to treat gastrointestinal and urinary tract infections.
- MeSH
- antibakteriální látky metabolismus farmakologie MeSH
- bakteriální polysacharidy metabolismus MeSH
- bakteriociny metabolismus farmakologie MeSH
- biofilmy účinky léků MeSH
- feces mikrobiologie MeSH
- kojenec MeSH
- Lactobacillales chemie genetika izolace a purifikace metabolismus MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- probiotika metabolismus farmakologie MeSH
- Pseudomonas aeruginosa účinky léků genetika fyziologie MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Detailed differentiation, classification, and phylogenetic analysis of the order Lactobacillales are performed using molecular techniques that involve the comparison of whole genomes, multilocus sequence analysis, DNA-DNA hybridisation, and 16S rRNA sequencing. Despite the wide application of the latter two techniques, issues associated with them are extensively discussed. Although complete genomic analyses are the most appropriate for phylogenetic studies, they are time-consuming and require high levels of expertise. Many phylogenetic/identification markers have been proposed for enterococci, lactobacilli, streptococci, and lactobacilli. However, none have been established for vagococci and some genera within the order Lactobacillales. The objective of the study was to find novel alternative housekeeping genes for classification, typing, and phylogenetic analysis of selected genera within the order Lactobacillales. We designed primers flanking variable regions of the infB (504 nt) and rpsB (333 nt) genes and amplified and sequenced them in 56 strains of different genera within the order Lactobacillales. Statistical analysis and characteristics of the gene regions suggested that they could be used for taxonomic purposes. Phylogenetic analyses, including assessment of (in)congruence between individual phylogenetic trees indicated the possibility of using the concatenation of the two genes as an alternative tool for the evaluation of phylogeny compared with the 16S rRNA gene representing the standard phylogenetic marker of prokaryotes. Moreover, infB, rpsB regions and their concatenate were phylogenetically consistent with two widely applied alternative genetic markers in taxonomy of particular Lactobacillales genera encoding the 60 kDa chaperonin protein (GroEL-hsp60) and phenylalanyl-tRNA synthetase, alpha subunit (pheS).
- MeSH
- bakteriální geny MeSH
- chaperon hsp60 genetika MeSH
- DNA bakterií chemie MeSH
- DNA primery MeSH
- esenciální geny MeSH
- fenylalanin-tRNA-ligasa genetika MeSH
- fylogeneze * MeSH
- genetické markery MeSH
- Lactobacillales klasifikace genetika MeSH
- multilokusová sekvenční typizace MeSH
- prokaryotický iniciační faktor 2 genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
Lactobacillus pobuzihii is a novel species which has been previously found in pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. However, the lactic acid bacteria (LAB) microflora in pobuzihi has not been studied in detail. In this study, LAB from pobuzihi were isolated, identified, and characterized. A total of 196 LAB were isolated; 79 cultures were isolated from the sample collected from a manufacturing factory, 38 from pobuzihi samples collected from 4 different markets, and 79 from 2 fresh cummingcordia samples. These isolates were characterized phenotypically and then divided into eight groups (A to H) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus plantarum was the most abundant LAB found in most samples during the fermentation of pobuzihi. On the other hand, Enterococcus casseliflavus and Weissella cibaria were, respectively, the major species found in the two fresh cummingcordia samples. A potential novel species or subspecies of lactococcal strain was found. In addition, seven L. plantarum and five W. cibaria strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T). This is the first report describing the distribution and varieties of LAB existing in the pobuzihi during its fermentation process and the final product on the market.
- MeSH
- biodiverzita * MeSH
- DNA bakterií chemie genetika MeSH
- fermentace MeSH
- Lactobacillales klasifikace genetika izolace a purifikace MeSH
- molekulární sekvence - údaje MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- potravinářská mikrobiologie * MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Taiwan MeSH
