scholarly journals Isolation and identification of bacteriocin-producing lactic acid bacteria from plants in Mount Makiling Forest Reserve, Philippines

2019 ◽  
pp. 1-15
Author(s):  
Emil Lat ◽  
Francisco Elegado ◽  
Ida Dalmacio
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bootstrap Method ◽  
16S Rrna Gene Sequencing ◽  
Indicator Strain ◽  
Homology Search ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Isolation And Identification ◽  
Forest Reserve

Sixty-three lactic acid bacteria (LAB) were isolated from trapped or stagnated water in parts of 14 different plant species in Mount Makiling Forest Reserve, Philippines and were screened for bacteriocin production. Thirteen of these isolates were found antagonistic to different indicator bacterial strains through direct assay. However, after screening of the pH neutralized culture supernatant using ‘spot-on-lawn’ method against Enterococcus faecium 79 as the indicator strain, only one isolate, designated as ML 258, was confirmed to be bacteriocin-producer. Its crude extract of bacteriocin exhibited inhibitory activity against E. faecium and Lactobacillus aciclophilus. The isolate was characterized and found to be gram-positive short rods that occurred in pairs, singly or in chains, catalase negative, microaerophilic, non-motile, non-spore forming and exhibited a heterofermentative type of fermentation. This isolate was obtained from fern (Microsorum longissimus) and was identified as Carnobacterium piscicola using API 50 CHL kit with good identification of 98%. However, the 16S rRNA gene sequencing and homology search from NCBI database using the BLAST program showed that the isolate is 93-95% similar with Carnobacterium maltaromaticum. On the other hand, phylogenetic analysis using MEGA 5 software that constructed a neighbor joining tree thru bootstrap method revealed that this isolate showed more similarity with Enterococcus sp.

scholarly journals Tracking of Intentionally Inoculated Lactic Acid Bacteria Strains in Yogurt and Probiotic Powder

2019 ◽  
Vol 8 (1) ◽  
pp. 5 ◽  
Author(s):  
Anshul Sharma ◽  
Jasmine Kaur ◽  
Sulhee Lee ◽  
Young-Seo Park
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Gene Sequence ◽  
Random Amplified Polymorphic Dna ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Sequence Alignments ◽  
Gene Sequence Analysis ◽  
Rrna Gene Sequencing

The present work aimed at tracking intentionally inoculated lactic acid bacteria (LAB) strains in yogurt and probiotic powder. Leuconostoc (Leu.) mesenteroides (11251), Lactobacillus (L.) brevis (B151), and Lactobacillus plantarum (LB41K) strains were tracked in yogurt, and L. plantarum (LB41P) was tracked in a commercial probiotic powder. The yogurt was intentionally inoculated with the selected bacterial strains. Two types of yogurt with known and unknown bacterial pools were utilized. The standard 16S rRNA gene sequencing was used to evaluate the initial screening. The molecular typing tools, random amplified polymorphic DNA (RAPD), repetitive element palindromic PCR (rep-PCR), and comparative gene sequence analysis of selected housekeeping loci were used to track the inoculated dubious strains. Out of 30 random selections for each inoculation, the developed method identified seven (11251), nine (B151), and five (LB41K) colonies in the yogurt. The validation was performed by identifying 7 colonies (LB41P) out of 30 in the probiotic powder. The DNA banding profiles and the gene sequence alignments led to the identification of the correct inoculated strains. Overall, the study summarizes the use of molecular tools to identify the deliberately inoculated LAB strains. In conclusion, the proposed polyphasic approach effectively tracked the intentionally inoculated strains: Leu. mesenteroides, L. brevis, and L. plantarum (LB41K) in yogurt and L. plantarum (LB41P) in probiotic powder. The study demonstrates how to track industrially relevant misused LAB strains in marketable food products.

Characterization of bacteria and antibiotic resistance in commercially-produced cheeses sold in China

2021 ◽  
Author(s):  
Jinghui Yao ◽  
Jing Gao ◽  
Jianming Guo ◽  
Hengan Wang ◽  
En Zhang ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Acquired Resistance ◽  
Antibiotic Resistance Genes ◽  
16S Rrna Gene Sequencing ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Rrna Gene ◽  
Rrna Gene Sequencing

The consumption of cheese in China is increasing rapidly. Little is known about the microbiota, the presence of antibiotic-resistant bacteria, or the distribution of antibiotic resistance genes (ARGs) in commercially-produced cheeses sold in China. These are important criteria for evaluating quality and safety. Thus, this study assessed the metagenomics of fifteen types of cheese using 16S rRNA gene sequencing. Fourteen bacterial genera were detected. Lactococcus , Lactobacillus , and Streptococcus were dominant based on numbers of sequence reads. Multidrug-resistant lactic acid bacteria were isolated from most of the types of cheese. The isolates showed 100% and 91.7% resistance to streptomycin and sulfamethoxazole, respectively, and genes involved in acquired resistance to streptomycin ( strB) and sulfonamides ( sul2) were detected with high frequency. To analyze the distribution of ARGs in the cheeses in overall, 309 ARGs from eight categories of ARG and nine transposase genes were profiled. A total of 169 ARGs were detected in the 15 cheeses; their occurrence and abundance varied significantly between cheeses. Our study demonstrates that there is various diversity of the bacteria and ARGs in cheeses sold in China. The risks associated with multidrug resistance of dominant lactic acid bacteria are of great concern.

scholarly journals Distribution and antimicrobial activity of lactic acid bacteria associated with lychee fruits

2020 ◽  
Vol 25 (6) ◽  
pp. 2079-2085
Author(s):  
LIN-HU NAN ◽  
◽  
YI-SHENG CHEN ◽  
HUI-CHUNG WU ◽  
YU-CHING SU ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
Novel Species ◽  
16S Rrna Gene Sequencing ◽  
Mass Spectrometry Analysis ◽  
Rrna Gene ◽  
Spectrometry Analysis ◽  
Gram Positive Bacteria ◽  
Rrna Gene Sequencing

Lychee is a popular fruit in China and southeastern Asia. Although it is very popular, the microbiota of lactic acid bacteria (LAB) associated with lychee remains poorly described. Lychee samples from seven different markets located in three cities in Taiwan were collected and a total of 104 LAB were isolated. Through RFLP analyses of 16S rDNA and rpoA genes for grouping and 16S rRNA gene sequencing, these isolates were finally divided into 6 groups (A to F). The most common genera of LAB in lychee samples were Weissella and Leuconostoc. Weissella confusa strain E was found to produce a bacteriocin active against Listeria monocytogenes and some other Gram-positive bacteria. Mass spectrometry analysis revealed the bacteriocin mass to be approximately 3426.77 Da, which is different to other known Weissella bacteriocins. In addition, strain MB7 included in the genus Leuconostoc was identified as potential novel species or subspecies on the basis of phylogenetic analysis of 16S rRNA, rpoA and pheS gene sequences. Thus, this is the first report describing the distribution and varieties of LAB associated with lychee fruits. In addition, one potential novel LAB species or subspecies and one potential novel bacteriocin were also reported in this study.

scholarly journals Evaluation of the probiotic and postbiotic potential of lactic acid bacteria from artisanal dairy products against pathogens

2021 ◽  
Vol 15 (01) ◽  
pp. 102-112
Author(s):  
Nazar Hussain ◽  
Muhammad Tariq ◽  
Per Erik Joakim Saris ◽  
Arsalan Zaidi
Keyword(s):  
Antibacterial Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
16S Rrna Gene Sequencing ◽  
Lactobacillus Delbrueckii ◽  
Human Consumption ◽  
Rrna Gene ◽  
Microdilution Method

Introduction: Probiotic and postbiotic potential of thirty-two strains of lactic acid bacteria (LAB), obtained earlier from artisanal dairy sources in Pakistan, have been investigated against major multi-drug resistant (MDR) and food borne pathogenic bacteria. Methodology: LAB strains were identified by 16S rRNA gene sequencing and their antibacterial activity was assessed by the microdilution method. Four LAB isolates, Weissella confusa PL6, Enterococcus faecium PL7, and Lactobacillus delbrueckii PL11 and PL13 were shortlisted. Their ability to degrade lactose and safety for human consumption in terms of hemolysis and antibiotic susceptibility were assessed in vitro. The antibacterial components in the cell-free supernatants (CFSs) of isolate cultures were characterized biochemically by HPLC. Results: Acid neutralization but not protease treatment abolished the antibacterial activity of CFSs. Lactic, acetic and propionic acids were the main acids in the CFSs, and acid production peaked in the stationary phase of growth. The antibacterial activity of the LAB cultures resulted from secretion of organic acids that lowered the pH. The strains exhibited variable ability to degrade lactose and were non-hemolytic and susceptible to the most common antibiotics. Conclusions: These LAB strains are probiotic candidates for further investigation of their postbiotic role in naturally preserving processed foods and for attenuation of lactose intolerance.

A PCR Assay for Detection of Acetic Acid–Tolerant Lactic Acid Bacteria in Acidic Food Products

2004 ◽  
Vol 67 (3) ◽  
pp. 610-615 ◽  
Author(s):  
SHIGERU NAKANO ◽  
ATSUSHI MATSUMURA ◽  
TOSHIHIRO YAMADA
Keyword(s):  
Acetic Acid ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Enrichment Culture ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Pcr Assay ◽  
Acid Tolerant

A PCR assay for the detection of acetic acid–tolerant lactic acid bacteria in the genera of Lactobacillus and Pediococcus was developed in this study. Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay. To determine the specificity of the assay, 56 different bacterial strains (of 33 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for most tested bacterial members of 16S rRNA gene–based phylogenetic groups (classi ed in the Lactobacillus casei and Pediococcus group), including Lactobacillus fructivorans, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus plantarum, and Lactobacillus paracasei. For all other bacterial strains and eukaryote tested, results were negative. Bacterial DNA for PCR was prepared with a simple procedure with the use of Chelex 100 resin from culture after growth in deMan Rogosa Sharpe broth (pH 6.0). To test this PCR assay for the monitoring of the acetic acid–tolerant lactic acid bacteria, L. fructivorans was inoculated into several acidic food as an indicator. Before the PCR, the inoculation of 10 to 50 CFU of bacteria per g of food was followed by a 28-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells. Including the enrichment culture step, the entire PCR detection process can be completed within 30 h.

scholarly journals Identification, stress tolerance, and antioxidant activity of lactic acid bacteria isolated from tropically grown fruits and leaves

2016 ◽  
Vol 62 (7) ◽  
pp. 550-561 ◽  
Author(s):  
Amandine Fessard ◽  
Emmanuel Bourdon ◽  
Bertrand Payet ◽  
Fabienne Remize
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antioxidant Activities ◽  
16S Rrna Gene Sequencing ◽  
Ldl Oxidation ◽  
Lactobacillus Species ◽  
Rrna Gene ◽  
Pineapple Juice ◽  
Functional Quality ◽  
Leuconostoc Pseudomesenteroides

From 6 samples of tropically grown fruits and leaves, 10 lactic acid bacteria belonging Leuconostoc, Weissella, and Lactobacillus species were isolated and identified by 16S rRNA gene sequencing and (GTG)5 fingerprinting. Acidification kinetics determined from BHI broth cultures showed genus-related patterns. In particular, Weissella cibaria appeared to act as a potent acidifier. Tolerance of isolates to acid, oxidative, or salt stress was highly variable and strain dependent. Isolate S14 (Leuconostoc pseudomesenteroides) growth was not affected by the presence of 0.05% H2O2, while Lactobacillus spp. isolates (S17 and S29) were the most tolerant to pH 4.5. The growth of 4 isolates, S5 (Leuconostoc mesenteroides), S14 and S10 (Leuconostoc pseudomesenteroides), and S27 (W. cibaria), was not affected by 5% NaCl. Nutritional beneficial properties were examined through measurement of antioxidant activities of short-term fermented pineapple juice, such as LDL oxidation and polyphenol content, and through exopolysaccharide formation from sucrose. Two isolates, S14 and S27, increased the antioxidant capacity of pineapple juice. The robust capacity of W. cibaria and of Leuconostoc pseudomesenteroides for vegetable lactic fermentation aimed to ameliorate food nutritional and functional quality was highlighted.

scholarly journals Diversity of lactic acid bacteria isolated during fermentation of indigenous cassava obtained from Sumba, East Nusa Tenggara, Indonesia

2021 ◽  
Vol 22 (7) ◽  
Author(s):  
IMMANUELLA BERLIANA ELVANI HUTAJULU ◽  
PERISKILA DINA KALI KULLA ◽  
Endah Retnaningrum
Keyword(s):  
Enzyme Activity ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Animal Feed ◽  
16S Rrna Gene Sequencing ◽  
Fermented Food ◽  
Rrna Gene ◽  
Acid Fermentation ◽  
Study Results ◽  
Food And Beverage

Abstract. Hutajulu IBE, Kulla PDK, Retnaningrum E. 2021. Diversity of lactic acid bacteria isolated during fermentation of indigenous cassava obtained from Sumba, East Nusa Tenggara, Indonesia. Biodiversitas 22: 2561-2570. Traditional fermented food products hold a great significance in the routine diet of Indonesian inhabitants. The present study investigated the diversity of lactic acid bacteria (LAB) found in fermented indigenous cassava, ubi karet busuk, produced in Sumba, East Nusa Tenggara, Indonesia. The isolated LAB was characterized for their phenotypic, biochemical, and molecular traits, which involved 16S rRNA gene sequencing, determination of lactic acid fermentation pathway, and measurement of b-glucosidase and tannase activity in the LAB strains. During fermentation, 15 LAB strains were obtained from the cassava. Among these, 93.33% strains were found to be homofermentative, whereas 6.73% strains displayed heterofermentative traits. BLASTn and phylogenetic analysis classified 15 LAB strains into six species, namely Enterococcus faecium, Enterococcus faecalis, Enterococcus casseliflavus, Leuconostoc mesenteroides, Lactococcus lactis, and Enterococcus sp. Among these, 10 LAB strains displayed b-glucosidase activity, with highest enzyme activity of 7.13 U/mL recorded in L. lactis F14IS3. Interestingly, two of these strains also showed expression of tannase, with F6IS1 strain displaying highest enzyme activity of 38.23 U/mL. Thus, the study results highlighted the suitability of these b-glucosidase and tannase-producing LAB strains to be used as a starter in the fermentation of cassava and other substrates such as vegetables, fruits, and legumes. These strains can also be utilized in animal feed, food, and beverage industries.

scholarly journals Culture-independent analysis of the bacterial communities in Chinese fermented vegetables and genomic analysis of lactic acid bacteria

2021 ◽  
Author(s):  
Jianming Zhang ◽  
Hye Seon Song ◽  
Chengcheng Zhang ◽  
Yeon Bee Kim ◽  
Seong Woon Roh ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bacterial Communities ◽  
High Throughput Sequencing ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Whole Genome Analysis ◽  
Sequencing Platform ◽  
Tuber Mustard ◽  
Fermented Vegetables

Abstract Six different fermented vegetables were collected from Zhejiang Province, China, to explore the associated bacterial communities using a high-throughput sequencing platform. A total of 24 phyla, 274 families and 569 genera were identified from six samples. Firmicutes and Proteobacteria were the main phyla in all of samples. Meanwhile, Brevibacterium was the major genus in Xiaoshan pickled radish. Lactobacillus-related genera and Vibrio were the major genera in fermented potherb mustard and its brine. Enterobacter and Cobetia were the major genera in fermented radish and its brine. Chromohalobacter was the major genus in the tuber mustard. These results indicated there were clear differences between the bacterial genera present in Xiaoshan pickled radish, fermented potherb mustard, fermented radish, and tuber mustard. This demonstrated the possible influences of raw materials and manufacturing processes. Furthermore, a large number of lactic acid bacteria were isolated and identified by culture-dependent and 16S rRNA gene sequence analysis, which accounted for more than 68% of all the isolates. In addition, whole genome analysis of Lactobacillus suantsaii, Lactobacillus sakei subsp. sakei, and Weissella cibaria showed they had large numbers of genes associated with carbohydrate metabolism. This may explain why these three bacterial strains can grow in fermented vegetable environments.

scholarly journals Starter cultures of lactic acid bacteria for special diet products

2017 ◽  
Vol 1 (1) ◽  
pp. 89-90 ◽  
Author(s):  
Hanna Aljaksandrauna Bareika ◽  
Anatasija Vyachaslavauna Sidarenka ◽  
Galina Ivanovna Novik
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Starter Cultures ◽  
Protein Profiling ◽  
Rrna Gene ◽  
Biochemical Characteristics ◽  
Special Diet ◽  
Rrna Gene Sequencing

Abstract From fresh and fermented vegetables (cucumber, sauercraut, eggplant), cereals (wheat, rice), legumes (soy) 37 cultures of lactic acid bacteria were isolated. Based on biochemical characteristics, MALDI-TOF MS protein profiling and 16S rRNA gene sequencing they were identified as representatives of Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Enterococcus genera. Six members of Lactobacillus genera and two members of Leuconostoc genera actively producing EPS were selected for further investigation as components of starters for production of special dietetic foodstuffs.

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