scholarly journals Large Plasmid Complement Resolved: Complete Genome Sequencing of Lactobacillus plantarum MF1298, a Candidate Probiotic Strain Associated with Unfavorable Effect

2019 ◽  
Vol 7 (8) ◽  
pp. 262
Author(s):  
McLeod ◽  
Fagerlund ◽  
Rud ◽  
Axelsson

Considerable attention has been given to the species Lactobacillus plantarum regarding its probiotic potential. L. plantarum strains have shown health benefits in several studies, and even nonstrain-specific claims are allowed in certain markets. L. plantarum strain MF1298 was considered a candidate probiotic, demonstrating in vitro probiotic properties and the ability to survive passage through the human intestinal tract. However, the strain showed an unfavorable effect on symptoms in subjects with irritable bowel syndrome in a clinical trial. The properties and the genome of this strain are thus of general interest. Obtaining the complete genome of strain MF1298 proved difficult due to its large plasmid complement. Here, we exploit a combination of sequencing approaches to obtain the complete chromosome and plasmid assemblies of MF1298. The Oxford Nanopore Technologies MinION long-read sequencer was particularly useful in resolving the unusually large number of plasmids in the strain, 14 in total. The complete genome sequence of 3,576,440 basepairs contains 3272 protein-encoding genes, of which 315 are located on plasmids. Few unique regions were found in comparison with other L. plantarum genomes. Notably, however, one of the plasmids contains genes related to vitamin B12 (cobalamin) turnover and genes encoding bacterial reverse transcriptases, features not previously reported for L. plantarum. The extensive plasmid information will be important for future studies with this strain.

2018 ◽  
Vol 26 (1) ◽  
pp. 47-55 ◽  
Author(s):  
Barbara Kazuń ◽  
Krzysztof Kazuń ◽  
Joanna Żylińska ◽  
Andrzej K. Siwicki

Abstract The presence of lactic acid bacteria (LAB) favors the stabilization of intestinal flora, facilitates digestion, improves the assimilability of fodder, and has an immunomodulatory effect on the immune system. According to current research, the application of LAB following antibiotic treatment prevents the development of opportunistic bacteria inhabiting the digestive tract. In the study the potential probiotic properties of Lactobacillus plantarum strains, which can be administered as an alternative to antibiotic treatment in aquaculture, were investigated under in vitro conditions. The strains of L. plantarum were characterized for important properties such as the ability to grow in the presence of 10% fish bile, a tolerance of low pH, and antagonism to pathogens dangerous for fish such as Aeromonas salmonicida and Pseudomonas fluorescens; therefore, they meeting the criteria for strains with probiotic properties. In view of currently increasing resistance to antibiotics and a decrease of their efficiency, probiotic bacteria can serve to support immunity to infections in the future.


2012 ◽  
Vol 57 (No. 11) ◽  
pp. 529-539 ◽  
Author(s):  
K. Saelim ◽  
N. Sohsomboon ◽  
S. Kaewsuwan ◽  
S. Maneerat

A bacteriocin-like substance (BLS) producing Enterococcus faecium CE5-1 was isolated from the gastrointestinal tract (GIT) of Thai indigenous chickens. Investigations of its probiotic potential were carried out. The competition between the BLS probiotic strain and antibiotic-resistant enterococci was also studied. Ent. faecium CE5-1 exhibited a good tolerance to pH 3.0 after 2 h and in 7% fresh chicken bile after 6 h, but the viability of Ent. faecium CE5-1 decreased by about 2–3 log CFU/ml after 2 h incubation in pH 2.5. It was susceptible to the antibiotics tested (tetracycline, erythromycin, penicillin G, and vancomycin). The maximum BLS production from Ent. faecium CE5-1 was observed at 15 h of cultivation. It showed activity against Listeria monocytogenes DMST17303, Pediococcus pentosaceus 3CE27, Lactobacillus sakei subsp. sakei JCM1157, and antibiotic-resistant enterococci. The detection by polymerase chain reaction (PCR) in the enterocin structural gene determined the presence of enterocin A gene in Ent. faecium CE5-1 only. Ent. faecium CE5-1 showed the highest inhibitory activity against two antibiotic-resistant Ent. faecalis VanB (from 6.68 to 4.29 log CFU/ml) and Ent. gallinarum VanC (from 6.76 to 4.31 log CFU/ml) after 12 h of co-cultivation. The results show the future possible use of Ent. faecium CE5-1 as a probiotic strain for livestock to control antibiotic-resistant enterococci.  


2021 ◽  
Vol 7 (10) ◽  
pp. 794
Author(s):  
Camilla I. dos Santos ◽  
Carmem D. L. Campos ◽  
Wallace R. Nunes-Neto ◽  
Monique S. do Carmo ◽  
Flávio A. B. Nogueira ◽  
...  

Limosilactobacillus fermentum (ATCC 23271) was originally isolated from the human intestine and has displayed antimicrobial activity, primarily against Candida species. Complete genome sequencing and comparative analyses were performed to elucidate the genetic basis underlying its probiotic potential. The ATCC 23271 genome was found to contain 2,193,335 bp, with 2123 protein-coding sequences. Phylogenetic analysis revealed that the ATCC 23271 strain shares 941 gene clusters with six other probiotic strains of L. fermentum. Putative genes known to confer probiotic properties have been identified in the genome, including genes related to adhesion, tolerance to acidic pH and bile salts, tolerance to oxidative stress, and metabolism and transport of sugars and other compounds. A search for bacteriocin genes revealed a sequence 48% similar to that of enterolysin A, a protein from Enterococcus faecalis. However, in vitro assays confirmed that the strain has inhibitory activity on the growth of Candida species and also interferes with their adhesion to HeLa cells. In silico analyses demonstrated a high probability of the protein with antimicrobial activity. Our data reveal the genome features of L. fermentum ATCC 23271, which may provide insight into its future use given the functional benefits, especially against Candida infections.


Author(s):  
Oluwabukola Atinuke Popoola ◽  
Abiodun A. Onilude ◽  
Hadiza Rasheed-Jada ◽  
Oyekanmi Nashiru

Forty eight isolates were randomly isolated from fermented red sorghum (Sorghum bicolor) and white sorghum-based (Sorghum vulgare) gruel and evaluated for their probiotic abilities. Forty five out of the forty eight isolates were Gram-positive, catalase negative and non-sporulating, which were further evaluated.  Twenty out of the forty five isolates showed tolerance to acid with their ability to survive extreme pH 2.0, 3.0 and 4.0 respectively at different levels. These were further screened for their in vitro inhibition ability against selected test pathogens (Escherichia coli, Salmonella Typhimurium, Bacillus sp., Pseudomonas aeruginosa, and Staphylococcus aureus). Fourteen out of the twenty isolates had antimicrobial activity against the test pathogens at different levels. They were further screened for their antibiotic susceptibility pattern, all the fourteen isolates showed susceptibility to different antibiotics at different levels. These isolates were screened for their bile tolerance ability, all the fourteen isolates showed tolerance to the bile salt concentration (0.3%, 0.5% and 1%) at different levels respectively. The fourteen isolates were also screened for their gastric transit tolerance ability; all the fourteen isolates were able to tolerate gastric transit at 0 minute, 90 minutes and 180 minutes respectively. They were further screened for their in vitro hydrophobicity ability, only six out of the fourteen isolates showed percentage hydrophobicity of 40% and above. These six isolates were identified at the sub-species level using 16S RRNA Sequencing for isolates identification and construction of phylogeny. They were identified as Lactobacillus plantarum strain ST1, Lactobacillus brevis strain SC4, Lactobacillus casei strain 0108, Lactobacillus plantarum strain HASOB9a, Pediococcus pentosaceus strain SH 740 and Lactobacillus plantarum ZJ316 respectively. Out of the six identified isolates, the complete genome of Lactobacillus plantarum ZJ316 had been sequenced and was retrieved alongside with the complete genome data of four other species of Lactobacillus plantarum of industrial importance available in public domain, which was used for the mini comparative genome analysis in this research, using bioinformatics and visual analytics tools (Tableau Desktop). It was revealed that the genome size of Lactobacillus plantarum ZJ316 is second to that of Lactobacillus plantarum 16 and higher than that of the others whose genome had been completely sequenced.


2021 ◽  
Vol 11 (13) ◽  
pp. 5765
Author(s):  
Joo-Yun Kim ◽  
Eun-Jung Choi ◽  
Jae-Ho Lee ◽  
Myeong-Seok Yoo ◽  
Keon Heo ◽  
...  

Vitamin B2, also known as riboflavin, is essential for maintaining human health. The purpose of this study was to isolate novel lactic acid bacteria that overproduce vitamin B2 and to validate their potential as probiotics. In this study, Lactobacillus plantarum HY7715 (HY7715) was selected among lactic acid bacteria isolated from Kimchi. HY7715 showed a very high riboflavin-producing ability compared to the control strain due to the high expression of ribA, ribB, ribC, ribH, and ribG genes. HY7715 produced 34.5 ± 2.41 mg/L of riboflavin for 24 h without consuming riboflavin in the medium under optimal growth conditions. It was able to produce riboflavin in an in vitro model of the intestinal environment. In addition, when riboflavin deficiency was induced in mice through nutritional restriction, higher levels of riboflavin were detected in plasma and urine in the HY7715 administration group than in the control group. HY7715 showed high survival rate in simulated gastrointestinal conditions and had antibiotic resistance below the cutoff MIC value suggested by the European Food Safety Authority; moreover, it did not cause hemolysis. In conclusion, HY7715 could be considered a beneficial probiotic strain for human and animal applications, suggesting that it could be a new alternative to address riboflavin deficiency.


2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Guesh Mulaw ◽  
Tesfaye Sisay Tessema ◽  
Diriba Muleta ◽  
Anteneh Tesfaye

Probiotics are live microorganisms which when consumed in large number together with a food promote the health of the consumer. The aim of this study was to evaluate in vitro probiotic properties of lactic acid bacteria (LAB) isolated from traditional Ethiopian fermented Teff injera dough, Ergo, and Kocho products. A total of 90 LAB were isolated, of which 4 (4.44%) isolates showed 45.35–97.11% and 38.40–90.49% survival rates at pH values (2, 2.5, and 3) for 3 and 6 h, in that order. The four acid-tolerant isolates were found tolerant to 0.3% bile salt for 24 h with 91.37 to 97.22% rate of survival. The acid-and-bile salt-tolerant LAB isolates were found inhibiting some food-borne test pathogenic bacteria to varying degrees. All acid-and-bile-tolerant isolates displayed varying sensitivity to different antibiotics. The in vitro adherence to stainless steel plates of the 4 screened probiotic LAB isolates were ranged from 32.75 to 36.30% adhesion rate. The four efficient probiotic LAB isolates that belonged to Lactobacillus species were identified to the strain level using 16S rDNA gene sequence comparisons and, namely, were Lactobacillus plantarum strain CIP 103151, Lactobacillus paracasei subsp. tolerans strain NBRC 15906, Lactobacillus paracasei strain NBRC 15889, and Lactobacillus plantarum strain JCM 1149. The four Lactobacillus strains were found to be potentially useful to produce probiotic products.


2020 ◽  
Vol 4 (4) ◽  
pp. 213-223
Author(s):  
Soumitra Nath ◽  
Jibalok Sikidar ◽  
Monisha Roy ◽  
Bibhas Deb

Abstract Objectives The screening of traditional fermented products is essential for the assessment of safety, security, and further development of functional foods for the well-being of human health. The aim of the present study was to isolate and identify bacteria from fermented raw milk samples that exhibit health benefits upon consumption. Methods In order to confirm the isolates as probiotics, several in vitro assays were conducted to assess the probiotic properties of isolated bacteria. The initial screening includes tolerance to acid, bile, pancreatin, and NaCl. The cell surface properties demonstrate their interaction with mucosal epithelium, which includes hydrophobicity and auto-aggregation assay. Safety assessment was done by performing haemolytic test and antibiotic susceptibility test. The antagonistic activity of probiotic strain was further evaluated against some pathogenic bacteria. Results Lactobacillus plantarum (L. plantarum) isolated from fermented raw milk was preliminarily identified by biochemical tests and further confirmed using 16S rRNA identification. The isolate designated as L. plantarum strain GCC_19M1 demonstrated significant tolerance to low pH, 0.3% bile, 0.5% pancreatin, and 5% NaCl. In the presence of simulated gastric juice (at pH 3), the isolate exhibited a survival rate of 93.48–96.97%. Furthermore, the development of ecological niches in the human gut and their successful accumulation have been revealed by auto-aggregation and hydrophobicity properties. Absence of haemolytic activity ensures the non-virulent nature of the strain. Lactobacillus plantarum strain GCC_19M1 showed susceptibility towards gentamicin, tetracycline, kanamycin, meropenem, and ceftriaxone and exhibited an antagonistic effect on pathogenic bacteria. Conclusion The obtained results conveyed that L. plantarum strain GCC_19M1 has strong probiotic potential, and its presence in the fermented raw milk products may serve as a potent functional probiotic food.


2008 ◽  
Vol 74 (19) ◽  
pp. 6032-6040 ◽  
Author(s):  
Anna Rosander ◽  
Eamonn Connolly ◽  
Stefan Roos

ABSTRACT The spread of antibiotic resistance in pathogens is primarily a consequence of the indiscriminate use of antibiotics, but there is concern that food-borne lactic acid bacteria may act as reservoirs of antibiotic resistance genes when distributed in large doses to the gastrointestinal tract. Lactobacillus reuteri ATCC 55730 is a commercially available probiotic strain which has been found to harbor potentially transferable resistance genes. The aims of this study were to define the location and nature of β-lactam, tetracycline, and lincosamide resistance determinants and, if they were found to be acquired, attempt to remove them from the strain by methods that do not genetically modify the organism before subsequently testing whether the probiotic characteristics were retained. No known β-lactam resistance genes was found, but penicillin-binding proteins from ATCC 55730, two additional resistant strains, and three sensitive strains of L. reuteri were sequenced and comparatively analyzed. The β-lactam resistance in ATCC 55730 is probably caused by a number of alterations in the corresponding genes and can be regarded as not transferable. The strain was found to harbor two plasmids carrying tet(W) tetracycline and lnu(A) lincosamide resistance genes, respectively. A new daughter strain, L. reuteri DSM 17938, was derived from ATCC 55730 by removal of the two plasmids, and it was shown to have lost the resistances associated with them. Direct comparison of the parent and daughter strains for a series of in vitro properties and in a human clinical trial confirmed the retained probiotic properties of the daughter strain.


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