Genomic Analysis of Limosilactobacillus fermentum ATCC 23271, a Potential Probiotic Strain with Anti-Candida Activity

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.

Download Full-text

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

Microorganisms ◽

10.3390/microorganisms7080262 ◽

2019 ◽

Vol 7 (8) ◽

pp. 262

Author(s):

McLeod ◽

Fagerlund ◽

Rud ◽

Axelsson

Keyword(s):

Lactobacillus Plantarum ◽

Complete Genome ◽

Probiotic Strain ◽

General Interest ◽

Probiotic Properties ◽

Large Plasmid ◽

Reverse Transcriptases ◽

Probiotic Potential ◽

Unfavorable Effect

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.

Download Full-text

Probiotic properties of Enterococcus faecium CE5-1 producing a bacteriocin-like substance and its antagonistic effect against antibiotic-resistant enterococci in vitro

Czech Journal of Animal Science ◽

10.17221/6386-cjas ◽

2012 ◽

Vol 57 (No. 11) ◽

pp. 529-539 ◽

Cited By ~ 10

Author(s):

K. Saelim ◽

N. Sohsomboon ◽

S. Kaewsuwan ◽

S. Maneerat

Keyword(s):

Enterococcus Faecium ◽

Probiotic Strain ◽

Antagonistic Effect ◽

Penicillin G ◽

Probiotic Properties ◽

Antibiotic Resistant ◽

Good Tolerance ◽

Probiotic Potential ◽

Enterocin A

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.  

Download Full-text

Lactiplantibacillus plantarum strains isolated from spontaneously fermented cocoa exhibit potential probiotic properties against Gardnerella vaginalis and Neisseria gonorrhoeae

BMC Microbiology ◽

10.1186/s12866-021-02264-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Nathan das Neves Selis ◽

Hellen Braga Martins de Oliveira ◽

Hiago Ferreira Leão ◽

Yan Bento dos Anjos ◽

Beatriz Almeida Sampaio ◽

...

Keyword(s):

Antimicrobial Activity ◽

Neisseria Gonorrhoeae ◽

Clinical Importance ◽

Gardnerella Vaginalis ◽

Haemolytic Activity ◽

Probiotic Properties ◽

Vaginal Infections ◽

Probiotic Potential

Abstract Background Probiotics are important tools in therapies against vaginal infections and can assist traditional antibiotic therapies in restoring healthy microbiota. Recent research has shown that microorganisms belonging to the genus Lactobacillus have probiotic potential. Thus, this study evaluated the potential in vitro probiotic properties of three strains of Lactiplantibacillus plantarum, isolated during the fermentation of high-quality cocoa, against Gardnerella vaginalis and Neisseria gonorrhoeae. Strains were evaluated for their physiological, safety, and antimicrobial characteristics. Results The hydrophobicity of L. plantarum strains varied from 26.67 to 91.67%, and their autoaggregation varied from 18.10 to 30.64%. The co-aggregation of L. plantarum strains with G. vaginalis ranged from 14.73 to 16.31%, and from 29.14 to 45.76% with N. gonorrhoeae. All L. plantarum strains could moderately or strongly produce biofilms. L. plantarum strains did not show haemolytic activity and were generally sensitive to the tested antimicrobials. All lactobacillus strains were tolerant to heat and pH resistance tests. All three strains of L. plantarum showed antimicrobial activity against the tested pathogens. The coincubation of L. plantarum strains with pathogens showed that the culture pH remained below 4.5 after 24 h. All cell-free culture supernatants (CFCS) demonstrated activity against the two pathogens tested, and all L. plantarum strains produced hydrogen peroxide. CFCS characterisation in conjunction with gas chromatography revealed that organic acids, especially lactic acid, were responsible for the antimicrobial activity against the pathogens evaluated. Conclusion The three strains of L. plantarum presented significant probiotic characteristics against the two pathogens of clinical importance. In vitro screening identified strong probiotic candidates for in vivo studies for the treatment of vaginal infections.

Download Full-text

Probiotic potential of Lactobacillus fermentum G-4 originating from the meconium of newborns

Journal of the Serbian Chemical Society ◽

10.2298/jsc181105015z ◽

2019 ◽

Vol 84 (4) ◽

pp. 365-376

Author(s):

Gordana Zavisic ◽

Sasa Petricevic ◽

Slavica Ristic ◽

Milena Rikalovic ◽

Natasa Jovanovic-Ljeskovic ◽

...

Keyword(s):

Antimicrobial Activity ◽

Bacterial Translocation ◽

Hemolytic Activity ◽

Lactobacillus Fermentum ◽

In Vitro Tests ◽

Rrna Gene ◽

Probiotic Properties ◽

Probiotic Potential

The present study was dedicated to determining probiotic potential of a human isolate G-4, originated from meconium. The isolate was identified using morphological, physiological and biochemical assays and molecular method based on 16S rRNA gene sequencing. In order to evaluate its probiotic properties in vitro tests were performed: the survival in simulated gastrointestinal conditions, adhesion to hexadecane, and antimicrobial activity. Safety aspects of the isolate were examined by testing toxicity, gastrointestinal tolerance and bacterial translocation in vivo, as well as hemolytic activity in vitro. The isolate G-4, identified as Lactobacillus fermentum, showed viability in artificial gastric and intestinal juice (low degree of cell viability reduction for 0.69 and 1.30 logCFU mL-1 units, respectively), moderate adhesion to hexadecane (39?2.1 %), and antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serotype Abony and Clostridium sporogenes, due to production of lactic acid (9.80 g L-1). No signs of toxicity, bacterial translocation, hemolytic activity, were observed.

Download Full-text

Probiotic and Functional Properties of Limosilactobacillus reuteri INIA P572

Nutrients ◽

10.3390/nu13061860 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1860

Author(s):

Patricia Diez-Echave ◽

Izaskun Martín-Cabrejas ◽

José Garrido-Mesa ◽

Susana Langa ◽

Teresa Vezza ◽

...

Keyword(s):

In Vitro Assays ◽

Immunomodulatory Activity ◽

Probiotic Properties ◽

Protective Effects ◽

Mice Model ◽

In Vivo Models ◽

Food Borne ◽

Gastrointestinal Conditions

Limosilactobacillus reuteri INIA P572 is a strain able to produce the antimicrobial compound reuterin in dairy products, exhibiting a protective effect against some food-borne pathogens. In this study, we investigated some probiotic properties of this strain such as resistance to gastrointestinal passage or to colonic conditions, reuterin production in a colonic environment, and immunomodulatory activity, using different in vitro and in vivo models. The results showed a high resistance of this strain to gastrointestinal conditions, as well as capacity to grow and produce reuterin in a human colonic model. Although the in vitro assays using the RAW 264.7 macrophage cell line did not demonstrate direct immunomodulatory properties, the in vivo assays using a Dextran Sulphate Sodium (DSS)-induced colitic mice model showed clear immunomodulatory and protective effects of this strain.

Download Full-text

Antimicrobial activity screening of rhizosphere soil bacteria from tomato and genome-based analysis of their antimicrobial biosynthetic potential

BMC Genomics ◽

10.1186/s12864-020-07346-8 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Lu Zhou ◽

Chunxu Song ◽

Zhibo Li ◽

Oscar P. Kuipers

Keyword(s):

Antimicrobial Activity ◽

Plant Growth ◽

Disease Control ◽

Pseudomonas Syringae ◽

Rhizosphere Soil ◽

Erwinia Carotovora ◽

Gene Clusters ◽

Vast Number ◽

Tomato Disease

Abstract Background Tomato plant growth is frequently hampered by a high susceptibility to pests and diseases. Traditional chemical control causes a serious impact on both the environment and human health. Therefore, seeking environment-friendly and cost-effective green methods in agricultural production becomes crucial nowadays. Plant Growth Promoting Rhizobacteria (PGPR) can promote plant growth through biological activity. Their use is considered to be a promising sustainable approach for crop growth. Moreover, a vast number of biosynthetic gene clusters (BGCs) for secondary metabolite production are being revealed in PGPR, which helps to find potential anti-microbial activities for tomato disease control. Results We isolated 181 Bacillus-like strains from healthy tomato, rhizosphere soil, and tomato tissues. In vitro antagonistic assays revealed that 34 Bacillus strains have antimicrobial activity against Erwinia carotovora, Pseudomonas syringae; Rhizoctonia solani; Botrytis cinerea; Verticillium dahliae and Phytophthora infestans. The genomes of 10 Bacillus and Paenibacillus strains with good antagonistic activity were sequenced. Via genome mining approaches, we identified 120 BGCs encoding NRPs, PKs-NRPs, PKs, terpenes and bacteriocins, including known compounds such as fengycin, surfactin, bacillibactin, subtilin, etc. In addition, several novel BGCs were identified. We discovered that the NRPs and PKs-NRPs BGCs in Bacillus species are encoding highly conserved known compounds as well as various novel variants. Conclusions This study highlights the great number of varieties of BGCs in Bacillus strains. These findings pave the road for future usage of Bacillus strains as biocontrol agents for tomato disease control and are a resource arsenal for novel antimicrobial discovery.

Download Full-text

Curcumin: Natural Antimicrobial and Anti Inflammatory Agent

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2020/v32i4331060 ◽

2021 ◽

pp. 1-8

Author(s):

Pehlivanović Belma ◽

Čaklovica Kenan ◽

Lagumdžija Dina ◽

Omerović Naida ◽

Žiga Smajić Nermina ◽

...

Keyword(s):

Antimicrobial Activity ◽

Protein Denaturation ◽

Dose Range ◽

In Vitro Assays ◽

Natural Antimicrobial ◽

In Vivo Models ◽

Vitro Assay ◽

Inflammatory Agent ◽

Anti Inflammatory

The pursuance of novel antimicrobial and anti-inflammatory agents has been expanding due to a significant need for more efficient pharmacotherapy of various infections and chronic diseases. During the last decade, pharmacokinetics, pharmacodynamics and pharmacological properties of curcumin have been extensively studied. The aim of the present study was to evaluate the antibacterial activity of curcumin against both Gram-positive and Gram-negative bacteria as well as its antifungal activity by using in vitro agar well diffusion assay. Moreover, the anti-inflammatory activity of curcumin was determined with in vitro assay of inhibition of protein denaturation. Results demonstrated wide antimicrobial activity of curcumin upon all of the test bacteria and fungi. The strongest activity of curcumin was observed at a concentration of 0.50 mg/ml against S. aureus, L. monocytogenes, E. coli, P. aeruginosa and C. albicans, resulting in a maximum zone of inhibition of 14.7 mm, 14.3 mm, 13.7 mm, 10.7 mm and 10.7 mm, respectively. Findings suggested that the antimicrobial activity of curcuminis dependent upon the concentrations. Furthermore, results demonstrated high effectiveness of curcumin compared to standard acetylsalicylic acid in inhibiting heat-induced protein denaturation, which activity is also depended upon the concentrations. The present study emphasises the potential application of curcumin as a natural antimicrobial and anti-inflammatory agent. However, findings of this study are restricted to in vitro assays and consideration should be given to conducting a study involving wider dose range test substances as well as including further research on in vivo models.

Download Full-text

Pseudomonas Lipopeptide-Mediated Biocontrol: Chemotaxonomy and Biological Activity

Molecules ◽

10.3390/molecules27020372 ◽

2022 ◽

Vol 27 (2) ◽

pp. 372

Author(s):

Feyisara Eyiwumi Oni ◽

Qassim Esmaeel ◽

Joseph Tobias Onyeka ◽

Rasheed Adeleke ◽

Cedric Jacquard ◽

...

Keyword(s):

Antimicrobial Activity ◽

In Vitro Assays ◽

Bacterial Physiology ◽

Lineage Specificity ◽

Microbe Interactions ◽

Host Microbe Interactions ◽

The One ◽

Taxonomic Groups ◽

Plant Pathogen Interactions

Pseudomonas lipopeptides (Ps-LPs) play crucial roles in bacterial physiology, host–microbe interactions and plant disease control. Beneficial LP producers have mainly been isolated from the rhizosphere, phyllosphere and from bulk soils. Despite their wide geographic distribution and host range, emerging evidence suggests that LP-producing pseudomonads and their corresponding molecules display tight specificity and follow a phylogenetic distribution. About a decade ago, biocontrol LPs were mainly reported from the P. fluorescens group, but this has drastically advanced due to increased LP diversity research. On the one hand, the presence of a close-knit relationship between Pseudomonas taxonomy and the molecule produced may provide a startup toolbox for the delineation of unknown LPs into existing (or novel) LP groups. Furthermore, a taxonomy–molecule match may facilitate decisions regarding antimicrobial activity profiling and subsequent agricultural relevance of such LPs. In this review, we highlight and discuss the production of beneficial Ps-LPs by strains situated within unique taxonomic groups and the lineage-specificity and coevolution of this relationship. We also chronicle the antimicrobial activity demonstrated by these biomolecules in limited plant systems compared with multiple in vitro assays. Our review further stresses the need to systematically elucidate the roles of diverse Ps-LP groups in direct plant–pathogen interactions and in the enhancement of plant innate immunity.

Download Full-text

In vitro study of Lactobacillus plantarum properties as a potential probiotic strain and an alternative method to antibiotic treatment of fish

Archives of Polish Fisheries ◽

10.2478/aopf-2018-0006 ◽

2018 ◽

Vol 26 (1) ◽

pp. 47-55 ◽

Cited By ~ 1

Author(s):

Barbara Kazuń ◽

Krzysztof Kazuń ◽

Joanna Żylińska ◽

Andrzej K. Siwicki

Keyword(s):

Lactobacillus Plantarum ◽

Antibiotic Treatment ◽

Intestinal Flora ◽

In Vitro Study ◽

Probiotic Strain ◽

Aeromonas Salmonicida ◽

Probiotic Properties ◽

Opportunistic Bacteria ◽

Fish Bile

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.

Download Full-text

Removal of Antibiotic Resistance Gene-Carrying Plasmids from Lactobacillus reuteri ATCC 55730 and Characterization of the Resulting Daughter Strain, L. reuteri DSM 17938

Applied and Environmental Microbiology ◽

10.1128/aem.00991-08 ◽

2008 ◽

Vol 74 (19) ◽

pp. 6032-6040 ◽

Cited By ~ 146

Author(s):

Anna Rosander ◽

Eamonn Connolly ◽

Stefan Roos

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Lactobacillus Reuteri ◽

Antibiotic Resistance Gene ◽

Genetically Modify ◽

Antibiotic Resistance Genes ◽

Probiotic Strain ◽

Probiotic Properties ◽

Food Borne

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.

Download Full-text