scholarly journals Genome analysis of two Lactobacillus plantarum strains, LLY-606 and pc-26, for evaluating their potential as probiotics

2019 ◽  
Author(s):  
Sudun ◽  
Lifeng Liang ◽  
Zhenzhen Li ◽  
Qingnan Lin ◽  
Can Peng ◽  
...  

AbstractLactobacillus plantarum was not only one of the most popular probiotics, but also one of the most versatile lactic acid bacteria. L. plantarum LLY-606 and L. plantarum pc-26 are strains isolated from human gut that are intended to be explored as probiotics. In this study, the genome sequences of LLY-606 and pc-26 were sequenced, and multiple genes related to probiotic properties were analyzed. First, the pathogenicity of these strains was evaluated, and antibiotic resistance genes were surveyed at the whole genome level to determine their primary safety. And then, genes for stress response, plantaricin (pln) biosynthesis, extracellular polysaccharide (EPS) biosynthesis, and bile salt hydrolase (BSH) were analyzed to evaluate their industrial utilization, adhesive capacity, and survival ability in gut, which were properties fundamental for probiotic strains. The physiological features assured by these genes were assayed in vitro. The strains were then evaluated in vivo for their ability to lower cholesterol, and they were both found to be effective in improving hypercholesterolemia in golden hamsters. In this study, a genetic pre-evaluation was conducted through genome analysis combined with in vitro physiological assay, and the probiotic properties of these strains were verified in vivo.

Author(s):  
Y. Zeng ◽  
Y. Li ◽  
Q. P. Wu ◽  
J. M. Zhang ◽  
X. Q. Xie ◽  
...  

This study investigated the antipathogenic activity and probiotic potential of indigenous lactic acid bacteria (LAB) isolated from Chinese homemade pickles. In total, 27 samples were collected from different sites in China. Fifty-nine yielded pure colonies were identified by 16S rRNA gene sequencing as LAB and were initially evaluated for the antibacterial activity in vitro. Initial screening yielded Lactobacillus plantarum GS083, GS086, and GS090, which showed a broad-spectrum antibacterial activity against food-borne pathogens, especially multidrug-resistant pathogens. Meanwhile, organic acids were mainly responsible for the antimicrobial activity of the LAB strains, and the most abundant of these was lactic acid (19.32 ± 0.95 to 24.79 ± 0.40 g/l). Additionally, three L. plantarum strains demonstrated several basic probiotic characteristics including cell surface hydrophobicity, autoaggregation, and survival under gastrointestinal (GI) tract conditions. The safety of these isolates was also evaluated based on their antibiotic susceptibility, hemolytic risk, bile salt hydrolase activity, and existence of virulence or antibiotic resistance genes. All strains were safe at both the genomic and phenotypic levels. Therefore, L. plantarum GS083, GS086, and GS090 are fairly promising probiotic candidates and may be favorable for use as preservatives in the food industry.


2019 ◽  
Author(s):  
Min Young Jung ◽  
Yangseon Kim ◽  
Jung-Ae Kim ◽  
Dae-Hyuk Kim

Abstract Background: The genus Bifidobacterium includes common healthy gut microbes in mammals and is widely acknowledged as a probiotics with health-promoting properties. Results: Bifidobacterium gallinarum CACC 514 was isolated from canine feces and its potential probiotic properties were characterized by comparative and functional genome analyses.The complete genome of strain CACC 514 was found to be 2.4 Mb, with a G + C content of 63.9 mol%. The strain possessed factors beneficial to mammalian health based on the presence of genes related to mucosal surface adhesion proteins, stress-related genes, and extracellular polysaccharide genes. A comparative genome analysis with other Bifidobacterium species revealed the unique characteristics of this species. These functional genomics have been confirmed for its superiority by probiotic properties, acid and bile tolerance, and adhesion to mucus. Conclusions: The genome analysis and in vitro probiotics characteristics revealed its superiority in the intestine. These results add to our comprehensive understanding of B. gallinarum and suggest that this strain has potential application in mammalian probiotics. Keywords: Bifidobacterium gallinarum, gut microbe, probiotics, genome analysis, canine, feces.


Foods ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 668
Author(s):  
Po-Lin Liao ◽  
Chien-Chen Wu ◽  
Tai-Ying Chen ◽  
Ying-Chieh Tsai ◽  
Wu-Shun Peng ◽  
...  

Probiotics are extensively available to consumers; however, the use of probiotics may not always be safe, and there are few reports on their side effects, including those of Lactobacillus. Lactobacillus plantarum strain PS128TM isolated from spontaneously fermented mustard greens in Taiwan was recently reported to exhibit probiotic properties. In this study, we aimed to assess the safety of strain PS128TM for use in humans via examining genotoxic and oral toxic effects using in vitro and in vivo testing. Five strains of Salmonella typhimurium were evaluated by the Ames test; no signs of increased reverse mutation were observed following exposure to PS128TM. Additional testing of Chinese hamster ovary (CHO) cells exposed to PS128TM revealed that the incidence of chromosomal aberrations in CHO cells had not increased. PS128TM treatment also did not affect the proportion of immature to total erythrocytes or the number of micronuclei in the immature erythrocytes of ICR mice. Moreover, following a 28 day study involving repeated oral dose toxicity tests (2400, 400, and 40 mg/kg body weight) utilizing an ICR mouse model, no observable adverse level (NOAEL) was found at any of the doses. PS128TM was sensitive to antibiotics; however, genes related to the production of biogenic amines were absent. While further research is required, these toxicological assessments suggest that PS128TM could be safe for human consumption.


Author(s):  
Dan Wang ◽  
Tiehua Zhang ◽  
Hongwei Hao ◽  
Hongxing Zhang ◽  
Haiqing Ye ◽  
...  

Background: Probiotics are a group of bacteria that play a critical role in intestinal microbiota homeostasis and may help adjunctively treat certain diseases like metabolic and immune disorders. Objective: We recently generated a space-flight mutated Lactobacillus plantarum SS18-50 with good in vitro probiotic characteristics. In the current research, we designed two in vivo experiments to evaluate whether L. plantarum SS18-50 had the ability to increase beneficial gut bacteria, regulate oxidative status and ameliorate inflammation in mice. Methods: Experiments I: the ICR mice were gavaged with L. plantarum SS18-50 or its wild type L. plantarum GS18 at 107 or 109 CFU/kg BW daily for one month, during which the body weight was recorded weekly. The feces were collected to determine the abundance of two main beneficial bacterial groups including Lactobacillus and Bifidobacterium by selective culturing, while the total triglycerides and cholesterols in sera were determined using commercial kits. Experiment II: the mice were gavaged with loperamide hydrochloride to develop oxidative stress and inflammation phenotypes. At the same time, the experimental mice were gavaged with L. plantarum SS18-50 or wild type L. plantarum GS18 at 107 or 109 CFU/kg BW daily for one month. At the end of experiment, oxidative indicators (SOD and MDA) and inflammatory cytokines (IL-17A and IL-10) were measured by commercial kits. Results: Results showed that L. plantarum SS18-50 increased the abundance of Lactobacillus and Bifidobacterium in mice after one month’s administration. L. plantarum SS18-50 also showed the anti-oxidant activity by increasing SOD and decreasing MDA, and exerted the anti-inflammatory effect by increasing IL-10 and decreasing IL-17A in Lop treated mice. Both the wild type stain and the space mutant had such biomedical effects, but L. plantarum SS18-50 was better in increasing gut beneficial bacteria and oxidative regulation than the wild type (P<0.05). Conclusion: we conclude that L. plantarum SS18-50 has a great potential to serve as a dietary functional probiotic supplement and/or adjunctive treatment strategy.


Nutrients ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1860
Author(s):  
Patricia Diez-Echave ◽  
Izaskun Martín-Cabrejas ◽  
José Garrido-Mesa ◽  
Susana Langa ◽  
Teresa Vezza ◽  
...  

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.


2001 ◽  
Vol 69 (6) ◽  
pp. 4079-4085 ◽  
Author(s):  
Sarah E. Cramton ◽  
Martina Ulrich ◽  
Friedrich Götz ◽  
Gerd Döring

ABSTRACT Products of the intercellular adhesion (ica) operon in Staphylococcus aureus and Staphylococcus epidermidis synthesize a linear β-1,6-linked glucosaminylglycan. This extracellular polysaccharide mediates bacterial cell-cell adhesion and is required for biofilm formation, which is thought to increase the virulence of both pathogens in association with prosthetic biomedical implants. The environmental signal(s) that triggers ica gene product and polysaccharide expression is unknown. Here we demonstrate that anaerobic in vitro growth conditions lead to increased polysaccharide expression in both S. aureus and S. epidermidis, although the regulation is less stringent inS. epidermidis. Anaerobiosis also dramatically stimulates ica-specific mRNA expression inica- and polysaccharide-positive strains of both S. aureus and S. epidermidis.These data suggest a mechanism whereby ica gene expression and polysaccharide production may act as a virulence factor in an anaerobic environment in vivo.


Author(s):  
Mihaela DUMITRU ◽  
Mihaela HĂBEANU ◽  
Cristina TABUC ◽  
Ștefana JURCOANE

This study aimed to evaluate some probiotic properties of Bacillus subtilis ATCC 6051a. The phenotypic profile, resistance to pH by simulated gastric juice (pH 2 and 3), bile salts by simulated intestinal fluid, survivability (%), heat and antibiotics tolerance were investigated. The strain is a Gram-positive, rod-shaped bacteria, arranged in short chains or in small irregular pairs with the ability to produce spores. Good viability at pH 2 and 3, with a survival of more than ≥80%, was found. In the presence of bile salts 0.3%, over 4 h, the strain exhibited a survival ≥85%. At 80°C, for 120 min., the strain showed good growth (9.04 log CFU/ml). Results were sensitive to most antibiotics, with a highly susceptible (between 16 – 25 mm) to erythromycin, clindamycin, amoxicillin, chloramphenicol, ciprofloxacin, amikacin and kanamycin. The strain was found to be sensitive to vancomycin, gentamicin, and tetracycline. The present research demonstrated that Bacillus subtilis ATCC 6051a can survive under gastrointestinal conditions, which involves them to future in vitro and in vivo probiotic studies.


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.


2017 ◽  
Vol 83 (4) ◽  
pp. 391-402
Author(s):  
Jhanina Rodríguez Carrasco ◽  
Paula García-Godos Alcázar

La investigación tuvo como objetivos aislar e identificar bacterias lácticas (BAL), evaluar la capacidad probiótica in vitro e in vivo de bacterias lácticas aisladas de chicha de molle, para ello se muestreó chichas de molle elaboradas artesanalmente de las provincias de Huanta y Huamanga, aislando 55 cepas BAL e identificando a Lactobacillus plantarum, Lactobacillus maltaromicus y Leuconostoc mesenteroides en base a la coloración Gram, producción de gas, gluconato y fermentación de azúcares. Para evaluar la capacidad probiótica in vitro se realizaron pruebas de antagonismo entre BAL con cuatro microorganismos patógenos (Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14028, Staphylococus aureus ATCC 25923 y Candida albicans ATCC 90028), mostrándose que 14 de las 55 cepas BAL producen sustancias inhibitorias de amplio espectro; asimismo, se evaluó la capacidad de tolerancia a condiciones gastrointestinales de cepas BAL, realizando ensayos a diferentes pHs , diferentes concentraciones de sales biliares y extracto gástrico artificial, resultando 25 cepas BAL con capacidad de tolerancia gastrointestinal y se seleccionaron cuatro cepas con mayor diámetro de halos de inhibición y cepas tolerantes a condiciones gastrointestinales siendo las cepas: BL-1 (Lactobacillus plantarum), BL-26 (Lactobacillus maltaromicus), BL-27 (Lactobacillus plantarum) y BL-53 (Lactobacillus maltaromicus), a las cuales se evaluaron la capacidad probiótica in vivo en 20 ratas para luego realizar recuento de BAL en el intestino a los 21 días, encontrándose en el grupo de estudio con BAL a 60x1019 UFC/ mL, mientras en el tratamiento con BAL más yacón a 25x1024 UFC/mL y los tratamientos de yacón y control a 50x1014 UFC/mL de BAL obteniéndose una de ganancia de peso en ratas en el grupo de estudio de BAL más yacón de 46 g, mientras con bacterias lácticas se tuvo 24 g y 16 g en el grupo control y extracto de yacón. En consecuencia esta investigación demuestra que la toma diaria de bebidas fermentadas tradicionales favorece el incremento de Lactobacillus en la microbiota intestinal.


2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S30-S31
Author(s):  
Gabriel Suarez ◽  
Bo Liu ◽  
Jeremy Herzog ◽  
Ryan Sartor

Abstract Sulfur metabolism is emerging as a signature of IBD gut microbiota. Overrepresentation of sulfur-reducing bacteria (SRB) in IBD results in SRB-derived epithelial toxic H2S production that can overwhelm the body’s detoxification capacity, leading to impaired cellular respiration by inhibiting oxygen binding to mitochondrial cytochrome-c-oxidase. Butyrate potently inhibits SRBs and H2S, yet IBD patients have reduced short chain fatty acid (SCFA) production. More critically, H2S blocks butyrate oxidation, the primary energy source of colonocytes; butyrate oxidation deficiency is a defining characteristic of IBD. Since cysteine is the preferred substrate for H2S production by SRBs, a cysteine-rich environment provided by either a high protein diet or local intestinal mucus degradation promotes ideal conditions for SRB establishment and proliferation. SRBs can catabolize other sulfur-containing compounds critical for immune homeostasis and cellular health, such as taurine-conjugated bile acids and the “master antioxidant” glutathione, leading to further toxic H2S production. However, the molecular underpinnings of sulfur metabolism by specific bacterial genera is understudied in IBD. Results: Using a combination of in-vivo and in-vitro screening to detect the relative induction of interleukin 10 (IL-10) and interferon g (IFNg) by 19 resident bacterial strains isolated from a healthy human donor, we identified 4 bacterial strains that induce a low IL-10/IFNg ratio. These 4 strains (low group), but not 3 bacterial strains that induce a high IL-10/IFNg ratio, induce colitis in selectively colonized gnotobiotic Il10-/- mice (Fig.1A). Two of these 4 disease-inducing strains, Clostridium perfringens (A12) and Clostridium bolteae (B6), produce high concentrations of H2S in monoassociated mice (Fig.1B). In-vitro H2S production by these strains is dependent on cysteine (Fig.1C). C. perfringens and C. bolteae each induce colitis in monoassociated Il10-/- mice (Fig.1D). We are dissecting the sulfur metabolic pathways in C. perfringens and C. bolteae and their contribution to inflammatory processes by interrupting key genes predicted to contribute to H2S production, cysteine catabolism and bile acid metabolism. We will use these mutants in both in-vitro and in-vivo Il10 -/- gnotobiotic mice models to characterize their metabolic and inflammatory profiles. We have created several mutants using Targetron gene editing, including the dissimilatory sulfate reductase (Δdsr), a putative sulfonate membrane transporter (ΔssuA), anaerobic sulfite reductase (ΔasrA) and bile salt hydrolase (Δbsh). Conclusions: H2S producing bacterial strains can induce experimental colitis. Our planned mechanistic studies will determine the metabolic routes for H2S production by specific aggressive bacteria to guide novel therapeutic or dietary interventions to improve IBD prognosis.


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