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Probiotic and anti-inflammatory attributes of an isolate Lactobacillus helveticus NS8 from Mongolian fermented koumiss.

Rong J, Zheng H, Liu M, Hu X, Wang T, Zhang X, Jin F, Wang L - BMC Microbiol. (2015)

Bottom Line: NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis.Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10.In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The Affiliated Hospital, School of Medicine, Hangzhou Normal University, Hangzhou, China. jjrong@hznu.edu.cn.

ABSTRACT

Background: Koumiss is a traditionally fermented mare's milk described with health-promoting potentials for decades. However, only a few studies focused on the probiotic strains isolated from koumiss. In this study, we collected koumiss samples from Inner Mongolian pasturing area of China and selected a promising strain of Lactobacillus helveticus, isolate NS8, based on the survival abilities in gastrointestinal tract (GIT) and adhesion to intestinal endothelial cells in vitro. As the ability to positively modulate host immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain, our study mainly focus on the immunomodulatory properties of L. helveticus NS8 by using in vivo and ex vivo analyses.

Results: L. helveticus NS8 was identified by molecular-typing methods, both at genus and species levels. As a typical food niche-specific bacteria, NS8 showed a moderate survival ability in GIT environment in vitro. However, an excellent binding capacity to the human intestinal epithelial cells, along with significant autoaggregation and cell-surface hydrophobicity was observed. Additionally, the presence of S-layer protein was responsible for the cell surface properties of this strain. NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis. In the meantime, co-culture with NS8 induced an increased level of secretion of anti-inflammatory cytokine IL-10 in peripheral blood mono-nuclear cells (PBMCs). Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10. Specially, adding of the purified S-layer protein didn't influence the production of IL-10. The specific ligand-host receptor interactions on the NS8 specific immune responses need to be learned further.

Conclusion: In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

No MeSH data available.


Related in: MedlinePlus

Molecular identification of L. helveticus NS8 isolate. a. PCR typing of NS8 isolate by using species-specific pepC/pepN/htrA primers (lane 2, pepC; lane 3, pepN; lane 4, htrA) and L. helveticus specific primers for amplification of partial 326 bp slpH gene (lane 5). There is no PCR product obtained by using primers targeted against L. acidophilus slpA (lane 6). Lane 1, DNA ladder. b. Morphology of NS8 under scanning electron microscope. NS8 are rod-shaped bacteria with high adhesion capacity to Caco-2 cells
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Fig1: Molecular identification of L. helveticus NS8 isolate. a. PCR typing of NS8 isolate by using species-specific pepC/pepN/htrA primers (lane 2, pepC; lane 3, pepN; lane 4, htrA) and L. helveticus specific primers for amplification of partial 326 bp slpH gene (lane 5). There is no PCR product obtained by using primers targeted against L. acidophilus slpA (lane 6). Lane 1, DNA ladder. b. Morphology of NS8 under scanning electron microscope. NS8 are rod-shaped bacteria with high adhesion capacity to Caco-2 cells

Mentions: In total, 12 typical colonies of lactic acid bacteria isolates were obtained from fermented koumiss based on morphological characteristics after Gram’s staining and catalase negative reaction. By testing the survival abilities in GIT environment and adherent potential to intestinal mucosa in vitro, only strain NS8 was selected as candidate for probiotic, which showed higher tolerance to low pH (70 % survival rate at pH 2) and toxic bile salts (65 % survival rate in 0.3 % bile). Then we identified this strain as L. helveticus NS8 by performing 16S rRNA gene sequencing (Genbank accession No. JQ013296.1) and BLAST search. Considering the wide inter-strain variability of L. helveticus and difficult discrimination between L. helveticus and closely related species, such as L. acidophilus and L. delbrueckii, L. helveticus specific primer pairs targeting the sequences of aminopeptidases C and N, and a trypsin-like serine protease gene (pepC, pepN, htrA) were applied in a species specific PCR assay [17]. The PCR fragments obtained had sizes of about 500, 700 and 900 bp respectively (Fig. 1a), which were in good accordance with the expected sizes.Fig. 1


Probiotic and anti-inflammatory attributes of an isolate Lactobacillus helveticus NS8 from Mongolian fermented koumiss.

Rong J, Zheng H, Liu M, Hu X, Wang T, Zhang X, Jin F, Wang L - BMC Microbiol. (2015)

Molecular identification of L. helveticus NS8 isolate. a. PCR typing of NS8 isolate by using species-specific pepC/pepN/htrA primers (lane 2, pepC; lane 3, pepN; lane 4, htrA) and L. helveticus specific primers for amplification of partial 326 bp slpH gene (lane 5). There is no PCR product obtained by using primers targeted against L. acidophilus slpA (lane 6). Lane 1, DNA ladder. b. Morphology of NS8 under scanning electron microscope. NS8 are rod-shaped bacteria with high adhesion capacity to Caco-2 cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4591576&req=5

Fig1: Molecular identification of L. helveticus NS8 isolate. a. PCR typing of NS8 isolate by using species-specific pepC/pepN/htrA primers (lane 2, pepC; lane 3, pepN; lane 4, htrA) and L. helveticus specific primers for amplification of partial 326 bp slpH gene (lane 5). There is no PCR product obtained by using primers targeted against L. acidophilus slpA (lane 6). Lane 1, DNA ladder. b. Morphology of NS8 under scanning electron microscope. NS8 are rod-shaped bacteria with high adhesion capacity to Caco-2 cells
Mentions: In total, 12 typical colonies of lactic acid bacteria isolates were obtained from fermented koumiss based on morphological characteristics after Gram’s staining and catalase negative reaction. By testing the survival abilities in GIT environment and adherent potential to intestinal mucosa in vitro, only strain NS8 was selected as candidate for probiotic, which showed higher tolerance to low pH (70 % survival rate at pH 2) and toxic bile salts (65 % survival rate in 0.3 % bile). Then we identified this strain as L. helveticus NS8 by performing 16S rRNA gene sequencing (Genbank accession No. JQ013296.1) and BLAST search. Considering the wide inter-strain variability of L. helveticus and difficult discrimination between L. helveticus and closely related species, such as L. acidophilus and L. delbrueckii, L. helveticus specific primer pairs targeting the sequences of aminopeptidases C and N, and a trypsin-like serine protease gene (pepC, pepN, htrA) were applied in a species specific PCR assay [17]. The PCR fragments obtained had sizes of about 500, 700 and 900 bp respectively (Fig. 1a), which were in good accordance with the expected sizes.Fig. 1

Bottom Line: NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis.Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10.In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The Affiliated Hospital, School of Medicine, Hangzhou Normal University, Hangzhou, China. jjrong@hznu.edu.cn.

ABSTRACT

Background: Koumiss is a traditionally fermented mare's milk described with health-promoting potentials for decades. However, only a few studies focused on the probiotic strains isolated from koumiss. In this study, we collected koumiss samples from Inner Mongolian pasturing area of China and selected a promising strain of Lactobacillus helveticus, isolate NS8, based on the survival abilities in gastrointestinal tract (GIT) and adhesion to intestinal endothelial cells in vitro. As the ability to positively modulate host immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain, our study mainly focus on the immunomodulatory properties of L. helveticus NS8 by using in vivo and ex vivo analyses.

Results: L. helveticus NS8 was identified by molecular-typing methods, both at genus and species levels. As a typical food niche-specific bacteria, NS8 showed a moderate survival ability in GIT environment in vitro. However, an excellent binding capacity to the human intestinal epithelial cells, along with significant autoaggregation and cell-surface hydrophobicity was observed. Additionally, the presence of S-layer protein was responsible for the cell surface properties of this strain. NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis. In the meantime, co-culture with NS8 induced an increased level of secretion of anti-inflammatory cytokine IL-10 in peripheral blood mono-nuclear cells (PBMCs). Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10. Specially, adding of the purified S-layer protein didn't influence the production of IL-10. The specific ligand-host receptor interactions on the NS8 specific immune responses need to be learned further.

Conclusion: In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.

No MeSH data available.


Related in: MedlinePlus