[국내논문]Probiotic Potential of Enterococcus faecium Isolated from Chicken Cecum with Immunomodulating Activity and Promoting Longevity in Caenorhabditis elegans원문보기
Sim, Insuk
(Department of Integrated Biomedical and Life Sciences, College of Health Science, Korea University)
,
Park, Keun-Tae
(Department of Research and Development Center, Milae Resources ML Co. Ltd.)
,
Kwon, Gayeung
(Department of Public Health Science (BK21 PLUS Program), Graduate School, Korea University)
,
Koh, Jong-Ho
(Department of Bio-Food Analysis and Processing, Bio-Campus Korea Polytechnic College)
,
Lim, Young-Hee
(Department of Integrated Biomedical and Life Sciences, College of Health Science, Korea University)
Probiotics, including Enterococcus faecium, confer a health benefit on the host. An Enterococcus strain was isolated from healthy chicken cecum, identified as E. faecium by 16S rDNA gene sequence analysis, and designated as E. faecium L11. To evaluate the potential of E. faecium L11 as a probiotic, ...
Probiotics, including Enterococcus faecium, confer a health benefit on the host. An Enterococcus strain was isolated from healthy chicken cecum, identified as E. faecium by 16S rDNA gene sequence analysis, and designated as E. faecium L11. To evaluate the potential of E. faecium L11 as a probiotic, the gastrointestinal tolerance, immunomodulatory activity, and lifespan extension properties of the strain were assayed. E. faecium L11 showed >66% and >62% survival in artificial gastric juice (0.3% pepsin, pH 2.5) and simulated small intestinal juice (0.5% bile salt and 0.1% pancreatin), respectively. Heat-killed E. faecium L11 significantly (p < 0.05) increased immune cell proliferation compared with controls, and stimulated the production of cytokines (IL-6 and $TNF-{\alpha}$) by activated macrophages obtained from ICR mice. In addition, E. faecium L11 showed a protective effect against Salmonella Typhimurium infection in Caenorhabditis elegans. In addition, feeding E. faecium L11 significantly (p < 0.05) extended the lifespan of C. elegans compared with the control. Furthermore, genes related to aging and host defense were upregulated in E. faecium L11-fed worms. In conclusion, E. faecium L11, which prolongs the lifespan of C. elegans, may be a potent probiotic supplement for livestock.
Probiotics, including Enterococcus faecium, confer a health benefit on the host. An Enterococcus strain was isolated from healthy chicken cecum, identified as E. faecium by 16S rDNA gene sequence analysis, and designated as E. faecium L11. To evaluate the potential of E. faecium L11 as a probiotic, the gastrointestinal tolerance, immunomodulatory activity, and lifespan extension properties of the strain were assayed. E. faecium L11 showed >66% and >62% survival in artificial gastric juice (0.3% pepsin, pH 2.5) and simulated small intestinal juice (0.5% bile salt and 0.1% pancreatin), respectively. Heat-killed E. faecium L11 significantly (p < 0.05) increased immune cell proliferation compared with controls, and stimulated the production of cytokines (IL-6 and $TNF-{\alpha}$) by activated macrophages obtained from ICR mice. In addition, E. faecium L11 showed a protective effect against Salmonella Typhimurium infection in Caenorhabditis elegans. In addition, feeding E. faecium L11 significantly (p < 0.05) extended the lifespan of C. elegans compared with the control. Furthermore, genes related to aging and host defense were upregulated in E. faecium L11-fed worms. In conclusion, E. faecium L11, which prolongs the lifespan of C. elegans, may be a potent probiotic supplement for livestock.
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제안 방법
The amplification conditions were 96℃ for 3 min, followed by 30 cycles of 94℃ for 1 min, annealing at 56℃ for 1 min, and extension at 72℃ for 2 min, and a final elongation at 72℃ for 7 min. Sequencing was performed using a BigDye Terminator cycle sequencing system (Applied Biosystems Inc., USA) and an ABI Prism 3730xl (Applied Biosystems Inc., USA). The sequence was deposited in GenBank under the accession number KM186186, and the Basic Local Alignment Search Tool was used to find homologous sequences.
After hatching, the worms were grown on E. coli OP50 until 3-day-old adult worms and then were allocated to a control group that was seeded with E. coli OP50 continually or a test group that was seeded with E. faecium L11 for 4 days. The 7-day-old worms were transferred to NGM agar plates with Salmonella Typhimurium lawns and incubated at 25℃.
To examine the beneficial effects of the newly isolated E. faecium L11 strain on resistance to pathogens, survival assays were performed. The lifespan of worms fed E.
대상 데이터
The scores of dead and live worms were checked every day [15]. Each test was assessed with at least 100 nematodes.
데이터처리
Statistical analysis was performed using SPSS ver. 10 (SPSS Inc., USA), and the difference between each sample and the control was evaluated for statistical significance using the Student's t-test.
이론/모형
faecium L11 (final concentrations of 1 × 106, 1 × 107, and 1 × 108 CFU/ml) at 37℃ for 48 h. Splenocyte stimulation was performed using the WST-1 assay.
The assays were carried out 3 times independently, and relative expression levels were estimated using the 2-ΔΔCT method [35].
, USA), and the difference between each sample and the control was evaluated for statistical significance using the Student's t-test. The C. elegans survival rate was estimated by the Kaplan-Meier method, and the significance of the differences in survival was evaluated using the log-rank test [36]. Differences were considered to be statistically significant when the p value was less than 0.
faecium L11. The proliferation of these cells was assessed using WST-1 test. The control was treated with PBS.
성능/효과
S. Typhimurium infection killed approximately 96% of C. elegans fed E. coli OP50 within 9 days after the worms were transferred to a Salmonella lawn plate, whereas 26% of worms fed E. faecium L11 survived (p < 0.001).
The results obtained in this study showed that E. faecium L11 induces primary macrophage maturation and is a potent inducer of IL-6 and TNF-α.
The lysozyme LYS-8 is under the control of dbl-1 and sma-3 [47]. In this study, worms fed E. faecium L11 had significantly increased resistance to S. Typhimurium compared with nematodes fed E. coli OP50. Expression levels of the pmk-1, sek-1, dbl-1, and sma-3 genes were significantly increased in worms fed E.
In conclusion, E. faecium L11 isolated from chicken cecum demonstrates digestive fluid tolerance, which is a prerequisite for probiotics. E.
Expression levels of genes related to lifespan extension, such as pmk-1, sek-1, dbl-1, sma-3, daf-2, and daf-16, were measured. The expression in C.
참고문헌 (50)
Wang Z, Burwinkel M, Chai W, Lange E, Blohm U, Breithaupt A, et al. 2014. Dietary Enterococcus faecium NCIMB 10415 and zinc oxide stimulate immune reactions to trivalent influenza vaccination in pigs but do not affect virological response upon challenge infection. PLoS One 9: e87007.
Tsai Y-T, Cheng P-C, Pan T-M. 2012. The immunomodulatory effects of lactic acid bacteria for improving immune functions and benefits. Appl. Microbiol. Biotechnol. 96: 853-862.
Signorini M, Soto L, Zbrun M, Sequeira G, Rosmini M, Frizzo L. 2012. Impact of probiotic administration on the health and fecal microbiota of young calves: a meta-analysis of randomized controlled trials of lactic acid bacteria. Res. Vet. Sci. 93: 250-258.
Salim H, Kang H, Akter N , Kim D , Kim J , Kim M, et al. 2013. Supplementation of direct-fed microbials as an alternative to antibiotic on growth performance, immune response, cecal microbial population, and ileal morphology of broiler chickens. Poult. Sci. 92: 2084-2090.
Cao G, Zeng X, Chen A, Zhou L, Zhang L , Xiao Y, et al. 2013. Effects of a probiotic, Enterococcus faecium, on growth performance, intestinal morphology, immune response, and cecal microflora in broiler chickens challenged with Escherichia coli K88. Poult. Sci. 92: 2949-2955.
Huang Y, Li Y, Huang Q , Cui Z , Yu D, Rajput IR, et al. 2012. Effect of orally administered Enterococcus faecium EF1 on intestinal cytokines and chemokines production of suckling piglets. Pak. Vet. J. 32: 81-84.
Scharek L, Guth J, Reiter K, Weyrauch K, Taras D, Schwerk P, et al. 2005. Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets. Vet. Immunol. Immunopathol. 105: 151-161.
Valeriano VD, Parungao-Balolong MM, Kang DK. 2014. In vitro evaluation of the mucin-adhesion ability and probiotic potential of Lactobacillus mucosae LM1. J. Appl. Microbiol. 117: 485-497.
Tellez G, Pixley C, Wolfenden R, Layton S , Hargis B. 2012. Probiotics/direct fed microbials for Salmonella control in poultry. Food Res. Int. 45: 628-633.
Lee J, Yun HS, Cho KW, Oh S, Kim SH, Chun T, et al. 2011. Evaluation of probiotic characteristics of newly isolated Lactobacillus spp.: immune modulation and longevity. Int. J. Food Microbiol. 148: 80-86.
Ye M, Sun L, Yang R, Wang Z, Qi K. 2017. The optimization of fermentation conditions for producing cellulase of Bacillus amyloliquefaciens and its application to goose feed. R. Soc. Open Sci. 4: 171012.
Messaoudi S, Kergourlay G, Rossero A, Ferchichi M, Prevost H, Drider D, et al. 2011. Identification of lactobacilli residing in chicken ceca with antagonism against Campylobacter. Int. Microbiol. 14: 103-110.
Anandharaj M, Sivasankari B. 2014. Isolation of potential probiotic Lactobacillus oris HMI68 from mother's milk with cholesterol-reducing property. J. Biosci. Bioeng. 118: 153-159.
Ricciardi A, Blaiotta G, Di Cerbo A, Succi M, Aponte M. 2014. Behaviour of lactic acid bacteria populations in Pecorino di Carmasciano cheese samples submitted to environmental conditions prevailing in the gastrointestinal tract: evaluation by means of a polyphasic approach. Int. J Food Microbiol. 179: 64-71.
Hwang JH, Yang HS, Ra KS, Park SS, Yu KW. 2013. Intestinal immune system-modulating activity through Peyer's patch of flavonoid glycoside purified from Citrus unshiu peel. J. Food Biochem. 37: 151-160.
Li N, Russell WM, Douglas-Escobar M, Hauser N, Lopez M, Neu J. 2009. Live and heat-killed Lactobacillus rhamnosus GG: effects on proinflammatory and anti-inflammatory cytokines/ chemokines in gastrostomy-fed infant rats. Pediatr. Res. 66: 203-207.
Thomas TP, Goonewardena SN, Majoros IJ, Kotlyar A, Cao Z, Leroueil PR, et al. 2011. Folate-targeted nanoparticles show efficacy in the treatment of inflammatory arthritis. Arthritis Rheum. 63: 2671-2680.
Stiernagle T. 1999. Maintenance of C. elegans, pp. 51-67. In Hope IA (ed.), C. elegans: A Practical Approach. Oxford University Press, New York.
Gruber J , Ng LF, Poovathingal S K, Halliwell B . 2009. Deceptively simple but simply deceptive- Caenorhabditis elegans lifespan studies: considerations for aging and antioxidant effects. FEBS Lett. 583: 3377-3387.
Burdine R D, Stern M J. 1996. Easy RNA isolation from C. elegans: a TRIZOL based method. Worm Breed. Gaz. 14: 10.
Greer E L, Dowlatshahi D , Banko MR, Villen J, Hoang K , Blanchard D, et al. 2007. An AMPK-FOXO pathway mediates longevity induced by a novel method of dietary restriction in C. elegans. Curr. Biol. 17: 1646-1656.
Untergasser A, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, et al. 2012. Primer3 - new capabilities and interfaces. Nucleic Acids Res. 40: e115.
Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the $2^{-{\Delta}{\Delta}CT}$ method. Methods 25: 402-408.
Zhao Y, Zhao L, Z heng X , Fu T, Guo H , Ren F. 2013. Lactobacillus salivarius strain FDB89 induced longevity in Caenorhabditis elegans by dietary restriction. J. Microbiol. 51: 183-188.
Joint FAO/WHO Working Group. 2002. Guidelines for the evaluation of probiotics in foods. Food and Agricultural Organization of the United Nations and World Health Organization FAO/WHO, London, Ontario, Canada.
Zhao T, Li Y, Liu B, Bronson RT, Halaweish I, Alam HB. 2014. Histone deacetylase III as a potential therapeutic target for the treatment of lethal sepsis. J. Trauma Acute Care Surg. 77: 913-919.
Renshaw M, Rockwell J, Engleman C, Gewirtz A, Katz J, Sambhara S. 2002. Cutting edge: impaired Toll-like receptor expression and function in aging. J. Immunol. 169: 4697-4701.
Radtke S, Wuller S, Yang X-P, Lippok BE, Mutze B, Mais C, et al. 2010. Cross-regulation of cytokine signalling: proinflammatory cytokines restrict IL-6 signalling through receptor internalisation and degradation. J. Cell Sci. 123: 947-959.
Niu X, Wang Y, Li W, Zhang H, Wang X, Mu Q, et al. 2015. Esculin exhibited anti-inflammatory activities in vivo and regulated TNF- ${\alpha}$ and IL-6 production in LPS-stimulated mouse peritoneal macrophages in vitro through MAPK pathway. Int. Immunopharmacol. 29: 779-786.
Engelmann I, Pujol N. 2010. Innate immunity in C. elegans, pp. 105-121. In Soderhall K (ed), Invertebrate Immunity. Advances in Experimental Medicine and Biology, Vol. 708. Springer, Boston, MA.
Araujo TF, Ferreira CLDLF. 2013. The genus Enterococcus as probiotic: safety concerns. Braz. Arch. Biol. Technol. 56: 457-466.
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