In this work, we wanted to develop a probiotic from famous longevity villages in Korea. We visited eight longevity villages in Korea to collect fecal samples from healthy adults who were aged above 80 years and had regular bowel movements, and isolated lactic-acid-producing bacteria from the samples...
In this work, we wanted to develop a probiotic from famous longevity villages in Korea. We visited eight longevity villages in Korea to collect fecal samples from healthy adults who were aged above 80 years and had regular bowel movements, and isolated lactic-acid-producing bacteria from the samples. Isolated colonies that appeared on MRS agar containing bromophenol blue were identified by means of 16S rRNA sequencing, and 102 of the isolates were identified as lactic-acid-producing bacteria (18 species). Lactobacillus fermentum was the most frequently found species. Eight isolates were selected on the basis of their ability to inhibit the growth of six intestinal pathogens (Escherichia coli O157:H7, Salmonella enterica subsp. enterica Typhimurium, Salmonella enterica subsp. enterica Enteritidis, Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes) and their susceptibility to 15 antimicrobial agents. Among these eight isolates, four Lactobacillus fermentum isolates were found not to produce any harmful enzymes or metabolites. Among them, Lactobacillus fermentum isolate no. 24 showed the strongest binding to intestinal epithelial cells, the highest immune-enhancing activity, anti-inflammation activity, and anti-oxidation activity as well as the highest survival rates in the presence of artificial gastric juice and bile solution. This isolate, designated Lactobacillus fermentum PL9988, has all the characteristics for a good probiotic.
In this work, we wanted to develop a probiotic from famous longevity villages in Korea. We visited eight longevity villages in Korea to collect fecal samples from healthy adults who were aged above 80 years and had regular bowel movements, and isolated lactic-acid-producing bacteria from the samples. Isolated colonies that appeared on MRS agar containing bromophenol blue were identified by means of 16S rRNA sequencing, and 102 of the isolates were identified as lactic-acid-producing bacteria (18 species). Lactobacillus fermentum was the most frequently found species. Eight isolates were selected on the basis of their ability to inhibit the growth of six intestinal pathogens (Escherichia coli O157:H7, Salmonella enterica subsp. enterica Typhimurium, Salmonella enterica subsp. enterica Enteritidis, Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes) and their susceptibility to 15 antimicrobial agents. Among these eight isolates, four Lactobacillus fermentum isolates were found not to produce any harmful enzymes or metabolites. Among them, Lactobacillus fermentum isolate no. 24 showed the strongest binding to intestinal epithelial cells, the highest immune-enhancing activity, anti-inflammation activity, and anti-oxidation activity as well as the highest survival rates in the presence of artificial gastric juice and bile solution. This isolate, designated Lactobacillus fermentum PL9988, has all the characteristics for a good probiotic.
Eight villages were selected for this study. Each village had at least more than 200 residents and higher percentages of seniors aged over 80 years.
Eight villages with more than 200 residents each were selected from the three Korean longevity counties, Suncheon, Jangsoo, and Kurea in Chulla Province. From 69 healthy adults who were aged above 80 years and had regular bowel movement, fecal samples were obtained and transferred to the laboratory on ice within 6 h of collection.
Eight villages with more than 200 residents each were selected from the three Korean longevity counties, Suncheon, Jangsoo, and Kurea in Chulla Province. From 69 healthy adults who were aged above 80 years and had regular bowel movement, fecal samples were obtained and transferred to the laboratory on ice within 6 h of collection. As soon as the samples arrived in the laboratory, they were diluted with sterile saline and inoculated on de Man Rogosa and Sharpe medium (BD, Sparks, MD, USA) containing 0.
이론/모형
Resistance to artificial gastric juice was assayed by the method of Cho et al. [6] with slight modification.
colonies that were gram-positive, catalase negative, and rod shaped were further identified by means of 16S rRNA sequencing by Bionics (Seoul, Korea). The generated sequences were compared with known sequences on the EzTaxon-e server (http://www.ezbiocloud.net/eztaxon) [21] using the BLAST algorithm at the National Center for Biotechnology Information Web server (http://www.ncbi.nlm.nih.gov).
The resistance of the LAB to superoxide anions induced by paraquat (1,1’-dimethyl-4,4;-bipyridinium dichloride) was determined by means of the diffusion assay method [22]. Overnight-cultured LAB cells were collected by centrifugation and dispersed in saline at a concentration of 107 CFU/ml.
The susceptibilities of LAB to antimicrobials were assayed by means of the liquid dilution method according to the ISO guideline [13]. The minimum inhibitory concentrations (MICs) of the following 15 antimicrobials were determined: ampicillin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, kanamycin, linezolid, neomycin, rifampicin, streptomycin, a quinupristin and dalfopristin combination (Synercid), tetracycline, trimethoprim, and vancomycin.
성능/효과
Several different colonies were collected from some people, while some had only one or no colony. A total 102 LAB were isolated from 69 people and these were identified as 18 LAB species by means of 16S rRNA sequencing (Table 1). L.
Each village had at least more than 200 residents and higher percentages of seniors aged over 80 years. A total of 102 fecal samples were collected from seniors aged over 80 years with regular bowl movement. Every colony with a different morphology on MRS-BPB agar plate was collected.
2013-0086153. The 16S rRNA sequence was deposited in GenBank (acc. no.
People used to select LAB that appeared on media containing antimicrobials. The MIC assay showed that only eight isolates (11.6%) among 69 LAB were susceptible to gentamicin, erythromycin, and clindamycin according to CLSI criteria and also susceptible to clindamycin, chloramphenicol, and ampicillin according to EUCAST criteria (Table 2). The high percentages of antimicrobialresistant LAB in these villages were not expected.
참고문헌 (47)
An SJ, Pae HO, Oh GS, Choi BM, Jeong S, Jang SI, et al. 2002. Inhibition of TNF-α, IL-1β, and IL-6 productions and NF-κB activation in lipopolysaccharide-activated RAW 264.7 macrophages by catalposide, an iridoid glycoside isolated from Catalpa ovata G. Don (Bignoniaceae). Int. Immunopharmacol. 2: 1173-1181.
Baccigalupi L, Di DA, Parlato M, Luongo D, Carbone V, Rossi M, et al. 2005. Small surface-associated factors mediate adhesion of a food-isolated strain of Lactobacillus fermentum to Caco-2 cells. Res. Microbiol. 156: 830-836.
Bruce-Keller AJ, Salbaum JM, Luo M, Blanchard E, Taylor CM, Welsh DA, Berthoud HR. 2015. Obese-type gut microbiota induce neurobehavioral changes in the absence of obesity. Biol. Psychiatry 77: 607-615.
Chadwick RW, George SE, Clawton LD. 1992. Role of the gastrointestinal mucosa and microflora in the bioactivation of dietary and environmental mutagens or carcinogens. Drug Metabol. Rev. 24: 425-492.
Cho IJ, Lee NK, Hahm YT. 2009. Characterization of Lactobacillus spp. isolated from the feces of breast-feeding piglets. J. Biosci. Bioeng. 108: 194-198.
D’Aimmo MR, Modesto M, Biavati B. 2007. Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp. isolated from dairy and pharmaceutical products. Int. J. Food Microbiol. 115: 35-42.
European Committee on Antimicrobial Susceptibility Testing (EUCAST). 2015. Clinical breakpoints. Available at http://www.eucast.org/clinical_breakpoints.
Foligné B, Grangette C, Pot B. 2005. Probiotics in IBD: mucosal and systemic routes of administration may promote similar effects. Gut 54: 727-728.
Haza AI, Zabala A, Morales P. 2004. Protective effect and cytokine production of a Lactobacillus plantarum strain isolated from ewes’ milk cheese. Int. Dairy J. 14: 29-38.
International Standard Organization. 2010. Milk and milk products ― Determination of the minimal inhibitory concentration (MIC) of antibiotics applicable to bifidobacteria and non-enterococcal lactic acid bacteria (LAB). ISO 10932:2010 (IDF 223:2010). Geneva, Switzerland.
Isenberg HD. 1992. Gelatin liquefaction test, pp. 1.19.42-1.19.43. Clinical Microbiology Procedures Handbook, Vol. 1. American Society of Microbiology, Washington, DC.
Isenberg HD. 1992. Hemolysis on sheep blood agar, pp. 1.20.16-1.20.17. Clinical Microbiology Procedures Handbook. Vol. 1. American Society of Microbiology, Washington D.C.
Isenberg HD. 1992. Indole test, pp. 1.19.13-1.19.15. Clinical Microbiology Procedures Handbook, Vol. 1. American Society of Microbiology, Washington DC.
Isenberg HD. 1992. Urease, pp. 2.6.8. Clinical Microbiology Procedures Handbook, Vol. 1. American Society of Microbiology, Washington, DC.
Ishibashi N, Yamazaki S. 2001. Probiotics and safety. Am. J. Clin. Nutr. 73: 465S-470S.
Khoruts A, Dicksved J, Jansson JK, Sadowsky MJ. 2010. Changes in the composition of the human fecal microbiome after bacteriotherapy for recurrent Clostridium difficile-associated diarrhea. J. Clin. Gastroenter. 44: 354-360.
Khoruts A, Sadowsky MJ, Hamilton MJ. 2015. Development of fecal microbiota transplantation suitable for mainstream medicine. Clin. Gastroenterol. Hepatol. 13: 246-250.
Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, et al. 2012. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int. J. Syst. Evol. Microbiol. 62: 716-721.
Kullisaar T, Zilmer M, Mikelsaar M, Vihalemm T, Annuk H, Kairane C, Kilk A. 2002. Two antioxidative lactobacilli strains as promising probiotics. Int. J. Food Microbiol. 72: 215-224.
Lee HY, Park JH, Seok SH, Baek MW, Kim DJ, Lee KE, et al. 2006. Human originated bacteria, Lactobacillus rhamnosus PL60, produce conjugated linoleic acid and show antiobesity effects in diet-induced obese mice. Biochim. Biophys. Acta 1761: 736-744.
Lee K, Paek K, Lee HY, Park JH, Lee Y. 2007. Antiobesity effect of trans-10, cis-12-conjugated linoleic acid-producing Lactobacillus plantarum PL62 on diet-induced obese mice. J. Appl. Microbiol. 103: 1140-1146.
Mathara JM, Schillinger U, Guigas C, Franz C, Kutima PM, Mbugua SK, et al. 2008. Functional characteristics of Lactobacillus spp. from traditional Maasai fermented milk products in Kenya. Int. J. Food Microbiol. 126: 57-64.
Nam H, Ha M, Bae O, Lee Y. 2002. Effect of Weissella confusa strain PL9001 on the adherence and growth of Helicobacter pylori. Appl. Environ. Microbiol. 9: 4642-4645.
Park JH, Lee Y, Moon E, Seok SH, Baek MW, Lee HY, et al. 2005. Safety assessment of Lactobacillus fermentum PL9005, a potential probiotic lactic acid bacterium, in mice. J. Microbiol. Biotechnol. 15: 603-608.
Park JH, Lee Y, Moon E, Seok SH, Cho SA, Baek MW, et al. 2005. Immunoenhancing effects of a new probiotic strain, Lactobacillus fermentum PL9005. J. Food Prot. 68: 571-576.
Song YL, Kato N, Matsumiya Y, Liu C, Kato H, Watanabe K. 1999. Identification of and hydrogen peroxide production by fecal and vaginal lactobacilli isolated from Japanese women and newborn infants. J. Clin. Microbiol. 37: 3062-3064.
Thumu SC, Halami PM. 2012. Presence of erythromycin and tetracycline resistance genes in lactic acid bacteria from fermented foods of Indian origin. Antonie Van Leeuwenhoek 102: 541-551.
Warren HS, Fitting C, Hoff E, Adib-Conquy M, Beasley-Topliffe L, Tesini B, et al. 2010. Resilience to bacterial infection: difference between species could be due to proteins in serum. J. Infect. Dis. 201: 223-232.
Wikler M, Clinical and Laboratory Standards Institute. 2007. Performance standards for antimicrobial susceptibility testing: seventeenth informational supplement. Clinical and Laboratory Standards Institute, Wayne, PA.
Yatsunenko T, Rey FE, Manary MJ, Trehan I, Dominguez-Bello MG, Contreras M, et al. 2012. Human gut microbiome viewed across age and geography. Nature 486: 222-227.
Yoon SB, Lee YJ, Park SK, Kim HC, Bae H, Kim HM, et al. 2009. Anti-inflammatory effects of Scutellaria baicalensis water extract on LPS-activated RAW264.7 macrophages. J. Ethnopharmacol. 125: 286-290.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.