期刊论文详细信息
Gut Pathogens
Metagenomic profile of gut microbiota in children during cholera and recovery
Munirul Alam2  Tetsuya Iida4  Sk Imran Ali2  Toshihiro Horii4  Takaaki Nakaya3  Nur Haque Alam2  Haruo Watanabe1  Kaori Izutsu4  Kazuyoshi Gotoh4  Shota Nakamura4  Shirajum Monira2 
[1] National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan;International Center for Diarrhoeal Disease Research, Bangladesh, 68 Shahid Tajjudin Ahmed, Sarani, Mohakhali, Dhaka 1212, Bangladesh;Department of Infectious Diseases, Kyoto Prefectural University of Medicine, 465 Kawaramachi-hirokoji, Kamigyo-ku, Kyoto, Japan;Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan
关键词: Children;    16S rDNA;    Gut;    Microbiota;    Cholera;   
Others  :  821745
DOI  :  10.1186/1757-4749-5-1
 received in 2013-01-03, accepted in 2013-01-21,  发布年份 2013
【 摘 要 】

Background

The diverse bacterial communities colonizing the gut (gastrointestinal tract) of infants as commensal flora, which play an important role in nutrient absorption and determining the state of health, are known to alter due to diarrhea.

Method

Bacterial community dynamics in children suffering from cholera and during recovery period were examined in the present study by employing metagenomic tool, followed by DNA sequencing and analysis. For this, bacterial community DNA was extracted from fecal samples of nine clinically confirmed cholera children (age 2–3 years) at day 0 (acute cholera), day 2 (antibiotic therapy), day 7 and, and day 28, and the variable region of 16S rRNA genes were amplified by universal primer PCR.

Results

454 parallel sequencing of the amplified DNA followed by similarity search of the sequenced data against an rRNA database allowed us to identify V. cholerae, the cause of cholera, in all nine children at day 0, and as predominant species in six children, accounting for 35% of the total gut microbiota on an average in all the nine children. The relative abundance (mean ± sem %) of bacteria belonging to phyla Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, was 55 ± 7, 18 ± 4, 13 ± 4, and 8 ± 4, respectively, at day 0, while these values were 12 ± 4, 43 ± 4, 33 ± 3, and 12 ± 2, respectively, at day 28. As antibiotic therapy began, V. cholerae count declined significantly (p< 0.001) and was found only in four children at day 2 and two children at day 7 with the relative abundance of 3.7% and 0.01%, respectively, which continued up to day 28 in the two children. Compared to acute cholera condition (day 0), the relative abundance of Escherichia coli, Enterococcus, and Veillonella increased at day 2 (antibiotic therapy) while Bifidobacterium, Bacteroides, and Ruminococcus decreased.

Conclusion

Cholera results expulsion of major commensal bacteria of phyla Bacteroidetes, Firmicutes, and Actinobacteria, and increase of harmful Proteobacteria to colonize the gut during acute and convalescence states. The observed microbiota disruption might explain the prevalent malnutrition in children of Bangladesh where diarrheal diseases are endemic.

【 授权许可】

   
2013 Monira et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Mai V, Draganov PV: Recent advances and remaining gaps in our knowledge of associations between gut microbiota and human health. World J Gastroenterol 2009, 15:81-85.
  • [2]Yan F, Polk DB: Commensal bacteria in the gut: learning who our friends are. Curr Opin Gastroenterol 2004, 20:565-571.
  • [3]Jumpertz R, Le DS, Turnbaugh PJ, Trinidad C, Bogardus C, Gordon JI, Krakoff J: Energy-balance studies reveal associations between gut microbes, caloric load, and nutrient absorption in humans. Am J Clin Nutr 2011, 94:58-65.
  • [4]Monira S, Alam NH, Suau A, Magne F, Nair GB, Karmakar PC, Rahman M, Pochart P, Desjeux JF: Time course of bacterial diversity in stool samples of malnourished children with cholera receiving treatment. J Pediatr Gastroenterol Nutr 2009, 48:571-578.
  • [5]Hopkins MJ, Sharp R, Macfarlane GT: Age and disease related changes in intestinal bacterial populations assessed by cell culture, 16S rRNA abundance, and community cellular fatty acid profiles. Gut 2001, 48:198-205.
  • [6]Gorbach SL, Banwell JG, Jacobs B, Chatterjee BD, Mitra R, Brigham KL, Neogy KN: Intestinal microflora in Asiatic cholera. I. “Rice-water” stool. J Infect Dis 1970, 121:32-37.
  • [7]Gorbach SL, Mahalanabis D, Brayton J, Jacobs B, Chatterjee BD, Neogy KN: Intestinal microflora in Asiatic cholera. 3. Studies in pediatric cholera. J Infect Dis 1970, 121:46-47.
  • [8]Albert MJ, Bhat P, Rajan D, Maiya PP, Pereira SM, Baker SJ: Faecal flora of south indian infants and young children in health and with acute gastroenteritis. J Med Microbiol 1978, 11:137-143.
  • [9]Tazume S, Takeshi K, Saidi SM, Ichoroh CG, Mutua WR, Waiyaki PG, Ozawa A: Ecological studies on intestinal microbial flora of Kenyan children with diarrhoea. J Trop Med Hyg 1990, 93:215-221.
  • [10]Tannock GW: Molecular methods for exploring the intestinal ecosystem. Br J Nutr 2002, 87:S199-S201.
  • [11]Suau A, Bonnet R, Sutren M, Godon JJ, Gibson GR, Collins MD, Dore J: Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut. Appl Environ Microbiol 1999, 65:4799-4807.
  • [12]Nakamura S, Maeda N, Miron IM, Yoh M, Izutsu K, Kataoka C, Honda T, Yasunaga T, Nakaya T, Kawai J: Metagenomic diagnosis of bacterial infections. Emerg Infect Dis 2008, 14:1784-1786.
  • [13]Monira S, Nakamura S, Gotoh K, Izutsu K, Watanabe H, Alam NH, Endtz HP, Cravioto A, Ali SI, Nakaya T: Gut microbiota of healthy and malnourished children in Bangladesh. Front Microbiol 2011, 2:228.
  • [14]Cohen R, Kalser MH, Arteaga I, Yawn E, Frazier D, Leite CA, Ahearn DG, Roth F: Microbial intestinal flora in acute diarrheal disease. JAMA 1967, 201:835-840.
  • [15]Ley RE, Backhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI: Obesity alters gut microbial ecology. Proc Natl Acad Sci U S A 2005, 102:11070-11075.
  • [16]Ley RE, Turnbaugh PJ, Klein S, Gordon JI: Microbial ecology: human gut microbes associated with obesity. Nature 2006, 444:1022-1023.
  • [17]Balamurugan R, Janardhan HP, George S, Raghava MV, Muliyil J, Ramakrishna BS: Molecular studies of fecal anaerobic commensal bacteria in acute diarrhea in children. J Pediatr Gastroenterol Nutr 2008, 46:514-519.
  • [18]Fujita K, Kaku M, Yanagase Y, Ezaki T, Furuse K, Ozawa A, Saidi SM, Sang WK, Waiyaki PG: Physicochemical characteristics and flora of diarrhoeal and recovery faeces in children with acute gastro-enteritis in Kenya. Ann Trop Paediatr 1990, 10:339-345.
  • [19]Sack DA, Sack RB, Nair GB, Siddique AK: Cholera. Lancet 2004, 363:223-233.
  • [20]Saha D, Karim MM, Khan WA, Ahmed S, Salam MA, Bennish ML: Single-dose azithromycin for the treatment of cholera in adults. N Engl J Med 2006, 354:2452-2462.
  • [21]Lindenbaum J, Greenough WB, Islam MR: Antibiotic therapy of cholera. Bull World Health Organ 1967, 36:871-883.
  • [22]Berg RD: Promotion of the translocation of enteric bacteria from the gastrointestinal tracts of mice by oral treatment with penicillin, clindamycin, or metronidazole. Infect Immun 1981, 33:854-861.
  • [23]Savage DC, Dubos R: Alterations in the mouse cecum and its flora produced by antibacterial drugs. J Exp Med 1968, 128:97-110.
  • [24]Ubeda C, Taur Y, Jenq RR, Equinda MJ, Son T, Samstein M, Viale A, Socci ND, van den Brink MR, Kamboj M, Pamer EG: Vancomycin-resistant Enterococcus domination of intestinal microbiota is enabled by antibiotic treatment in mice and precedes bloodstream invasion in humans. J Clin Invest 2010, 120:4332-4341.
  • [25]Looft T, Johnson TA, Allen HK, Bayles DO, Alt DP, Stedtfeld RD, Sul WJ, Stedtfeld TM, Chai B, Cole JR: In-feed antibiotic effects on the swine intestinal microbiome. Proc Natl Acad Sci U S A 2012, 109:1691-1696.
  • [26]Broderick NA, Raffa KF, Goodman RM, Handelsman J: Census of the bacterial community of the gypsy moth larval midgut by using culturing and culture-independent methods. Appl Environ Microbiol 2004, 70:293-300.
  • [27]Flint HJ, Duncan SH, Scott KP, Louis P: Interactions and competition within the microbial community of the human colon: links between diet and health. Environ Microbiol 2007, 9:1101-1111.
  • [28]Turnbaugh PJ, Ridaura VK, Faith JJ, Rey FE, Knight R, Gordon JI: The effect of diet on the human gut microbiome: a metagenomic analysis in humanized gnotobiotic mice. Sci Transl Med 2009, 1:6ra14.
  • [29]Harmsen HJ, Wildeboer-Veloo AC, Raangs GC, Wagendorp AA, Klijn N, Bindels JG, Welling GW: Analysis of intestinal flora development in breast-fed and formula-fed infants by using molecular identification and detection methods. J Pediatr Gastroenterol Nutr 2000, 30:61-67.
  • [30]Marteau P, Lepage P, Mangin I, Suau A, Dore J, Pochart P, Seksik P: Review article: gut flora and inflammatory bowel disease. Aliment Pharmacol Ther 2004, 20(Suppl 4):18-23.
  • [31]Kau AL, Ahern PP, Griffin NW, Goodman AL, Gordon JI: Human nutrition, the gut microbiome and the immune system. Nature 2011, 474:327-336.
  • [32]Monira S, Hoq MM, Chowdhury AK, Suau A, Magne F, Endtz HP, Alam M, Rahman M, Pochart P, Desjeux JF, Alam NH: Short-chain fatty acids and commensal microbiota in the faeces of severely malnourished children with cholera rehydrated with three different carbohydrates. Eur J Clin Nutr 2010, 64:1116-1124.
  • [33]Magne F, Abely M, Boyer F, Morville P, Pochart P, Suau A: Low species diversity and high interindividual variability in faeces of preterm infants as revealed by sequences of 16S rRNA genes and PCR-temporal temperature gradient gel electrophoresis profiles. FEMS Microbiol Ecol 2006, 57:128-138.
  • [34]Andersson AF, Lindberg M, Jakobsson H, Backhed F, Nyren P, Engstrand L: Comparative analysis of human gut microbiota by barcoded pyrosequencing. PLoS One 2008, 3:e2836.
  • [35]Thompson JR, Marcelino LA, Polz MF: Heteroduplexes in mixed-template amplifications: formation, consequence and elimination by ‘reconditioning PCR’. Nucleic Acids Res 2002, 30:2083-2088.
  • [36]Goto N, Prins P, Nakao M, Bonnal R, Aerts J, Katayama T: BioRuby: bioinformatics software for the Ruby programming language. Bioinformatics 2010, 26:2617-2619.
  • [37]Federhen S: The NCBI Taxonomy database. Nucleic Acids Res 2012, 40:D136-D143.
  • [38]Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J, Glockner FO: SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res 2007, 35:7188-7196.
  • [39]Sun Y, Cai Y, Liu L, Yu F, Farrell ML, McKendree W, Farmerie W: ESPRIT: estimating species richness using large collections of 16S rRNA pyrosequences. Nucleic Acids Res 2009, 37:e76.
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