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Multilocus sequence typing breathes life into a microbial metagenome
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Mahenthiralingam, Eshwar, Baldwin, Adam, Drevinek, Pavel, Vanlaere, Elke, Vandamme, Peter, LiPuma, John J. and Dowson, Christopher G.. (2006) Multilocus sequence typing breathes life into a microbial metagenome. PLoS One, Vol.1 (No.1). ISSN 1932-6203
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Official URL: http://dx.doi.org/10.1371/journal.pone.0000017
Abstract
Shot-gun sequencing of DNA isolated from the environment and the assembly of metagenomes from the resulting data has considerably advanced the study of microbial diversity. However, the subsequent matching of these hypothetical metagenomes to cultivable microorganisms is a limitation of such cultivation-independent methods of population analysis. Using a nucleotide sequence-based genetic typing method, multilocus sequence typing, we were able for the first time to match clonal cultivable isolates to a published and controversial bacterial metagenome, Burkholderia SAR-1, which derived from analysis of the Sargasso Sea. The matching cultivable isolates were all associated with infection and geographically widely distributed; taxonomic analysis demonstrated they were members of Burkholderia cepacia complex Group K. Comparison of the Burkholderia SAR-1 metagenome to closely related B. cepacia complex genomes indicated that it was greater than 98% intact in terms of conserved genes, and it also shared complete sequence identity with the cultivable isolates at random loci beyond the genes sampled by the multilocus sequence typing. Two features of the extant cultivable clones support the argument that the Burkholderia SAR-1 sequence may have been a contaminant in the original metagenomic survey: (i) their growth in conditions reflective of sea water was poor, suggesting the ocean was not their preferred habitat, and (ii) several of the matching isolates were epidemiologically linked to outbreaks of infection that resulted from contaminated medical devices or products, indicating an adaptive fitness of this bacterial strain towards contamination-associated environments. The ability to match identical cultivable strains of bacteria to a hypothetical metagenome is a unique feature of nucleotide sequence-based microbial typing methods; such matching would not have been possible with more traditional methods of genetic typing, such as those based on pattern matching of genomic restriction fragments or amplified DNA fragments. Overall, we have taken the first steps in moving the status of the Burkholderia SAR-1 metagenome from a hypothetical entity towards the basis for life of cultivable strains that may now be analysed in conjunction with the assembled metagenomic sequence data by the wider scientific community.
| Item Type: | Journal Article |
|---|---|
| Subjects: | Q Science > QR Microbiology |
| Divisions: | Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) |
| Library of Congress Subject Headings (LCSH): | Nucleotide sequence, Gene mapping, Marine microbiology |
| Journal or Publication Title: | PLoS One |
| Publisher: | Public Library of Science |
| ISSN: | 1932-6203 |
| Date: | 20 December 2006 |
| Volume: | Vol.1 |
| Number: | No.1 |
| Number of Pages: | 8 |
| Identification Number: | 10.1371/journal.pone.0000017 |
| Status: | Peer Reviewed |
| Publication Status: | Published |
| Access rights to Published version: | Open Access |
| Funder: | Natural Environment Research Council (Great Britain) (NERC), Wellcome Trust (London, England), Stichting Kapteinfonds, Nederlandse Cystic Fibrosis Stichting, Cystic Fibrosis Foundation |
| Grant number: | 072853 (WT), 075586 (WT), NER/T/S/2001/0299 (NERC) |
| References: | 1. Handelsman J (2004) Metagenomics: application of genomics to uncultured microorganisms. Microbiol Mol Biol Rev 68: 669–685. 2. DeLong EF (2005) Microbial community genomics in the ocean. Nat Rev Microbiol 3: 459–469. 3. Venter JC, Remington K, Heidelberg JF, Halpern AL, Rusch D, et al. (2004) Environmental genome shotgun sequencing of the Sargasso Sea. Science 304: 66–74. 4. Coenye T, Vandamme P (2003) Diversity and significance of Burkholderia species occupying diverse ecological niches. Environ Microbiol 5: 719–729. 5. Leff LG, Kernan RM, McArthur JV, Shimkets LJ (1995) Identification of aquatic Burkholderia (Pseudomonas) cepacia by hybridization with species-specific rRNA gene probes. Appl Environ Microbiol 61: 1634–1636. 6. O’Sullivan L A, Mahenthiralingam E (2005) Biotechnological potential within the genus Burkholderia. Lett Appl Microbiol 41: 8–11. 7. Mahenthiralingam E, Urban TA, Goldberg JB (2005) The multifarious, multireplicon Burkholderia cepacia complex. Nat Rev Microbiol 3: 144–156. 8. Holden MT, Titball RW, Peacock SJ, Cerdeno-Tarraga AM, Atkins T, et al. (2004) Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei. Proc Natl Acad Sci U S A 101: 14240–14245. 9. Coenye T, Vandamme P, Govan JR, LiPuma JJ (2001) Taxonomy and identification of the Burkholderia cepacia complex. J Clin Microbiol 39: 3427–3436. 10. Mahenthiralingam E, Campbell ME, Henry DA, Speert DP (1996) Epidemiology of Burkholderia cepacia infection in patients with cystic fibrosis: Analysis by randomly amplified polymorphic DNA fingerprinting. J Clin Microbiol 34: 2914–2920. 11. Baldwin A, Mahenthiralingam E, Thickett KM, Honeybourne D, Maiden MC, et al. (2005) Multilocus sequence typing scheme that provides both species and strain differentiation for the Burkholderia cepacia complex. J Clin Microbiol 43: 4665–4673. 12. Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, et al. (1998) Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci U S A 95: 3140–3145. 13. Enright MC, Spratt BG (1999) Multilocus sequence typing. Trends Microbiol 7: 482–487. 14. LiPuma JJ, Spilker T, Gill LH, Campbell PW 3rd, Liu L, et al. (2001) Disproportionate distribution of Burkholderia cepacia complex species and transmissibility markers in cystic fibrosis. Am J Respir Crit Care Med 164: 92–96. 15. Mahenthiralingam E, Bischof J, Byrne SK, Radomski C, Davies JE, et al. (2000) DNA-Based diagnostic approaches for identification of Burkholderia cepacia complex, Burkholderia vietnamiensis, Burkholderia multivorans, Burkholderia stabilis, and Burkholderia cepacia genomovars I and III. J Clin Microbiol 38: 3165–3173. 16. Payne GW, Vandamme P, Morgan SH, LiPuma JJ, Coenye T, et al. (2005) Development of a recA Gene-Based Identification Approach for the Entire Burkholderia Genus. Appl Environ Microbiol 71: 3917–3927. 17. Vandamme P, Mahenthiralingam E, Holmes B, Coenye T, Hoste B, et al. (2000) Identification and population structure of Burkholderia stabilis sp nov (formerly Burkholderia cepacia genomovar IV). J Clin Microbiol 38: 1042–1047. 18. Vermis K, Coenye T, Mahenthiralingam E, Nelis HJ, Vandamme P (2002) Evaluation of species-specific recA-based PCR tests for genomovar level identification within the Burkholderia cepacia complex. J Med Microbiol 51: 937–940. 19. Anonymous (2004) Notice to readers: manufacturer’s recall of nasal spray contaminated with Burkholderia cepacia complex. Morbid Mortality Weekly Repor t : ht t p : / /www.cdc .gov/mmwr/pr e view/mmwrhtml/ mm5311a5318.htm.. 20. Souza AV, Moreira CR, Pasternak J, Hirata Mde L, Saltini DA, et al. (2004) Characterizing uncommon Burkholderia cepacia complex isolates from an outbreak in a haemodialysis unit. J Med Microbiol 53: 999–1005. 21. Lewis DA, Jones A, Parkhill J, Speert DP, Govan JR, et al. (2005) Identification of DNA Markers for a Transmissible Pseudomonas aeruginosa Cystic Fibrosis Strain. Am J Respir Cell Mol Biol 33: 56–64. 22. Mahenthiralingam E, Coenye T, Chung JW, Speert DP, Govan JR, et al. (2000) Diagnostically and experimentally useful panel of strains from the Burkholderia cepacia complex. J Clin Microbiol 38: 910–913. 23. Stanier RY, Palleroni NJ, Doudoroff M (1966) The aerobic pseudomonads: a taxonomic study. J Gen Microbiol 43: 159–271. 24. Wolfgang MC, Kulasekara BR, Liang X, Boyd D, Wu K, et al. (2003) Conservation of genome content and virulence determinants among clinical and environmental isolates of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 100: 8484–8489. 25. Kimata N, Nishino T, Suzuki S, Kogure K (2004) Pseudomonas aeruginosa isolated from marine environments in Tokyo Bay. Microb Ecol 47: 41–47. 26. Anderson RL, Vess RW, Carr JH, Bond WW, Panlilio AL, et al. (1991) Investigations of intrinsic Pseudomonas cepacia contamination in commercially manufactured povidone-iodine. Infect Control Hosp Epidemiol 12: 297–302. 27. Perry B (2001) Cometic microbiology. Microbiology Today 28: 185–187. 28. Shiraki M, Shimada T, Tatsumichi M, Saito T (1995) Purification and characterization of extracellular poly(3-hydroxybutyrate) depolymerases. J Polymers & Environ 3: 13–21. 29. Hicks CL, Kinoshita R, Ladds PW (2000) Pathology of melioidosis in captive marine mammals. Aust Vet J 78: 193–195. 30. Baldwin A, Sokol PA, Parkhill J, Mahenthiralingam E (2004) The Burkholderia cepacia Epidemic Strain Marker Is Part of a Novel Genomic Island Encoding Both Virulence and Metabolism-Associated Genes in Burkholderia cenocepacia. Infect Immun 72: 1537–1547. 31. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor, N.Y.: Cold Spring Harbor Press. 32. Carver TJ, Rutherford KM, Berriman M, Rajandream MA, Barrell BG, et al. (2005) ACT: the Artemis Comparison Tool. Bioinformatics 21: 3422–3423. 33. Hareland WA, Crawford RL, Chapman PJ, Dagley S (1975) Metabolic function and properties of 4-hydroxyphenylacetic acid 1-hydroxylase from Pseudomonas acidovorans. J Bacteriol 121: 272–285. 34. Berriatua E, Ziluaga I, Miguel-Virto C, Uribarren P, Juste R, et al. (2001) Outbreak of Subclinical Mastitis in a Flock of Dairy Sheep Associated with Burkholderia cepacia Complex Infection. J Clin Microbiol 39: 990–994. 35. De Boeck K, Malfroot A, Van Schil L, Lebecque P, Knoop C, et al. (2004) Epidemiology of Burkholderia cepacia complex colonisation in cystic fibrosis patients. Eur Respir J 23: 851–856. 36. Campana S, Taccetti G, Ravenni N, Favari F, Cariani L, et al. (2005) Transmission of Burkholderia cepacia complex: evidence for new epidemic clones infecting cystic fibrosis patients in Italy. J Clin Microbiol 43: 5136–5142. |
| URI: | http://wrap.warwick.ac.uk/id/eprint/32621 |
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