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PMID: 18797955 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular analysis of microbiota along the digestive tract of juvenile Atlantic salmon (Salmo salar L.).

Microbial ecology ·Vol. 57 ·No. 3 ·2009-04-00 ·Pages 550-61

Navarrete P, Espejo RT, Romero J

Abstract

Dominant bacterial microbiota of the gut of juvenile farmed Atlantic salmon was investigated using a combination of molecular approaches. Bacterial community composition from the stomach, the pyloric caeca, and the intestine was assessed by extracting DNA directly from each gut compartment. Temporal temperature gradient gel electrophoresis (TTGE) analysis of 16S ribosomal DNA (rDNA) amplicons showed very similar bacterial compositions throughout the digestive tract. Band sequencing revealed a narrow diversity of species with a dominance of Pseudomonas in the three compartments. However, cloning revealed more diversity among the Pseudomonas sequences. To confirm these results, we analyzed the bacterial community by amplifying the variable 16S-23S rDNA intergenic spacer region (ITS). Similar ITS profiles were observed among gastrointestinal compartments of salmon, confirming the TTGE results. Moreover, the dominant ITS band at 650 bp, identified as Pseudomonas, was observed in the ITS profile from fish collected in two seasons (July 2003 and 2004). In contrast, aerobic culture analysis revealed Shewanella spp. as the most prevalent isolate. This discrepancy was resolved by evaluating 16S rDNA and ITS polymerase chain reaction amplification efficiency from both Shewanella and Pseudomonas isolates. Very similar efficiencies were observed in the two bacteria. Hence, this discrepancy may be explained by preferential cultivation of Shewanella spp. under the experimental conditions. Also, we included analyses of pelleted feed and the water influent to explore environmental influences on the bacterial composition of the gut microbiota. Overall, these results indicate a homogeneous composition of the bacterial community composition along the gastrointestinal tract of reared juvenile salmon. This community is mainly composed of Pseudomonas spp., which could be derived from water influent and may be selectively associated with salmon in this hatchery.

MeSH Terms
Animals Colony Count, Microbial DNA, Bacterial/genetics DNA, Intergenic/genetics Ecosystem Gastrointestinal Tract/microbiology Polymorphism, Restriction Fragment Length Pseudomonas/classification,genetics,isolation & purification RNA, Ribosomal, 16S/genetics Salmo salar/microbiology Sequence Analysis, DNA Shewanella/classification,genetics,isolation & purification
Chemicals
DNA, Bacterial DNA, Intergenic RNA, Ribosomal, 16S
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Navarrete P
Laboratorio de Biotecnología, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
Espejo R T
Romero J
References (36)
36 references, click to expand
  1. Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota.
    Proc Natl Acad Sci U S A. 2004 Mar 30;101(13):4596-601 PMID: 15070763
  2. Phylogenetic analysis of intestinal microflora indicates a novel Mycoplasma phylotype in farmed and wild salmon.
    Microb Ecol. 2002 Aug;44(2):175-85 PMID: 12082453
  3. PAGE analysis of the heteroduplexes formed between PCR-amplified 16S rRNA genes: estimation of sequence similarity and rDNA complexity.
    Microbiology (Reading). 1998 Jun;144 ( Pt 6):1611-1617 PMID: 9639932
  4. A genomic view of our symbiosis with members of the gut microbiota.
    J Pediatr Gastroenterol Nutr. 2005 Apr;40 Suppl 1:S28 PMID: 15805839
  5. Phylogenetic analysis of bacterial communities associated with larvae of the Atlantic halibut propose succession from a uniform normal flora.
    Syst Appl Microbiol. 2004 Nov;27(6):728-36 PMID: 15612631
  6. Bacterial 16S rRNA gene analysis revealed that bacteria related to Arcobacter spp. constitute an abundant and common component of the oyster microbiota (Tiostrea chilensis).
    Microb Ecol. 2002 Nov;44(4):365-71 PMID: 12399898
  7. Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis.
    FEMS Microbiol Rev. 1997 Nov;21(3):213-29 PMID: 9451814
  8. The ribosomal database project (RDP-II): introducing myRDP space and quality controlled public data.
    Nucleic Acids Res. 2007 Jan;35(Database issue):D169-72 PMID: 17090583
  9. Phylogenetic identification and in situ detection of individual microbial cells without cultivation.
    Microbiol Rev. 1995 Mar;59(1):143-69 PMID: 7535888
  10. Construction of evolutionary distance trees with TREECON for Windows: accounting for variation in nucleotide substitution rate among sites.
    Comput Appl Biosci. 1997 Jun;13(3):227-30 PMID: 9183524
  11. The bacterial microflora of fish, revised.
    ScientificWorldJournal. 2006 Aug 11;6:931-45 PMID: 16906326
  12. Genetic and phenotypic adaptation of intestinal nutrient transport to diet in fish.
    J Physiol. 1987 Dec;393:261-81 PMID: 3446799
  13. Evolution of mammals and their gut microbes.
    Science. 2008 Jun 20;320(5883):1647-51 PMID: 18497261
  14. Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms.
    Appl Environ Microbiol. 1993 Apr;59(4):945-52 PMID: 8476298
  15. Polymorphism in repeated 16S rRNA genes is a common property of type strains and environmental isolates of the genus Vibrio.
    Microbiology (Reading). 2002 Apr;148(Pt 4):1233-1239 PMID: 11932467
  16. Reciprocal gut microbiota transplants from zebrafish and mice to germ-free recipients reveal host habitat selection.
    Cell. 2006 Oct 20;127(2):423-33 PMID: 17055441
  17. Bacterial flora of fishes: A review.
    Microb Ecol. 1990 Jan;19(1):21-41 PMID: 24196252
  18. Molecular ecological analysis of dietary and antibiotic-induced alterations of the mouse intestinal microbiota.
    J Nutr. 2001 Jun;131(6):1862-70 PMID: 11385080
  19. The intestines of carnivorous fish: structure and functions and the relations with diet.
    Acta Physiol Scand Suppl. 1997;638:67-80 PMID: 9421581
  20. 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 Jul;57(1):128-38 PMID: 16819956
  21. The gut microbiota as an environmental factor that regulates fat storage.
    Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15718-23 PMID: 15505215
  22. Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut.
    Appl Environ Microbiol. 1999 Nov;65(11):4799-807 PMID: 10543789
  23. Getting a grip on things: how do communities of bacterial symbionts become established in our intestine?
    Nat Immunol. 2004 Jun;5(6):569-73 PMID: 15164016
  24. Pyloric ceca of fish: a "new" absorptive organ.
    Am J Physiol. 1987 Jan;252(1 Pt 1):G65-76 PMID: 3812690
  25. Detection of HIV1 DNA by a simple procedure of polymerase chain reaction, using "primer-dimer" formation as an internal control of amplification.
    Res Virol. 1993 May-Jun;144(3):243-6 PMID: 8356345
  26. Probiotic bacteria as biological control agents in aquaculture.
    Microbiol Mol Biol Rev. 2000 Dec;64(4):655-71 PMID: 11104813
  27. Microbial diversity of intestinal contents and mucus in rainbow trout (Oncorhynchus mykiss).
    J Appl Microbiol. 2007 Jun;102(6):1654-64 PMID: 17578431
  28. The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis.
    Nucleic Acids Res. 2005 Jan 1;33(Database issue):D294-6 PMID: 15608200
  29. 16S rDNA-based analysis of dominant bacterial populations associated with early life stages of coho salmon (Oncorhynchus kisutch).
    Microb Ecol. 2006 May;51(4):422-30 PMID: 16598631
  30. Bacterial Interactions in Early Life Stages of Marine Cold Water Fish.
    Microb Ecol. 1999 Jul;38(1):1-26 PMID: 10384006
  31. Bacterial community in copper sulfide ores inoculated and leached with solution from a commercial-scale copper leaching plant.
    Appl Environ Microbiol. 1997 Apr;63(4):1344-8 PMID: 16535570
  32. In vivo imaging and genetic analysis link bacterial motility and symbiosis in the zebrafish gut.
    Proc Natl Acad Sci U S A. 2007 May 1;104(18):7622-7 PMID: 17456593
  33. DNA-DNA hybridization values and their relationship to whole-genome sequence similarities.
    Int J Syst Evol Microbiol. 2007 Jan;57(Pt 1):81-91 PMID: 17220447
  34. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA.
    Appl Environ Microbiol. 1993 Mar;59(3):695-700 PMID: 7683183
  35. Yeasts isolated from the intestine of rainbow trout adhere to and grow in intestinal mucus.
    Mol Mar Biol Biotechnol. 1998 Jun;7(2):115-26 PMID: 9628007
  36. Distinct signals from the microbiota promote different aspects of zebrafish gut differentiation.
    Dev Biol. 2006 Sep 15;297(2):374-86 PMID: 16781702
Article Info
Journal
Microbial ecology
Abbr.
Microb Ecol
ISSN
1432-184X
Published
2009-04-00
Epub
2008-00-17
Pages
550-61
Language
English
Region
United States
NLM ID
7500663
Subset
IM
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