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      Pyrosequencing of 16S rRNA gene amplicons to study the microbiota in the gastrointestinal tract of carp ( Cyprinus carpio L.)

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          Abstract

          The microbes in the gastrointestinal (GI) tract are of high importance for the health of the host. In this study, Roche 454 pyrosequencing was applied to a pooled set of different 16S rRNA gene amplicons obtained from GI content of common carp ( Cyprinus carpio) to make an inventory of the diversity of the microbiota in the GI tract. Compared to other studies, our culture-independent investigation reveals an impressive diversity of the microbial flora of the carp GI tract. The major group of obtained sequences belonged to the phylum Fusobacteria. Bacteroidetes, Planctomycetes and Gammaproteobacteria were other well represented groups of micro-organisms. Verrucomicrobiae, Clostridia and Bacilli (the latter two belonging to the phylum Firmicutes) had fewer representatives among the analyzed sequences. Many of these bacteria might be of high physiological relevance for carp as these groups have been implicated in vitamin production, nitrogen cycling and (cellulose) fermentation.

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          Most cited references41

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          Reciprocal gut microbiota transplants from zebrafish and mice to germ-free recipients reveal host habitat selection.

          The gut microbiotas of zebrafish and mice share six bacterial divisions, although the specific bacteria within these divisions differ. To test how factors specific to host gut habitat shape microbial community structure, we performed reciprocal transplantations of these microbiotas into germ-free zebrafish and mouse recipients. The results reveal that communities are assembled in predictable ways. The transplanted community resembles its community of origin in terms of the lineages present, but the relative abundance of the lineages changes to resemble the normal gut microbial community composition of the recipient host. Thus, differences in community structure between zebrafish and mice arise in part from distinct selective pressures imposed within the gut habitat of each host. Nonetheless, vertebrate responses to microbial colonization of the gut are ancient: Functional genomic studies disclosed shared host responses to their compositionally distinct microbial communities and distinct microbial species that elicit conserved responses.
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            Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota.

            Animals have developed the means for supporting complex and dynamic consortia of microorganisms during their life cycle. A transcendent view of vertebrate biology therefore requires an understanding of the contributions of these indigenous microbial communities to host development and adult physiology. These contributions are most obvious in the gut, where studies of gnotobiotic mice have disclosed that the microbiota affects a wide range of biological processes, including nutrient processing and absorption, development of the mucosal immune system, angiogenesis, and epithelial renewal. The zebrafish (Danio rerio) provides an opportunity to investigate the molecular mechanisms underlying these interactions through genetic and chemical screens that take advantage of its transparency during larval and juvenile stages. Therefore, we developed methods for producing and rearing germ-free zebrafish through late juvenile stages. DNA microarray comparisons of gene expression in the digestive tracts of 6 days post fertilization germ-free, conventionalized, and conventionally raised zebrafish revealed 212 genes regulated by the microbiota, and 59 responses that are conserved in the mouse intestine, including those involved in stimulation of epithelial proliferation, promotion of nutrient metabolism, and innate immune responses. The microbial ecology of the digestive tracts of conventionally raised and conventionalized zebrafish was characterized by sequencing libraries of bacterial 16S rDNA amplicons. Colonization of germ-free zebrafish with individual members of its microbiota revealed the bacterial species specificity of selected host responses. Together, these studies establish gnotobiotic zebrafish as a useful model for dissecting the molecular foundations of host-microbial interactions in the vertebrate digestive tract.
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              Learning how to live together: genomic insights into prokaryote-animal symbioses.

              Our understanding of prokaryote-eukaryote symbioses as a source of evolutionary innovation has been rapidly increased by the advent of genomics, which has made possible the biological study of uncultivable endosymbionts. Genomics is allowing the dissection of the evolutionary process that starts with host invasion then progresses from facultative to obligate symbiosis and ends with replacement by, or coexistence with, new symbionts. Moreover, genomics has provided important clues on the mechanisms driving the genome-reduction process, the functions that are retained by the endosymbionts, the role of the host, and the factors that might determine whether the association will become parasitic or mutualistic.
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                Author and article information

                Journal
                AMB Express
                AMB Express
                Springer
                2191-0855
                2011
                18 November 2011
                : 1
                : 41
                Affiliations
                [1 ]Department of Microbiology, IWWR, Radboud University Nijmegen, Heyendaalseweg 135, NL-6525 AJ Nijmegen, the Netherlands
                [2 ]Department of Animal Physiology, IWWR, Radboud University Nijmegen, Heyendaalseweg 135, NL-6525 AJ Nijmegen, the Netherlands
                [3 ]Center for Molecular and Biomolecular Informatics, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, Geert Grooteplein 28, NL-6525 GA Nijmegen, the Netherlands
                [4 ]Departments of Computer Science and Biology, San Diego State University, 5500 Campanile Drive, San Diego CA 92182, USA
                [5 ]Department of Human Genetics, Radboud University Nijmegen Medical Centre, Geert Grooteplein 10, NL-6525 GA Nijmegen, the Netherlands
                Article
                2191-0855-1-41
                10.1186/2191-0855-1-41
                3226434
                22093413
                5c3176ac-a7f1-47d7-b994-72fad30018b4
                Copyright ©2011 van Kessel et al; licensee Springer.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 3 November 2011
                : 18 November 2011
                Categories
                Original

                Biotechnology
                pyrosequencing,intestinal tract,biodiversity,16s rrna,carp,aquaculture
                Biotechnology
                pyrosequencing, intestinal tract, biodiversity, 16s rrna, carp, aquaculture

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