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      Nearly (?) sterile avian egg in a passerine bird

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          Abstract

          During early ontogeny, microbiome affects development of the gastrointestinal tract, immunity, and survival in vertebrates. Bird eggs are thought to be (1) initially sterile ( sterile egg hypothesis) and (2) colonized after oviposition through horizontal trans-shell migration, or (3) initially seeded with bacteria by vertical transfer from mother oviduct. To date, however, little empirical data illuminate the contribution of these mechanisms to gut microbiota formation in avian embryos. We investigated microbiome of the egg content (day 0; E0-egg), embryonic gut at day 13 (E13) and female faeces in a free-living passerine, the great tit ( Parus major), using a methodologically advanced procedure combining 16S rRNA gene sequencing and microbe-specific qPCR assays. Our metabarcoding revealed that the avian egg is (nearly) sterile, but acquires a slightly richer microbiome during the embryonic development. Of the three potentially pathogenic bacteria targeted by qPCR, only Dietzia was found in E0-egg (yet also in negative controls), E13 gut and female samples, which might indicate possible vertical transfer. Unlike in poultry, we have shown that major bacterial colonization of the gut in passerines does not occur before hatching. We emphasize that protocols that carefully check for environmental contamination are critical in studies with low-bacterial biomass samples.

          Abstract

          The eggs of great tits are nearly sterile when they are laid and are first colonised during embryonic development. Our results indicate that diversified microbial communities form in passerine birds after hatching.

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            DADA2: High resolution sample inference from Illumina amplicon data

            We present DADA2, a software package that models and corrects Illumina-sequenced amplicon errors. DADA2 infers sample sequences exactly, without coarse-graining into OTUs, and resolves differences of as little as one nucleotide. In several mock communities DADA2 identified more real variants and output fewer spurious sequences than other methods. We applied DADA2 to vaginal samples from a cohort of pregnant women, revealing a diversity of previously undetected Lactobacillus crispatus variants.
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              The SILVA ribosomal RNA gene database project: improved data processing and web-based tools

              SILVA (from Latin silva, forest, http://www.arb-silva.de) is a comprehensive web resource for up to date, quality-controlled databases of aligned ribosomal RNA (rRNA) gene sequences from the Bacteria, Archaea and Eukaryota domains and supplementary online services. The referred database release 111 (July 2012) contains 3 194 778 small subunit and 288 717 large subunit rRNA gene sequences. Since the initial description of the project, substantial new features have been introduced, including advanced quality control procedures, an improved rRNA gene aligner, online tools for probe and primer evaluation and optimized browsing, searching and downloading on the website. Furthermore, the extensively curated SILVA taxonomy and the new non-redundant SILVA datasets provide an ideal reference for high-throughput classification of data from next-generation sequencing approaches.
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                Author and article information

                Contributors
                Journal
                FEMS Microbiol Ecol
                FEMS Microbiol Ecol
                femsec
                FEMS Microbiology Ecology
                Oxford University Press
                0168-6496
                1574-6941
                January 2024
                19 December 2023
                19 December 2023
                : 100
                : 1
                : fiad164
                Affiliations
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Institute of Vertebrate Biology, v.v.i., The Czech Academy of Sciences , Květná 8, Brno 603 65, Czech Republic
                Institute of Paleonatomy, Domestification Research and History of Veterinary Medicine, Ludwig Maxmilian University of Munich , Kaulbachstr. 37 III, 80539 Munich, Germany
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Institute of Vertebrate Biology, v.v.i., The Czech Academy of Sciences , Květná 8, Brno 603 65, Czech Republic
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Faculty of Science, Department of Parasitology, Campus Universitario de Fuentenueva, University of Granada , Profesor Adolfo Rancano, 18071 Granada, Spain
                Division of Air Quality, Czech Hydrometeorological Institute , Tušimice Observatory , Tušimice 6, 432 01 Kadaň, Czech Republic
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Faculty of Science, Department of Zoology, Charles University , Viničná 7, 128 43 Prague, Czech Republic
                Author notes
                Corresponding author. Faculty of Science, Department of Zoology, Charles University, Viničná 7, 128 43 Prague, Czech Republic, EU. Tel: +420221951845; Fax: +420221951841; E-mail: martin.tesicky@ 123456natur.cuni.cz

                Present addresses: Institute of Vertebrate Biology, v.v.i., The Czech Academy of Sciences, Květná 8, Brno 603 65, Czech Republic and Institute of Paleonatomy, Domestification Research and History of Veterinary Medicine, Ludwig Maxmilian University of Munich, Kaulbachstr. 37 III, 80539 Munich, Germany

                Author information
                https://orcid.org/0000-0001-8097-5331
                Article
                fiad164
                10.1093/femsec/fiad164
                10791042
                38115624
                41d0c268-ca1e-4659-9108-4b70588e1c2d
                © The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@ 123456oup.com

                History
                : 14 February 2023
                : 02 December 2023
                : 18 December 2023
                : 16 January 2024
                Page count
                Pages: 13
                Funding
                Funded by: Charles University, DOI 10.13039/100007397;
                Award ID: GAUK 1158217
                Award ID: UNCE 204069
                Award ID: START/SCI/113
                Funded by: MEYS, DOI 10.13039/501100001823;
                Award ID: LM2015091
                Categories
                Research Article
                AcademicSubjects/SCI01150

                Microbiology & Virology
                egg microbiome,embryo,gastrointestinal tract microbiota,passerine bird,pathogenic bacteria,sterile egg

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