An observational field study of the cloacal microbiota in adult laying hens with and without access to an outdoor range

Janneke Schreuder1, Francisca C. Velkers1, Ruth Bouwstra2, Nancy Beerens3, J.A. Stegeman1, Willem F. de Boer4, Pim van Hooft4, A.R.W. Elbers5, Alex Bossers6, Stephanie D. Jurburg6
1Faculty of Veterinary Medicine, Department Population Health Sciences, Utrecht University, Yalelaan 7, 3584, CL, Utrecht, The Netherlands
2GD Animal Health, Deventer, The Netherlands
3Department of Virology, Wageningen Bioveterinary Research, Lelystad, the Netherlands
4Wildlife Ecology and Conservation Group, Wageningen University & Research, Wageningen, The Netherlands
5Department of Bacteriology and Epidemiology, Wageningen Bioveterinary Research, Lelystad, the Netherlands
6Department of Infection Biology, Wageningen Bioveterinary Research, Lelystad, the Netherlands

Tóm tắt

Abstract Background Laying hens with access to outdoor ranges are exposed to additional environmental factors and microorganisms, including potential pathogens. Differences in composition of the cloacal microbial community between indoor- and outdoor-housed layers may serve as an indicator for exposure to the outdoor environment, including its pathogens, and may yield insights into factors affecting the chickens’ microbiota community dynamics. However, little is known about the influence of outdoor housing on microbiota community composition in commercial layer flocks. We performed a cross-sectional field study to evaluate differences in the cloacal microbiota of indoor- vs outdoor-layers across farms. Eight layer flocks (four indoor, four outdoor) from five commercial poultry farms were sampled. Indoor and outdoor flocks with the same rearing flock of origin, age, and breed were selected. In each flock, cloacal swabs were taken from ten layers, and microbiota were analysed with 16S rRNA gene amplicon sequencing. Results Housing type (indoor vs outdoor), rearing farm, farm and poultry house within the farm all significantly contributed to bacterial community composition. Poultry house explained most of the variation (20.9%), while housing type only explained 0.2% of the variation in community composition. Bacterial diversity was higher in indoor-layers than in outdoor-layers, and indoor-layers also had more variation in their bacterial community composition. No phyla or genera were found to be differentially abundant between indoor and outdoor poultry houses. One amplicon sequence variant was exclusively present in outdoor-layers across all outdoor poultry houses, and was identified as Dietzia maris. Conclusions This study shows that exposure to an outdoor environment is responsible for a relatively small proportion of the community variation in the microbiota of layers. The poultry house, farm, and rearing flock play a much greater role in determining the cloacal microbiota composition of adult laying hens. Overall, measuring differences in cloacal microbiota of layers as an indicator for the level of exposure to potential pathogens and biosecurity seems of limited practical use. To gain more insight into environmental drivers of the gut microbiota, future research should aim at investigating community composition of commercial layer flocks over time.

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Tài liệu tham khảo

Bestman MWP, Wagenaar JP. Farm level factors associated with feather pecking in organic laying hens. Livest Prod Sci. 2003. https://doi.org/10.1016/S0301-6226(02)00314-7.

Green LE, Lewis K, Kimpton A, Nicol CJ. Cross-sectional study of the prevalence of feather pecking in laying hens in alternative systems and its associations with management and disease. Vet Rec. 2000. https://doi.org/10.1136/vr.147.9.233.

Miao ZH, Glatz PC, Ru YJ. Free-range poultry production - a review. Asian-Australas J Anim Sci. 2005. https://doi.org/10.5713/ajas.2005.113.

Koch G, Elbers ARW. Outdoor ranging of poultry: a major risk factor for the introduction and development of high-Pathogenecity avian influenza. NJAS - Wageningen J Life Sci. 2006. https://doi.org/10.1016/S1573-5214(06)80021-7.

Bouwstra R, Gonzales JL, de Wit S, Stahl J, Fouchier RAMM, Elbers ARWW. Risk for low pathogenicity avian influenza virus on poultry farms, the Netherlands, 2007–2013. Emerg Infect Dis. 2017. https://doi.org/10.3201/eid2309.170276.

Brown JD, Swayne DE, Cooper RJ, Burns RE, Stallknecht DE. Persistence of H5 and H7 avian influenza viruses in water. Avian Dis. 2007. https://doi.org/10.1637/7636-042806R.1.

Rohani P, Breban R, Stallknecht DE, Drake JM. Environmental transmission of low pathogenicity avian influenza viruses and its implications for pathogen invasion. Proc Natl Acad Sci. 2009. https://doi.org/10.1073/pnas.0809026106.

Kers JG, Velkers FC, Fischer EAJ, Hermes GDA, Stegeman JA, Smidt H. Host and environmental factors affecting the intestinal microbiota in chickens. Front Microbiol. 2018. https://doi.org/10.3389/fmicb.2018.00235.

Best AA, Porter AL, Fraley SM, Fraley GS. Characterization of gut microbiome dynamics in developing Pekin ducks and impact of management system. Front Microbiol. 2017;7. https://doi.org/10.3389/fmicb.2016.02125.

Ocejo M, Oporto B, Hurtado A. 16S rRNA amplicon sequencing characterization of caecal microbiome composition of broilers and free-range slow-growing chickens throughout their productive lifespan. Sci Rep. 2019. https://doi.org/10.1038/s41598-019-39323-x.

Hubert SM, Al-Ajeeli M, Bailey CA, Athrey G. The role of housing environment and dietary protein source on the gut microbiota of chicken. Animals. 2019. https://doi.org/10.3390/ani9121085.

Xu Y, Yang H, Zhang L, Su Y, Shi D, Xiao H, et al. High-throughput sequencing technology to reveal the composition and function of cecal microbiota in Dagu chicken. BMC Microbiol. 2016. https://doi.org/10.1186/s12866-016-0877-2.

Chen S, Xiang H, Zhu X, Zhang H, Wang D, Liu H, et al. Free dietary choice and free-range rearing improve the product quality, gait score, and microbial richness of chickens. Animals. 2018. https://doi.org/10.3390/ani8060084.

Basic local alignment search tool. https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE_TYPE=BlastSearch&BLAST_SPEC=TargLociBlast. Accessed 30 Jan 2020.

Rainey FA, Klatte S, Kroppenstedt RM, Stackebrandt E. Dietzia, new genus including Dietzia maris comb. nov., formerly Rhodococcus maris. Int J Syst Evol Microbiol. 1995. https://doi.org/10.1099/00207713-45-1-32.

Schreuder J, Velkers FC, Bouwstra RJ, Beerens N, Stegeman JA, de Boer WF, et al. Limited changes in the fecal microbiome composition of laying hens after oral inoculation with wild duck feces. Poult Sci. 2019. https://doi.org/10.3382/ps/pez526.

Videnska P, Sedlar K, Lukac M, Faldynova M, Gerzova L, Cejkova D, et al. Succession and replacement of bacterial populations in the caecum of egg laying hens over their whole life. PLoS One. 2014. https://doi.org/10.1371/journal.pone.0115142.

Gong J, Yu H, Liu T, Gill JJ, Chambers JR, Wheatcroft R, et al. Effects of zinc bacitracin , bird age and access to range on bacterial microbiota in the ileum and caeca of broiler chickens; 2008. https://doi.org/10.1111/j.1365-2672.2007.03699.x.

Han Z, Willer T, Li L, Pielsticker C, Rychlik I, Velge P, et al. Influence of the gut microbiota composition on campylobacter jejuni colonization in chickens. Infect Immun. 2017. https://doi.org/10.1128/IAI.00380-17.

Lawley TD, Walker AW. Intestinal colonization resistance. Immunology. 2013. https://doi.org/10.1111/j.1365-2567.2012.03616.x.

van Niekerk TGCM, Leenstra FR. Inventory of the effects of free-range poultry farming on soil, water and air quality [in Dutch]. Wageningen; 2016. https://edepot.wur.nl/385390. Accessed on 6 February 2020.

Hegelund L, Sørensen JT, Kjær JB, Kristensen IS. Use of the range area in organic egg production systems: effect of climatic factors, flock size, age and artificial cover. Br Poult Sci. 2005. https://doi.org/10.1080/00071660400023813.

Gebhardt-Henrich SG, Toscano MJ, Fröhlich EKF. Use of outdoor ranges by laying hens in different sized flocks. Appl Anim Behav Sci. 2014. https://doi.org/10.1016/j.applanim.2014.03.010.

Bestman MWP. Lessons learnt - agroforestry for organic and free-range egg production in the Netherlands. Wageningen; 2017. http://www.louisbolk.org/downloads/3310.pdf. Accessed on 6 February 2020.

Cui Y, Wang Q, Liu S, Sun R, Zhou Y, Li Y. Age-related variations in intestinal microflora of free-range and caged hens. Front Microbiol. 2017. https://doi.org/10.3389/fmicb.2017.01310.

Nordentoft S, Mølbak L, Bjerrum L, De Vylder J, Van Immerseel F, Pedersen K. The influence of the cage system and colonisation of salmonella Enteritidis on the microbial gut flora of laying hens studied by T-RFLP and 454 pyrosequencing. BMC Microbiol. 2011. https://doi.org/10.1186/1471-2180-11-187.

Schouwenburg H. Regulations Dutch quality assurance scheme IKB egg layer farms, aviary-systems [in Dutch]; 2019. Nieuwegein; https://ikbei.nl/wp-content/uploads/2019/05/OVO-IKBEI-BIJL-100-07-20190601-Voorschriften-primaire-bedrijven-Voorschriften-legeind-scharrel.pdf. Accessed on 15 Jan 2020.

Laukens D, Brinkman BM, Raes J, De Vos M, Vandenabeele P. Heterogeneity of the gut microbiome in mice: guidelines for optimizing experimental design. FEMS Microbiol Rev. 2016. https://doi.org/10.1093/femsre/fuv036.

Stanley D, Geier MS, Chen H, Hughes RJ, Moore RJ. Comparison of fecal and cecal microbiotas reveals qualitative similarities but quantitative differences. BMC Microbiol. 2015. https://doi.org/10.1186/s12866-015-0388-6.

Oakley BB, Kogut MH. Spatial and temporal changes in the broiler chicken cecal and fecal microbiomes and correlations of bacterial taxa with cytokine gene expression. Front Vet Sci. 2016. https://doi.org/10.3389/fvets.2016.00011.

Kers JG, Velkers FC, Fischer EAJ, Hermes GDA, Lamot DM, Stegeman JA, et al. Take care of the environment: housing conditions affect the interplay of nutritional interventions and intestinal microbiota in broiler chickens. Anim Microbiome. 2019. https://doi.org/10.1186/s42523-019-0009-z.

Parker KD, Albeke SE, Gigley JP, Goldstein AM, Ward NL. Microbiome composition in both wild-type and disease model mice is heavily influenced by mouse facility. Front Microbiol. 2018. https://doi.org/10.3389/fmicb.2018.01598.

Schouwenburg H. Regulations Dutch quality assurance scheme IKB egg layer farms, free-range and organic systems [in Dutch]; 2019. Nieuwegein; https://ikbei.nl/wp-content/uploads/2019/05/OVO-IKBEI-BIJL-100-07-20190601-Voorschriften-primaire-bedrijven-Voorschriften-legeind-uitloop-en-biologisch.pdf. Accessed on 15 January 2020.

R Core Team. R: a language and environment for statistical computing. R Foundation for statistical computing; 2013. http://www.r-project.org/.

Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods. 2016; http://benjjneb.github.io/dada2/.

Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 2012;41:D590–6.

McMurdie PJ, Holmes S. Phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS One. 2013. https://doi.org/10.1371/journal.pone.0061217.

Oksanen J, Blanchet FG, Kindt R, Legendre P, Minchin PR, O’Hara RB, et al. The vegan package. Community Ecol. 2007; https://cran.r-project.org/.

Pielou EC. The measurement of diversity in different types of biological collections. J Theor Biol. 1966. https://doi.org/10.1016/0022-5193(66)90013-0.

Bray JR, Curtis JT. An ordination of the upland forest communities of southern Wisconsin. Ecol Monogr. 1957. https://doi.org/10.2307/1942268.

Anderson MJ. A new method for non-parametric multivariate analysis of variance. Austral Ecol. 2001. https://doi.org/10.1111/j.1442-9993.2001.01070.pp.x.

Borcard D, Legendre P, Drapeau P. Partialling out the spatial component of ecological variation. Ecology. 1992. https://doi.org/10.2307/1940179.

Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014. https://doi.org/10.1186/s13059-014-0550-8.

Warton DI, Wright ST, Wang Y. Distance-based multivariate analyses confound location and dispersion effects. Methods Ecol Evol. 2012. https://doi.org/10.1111/j.2041-210X.2011.00127.x.