Abstract

Free-range pullets are reared indoors but the adult hens can go outside which is a mismatch that may reduce adaptation in the laying environment. Rearing enrichments might enhance pullet development and adaptations to subsequent free-range housing with impact on behavior and health measures including gut microbiota. Adult free-range hens vary in range use which may also be associated with microbiota composition. A total of 1,700 Hy-Line Brown® chicks were reared indoors across 16 weeks with three enrichment treatment groups: “control” with standard litter housing, “novelty” with weekly changed novel objects, and “structural” with custom-designed perching structures in the pens. At 15 weeks, 45 pullet cecal contents were sampled before moving 1,386 pullets to the free-range housing system. At 25 weeks, range access commenced, and movements were tracked via radio-frequency identification technology. At 65 weeks, 91 hens were selected based on range use patterns (“indoor”: no ranging; “high outdoor”: daily ranging) across all rearing enrichment groups and cecal contents were collected for microbiota analysis via 16S rRNA amplicon sequencing at V3-V4 regions. The most common bacteria in pullets were unclassified Barnesiellaceae, Prevotella, Blautia and Clostridium and in hens Unclassified, Ruminococcus, unclassified Lachnospiraceae, unclassified Bacteroidales, unclassified Paraprevotellaceae YRC22, and Blautia. The microbial alpha diversity was not significant within the enrichment/ranging groups (pullets: P ≥ 0.17, hen rearing enrichment groups: P ≥ 0.06, hen ranging groups: P ≥ 0.54), but beta diversity significantly varied between these groups (pullets: P ≤ 0.002, hen rearing enrichment groups: P ≤ 0.001, hen ranging groups: P ≤ 0.008). Among the short-chain fatty acids (SCFAs), the propionic acid content was higher (P = 0.03) in the novelty group of pullets than the control group. There were no other significant differences in the SCFA contents between the rearing enrichment groups (all P ≥ 0.10), and the ranging groups (all P ≥ 0.17). Most of the genera identified were more abundant in the indoor than high outdoor hens. Overall, rearing enrichments affected the cecal microbiota diversity of both pullets and adult hens and was able to distinguish hens that remained inside compared with hens that ranging daily for several hours.

Highlights

  • Numerous species of bacteria are sheltered in the gastro-intestinal (GI) tract of chickens, known as gut microbiota (Rychlik, 2020; Madlala et al, 2021), which play an important role in physiological processes including digestion, absorption, health, and production (Pourabedin and Zhao, 2015)

  • The Linear Discriminant (LDA) Effect Size (LEfSe) analysis (Figure 2A) showed the differentially abundant explanatory microbiota genera in the structural group included unclassified Barnesiellaceae, Phascolarctobacterium, Adlercreutzia, and AF12 with Prevotella, Blautia and Eubacterium predominant in the novelty group, and Methanobrevibacter and unclassified Rikenellaceae explanatory in the control group

  • The Blautia was less abundant in the structural group than in both novelty and control pullets (P ≤ 0.01), but there was no difference in the Blautia abundance between the control and novelty pullets

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Summary

Introduction

Numerous species of bacteria are sheltered in the gastro-intestinal (GI) tract of chickens, known as gut microbiota (Rychlik, 2020; Madlala et al, 2021), which play an important role in physiological processes including digestion, absorption, health, and production (Pourabedin and Zhao, 2015). Fu et al (2018) explored the gut microbiota diversity of laying hens based on 16S rRNA sequencing in China and found the Bacterioidetes, Firmicutes phyla were the most predominant among the cecal microbes. Actinobacteria and Proteobacteria are typically represented in the ceca of most adult chickens there can be high variation in microbial composition between individuals (Rychlik, 2020). The most common genera of microbiota in the ceca of laying hens have been shown to be the Bacteroides and Prevotella (Yan et al, 2017). Identifying the composition of microbes within chickens is important for understanding their function and how they may be affected by internal and external environmental parameters

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