Abstract
The digestive system of the chicken plays an important role in metabolism, immunity, and chicken health and production performance. The chicken ceca harbor a diverse microbial community and play a crucial role in the microbial fermentation and production of energy-rich short-chain fatty acids (SCFA). For humans, dogs, and piglets in vitro digestive system models have been developed and are used to study the microbiota composition and metabolism after intervention studies. For chickens, most research on the cecal microbiota has been performed in in vivo experiments or in static in vitro models that may not accurately resemble the in vivo situations. This paper introduces an optimized digestive system model that simulates the conditions in the ceca of the chicken, i.e., the Chicken ALIMEntary tRact mOdel-2 (CALIMERO-2). The system is based on the well-validated TNO in vitro model of the colon-2 (TIM-2) and is the first dynamic in vitro digestion model for chickens species. To validate this model, the pH, temperature, and different types of microbial feeding were compared and analyzed, to best mimic the conditions in the chicken ceca. The bacterial composition, as well as the metabolite production at 72 h, showed no significant difference between the different microbial feedings. Moreover, we compared the CALIMERO-2 digestive samples to the original inoculum and found some significant shifts in bacterial composition after the fermentation started. Over time the bacterial diversity increased and became more similar to the original inoculum. We can conclude that CALIMERO-2 is reproducible and can be used as a digestive system model for the chicken ceca, in which the microbial composition and activity can be maintained and shows similar results to the in vivo cecum. CALIMERO-2 can be used to study effects on composition and activity of the chicken cecum microbiota in response to in-feed interventions.
Highlights
MATERIALS AND METHODSThe digestive system of the chicken plays a pivotal role in metabolism, immunity, and therewith in the health and production performance (Gong et al, 2002; Round and Mazmanian, 2009; Xiao et al, 2017; Borda-Molina et al, 2018)
The bacterial composition of the lumen samples from runs with the different types of microbial feeding, i.e., Standard Ileal Effluent Media (SIEM), Modified SIEM-I, and Modified SIEM-II, which served as microbial growth medium, were analyzed and compared to the original inoculum, i.e., the pooled and standardized sample before inoculation into the system at time point -16 h
CALIMERO-2 can be used as a digestion system model for the chicken ceca, in which the microbial composition and activity can be maintained in a similar manner to the in vivo cecum
Summary
MATERIALS AND METHODSThe digestive system of the chicken plays a pivotal role in metabolism, immunity, and therewith in the health and production performance (Gong et al, 2002; Round and Mazmanian, 2009; Xiao et al, 2017; Borda-Molina et al, 2018). Previous studies have shown that prebiotics can stimulate the growth of beneficial endogenous microbes by providing nutrients to beneficial bacteria (Gibson et al, 2004; Adhikari et al, 2020), and can lead to better growth and health of the chickens (Chambers and Gong, 2011). Most of the current research on the microbiota in the chicken GIT has been performed in in vivo experiments or field studies (De Carvalho et al, 2021). In vivo experiments have many downsides, including that these methods are invasive for the animals or in case of non-invasive methods like cloacal swabs, these might not completely represent the composition of the ceca (Minekus et al, 1999; Gibson et al, 2004). There is a need for in vitro digestive system models to study the behavior of microbiota, with high predictive value for in vivo animal trials, to gain in-depth knowledge of the effect and possible mechanisms of action of dietary interventions on the chicken cecal microbiota
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