High throughput accurate method for estimating in vitro dry matter digestibility of maize silage

Abstract Background Since the introduction of studies on maize silage digestibility at the end of the nineteenth century, protocols to estimate dry matter digestibility have not stopped evolving. Since the early 1980s, the protocol developed by Aufrère became a benchmark in many laboratories to estimate in vitro dry matter digestibility. In order to increase its throughput, to facilitate its execution and to decipher the impact of the different parameters of the protocol we decided to test the combination of 7 parameters in 21 different protocols. Results We thus tested the impact of (1) the presence or absence of pepsin in HCl solution, (2) the temperature of incubation during enzymatic hydrolysis, (3) the presence or absence of a gelatinization step, (4) washing/rinsing versus neutralization step, (5) the presence or absence of α-amyloglucosidase in enzymatic solution, (6) the duration of cellulase incubation, and (7) the concentration of the cellulase solution. The major result of our work highlighted that it was essential to carry out a gelatinization step to correctly estimate the in vitro dry matter digestibility of maize silage. Conclusions The proposed protocol in this paper is innovative, reliable, highthroughput and easy to implement in many laboratories to accurately quantity in vitro dry matter digestibility