Ozone Decreased Enteric Methane Production by 20% in an in vitro Rumen Fermentation System

Ozone (O₃) is volatile, highly oxidative, and has theoretical potential to reduce ruminant enteric methanogenesis by interactions between archaea and bacteria, and substrate and oxygen. The effects of O₃ on the rumen microbiota, fermentation parameters, and CH₄ emissions were studied through in vitro fermentation using a RUSITEC apparatus with O₃ dissolved in the salivary buffer. The substrate consisted of maize silage or grain concentrates, and the treatments were (1) control (no O₃) and (2) O₃ at 0.07 ± 0.022 mg/L in the buffer. A 4-day adaptation period followed by a 6-day experimental period was used for measuring gas production and composition, as well as fermentation characteristics, which included ruminal volatile fatty acids (VFA) and liquid- and solid-associated microbial communities. Ozone treatment decreased total gas production by 15.4%, most notably CH₄ production by 20.4%, and CH₄ gas concentration by 5.8%, without compromising dry matter digestibility (DMD) of either maize silage or grain concentrates. There were no significant effects of O₃ treatment on VFA production or pH. Ozone treatment reduced the relative abundance of methanogens, particularly Methanomicrobium. This study demonstrates the potential use of O₃ as a method to reduce ruminant enteric methanogenesis.