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Ozone (O 3 ) 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 O 3  on the rumen microbiota, fermentation parameters, and CH 4  emissions were studied through  in vitro  fermentation using a RUSITEC apparatus with O 3  dissolved in the salivary buffer. The substrate consisted of maize silage or grain concentrates, and the treatments were (1) control (no O 3 ) and (2) O 3  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 CH 4  production by 20.4%, and CH 4  gas concentration by 5.8%, without compromising dry matter digestibility (DMD) of either maize silage or grain concentrates. There were no significant effects of O 3  treatment on VFA production or pH. Ozone treatment reduced the relative abundance of methanogens, particularly  Methanomicrobium . This study demonstrates the potential use of O 3  as a method to reduce ruminant enteric methanogenesis.

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