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Background  The awareness of environmental and socio-economic impacts caused by greenhouse gas emissions from the livestock sector leverages the adoption of strategies to counteract it. Feed supplements can play an important role in the reduction of the main greenhouse gas produced by ruminants—methane (CH 4 ). In this context, this study aims to assess the effect of two biochar sources and inclusion levels on rumen fermentation parameters  in vitro .    Methods  Two sources of biochar (agro-forestry residues, AFB, and potato peel, PPB) were added at two levels (5 and 10%, dry matter (DM) basis) to two basal substrates (haylage and corn silage) and incubated 24-h with rumen inocula to assess the effects on CH 4  production and main rumen fermentation parameters  in vitro .    Results  AFB and PPB were obtained at different carbonization conditions resulting in different apparent surface areas, ash content, pH at the point of zero charge (pHpzc), and elemental analysis. Relative to control (0% biochar), biochar supplementation kept unaffected total gas production and yield (mL and mL/g DM,  p  = 0.140 and  p  = 0.240, respectively) and fermentation pH ( p  = 0.666), increased CH 4 production and yield (mL and mL/g DM, respectively,  p  = 0.001) and ammonia-N (NH 3 -N,  p  = 0.040), and decreased total volatile fatty acids (VFA) production ( p  < 0.001) and H 2  generated and consumed ( p  ≤ 0.001). Biochar sources and inclusion levels had no negative effect on most of the fermentation parameters and efficiency. Acetic:propionic acid ratio ( p  = 0.048) and H 2  consumed ( p  = 0.019) were lower with AFB inclusion when compared to PPB. Biochar inclusion at 10% reduced H 2  consumed ( p  < 0.001) and tended to reduce total gas production ( p  = 0.055). Total VFA production ( p  = 0.019), acetic acid proportion ( p  = 0.011) and H 2  generated ( p  = 0.048) were the lowest with AFB supplemented at 10%, no differences being observed among the other treatments. The basal substrate affected most fermentation parameters independently of biochar source and level used.    Discussion  Biochar supplementation increased NH 3 -N content,  iso -butyric,  iso -valeric and valeric acid proportions, and decreased VFA production suggesting a reduced energy supply for microbial growth, higher proteolysis and deamination of substrate N, and a decrease of NH 3 -N incorporation into microbial protein. No interaction was found between substrate and biochar source or level on any of the parameters measured. Although AFB and PPB had different textural and compositional characteristics, their effects on the rumen fermentation parameters were similar, the only observed effects being due to AFB included at 10%. Biochar supplementation promoted CH 4  production regardless of the source and inclusion level, suggesting that there may be other effects beyond biomass and temperature of production of biochar, highlighting the need to consider other characteristics to better identify the mechanism by which biochar may influence CH 4  production.

Assessment of potato peel and agro-forestry biochars supplementation on in vitro ruminal fermentation