Attenuation of ruminal methanogenesis

Ruminal methanogens reduce carbon dioxide to methane (CH 4 ), thereby preventing hydrogen use by bacteria for VFA synthesis resulting in a 2 to 12% loss in feed gross energy. Methane is a greenhouse gas that contributes to global warming. The objectives of this work were to determine: (1) the extent to which ruminal cultures acquire resistance to a nitrofuranyl derivative of para-aminobenzoate (NFP) and an extract from the plant Yucca shidigera (Yucca); (2) the effect of distillers dried grains plus solubles (DDGS) on ruminal CH4 production; (3) the effect of brome hay-based diets, corn-based diets, and in vivo 2-bromoethansulfonate treatment on ruminal methane (CH4 ) production; and (4) the effect of the above treatments on the methanogen population. Ruminal cultures treated with NFP for 90 d maintained a diminished capacity to generate CH4 , but cultures became resistant to the inhibitory effects of Yucca treatment within 10 d. Both treatments decreased (P < 0.01) the relative abundance of total Archaea and the order Methanomicrobiales, but Yucca treatment increased (P < 0.01) the relative abundance of the order Methanobacteriales. The replacement of brome hay and corn with DDGS in lamb diets decreased (P < 0.01) and increased (P < 0.05), respectively, the amount of CH4 produced per unit of digested DM. The substitution of DDGS for brome hay increased (P < 0.01) the relative abundance of the order Methanomicrobiales. The replacement of brome hay with corn decreased (P < 0.05) the amount of CH4 produced per unit of digested DM, and also decreased (P < 0.05) the relative abundance of both Archaea and the order Methanomicrobiales. However, the abundance of the order Methanobacteriales increased (P < 0.05) as corn replaced brome hay. Intraruminal administration of 2-bromoethansulfonate decreased (P < 0.05) CH4 emissions, and decreased (P < 0.05) the relative abundance of Archaea and Methanobacteriales. In conclusion, NFP may be efficacious for chronically inhibiting ruminal methanogenesis, and the replacement of dietary forage with DDGS attenuates CH4 emissions from ruminant animals. Changes in domain- and order-specific ribosomal DNA indicators of methanogens are not consistently correlated with changes in CH4 production.

INFLUENCE OF TYPE 2 BOVINE VIRAL DIARRHEA VIRUS NPRO ON ENHANCEMENT OF BOVINE RESPIRATORY SYNCYTIAL VIRUS REPLICATION MEDIATED BY ANTAGONISM OF HOST CELL INTERFERON TYPE I RESPONSES

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, Family Flaviviridae. The virus can infect many species of animals of the order Artiodactyla. The BVDV genome encodes an auto protease, Npro, that degrades interferon regulatory factor-3 (IRF-3) reducing type I interferon (IFN-I) production from host cells. Bovine respiratory syncytial virus (BRSV) is a member of the genus Pneumovirus, Family Paramyxoviridae. Concurrent infection with BVDV and BRSV causes more severe respiratory and enteric disease than infection with either virus alone. Our hypothesis was that Npro modulates the innate immune responses to BVDV infection and enhances replication of BVDV or BRSV co-infection. The noncytopathic BVDV2 viruses NY93/c N- Npro 18 EGFP (a mutant with modified Npro fused with enhanced green fluorescent protein), NY93 infectious clone (NY93/c), wild-type NY93-BVDV2 (NY93-wt), and BRSV were evaluated in this study. The objectives of this study were: (1) to characterize the replication kinetics and IFN-I induction in Madin-Darby bovine kidney (MDBK) cells following infection with each of the BVDV isolates, and (2) to characterize the influence of BVDV-mediated IFN-I antagonism on enhancement of BRSV replication in bovine turbinate (BT) cells. NY93/c N- Npro 18 EGFP replicated 0.4 – 1.6 TCID50 logs lower than NY93-wt in MDBK cells. NY93/c N- Npro 18 EGFP-infected MDBK cells synthesized IFN-I significantly higher than NY93/c- and NY93-wt-infected MDBK cells. BT cells co-infected with NY93/c N- Npro 18 EGFP/BRSV or NY93-wt/BRSV were evaluated to determine the effects of co-infection on BRSV replication and IFN-I induction in BT cells. BRSV RNA levels in NY93-wt/BRSV co-infected BT cells were 2.49, 2.79, and 2.89 copy number logs significantly greater than in NY93/c N- Npro 18 EGFP/BRSV co-infected BT cells on days 5, 7, and 9 post-infection, respectively. BVDV RNA levels in NY93/c N- Npro 18 EGFP-infected BT cells were 1.64 – 4.38 copy number logs lower than in NY93-wt-infected BT cells. NY93/c N- Npro 18 EGFP single and co-infected BT cells synthesized IFN-I significantly higher than NY93-wt single and co-infected BT cells. In summary, these findings suggest: (1) NY93/c N- Npro 18 EGFP BVDV2 induced higher levels of IFN-I than BVDV2-wt and may be useful as a safer, replicating BVDV vaccine, and (2) Enhancement of BRSV infection by BVDV co-infection is mediated by antagonism of IFN-I.