A. B. C. Succiniclasticum. Paraprevotella. Control DPA EPA DHA Control DPA EPA DHA a b. a a. b c.

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1 389 Session 2: Miroil eosystem nd herivore nutrition Effet of dietry ddition of EPA, DPA nd DHA on rumen teril ommunity in ows nd ewes. An in vitro pproh Dvid Crreño 1, Álvro Belenguer 1, Eri Pinlohe 2, Dvid Yáñez-Ruiz 3, Plo G. Torl 1, Gonzlo Hervás 1, Neil R. MEwn 2,4, C. Jmie Newold 2,5, Frutos Pilr 1 1 Instituto de Gnderí de Montñ (CSIC-ULE), León, Spin, 2 IBERS, Aerystwyth University, Aerystwyth, UK, 3 Estión Experimentl del Zidin (CSIC), Grnd, Spin, 4 Roert Gordon University, Aerdeen, UK, 5 Sotlnd s Rurl College, Edinurgh, UK E-mil: p.frutos@si.es Tke home messge Dietry ddition of EPA, DPA, nd DHA ltered the undne of severl teril gener, suh s Prevotell, Hllell, Prprevotell nd Suinilstium, tht might prtiipte in rumen iohydrogention. Introdution The role of mrine lipids s modultors of rumen iohydrogention (BH) of unsturted ftty ids (FA) is likely to e due to the impt of their n-3 polyunsturted FA on the miroiot (Torl et l., 2017). However, the effet of individul FA, nmely 20:5 n-3 (EPA), 22:5 n-3 (DPA), nd 22:6 n-3 (DHA) hs rrely een studied nd it is still unertin whih rumen teri re involved in the BH proess. Moreover, despite interspeies differenes in the rumen teril omposition (Torl et l., 2016), we re not wre of ny reports ompring the effets of these FA in ows nd sheep. Therefore, rumen inoul from these speies were used to exmine in vitro the effet of EPA, DPA nd DHA on the teril ommunity. A onomitnt study hd nlysed their influene on ruminl fermenttion nd BH (Torl et l., 2017). Mterils & methods Bth ultures of rumen miroorgnisms were onduted using inoul from 2 ruminnt speies (i.e., 2 nnulted ows nd 2 nnulted ewes fed TMR with forge:onentrte rtio 50:50) nd 4 tretments (ontrol the TMR without supplementtion, nd the TMR plus 2% DM of EPA, DPA or DHA). Inutions lsted for 24 h nd were repeted on 3 onseutive dys. DNA ws extrted from freeze dried smples nd teril 16S rrna (V1-V2) mplion ws nlysed y Ion Torrent PGM next genertion sequening (de l Fuente et l., 2014). The effet of niml speies nd tretments on the tx s reltive undne ws nlysed y ANOVA, with the MIXED proedure of the SAS 9.4. Results & disussion Only effets on teri tht were previously suggested to e somehow relted to rumen BH re reported here. Most hnges were omprle in oth speies, ut some groups shifted with EPA nd/or DHA only in ttle (e.g., inreses in Hllell, Ruminoous nd Ruminoter) or only in sheep (inrements of Prprevotell nd dereses in Osilliter). Prevotell undne ws redued y ll tretments, espeilly EPA nd DHA, in the two niml speies (Figure 1A). This genus hs een relted to 18:0 formtion (Huws et l., 2011), whih is onsistent with oservtions y Torl et l. (2017). Aundnes of Hllell in ttle nd Prprevotell in sheep inresed similrly with EPA nd DHA (Figure 1B). These miroorgnisms form suinte, preursor of propionte for Suinilstium spp., whose undne rised with ll tretments in ttle nd with DHA in sheep (Figure 1C). A puttive ssoition etween propionte formtion nd shift in BH pthwys ws previously speulted in Torl et l. (2017) nd suggest tht teri relted to propionte metolism might ply prt in those BH routes Prevotell Cttle Hllell Prprevotell Cttle A. B. C d Suinilstium Cttle Figure 1 Reltive undnes of Prevotell (A), Hllell nd Prprevotell (B), nd Suinilstium (C) in ruminl digest of ttle nd sheep fter 24 h-inutions without FA supplementtion (Control) or with DPA, EPA or DHA. Conlusion Dietry ddition of 2% DM of EPA, DPA nd DHA in in vitro th ultures lter some teril gener potentilly involved in lipid metolism, suh s Prevotell, Hllell, Prprevotell nd Suinilstium. Most hnges were omprle in sheep nd ows, ut there were lso vritions exlusive to eh ruminnt speies. The effet of DPA ws less pronouned thn tht of EPA or DHA. Aknowledgements Support y the Spnish MINECO (AGL nd AGL ) nd the Europen Regionl Development Fund is knowledged. D. Crreño enefits from FPI nd P.G. Torl from Rmón y Cjl reserh ontrt. Referenes De l Fuente G, Belnhe A, Girwood SE, Pinlohe E, Wilkinson T nd Newold CJ Plos One 9, Huws SA, Kim EJ, Lee MRF, Sott MB, Tweed JKS, Pinlohe E, Wlle R J nd Solln ND Environmentl Miroiology 13, Torl PG, Bernrd L, Belenguer A, Rouel J, Hervás G, Chillird Y nd Frutos P, Journl of Diry Siene 99, Torl PG, Hervás G, Crreño D, Leskinen H, Belenguer A, Shingfield KJ nd Frutos P Journl of Diry Siene 100,

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3 (Generl Seretry) (Chir) (Chir) (Chir)

4 Herivore nutrition supporting sustinle intensifition nd Advnes in Animl Biosienes gro-eologil pprohes niml VOLUME 9 ISSUE 3 Pges British Soiety of Animl Siene (Pulished April 2018) Interntionl Bull Fertility (Pulished My 2018) Interntionl Symposium on the Nutrition of Herivores EU Cost Ation IPEMA Biology of Lttion in Frm Animls Interntionl Symposium on Digestive Physiology of Pigs British Soiety of Animl Siene Interntionl Symposium on Ruminnt Physiology SEPTEMBER 2018 ISSUE 3 Current nd Upoming Conferenes in Advnes in Animl Biosienes nd Animl VOLUME 9 Herivore Herivore nutrition nutrition supporting supporting sustinle intensifi sustinle intensifition tion nd nd gro-eologil gro-eologil pprohes pprohes SEPTEMBER 2018 SEPTEMBER 2018 VOLUME 9 ISSUE 3 ISSN dvnes in niml iosienes Advnes in Animl Biosienes Herivore nutrition supporting sustinle intensifition nd gro-eologil pprohes Proeedings of the 10th Interntionl Symposium on the Nutrition of Herivores ISNH 2018 Editors René Bumont Mthieu Silererg Iselle Cssr-Mlek

5 Effet of dietry ddition of EPA, DPA nd DHA on rumen teril ommunity in ows nd ewes. An in vitro pproh D. Crreño1, A. Belenguer1, E. Pinlohe2, D.R. Yáñez-Ruiz3, P.G. Torl1, G. Hervás1, N.R. MEwn2, C.J. Newold2 nd P. Frutos1 1Instituto de Gnderí de Montñ (CSIC ULE), Fin Mrzns s/n, Grulleros (León), Spin 2Institute of Biologil, Environmentl nd Rurl Sienes, Aerystwyth University, Aerystwyth SY23 3DA, United Kingdom 3Instituto de Nutriión Animl, Estión Experimentl del Zidín (CSIC), Grnd, Spin TAKE-HOME MESSAGE Dietry ddition of EPA, DPA, nd DHA ltered the undne of severl teril gener, suh s Prevotell, Hllell, Prprevotell nd Suinilstium, tht might prtiipte in rumen iohydrogention. RESULTS The role of mrine lipids s modultors of rumen iohydrogention (BH) of unsturted ftty ids (FA) is likely to e due to the impt of their n-3 polyunsturted FA on the miroiot. However, the effet of individul FA, nmely 20:5 n-3 (EPA), 22:5 n-3 (DPA), nd 22:6 n-3 (DHA), hs rrely een studied nd it is still unertin whih rumen teri re involved in BH. Despite interspeies differenes in the rumen teril omposition (Torl et l., 2016), we re not wre of ny reports ompring the effets of these FA in ows nd sheep. Only effets on teri previously suggested to e relted to rumen BH re reported. Most hnges were omprle in oth speies, ut some groups shifted with EPA nd/or DHA only in ttle (e.g., Hllell, Ruminoous nd Ruminoter) or only in sheep (e.g., Prprevotell nd Osilliter). Therefore, rumen inoul from ttle nd ewes were used to exmine in vitro the effet of EPA, DPA nd DHA on the teril ommunity. A onomitnt study hd nlysed their influene on ruminl fermenttion nd BH (Torl et l., 2017). % totl sequenes INTRODUCTION Prevotell Cttle MATERIAL AND METHODS Bth ultures of rumen miroorgnisms Sustrte - TMR with forge:onentrte rtio 50:50 % totl sequenes Prevotell undne ws redued y ll tretments, espeilly EPA nd DHA, in oth speies. This genus hs een relted to 18:0 formtion (Huws et l., 2011) Inoulum - Rumen fluid from nnulted nimls (3 runs on different dys) Inution time - 24h Speies - Cttle (2 ows) nd sheep (2 ewes) Hllell TMR + 2% DM of 22:5 n-3 (DPA) TMR + 2% DM of 22:6 n-3 (DHA) Cttle Aundnes of Hllell in ttle nd Prprevotell in sheep inresed similrly with EPA nd DHA. Suinilstium % totl sequenes Tretments TMR without supplementtion (Control) TMR + 2% DM of 20:5 n-3 (EPA) Prprevotell d Cttle Hllell nd Prprevotell form suinte, preursor of propionte for Suinilstium spp., whose undne rised with ll tretments in ttle nd with DHA in sheep. DNA extrtion nd sequening Bteril 16S rrna (V1-V2) Ion Torrent PGM next genertion sequening An ssoition etween propionte formtion nd shift in BH pthwys ws previously speulted (Torl et l. 2017), suggesting tht teri relted to propionte metolism might ply prt in those BH routes. CONCLUSION Dietry ddition of 2% DM of EPA, DPA nd DHA in in vitro th ultures lter some teril gener potentilly involved in lipid metolism (e.g., Prevotell, Hllell, Prprevotell nd Suinilstium). Most hnges were omprle in sheep nd ows, ut there were lso vritions exlusive to eh ruminnt speies. The effet of DPA ws less pronouned thn tht of EPA or DHA. Huws S A, Kim E J, Lee M R F, Sott M B, Tweed J K S, Pinlohe E, Wlle R J, Solln N D 2011 Environmentl Miroiology 13, Torl P G, Bernrd L, Belenguer A, Rouel J, Hervás G, Chillird Y, Frutos P 2016 Journl of Diry Siene 99, Torl P G, Hervás G, Crreño D, Leskinen H, Belenguer A, Shingfield K J, Frutos P 2017 Journl of Diry Siene 100, th Interntionl Symposium on the Nutrition of Herivores 2018 (Clermont-Ferrnd, Frne) Aknowledgments: Spnish MINECO (AGL nd AGL , nd FPI nd Rmon y Cjl progrmmes) nd Europen Regionl Development Fund