Long-term high-grain diet altered the ruminal pH, fermentation, and composition and functions of the rumen bacterial community, leading to enhanced lactic acid production in Japanese Black beef cattle during fattening

Autoři: Toru Ogata aff001;  Hiroki Makino aff002;  Naoki Ishizuka aff002;  Eiji Iwamoto aff003;  Tatsunori Masaki aff003;  Kentaro Ikuta aff004;  Yo-Han Kim aff002;  Shigeru Sato aff001
Působiště autorů: United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan aff001;  Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, Morioka, Iwate, Japan aff002;  Hyogo Prefectural Technology Center of Agriculture, Forestry and Fisheries, Hyogo, Japan aff003;  Awaji Agricultural Technology Center, Minami-Awaji, Hyogo, Japan aff004
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225448


To increase intramuscular fat accumulation, Japanese Black cattle are commonly fed a high-grain diet from 10 to 30 months of age although it can result in the abnormal accumulation of organic acids in the rumen. We explored the effect of long-term high-concentrate diet feeding on ruminal pH and fermentation, and its effect on the rumen bacterial community in Japanese Black beef cattle during a 20-month fattening period. Nine castrated and fistulated Japanese Black beef cattle were housed with free access to food and water throughout the study period (10–30 months of age). The fattening stages included Early, Middle, and Late stages (10–14, 15–22, and 23–30 months of age, respectively). Cattle were fed high-concentrate diets for the experimental cattle during fattening. The body weight of the cattle was 439 ± 7.6, 561 ± 11.6, and 712 ± 18.5 kg (mean ± SE) during the Early, Middle, and Late stages, respectively. Ruminal pH was measured continuously during the final 7 days of each stage, and rumen fluid and blood samples were collected on day 4 (fourth day during the final 7 days of the pH measurements). The 24-h mean ruminal pH during the Late stage was significantly lower than that during the Early stage. Total volatile fatty acid (VFA) during the Late stage was significantly lower than during the Early and Middle stages, but no changes were noted in individual VFA components. The lactic acid concentration during the Late stage was significantly higher than that during the Early and Middle stages. The bacterial richness indices decreased significantly during the Late stage in accordance with the 24-h mean ruminal pH. Among the 35 bacterial operational taxonomic units (OTUs) shared by all samples, the relative abundances of OTU8 (Family Ruminococcaceae) and OTU26 (Genus Butyrivibrio) were positively correlated with the 24-h mean ruminal pH. Total VFA concentration was negatively correlated with OTU167 (Genus Intestinimonas), and lactic acid concentration was correlated positively with OTU167 and OTU238 (Family Lachnospiraceae). These results suggested that long-term high-grain diet feeding gradually lowers ruminal pH and total VFA production during the Late fattening stage. However, the ruminal bacterial community adapted to feeding management and the lower pH during the Late stage by preserving their diversity or altering their richness, composition, and function, to enhance lactic acid production in Japanese Black beef cattle.

Klíčová slova:

Beef – Cattle – Diet – Fermentation – Lactic acid – Ruminococcus – Sequence databases – Shannon index


1. Allen MS. Relationship between fermentation acid production in the rumen and the requirement for physically effective fiber. J. Dairy Sci. 1997;80: 1447–1462. doi: 10.3168/jds.S0022-0302(97)76074-0 9241607

2. Nagaraja TG, Titgemeyer EC. Ruminal acidosis in beef cattle: the current microbiological and nutritional outlook. J. Dairy Sci. 2007;90: E17–E38. doi: 10.3168/jds.2006-478 17517750

3. Nagata R, Kim YH, Ohkubo A, Kushibiki S, Ichijo T, Sato S. Effects of repeated subacute ruminal acidosis challenges on the adaptation of the rumen bacterial community in Holstein bulls. J. Dairy Sci. 2018;101: 4424–4436. doi: 10.3168/jds.2017-13859 29477528

4. Khafipour E, Li S, Plaizier JC, Krause DO. Rumen microbiome composition determined using two nutritional models of subacute ruminal acidosis. Appl. Environ. Microbiol. 2009;75: 7115–7124. doi: 10.1128/AEM.00739-09 19783747

5. Petri RM, Schwaiger T, Penner GB, Beauchemin KA, Forster RJ, McKinnon JJ, et al. Characterization of the core rumen microbiome in cattle during transition from forage to concentrate as well as during and after an acidotic challenge. PloS one 2013;8: e83424. doi: 10.1371/journal.pone.0083424 24391765

6. Plaizier JC, Li S, Tun HM, Khafipour E. Nutritional models of experimentally-induced subacute ruminal acidosis (SARA) differ in their impact on rumen and hindgut bacterial communities in dairy cows. Front. Microbiol. 2017;7: 2128. doi: 10.3389/fmicb.2016.02128 28179895

7. Mao SY, Zhang RY, Wang DS, Zhu WY. Impact of subacute ruminal acidosis (SARA) adaptation on rumen microbiota in dairy cattle using pyrosequencing. Anaerobe 2013;24: 12–19. doi: 10.1016/j.anaerobe.2013.08.003 23994204

8. Huo W, Zhu W, Mao S. Impact of subacute ruminal acidosis on the diversity of liquid and solid-associated bacteria in the rumen of goats. World J. Microbiol. Biotech. 2014;30: 669–680. doi: 10.1007/s11274-013-1489-8 24068532

9. Cameron PJ, Zembayashi M, Lunt DK, Mitsuhashi T, Mitsumoto M, Ozawa S, et al. Relationship between Japanese beef marbling standard and intramuscular lipid in the M. longissimus thoracis of Japanese Black and American Wagyu cattle. Meat Sci. 1994;38: 361–364. doi: 10.1016/0309-1740(94)90125-2 22059673

10. Oka A, Maruo Y, Miki T, Yamasaki T, Saito T. Influence of vitamin A on the quality of beef from the Tajima strain of Japanese Black cattle. Meat Sci. 1998;48: 159–167. doi: 10.1016/s0309-1740(97)00086-7 22062888

11. Ogata T, Kim YH, Masaki T, Iwamoto E, Ohtani Y, Orihashi T, et al. Effects of an increased concentrate diet on rumen pH and the bacterial community in Japanese Black beef cattle at different fattening stages. J. Vet. Med. Sci. 2019;81: 968–974. doi: 10.1292/jvms.19-0077 31118356

12. Ministry of Agriculture, Forestry and Fisheries (MAFF). [Internet]. Establishment of the methods of analysis in feeds and feed additives (notification no. 19‐shoan‐14729); 2008. [cited 2019 Sep 10]. http://www.famic.go.jp/ffis/feed/tuti/19_14729.html. Japanese

13. National Agriculture and Food Research Organization (NARO). Japanese Feeding Standard for Beef Cattle. 2008 ed. Tokyo, Japan: Japan Livestock Industry Association; 2009. Japanese

14. Sato S, Kimura A, Anan T, Yamagishi N, Okada K, Mizuguchi H, et al. A radio transmission pH measurement system for continuous evaluation of fluid pH in the rumen of cows. Vet. Res. Commun. 2012;36: 85–89. doi: 10.1007/s11259-012-9518-x 22281863

15. Hirabayashi H, Kawashima K, Okimura T, Tateno A, Suzuki A, Asakuma S, et al. Effect of nutrient levels during the far-off period on postpartum productivity in dairy cows. Anim. Sci. J. 2017;88: 1162–1170. doi: 10.1111/asj.12743 27957823

16. Kim Y-H, Nagata R, Ohtani N, Ichijo T, Ikuta K, Sato S. Effects of dietary forage and calf starter diet on ruminal pH and bacteria in Holstein calves during weaning transition. Front. Microbiol. 2016a;7: 1575. doi: 10.3389/fmicb.2016.01575 27818645

17. Amplicon PCR, Clean-Up PCR, Index PCR [Internet]. 16S Metagenomic Sequencing Library Preparation; 2013 [cited 2019 Sep 10]. https://emea.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/16s/16s-metagenomic-library-prep-guide-15044223-b.pdf.

18. Klindworth A, Pruesse E, Schweer T, Peplies J, Quast C, Horn M, et al. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res. 2013;41: e1. doi: 10.1093/nar/gks808 22933715

19. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, et al. Introducing mothur: Open-source, platform- independent, community-supported software for describing and comparing microbial communities. Appl. Environ. Microbiol. 2009;75: 7537–7541. doi: 10.1128/AEM.01541-09 19801464

20. Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD. Development of a dual-index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl. Environ. Microbiol. 2013;79: 5112–5120. doi: 10.1128/AEM.01043-13 23793624

21. Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J, et al. SILVA: A comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 2007;35: 7188–7196. doi: 10.1093/nar/gkm864 17947321

22. Iwai S, Weinmaier T, Schmidt BL, Albertson DG, Poloso NJ, Dabbagh K, et al. Piphillin: improved prediction of metagenomic content by direct inference from human microbiomes. PloS one 2016;11: e0166104. doi: 10.1371/journal.pone.0166104 27820856

23. Katamoto H, Yamada Y, Nishizaki S, Hashimoto T. Seasonal changes in serum vitamin A, vitamin E and β-carotene concentrations in Japanese Black breeding cattle in Hyogo prefecture. J. Vet. Med. Sci. 2003;65: 1001–1002. doi: 10.1292/jvms.65.1001 14532693

24. Takemura K, Shingu H, Mizuguchi H, Kim YH, Sato S, Kushibiki S. Effects of forage feeding on rumen fermentation, plasma metabolites, and hormones in Holstein calves during pre-and post-weaning periods. J. Anim. Sci. 2019;97: 2220–2229. doi: 10.1093/jas/skz088 30873561

25. Plaizier JC, Krause DO, Gozho GN, McBride BW. Subacute ruminal acidosis in dairy cows: The physiological causes, incidence and consequences. Vet. J. 2008;176:21–31. doi: 10.1016/j.tvjl.2007.12.016 18329918

26. Watanabe Y, Kim YH, Kushibiki S, Ikuta K, Ichijo T, Sato S. Effects of active dried Saccharomyces cerevisiae on ruminal fermentation and bacterial community during the short-term ruminal acidosis challenge model in Holstein calves. J. Dairy Sci. 2019;102: 6518–6531. doi: 10.3168/jds.2018-15871 31030914

27. Kimura A, Sato S, Kato T, Ikuta K, Yamagishi N, Okada K, et al. Relationship between pH and temperature in the ruminal fluid of cows, based on a ratio-transmission pH-measurement system. J. Vet. Med. Sci. 2012;74: 1023–1028. doi: 10.1292/jvms.12-0084 22516695

28. Schlau N, Guan NN, Oba M. The relationship between rumen acidosis resistance and expression of genes involved in regulation of intracellular pH and butyrate metabolism of ruminal epithelial cells in steers. J. Dairy Sci. 2012;95: 5866–5875. doi: 10.3168/jds.2011-5167 22863095

29. Kim YH, Toji N, Kizaki K, Kushibiki S, Ichijo T, Sato S. Effects of dietary forage and calf starter on ruminal pH and transcriptomic adaptation of the rumen epithelium in Holstein calves during the weaning transition. Physiol. Genomics 2016b;48: 803–809.

30. Abaker JA, Xu TL, Jin D, Chang GJ, Zhang K, Shen XZ. Lipopolysaccharide derived from the digestive tract provokes oxidative stress in the liver of dairy cows fed a high-grain diet. J. Dairy Sci. 2017;100: 666–678. doi: 10.3168/jds.2016-10871 27865500

31. Guo J, Chang G, Zhang K, Xu L, Jin D, Bilal MS, et al. Rumen-derived lipopolysaccharide provoked inflammatory injury in the liver of dairy cows fed a high-concentrate diet. Oncotarget 2017;8: 46769. doi: 10.18632/oncotarget.18151 28596485

32. Neubauer V, Petri RM, Humer E, Kröger I, Mann E, Reisinger N, et al. High-grain diets supplemented with phytogenic compounds or autolyzed yeast modulate ruminal bacterial community and fermentation in dry cows. J. Dairy Sci. 2018;101: 2335–2349. doi: 10.3168/jds.2017-13565 29331466

33. Zhang RY, Jin W, Feng PF, Liu JH, Mao SY. High-grain diet feeding altered the composition and functions of the rumen bacterial community and caused the damage to the laminar tissues of goats. Animal 2018;12: 2511–2520. doi: 10.1017/S175173111800040X 29553005

34. Golder HM, Denman SE, McSweeney C, Celi P, Lean IJ. Ruminal bacterial community shifts in grain-, sugar-, and histidine-challenged dairy heifers. J. Dairy Sci. 2014;97: 5131–5150. doi: 10.3168/jds.2014-8003 24881800

35. Aikman PC, Reynolds CK, Beever DE. Diet digestibility, rate of passage, and eating and rumination behavior of Jersey and Holstein cows. J. Dairy Sci. 2008;91: 1103–1114. doi: 10.3168/jds.2007-0724 18292266

36. Roche JF. The effect of nutritional management of the dairy cow on reproductive efficiency. Anim. Reprod. Sci. 2006;96:282–296. doi: 10.1016/j.anireprosci.2006.08.007 16996705

37. Biddle A, Stewart L, Blanchard J, Leschine S. Untangling the genetic basis of fibrolytic specialization by Lachnospiraceae and Ruminococcaceae in diverse gut communities. Diversity 2013;5: 627–640. doi: 10.3390/d5030627

38. Hook SE, Steele MA, Northwood KS, Dijkstra J, France J, Wright ADG, et al. Impact of subacute ruminal acidosis (SARA) adaptation and recovery on the density and diversity of bacteria in the rumen of dairy cows. FEMS Microbiol. Ecol. 2011;78: 275–284. doi: 10.1111/j.1574-6941.2011.01154.x 21692816

39. Seo B, Yoo JE, Lee YM, Ko G. Merdimonas faecis gen. nov., sp. nov., isolated from human faeces. Int. J. Syst. Evol. Microbiol. 2017;67: 2430–2435. doi: 10.1099/ijsem.0.001977 28741995

40. Kläring K, Hanske L, Bui N, Charrier C, Blaut M, Haller D, et al. Intestinimonas butyriciproducens gen. nov., sp. nov., a butyrate-producing bacterium from the mouse intestine. Int. J. Syst. Evol. Microbiol. 2013;63: 4606–4612. doi: 10.1099/ijs.0.051441-0 23918795

Článek vyšel v časopise


2019 Číslo 11