Enhancing Mulberry Leaf Meal with Urea by Pelleting to Improve Rumen Fermentation in Cattle

  • Tan, N.D. (Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Wanapat, M. (Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Uriyapongson, S. (Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Cherdthong, A. (Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Pilajun, R. (Tropical Feed Resources Research and Development Center (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University)
  • Received : 2011.08.07
  • Accepted : 2011.10.27
  • Published : 2012.04.01


Four, ruminally fistulated crossbred (Brahman${\times}$native) beef cattle with initial body weight of $420{\pm}15kg$ were randomly assigned according to a $4{\times}4$ Latin square design. The dietary treatments were mulberry leaf pellet (MUP) supplementation at 0, 200, 400 and 600 g/hd/d with rice straw fed to allow ad libitum intake. All steers were kept in individual pens and supplemented with concentrate at 5 g/kg of body weight daily. The experiment was 4 periods, and each lasted 21 d. During the first 14 d, all steers were fed their respective diets ad libitum and during the last 7 d, they were moved to metabolism crates for total urine and fecal collection. It was found that increasing MUP levels resulted in linearly increasing rice straw and total intakes (p<0.05). Ruminal temperature and pH were not significantly affected by MUP supplementation while $NH_3$-N concentration was increased (p<0.05) and maintained at a high level (18.5 mg/dl) with supplementation of MUP at 600 g/hd/d. Similarly, viable total bacteria in the rumen and cellulolytic bacteria were enriched by MUP supplementation at 600 g/hd/d. However, the rumen microbial diversity determined with a PCR-DGGE technique showed similar methanogenic diversity between treatments and sampling times and were similar at a 69% genetic relationship as determined by a UPGMA method. Based on this study, it could be concluded that supplementation of MUP at 600 g/hd/d improved DM intake, ruminal $NH_3$-N, and cellulolytic bacteria thus iimproving rumen ecology in beef cattle fed with rice straw.


  1. Anbarasu, C., N. Dutta, K. Sharma and M. Rawat. 2004. Response of goats to partial replacement of dietary protein by a leaf meal mixture containing Leucaena leucocephala, Morus alba and Tectona grandis. Small Rumin. Res. 51:47-56.
  2. AOAC. 1995. Official method of analysis, 16th ed. Animal Feeds: Association of Official Analytical Chemists, VA, USA
  3. Bakshi, M. P. S. and M. Wadhwa. 2007. Tree leaves as complete feed for goat bucks. Small Rumin. Res. 69:74-78.
  4. Bano, N., S. Ruffin, B. Ransom and J. T. Hollibaugh. 2004. Phylogenetic composition of arctic ocean archaeal assemblages and comparison with Antarctic assemblages. Appl. Environ. Microbiol. 70:781-789.
  5. Ba, N. X., V. D. Giang and L. D. Ngoan. 2005. Ensiling of mulberry foliage (Morus alba) and the nutritive value of mulberry foliage silage for goats in central Vietnam. Livestock Research for Rural Development. 17 (15).
  6. Calsamiglia, S., P. W. Cardozo, A. Ferret and A. Bach. 2008. Changes in rumen microbial fermentation are due to a combined effect of type of diet and pH. J. Anim. Sci. 86:702-711.
  7. Chen, X. B. and M. J. Gomes. 1995. Estimation of microbial protein supply to sheep and cattle based on urinary excretion of purine derivatives-an overview of the technical details. Occasional Publication 1992. International Feed Resources Unit, Rowett Research Institute, Aberdeen, UK.
  8. Cherdthong, A. and M. Wanapat. 2010. Development of urea products as rumen slow-release feed on ruminant production: A review. Aust. J. Basic Appl. Sci. 4:2232-2241.
  9. Cherdthong, A., M. Wanapat, P. Kongmun, R. Pilajun and P. Khekornsart. 2010. Rumen fermentation, microbial protein synthesis and cellulolytic bacterial population of swamp buffaloes as affected by roughage to concentrate ratio. J. Anim. Vet. Adv. 9:1667-1675.
  10. Cherdthong, A., M. Wanapat and C. Wachirapakorn. 2011a. Influence of urea calcium mixture supplementation on ruminal fermentation characteristics of beef cattle fed on concentrates containing high levels of cassava chips and rice straw. Anim. Feed Sci. Technol. 163:43-51.
  11. Cherdthong, A., M. Wanapat and C. Wachirapakorn. 2011b. Effects of urea-calcium mixture in concentrate containing high cassava chip on feed intake, rumen fermentation and performance of lactating dairy cows fed on rice straw. Livest. Sci. 136:76-84.
  12. Cherdthong, A., M. Wanapat and C. Wachirapakorn. 2011c. Influence of urea-calcium mixtures as rumen slow-release feed on in vitro fermentation using gas production technique. Arch. Anim. Nutr. 65:242-254.
  13. Chumpawadee, S., K. Sommart, T. Vongpralub and V. Pattarajinda. 2006. Effects of synchronizing the rate of dietary energy and nitrogen release on ruminal fermentation, microbial protein synthesis, blood urea nitrogen and nutrient digestibility in beef cattle. Asian-Aust. J. Anim. Sci. 19:181-188.
  14. Denman, S. E. and C. S. McSweeney. 2006. Developmentofa real-time PCR assay formonitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS Microbiol. Ecol. 58:572-582.
  15. Foiklang, S., M. Wanapat and W. Toburan. 2011. Effects of various plant protein sources in high-quality feed block on feed intake, rumen fermentation, and microbial population in swamp buffalo. Trop. Anim. Health Prod. 43:1517-1524.
  16. Galyean, M. 1989. Laboratory procedure in animal nutrition research. Department of Animal and Range Sciences. NewMexico State University, USA, pp. 107-122.
  17. Hristov, A. N., M. Vander Pol, M. Agle, S. Zaman, C. Schneider, P. Ndegwa, V. K. Vaddella, K. Johnson, K. J. Shingfield and S. K. R. Karnati. 2009. Effect of lauric acid and coconut oil on ruminal fermentation, digestion, ammonia losses from manure, and milk fatty acid composition in lactating cows. J. Dairy Sci. 92:5561-5582.
  18. Hungate, R. E. 1969. A roll tube method for cultivation of strict anaerobes. In: Methods in Microbiology (Ed. J. R. Norris and D. W. Ribbons). Academic, New York, pp. 117-313.
  19. Javaid, A., M. Aasif Shahzad, M. Nisa and M. Sarwar. 2011. Ruminal dynamics of ad libitum feeding in buffalo bulls receiving different level of rumen defradable protein. Livest. Sci. 135:98-102.
  20. Kandylis, K., I. Hadjigeorgiou and P. Harizanis. 2009. The nutritive value of mulberry leaves (Morus alba) as a feed supplement for sheep. Trop. Anim. Health Prod. 41:17-24.
  21. Koike, S. and Y. Kobayashi. 2001. Development and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus Favefaciens. FEMS Microbiol. Lett. 204:361-366.
  22. Kongmun, P., M. Wanapat, P. Pakdee and C. Navanukraw. 2010. Effect of coconut oil and garlic powder on in vitro fermentation using gas production technique. Livest. Sci. 127: 38-44.
  23. Mathis, C. P., R. C. Cochran, J. S. Heldt, B. C. Woods, I. E. Abdelgadir, K. C. Olson, E. C. Titgemeryer and E. S. Vanzant. 2000. Effec of supplemetal degradable intake protein on utilization of medium-to low-quality forage. J. Anim. Sci. 78:224-232.
  24. Moon, Y. H., J. U. Ok, S. J. Lee, J. K. Ha and S. S. Lee. 2010. A comparative study on the rumen microbial population, hydrolytic enzyme activities and dry matter degradability between different species of ruminant. Anim. Sci. J. 81:642-647.
  25. Orskov, E. R. 1994. Recent advances in understanding of microbial transformation in ruminants. Livest. Prod. Sci. 39: 53-60.
  26. Pilajun, R. and M. Wanapat. 2011. Methane production and methanogen population in rumen loquor of swamp buffalo as influenced by coconut oil and mangosteen peel powder supplementation. J. Anim. Vet. Adv. 10:2523-2527.
  27. Poungchompu, O., M. Wanapat, C. Wachirapakorn, S. Wanapat and A. Cherdthong. 2009. Manipulation of ruminal fermentation and methane production by dietary saponins and tannins from mangosteen peel and soapberry fruit. Arch. Anim. Nutr. 63:389-400.
  28. Ribeiro, S. S., J. T. Vasconcelos, M. C. Morais, C. B. C. F. Ítavo and G. L. Franco. 2011. Effect of ruminal infusion of slow-release polymer-coated urea of conventional urea on apparent nutrient digestibility, in situ degradability, and rumen parameters in cattle fed low-quality hay. Anim. Feed Sci. Technolo. 164:53-61.
  29. Salisbury, M. W., C. R. Krehbiel, T. T. Ross, C. L. Schultz and L. L. Melton. 2004. Effects of supplemental protein type on intake, nitrogen balance, and site, and extent of digestion in whiteface wethers consuming low-quality grass hay. J. Anim. Sci. 82:3567-3576.
  30. Samuel, M., S. Sagathewan, J. Thomas and G. Mathen. 1997. An HPLC method for estimation of volatile fatty acids of ruminal fluid. Indian J. Anim. Sci. 67:805-807.
  31. SAS. 1998. SAS/STAT user's guide. Version 6.12. SAS Inst. Inc., Cary, NC.
  32. Satter, L. D. and L. L. Slyter. 1974. Effect of ammonia concentration on rumen microbial protein production in-vitro. Br. J. Nutr. 32:199-208.
  33. Seo, J. K., S. W. Kim, M. H. Kim, S. D. Upadhaya, D. K. Kam and J. K. Ha. 2010. Direct-fed microbials for ruminant animals. Asian-Aust. J. Anim. Sci. 23:1657-1667.
  34. Steel, R. G. D. and J. H. Torrie. 1980. Principles and procedures of statistics: A Biometerial Approach. (2nd Ed.). Mc Graw-Hill, New York, USA.
  35. Van Soest, P. J., J. B. Robertson and B. A. Lewis. 1991. Method for dietary fiber, neutral detergent fiber and non-starch polysaccharide in relation to animal nutrition. J. Dairy Sci. 74: 3583-3597.
  36. Wanapat, M. 2009. Potential uses of local feed resources for ruminants. Trop. Anim. Health Prod. 42:1035-1049.
  37. Wanapat, M. and A. Cherdthong. 2009. Use of real-time PCR technique in studying rumen cellulolytic bacteria population as affected by level of roughage in swamp buffalo. Curr. Microbiol. 58:294-299.
  38. Wanapat, M., A. Cherdthong, P. Pakdee and S. Wanapat. 2008. Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citrates Stapf) powder supplementation. J. Anim. Sci. 86:3497-3503.
  39. Wanapat, M., C. Promkot and S. Wanapat. 2006. Effect of cassoy-urea pellet as a protein source in concentrate on ruminal fementation and digestibility in cattle. Asian-Aust. J. Anim. Sci. 19:1004-1009.
  40. Wanapat, M., R. Pilajun and P. Kongmun. 2009a. Ruminal ecology of swamp buffalo as influenced by dietary sources. Anim. Feed Sci. Technol. 151:205-214.
  41. Wanapat, M., S. Polyorach, K. Boonnop, C. Mapato and A. Cherdthong. 2009b. Effects of treating rice straw with urea or urea and calcium hydroxide upon intake, digestibility, rumen fermentation and milk yield of dairy cows. Livest. Sci. 125:238-243.
  42. Wright, A. D. G., C. H. Auckland and D. H. Lynn. 2007. Molecular diversity of methanogens in feedlot cattle from ontario and Prince Edward Island, Canada. Appl. Environ. Microbiol. 73: 4206-4210.
  43. Xin, H. S., D. M. Schaefer, Q. P. Liu, D. E. Axe and Q. X. Meng. 2010. Effects of polyurethane coated urea supplement on in vitro ruminal fermentation, ammonia release dynamics and lactating performance of Holstein dairy cows fed a steam-flaked corn-based diet. Asian-Aust. J. Anim. Sci. 23:491-500.
  44. Yang, J. Y., J. Seo, H. J. Kim, S. Seo and J. K. Ha. 2010. Nutrient synchrony: Is it a suitable strategy to improve nitrogen utilization and animal performance? Asian-Aust. J. Anim. Sci. 23:972-979.
  45. Yu, Z. and M. Morrison. 2004. Improved extraction of PCR-quality community DNA from digesta and fecal sample. BioTechniques 36:808-812.
  46. Zhu, Z., R. B. Hinson, L. Ma, D. Li and G. L. Allee. 2010. Growth performance of nursery pigs fed 30% distillers dried grain with solubles (DDGS) and the effects of pelleting on performance and nutrient digestibility. Asian-Aust. J. Anim. Sci. 23:792-798.

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