Rumen fermentation and bacterial communities in weaned Chahaer lambs on diets with different protein levels

doi:10.1016/S2095-3119(15)61217-5

摘要/Abstract

Abstract: We evaluated the effects of diets with different crude protein (CP) levels on growth performance, rumen fermentation and bacterial communities in weaned Chahaer lambs. 128 weaned Chahaer lambs ((20.56±1.43) kg body weight; ram:ewe 1:1) aged (61±1.85) d were randomly allotted to one of four diets with CP content of 11.17% (T1), 12.06% (T2), 13.40% (T3) or 14.36% (T4). Ruminal fermentation parameters were measured and bacterial communities were analysed using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative PCR. The average daily gain and feed utilization efficiency in T3 were higher than those in the other groups (P<0.05), although the dry matter intake and metabolizable energy intake were similar. Total volatile fatty acid concentration in the ruminal fluid of T3 was lower than that of T1 (P=0.011), T2 (P=0.008) or T4 (P=0.309). The ammonia nitrogen concentration and acetate/propionate ratio of ruminal fluid were significantly higher in lambs fed the higher CP diets, whereas the molar concentrations of propionate and butyrate of ruminal fluid were lower. The rumen bacterial community was similar in T2 and T3 which shown more stable and diverse rumen microbes ecosystem compared with the other groups. The DGGE profiles and phylogenetic tree indicated that Bacteroides uniformis, Clostridium alkalicellulosi, Alkalibaculum bacchi and Saccharofermentans sp. were common bacterium of Chahaer lamb rumen. B. uniformis, C. alkalicellulosi, Saccharofermentans sp. and Gracilibacter thermotolerans, which belong to the Bacteroidetes and Firmicutes phyla, were the dominant species in the rumen of lambs fed 13.40% CP. However, Ruminococcus albus, Ruminococcus flavefaciens and Butyrivibrio fibrisolvens were not different in lambs fed different CP diets. Therefore, it could be concluded that B. uniformis, C. alkalicellulosi, A. bacchi and Saccharofermentans sp. were common bacteria of Chahaer lamb rumen. Furthermore, the dietary CP of 13.04% could improve performance and change rumen fermentation model by increasing the dominant species’ peak intensities of B. uniformis, C. alkalicellulosi, Saccharofermentans sp. and Gracilibacter thermotolerans and stabilizing rumen microbial ecosystem.

Key words: dietary crude protein , rumen fermentation , bacterial diversity , weaned lamb , PCR-DGGE

YANG Chun-tao, SI Bing-wen, DIAO Qi-yu, JIN Hai, ZENG Shu-qin, TU Yan. Rumen fermentation and bacterial communities in weaned Chahaer lambs on diets with different protein levels[J]. Journal of Integrative Agriculture, 2016, 15(7): 1564-1574.

参考文献

complete rations with variable protein and energy levels prepared using by-products of pulses and oilseeds on carcass characteristics, meat and meat ball quality of goats. Asian Australasian Journal of Animal Sciences, 19, 1437–1449.

Askar A R, Salama R, El-Shaer H M, Safwat M A, Poraei M, Nassar M S, Badawy H S, Raef O. 2014. Evaluation of the use of arid-area rangelands by grazing sheep: Effect of season and supplementary feeding. Small Ruminant Research, 121, 262–270.

Broderick G A, Muck R E. 2009. Effect of alfalfa silage storage structure and rumen-protected methionine on production in lactating dairy cows. Journal of Dairy Science, 92, 1281–1289.

Chanthakhoun V, Wanapat M. 2010. Effect of legume (Phaseolus calcaratus) hay supplementation on rumen cellulolytic bacterial populations in swamp buffaloes investigated by the real-time PCR technique. Journal of Animal and Veterinary Advances, 9, 1654–1659.

Chanthakhoun V, Wanapat M, Berg J. 2012. Level of crude protein in concentrate supplements influenced rumen characteristics, microbial protein synthesis and digestibility in swamp buffaloes (Bubalus bubalis). Livestock Science, 144, 197–204.

Chegeni A, Li Y L, Deng K D, Jiang C G, Diao Q Y. 2013. Effect of dietary polymer-coated urea and sodium bentonite on digestibility, rumen fermentation, and microbial protein yield in sheep fed high levels of corn stalk. Livestock Science, 157, 141–150.

Chen S, Niu L, Zhang Y. 2010. Saccharofermentans acetigenes gen. nov., sp. nov., an anaerobic bacterium isolated from sludge treating brewery wastewater. International Journal of Systematic and Evolutionary Microbiology, 60, 2735–2738.

Cherdthong A, Wanapat M, Wachirapakorn C. 2011. 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. Livestock Science, 136, 76–84.

Collins M D, Lawson P A, Willems A, Cordoba J J, Fernandez-Garayzabal J, Garcia P, Cai J, Hippe H, Farrow J. 1994. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. International Journal of Systematic Bacteriology, 44, 812–826.

Davidson S, Hopkins B A, Diaz D E, Bolt S M, Brownie C, Fellner V, Whitlow L W. 2003. Effects of amounts and degradability of dietary protein on lactation, nitrogen utilization, and excretion in early lactation Holstein cows. Journal of Dairy Science, 86, 1681–1689.

Denman S E, McSweeney C S. 2006. Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS Microbiology Ecology, 58, 572–582.

Dutta T K, Agnihotri M K, Sahoo P K, Rajkumar V, Das A K. 2009. Effect of different protein-energy ratio in pulse by-products and residue based pelleted feeds on growth, rumen fermentation, carcass and sausage quality in Barbari kids. Small Ruminant Research, 85, 34–41.

El-Shaer H M. 2010. Halophytes and salt-tolerant plants as potential forage for ruminants in the Near East region. Small Ruminant Research, 91, 3–12.

Fahmy A A, Youssef K M, El Shaer H M. 2010. Intake and nutritive value of some salt-tolerant fodder grasses for sheep under saline conditions of South Sinai, Egypt. Small Ruminant Research, 91, 110–115.

Fawcett J K, Scott J. 1960. A rapid and precise method for the determination of urea. Journal of Clinical Pathology, 13, 156–159.

Fernando S C, Purvis H T, Najar F Z, Sukharnikov L O, Krehbiel C R, Nagaraja T G, Roe B A, DeSilva U. 2010. Rumen microbial population dynamics during adaptation to a high-grain diet. Applied and Environmental Microbiology, 76, 7482–7490.

Firkins J L, Yu Z, Morrison M. 2007. Ruminal nitrogen metabolism: Perspectives for integration of microbiology and nutrition for dairy. Journal of Dairy Science, 90, E1–E16.

Fromin N, Hamelin J, Tarnawski S, Roesti D, Jourdain Miserez K, Forestier N, Teyssier Cuvelle S, Gillet F, Aragno M, Rossi P. 2002. Statistical analysis of denaturing gel electrophoresis (DGE) fingerprinting patterns. Environmental Microbiology, 4, 634–643.

Hobson P N, Stewart C S. 1997. The Rumen Microbial Ecosystem. Springer, The Netherlands.

Kabir F, Sultana M S, Shahjala M, Khan M J, Alam M Z. 2004. Effect of protein supplementation on growth performance in female goats and sheep under grazing condition. Pakistan Journal of Nutrition, 3, 237–239.

Kang S, Wanapat M, Pakdee P, Pilajun R, Cherdthong A. 2012. Effects of energy level and Leucaena leucocephala leaf meal as a protein source on rumen fermentation efficiency and digestibility in swamp buffalo. Animal Feed Science and Technology, 174, 131–139.

Koike S, Yoshitani S, Kobayashi Y, Tanaka K. 2003. Phylogenetic analysis of fiber-associated rumen bacterial community and PCR detection of uncultured bacteria. FEMS Microbiology Letters, 229, 23–30.

Kolver E S, De Veth M J. 2002. Prediction of ruminal pH from pasture-based diets. Journal of Dairy Science, 85, 1255–1266.

Kongmun P, Wanapat M, Pakdee P, Navanukraw C, Yu Z. 2011. Manipulation of rumen fermentation and ecology of swamp buffalo by coconut oil and garlic powder supplementation. Livestock Science, 135, 84–92.

Li M, Penner G B, Hernandez Sanabria E, Oba M, Guan L L. 2009. Effects of sampling location and time, and host animal on assessment of bacterial diversity and fermentation parameters in the bovine rumen. Journal of Applied Microbiology, 107, 1924–1934.

Li M, Zhou M, Adamowicz E, Basarab J A, Guan L L. 2012. Characterization of bovine ruminal epithelial bacterial communities using 16S rRNA sequencing, PCR-DGGE, and qRT-PCR analysis. Veterinary Microbiology, 155, 72–80.

Ma T, Chen D D, Tu Y, Zhang N F, Si B W, Deng K D, Diao Q Y. 2014. Effect of dietary supplementation with resveratrol on nutrient digestibility, methanogenesis and ruminal microbial flora in sheep. Journal of Animal Physiology and Animal Nutrition, 99, 676–683.

Mcdonald P. 2002. Animal Nutrition. Pearson Education, Prentice Hall, Harlow.

Muyzer G, De Waal E C, Uitterlinden A G. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology, 59, 695–700.

NRC (National Research Council). 2001. Nutrient Requirements of Dairy Cattle. 7th ed. National Academy Press, Washington, D.C.

Odum E P, Odum H T, Andrews J. 1971. Fundamentals of Ecology. Saunders College Publishing, Philadelphia, PA.

Oliveira J S D, Queiroz A C D, Mantovani H C, Melo M R D, Detmann E, Santos E. M, Bayão G F V. 2011. Effect of propionic and lactic acids on in vitro ruminal bacteria growth. Revista Brasileira de Zootecnia, 40, 1121–1127.

Pimentel P, Vilela F, Nguluve D W, Muir J P, José A. 2011. Supplementary feeding increases live weight gain of Angoni cattle during the dry season in Mozambique. Livestock Research for Rural Development, 23, 124.

Prakash B, Dutta T K, Siddiqui I A. 2006. Effect of plane of nutrition on nutrient utilization and performance of Barbari kids. Indian Journal Animal Nutrition, 23, 29–33.

Reynal S M, Broderick G A. 2005. Effect of dietary level of rumen-degraded protein on production and nitrogen metabolism in lactating dairy cows. Journal of Dairy Science, 88, 4045–4064.

Robinson P H, McQueen R E, Burgess P L. 1991. Influence of rumen undegradable protein levels on feed intake and milk production of dairy cows. Journal of Dairy Science, 74, 1623–1631.

Ryan S M, Unruh J A, Corrigan M E, Drouillard J S, Seyfert M. 2007. Effects of concentrate level on carcass traits of Boer crossbred goats. Small Ruminant Research, 73, 67–76.

Salyers A A. 1995. Fermentation of polysaccharides by human colonic anaerobes. In: Dietary Fibre, Mechanisms of Action in Human Physiology and Metabolism. John Libbey Eurotext, Paris.

Sarwar M, Nisa M, Javaid A, Shahzad M A. 2010. Ruminal characteristics, blood pH, blood urea nitrogen and nitrogen balance in Nili-Ravi buffalo (Bubalus bubalis) bulls fed diets containing various level of ruminally degradable protein. Italian Journal of Animal Science, 6, 441–445.

Shahjalal M, Bishwas M A, Tareque A M, Dohi H. 2000. Growth and carcass characteristics of goats given diets varying protein concentration and feeding level. Asian Australasian Journal of Animal Sciences, 13, 613–618.

Steele M A, Dionissopoulos L, AlZahal O, Doelman J, McBride B W. 2012. Rumen epithelial adaptation to ruminal acidosis in lactating cattle involves the coordinated expression of insulin-like growth factor-binding proteins and a cholesterolgenic enzyme. Journal of Dairy Science, 95, 318–327.

Stewart C S, Flint H J, Bryant M P. 1997. The rumen bacteria. In: The Rumen Microbial Ecosystem. Springer, The Netherlands.

Sun P, Wang J Q, Zhang H T. 2011. Effects of supplementation of Bacillus subtilis natto Na and N1 strains on rumen development in dairy calves. Animal Feed Science and Technology, 164, 154–160.

Vinh N T, Wanapat M, Khejornsart P, Kongmun P. 2011. Studies of diversity of rumen microorganisms and fermentation in swamp buffalo fed different diets. Journal of Animal Veterinary Advances, 10, 406–414.

Wang D, Ba L. 2008. Ecology of meadow steppe in northeast China. The Rangeland Journal, 30, 247–254.

Yang S, Ma S, Chen J, Mao H, He Y, Xi D, Yang L, He T, Deng W. 2010. Bacterial diversity in the rumen of Gayals (Bos frontalis), Swamp buffaloes (Bubalus bubalis) and Holstein cow as revealed by cloned 16S rRNA gene sequences. Molecular Biology Reports, 37, 2063–2073.