Evaluation of Tea and Spent Tea Leaves As Additives for Their Use in Ruminant Diets School of Agriculture, Food, and Rural Deve

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Evaluation of Tea and Spent Tea Leaves As Additives for Their Use in Ruminant Diets School of Agriculture, Food, and Rural Deve Evaluation of tea and spent tea leaves as additives for their use in ruminant diets The thesis submitted for the degree of Doctor of Philosophy by Diky Ramdani Bachelor of Animal Husbandry, Universitas Padjadjaran, Indonesia Master of Animal Studies, University of Queensland, Australia School of Agriculture, Food, and Rural Development Newcastle University Newcastle upon Tyne, United Kingdom October, 2014 Declaration I confirm that the work undertaken and written in this thesis is my own work that it has not been submitted in any previous degree application. All quoted materials are clearly distinguished by citation marks and source of references are acknowledged. The articles published in a peer review journal and conference proceedings from the thesis are listed below: Journal Ramdani, D., Chaudhry, A.S. and Seal, C.J. (2013) 'Chemical composition, plant secondary metabolites, and minerals of green and black teas and the effect of different tea-to-water ratios during their extraction on the composition of their spent leaves as potential additives for ruminants', Journal of Agricultural and Food Chemistry, 61(20): 4961-4967. Proceedings Ramdani, D., Seal, C.J. and Chaudhry, A.S., (2012a) ‘Simultaneous HPLC analysis of alkaloid and phenolic compounds in green and black teas (Camellia sinensis var. Assamica)’, Advances in Animal Biosciences, Proceeding of the British Society of Animal Science Annual Conference, Nottingham University, UK, April 2012, p. 60. Ramdani, D., Seal, C.J. and Chaudhry, A.S., (2012b) ‘Effect of different tea-to-water ratios on proximate, fibre, and secondary metabolite compositions of spent tea leaves as a potential ruminant feed additive’, Advances in Animal Biosciences, Proceeding of the British Society of Animal Science Annual Conference, Nottingham University, UK, April 2012, p. 62. Ramdani, D., Chaudhry, A.S. and Seal, C.J., (2013a) ‘Effect of spent tea leaves on in-vitro total gas production from rice straw-based ruminant diets’, Advances in Animal Biosciences, Proceeding of the British Society of Animal Science Annual Conference, Nottingham University, UK, April 2013, p. 49. Ramdani, D., Chaudhry, A.S. and Seal, C.J., (2013b) ‘Using spent tea leaves to improve in-vitro degradability but reduce rumen ammonia from rice straw-based diets’, Advances in Animal Biosciences, Proceeding of the British Society of Animal Science Annual Conference, Nottingham University, UK, April 2013, p. 121. i Ramdani, D., Chaudhry, A.S. and Seal, C.J., (2013c) ‘Tea leaves improve in-vitro degradability but reduce rumen ammonia from rice-straws based diet’, Book of Abstract of the 64th Annual Meeting of the European Federation of Animal Science, Nantes, France, August 2013, p. 582. Ramdani, D., Chaudhry, A.S. and Seal, C.J., (2014a) ‘Potential use of tea leaves to reduce rumen ammonia and methane productions’, Advances in Animal Biosciences, Proceeding of the British Society of Animal Science Annual Conference, Nottingham University, UK, April 2014, p. 83. ii Acknowledgment Firstly, all praise is due to Almighty God, the Creator and Lord of the universe, who enabled me to complete this thesis. My deepest and warmest thanks and appreciation to my main supervisor, Dr. Abdul S. Chaudhry for his endless encouragement, kindness, support, and friendship throughout the years of my PhD study. A debt of gratitude is owed to Professor Chris Seal as my second supervisor for his continues support and encouragement to complete my PhD thesis. It has been an honour for me to work with both of you. I also would like to thank Professor Sandra Edwards (internal examiner) and Dr. Gilbert Pellikaan (external examiner, Wageningen University) for their efforts to improve the quality of this thesis. I gratefully acknowledge Directorate General of Higher Education, Indonesian Ministry of Education and Culture for the financial support through a PhD scholarship scheme. I am also grateful to my employer, Faculty of Animal Husbandry, Universitas Padjadjaran, Indonesia for moral supports during my study. I would like to to thank Dr. Mohammad M.H Khan, Nuhu Bello Rano, Dr. Sujiang Zhang, Dr. Socrates Stergiadis, and Eleni Chatzidimitriou for their helps in different experiments and laboratory analysis. I also thank Peter Shotton, Chris Bulman, Fiona Maclachan, Wendy Bal, Roy Lamb, Craig Oliver, and other staff members of the School of Agriculture, Food, and Rural Development for their technical assistance during my study. Thanks to Jim Wightman, David Watson, and Angela Fogerty and sons for their assistance during the Farm trial. In addition, I would like to give special thanks to PT. Kabepe Chakra for providing green and black tea leaves, PT. Coca-Cola Bottling Indonesia for providing green and black spent tea leaves, and Linden Foods Ltd. for rumen fluid, fat, and carcass data collections of the experimental lambs. Lastly, I would like to express my gratitude to my beloved and precious mother, dad, brothers, wife, and daughters who always give me love, patience, encouragement, support, and their prayers which are valuable for the successful completion of my PhD study. iii Abbreviations ADG, average daily gain ADF, acid detergent fibre ADIC, acid detergent insoluble carbon ADIN, acid detergent insoluble nitrogen ADIP, acid detergent insoluble protein A:P, acetate to propionate ratio ATP, adenosine triphosphate BS, barley straws BSP, buffer soluble protein BTL, black tea leaves C, carbon C, catechin CG, catechin gallate CH4, methane CLA, conjugated linoleic acid CO2, carbon dioxide CON, concentrate mix diet CP, crude protein CSBTL, company spent black tea leaves CSGTL, company spent green tea leaves CT, condensed tannins DM, dry matter DMI, dry matter intake tDMI, total dry matter intake DNA, deoxyribonucleic acid EC, epicatechin ECG, epicatechin gallate EE, ether extract EGC, epigallocatechin EGCG, epigallocatechin gallate EO, essential oils EU, European union FA, fatty acids iv FAME, fatty acid methyl esters FCR, feed conversion ratio GC, gas chromatography GC, gallocatechin GCG, gallocatechin gallate GC-MS, gas chromatography- mass spectroscopy GE, gross energy tGP, total gas production GTL, green tea leaves H2, hydrogen HCl, hydrochloric acid HiCON, high concentrate diet HPLC, high performance liquid chromatography ICP-OES, inductively coupled plasma-optical emission spectroscopy IVCPD, in-vitro crude protein degradability IVDMD, in-vitro dry matter degradability IVOMD, in-vitro organic matter degradability KCl, potassium chloride KJ, kilo joule LoCON, low concentrate diet ME, metabolisable energy MJ, mega joule MUFA, monounsaturated fatty acids N, nitrogen NDF, neutral detergent fibre NDIC, neutral detergent insoluble carbon NDIN, neutral detergent insoluble nitrogen NDIP, neutral detergent insoluble protein NH3, ammonia NS, non-significant n3:n6, omega 3 to omega 6 ratio O2, Oxygen OM, organic matter PRS, perennial ryegrass silage PUFA, polyunsaturated fatty acids v PEG, polyethylene glycol RH, ryegrass hay RNA, ribonucleic acid RS, rice straws S, sulphur SBM, soybean meal SBTL, spent black tea teaves SD, standard deviation SEM, standard error of mean SFA, saturated fatty acids SGTL, spent green tea leaves STL, spent tea leaves TF, theaflavin TF-G, theaflavin gallate TP, total phenols TS, total saponins TT, total tannins VFA, volatile fatty acids tVFA, total volatile fatty acids WS, wheat straws WSC, water soluble carbohydrate vi Abstract Animal scientists have been challenged to improve animal production systems with respect not only to competitiveness and efficiency but at the same time producing products which are healthy for the consumers and friendly to the environment. Plant secondary metabolites such as tannins, saponins, and essential oils have been investigated for their advantageous outcomes as ‘natural’ additives to manipulate rumen fermentation via decreased ammonia (NH3) and methane (CH4) production, improved animal health and vitality, and increased meat quality. Tea leaves is one of native plants being rich in secondary metabolites and widely known to have health benefits for human consumption. However, the information on chemical characteristics of tea leaves and their spent tea leaves (STL) as residues along with their prospective as additives for ruminants is still inadequate. Therefore, a series of four studies aimed to evaluate chemical characteristics of tea and their STL as additives for their use in ruminant diets through in-vitro and in-vivo experiments. The first study aimed to (1) characterize chemical composition, plant secondary metabolites, minerals, and fatty acid profiles in green (GTL) and black (BTL) tea leaves as well as their STL, and to (2) test the hypothesis that a higher tea-to-water ratio would affect the extraction of the chemical compounds from tea leaves into water to yield a more nutrient-rich tea drink and STL. Green (SGTL) and black (SBTL) STL were obtained following a 3 x 2 factorial arrangement by extracting 3 different amounts (T1= 2.8 g, T2= 5.6 g and T3= 11.2 g) of the 2 tea types (green and black) in a fixed volume of 300 ml boiling water for 5 minutes. GTL and BTL had similar (g/kg DM or as stated otherwise) dry matter (DM, g/kg), organic matter (OM), crude protein (CP), ash, and total monounsaturated fatty acids (MUFA, %) but GTL had significantly higher ether extract (EE), total phenols (TP), total tannins (TT), condensed tannins (CT), total saponins (TS), alkaloids, catechins, and total polyunsaturated fatty acids (PUFA, %) with lower neutral (NDF) and acid (ADF) detergent fibres, theaflavins, and total saturated fatty acids (SFA, %) compared with BTL. There was no significant difference between GTL and BTL for most mineral components (mg/kg DM) except Mn, which was significantly higher in GTL, and Na and Cu which were significantly higher in BTL. Company SGTL (CSGTL) had the same CP, NDF, CT, alkaloids, SFA, MUFA, PUFA, Mg, Cu, and Cd but higher EE, ash, TP, TT, TS, catechins, Ca, K, P, Mn, Fe, Cr, and Pb with lower DM, OM, ADF, and theaflavins compared with company SBTL (CSBTL). In addition, a higher tea-to-water ratio during extraction significantly reduced the loss of soluble compounds into water and hence yielded a more nutrient-rich STL.
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