Optimizing alfalfa silage with rumen-focused tannin dynamics: Toward enhanced protein retention

Articles in Press

Document Type : Research Article (Regular Paper)

Author

Special Domestic Animals Institute, Research Institute of Zabol, Zabol, Iran

Abstract

The effects of tannic acid (TA) and purified tannins from pistachio by-products (PB) and pomegranate pulp (PP) on the chemical composition, in vitro gas production and in situ ruminal disappearance of crude protein (CP) and tannins in alfalfa silage (AS) were investigated. The experimental treatments (on a dry matter [DM] basis) included: 1) control silage (CS), 2) alfalfa silage treated with 2% TA (TAS), 3) alfalfa silage treated with 2% PB tannin (PBS), and 4) alfalfa silage treated with 2% PP tannin (PPS). Four replicates of each treatment were prepared and ensiled for 60 days. The results showed that the treatment of AS with PB and PP tannins significantly lowered the pH of silage compared to the control (P<0.05). In addition, all tannin sources significantly decreased the ammonia nitrogen (NH₃-N) content in the silages (P<0.05). The treatment of AS with all sources of tannins decreased the A fraction of CP (non-protein nitrogen compounds; NPN), while it increased the B1 fraction (true soluble protein). Potential gas production (mL) was significantly lower in PB- and PP-treated silages compared to control and TA-treated silages (P<0.05); however, the rate constant of gas production was only reduced in TA-treated silage compared to control (P<0.05). Organic matter digestibility and metabolizable energy were also significantly reduced by all tannin sources (P<0.05). Compared to the control, the in situ degradability of the rapidly soluble fraction (a) of CP decreased in the AS treated with PB and PP tannins, while the slowly degradable fraction (b) increased (P<0.05). The degradability rate was higher in the PBS than in the TA-treated and control silages (P<0.05). No significant differences were found among treatments in ruminal tannin disappearance, with approximately 73% of tannins disappearing at the time zero in all silages. In conclusion, incorporation of 2% tannins from PB or PP into AS, compared to TA and the control, effectively reduced NH₃-N and the soluble protein fraction in the rumen without significantly altering the potential and effective degradability of protein.

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References

AOAC, 2005. Official Methods of Analysis. 18th ed. Association of Official Analytical Chemists, Washington, DC. USA.
Bae, H.D., McAllister, T.A., Yanke, L.J., Cheng, K.J., Muir, A.D., 1993. Effect of condensed tannins on endoglucanase activity and filter paper digestion by Fibrobacter succinogenes S85. Applied Environmental Microbiology 59, 2132-2138.
Bagheripour, E., Rouzbehan, Y., Alipour, D., 2008. Effects of ensiling, air-drying and addition of polyethylene glycol on in vitro gas production of pistachio by-products. Animal Feed Science and Technology 146, 327-336.
Battelli, M., Colombini, S., Parma, P., Galassi, G., Crovetto, G.M., Spanghero, M., Pravettoni, D., Zanzani, S.A., Manfredi, M.T., Rapetti, L., 2023. In vitro effects of different levels of quebracho and chestnut tannins on rumen methane production, fermentation parameters, and microbiota. Frontiers in Veterinary Science 10, 1178288.
Bernardes, T.F., Gervásio, J.R.S., De Morais, G., Casagrande, D.R., 2019. A comparison of methods to determine pH in silages. Journal of Dairy Science 102, 9039-9042.
Bhatta, R., Saravanan, M., Baruah, L., Sampath, K.T., 2015. Nutritional and anti-nutritional composition of unconventional feed resources and their effects on rumen fermentation. Tropical Animal Health and Production 47, 437-445.
Chen, L., Bao, X., Guo, G., Huo, W., Xu, Q., Wang, C., Liu, Q., 2021a. Effects of hydrolyzable tannin with or without condensed tannin on alfalfa silage fermentation characteristics and in vitro ruminal methane production, fermentation patterns, and microbiota. Animals 11, 1967.
Chen, L., Bao, X., Guo, G., Huo, W., Xu, Q., Wang, C., Liu, Q., 2021b. Treatment of alfalfa silage with tannin acid at different levels modulates ensiling characteristics, methane mitigation, ruminal fermentation patterns and microbiota. Animal Feed Science and Technology 278, 114997.
Dentinho, M.T.P., Paulos, K., Portugal, P.V., Moreira, O.C., Santos‐Silva, J., Bessa, R.J., 2019. Proteolysis and in situ ruminal degradation of lucerne ensiled with Cistus ladanifer tannins. Grass and Forage Science 74, 78-85.
Esen, S., 2023. Optimizing ruminant nutrition: Insights from a comprehensive analysis of silage composition and in vitro gas production dynamics using nonlinear models. Biosystems 234, 105062.
Fijałkowska, M., Pysera, B., Lipiński, K., Strusińska, D., 2015. Changes of nitrogen compounds during ensiling of high protein herbages–a review. Annals of Animal Science 15, 289-305.
Fagundes, G.M., Benetel, G., Santos, K.C., Welter, K.C., Melo, F.A., Muir, J.P., Bueno, I.C.S., 2020. Tannin-rich plants as natural manipulators of rumen fermentation in the livestock industry. Molecules 25, 2943.
Fonseca, N.V.B., Cardoso, A.D.S., Bahia, A.S.R.D.S., Messana, J.D., Vicente, E.F., Reis, R.A., 2023. Additive tannins in ruminant nutrition: An alternative to achieve sustainability in animal production. Sustainability 15, 4162.
Givens, D.I., Rulquin, H., 2004. Utilisation by ruminants of nitrogen compounds in silage-based diets. Animal Feed Science and Technology 114, 1-18.
Guo, X., Chen, D., Huang, P., Gao, L., Zhou, W., Zhang, J., Zhang, Q., 2024. Effects of tannin‐tolerant lactic acid bacteria in combination with tannic acid on the fermentation quality, protease activity and bacterial community of stylo silage. Journal of the Science of Food and Agriculture https://doi.org/10.1002/jsfa.14027.
He, L., Lv, H., Chen, N., Wang, C., Zhou, W., Chen, X., Zhang, Q., 2020. Improving fermentation, protein preservation and antioxidant activity of Moringa oleifera leaves silage with gallic acid and tannin acid. Bioresource Technology 297, 122390.
Henderson, N., 1993. Silage additives. Animal Feed Science and Technology 45(1), 35–56.
Iranian Council of Animal Care, 1995. Guide to the Care and Use of Experimental Animals. Vol. 1. Isfahan: Isfahan University of Technology.
Jayanegara, A., Sujarnoko, T.U., Ridla, M., Kondo, M., Kreuzer, M., 2019. Silage quality as influenced by concentration and type of tannins present in the material ensiled: A meta-analysis. Journal of Animal Physiology and Animal Nutrition 103, 456-465.
Ke, W., Zhang, H., Li, S., Xue, Y., Wang, Y., Dong, W., Cai, Y., Zhang, G., 2022. Influence of condensed and hydrolyzable tannins on the bacterial community, protein degradation, and fermentation quality of alfalfa silage. Animals 12, 831.
Lagrange, S., Lobón, S., Villalba, J.J., 2019. Gas production kinetics and in vitro degradability of tannin-containing legumes, alfalfa and their mixtures. Animal Feed Science and Technology 253, 56-64.
Li, X., Tian, J., Zhang, Q., Jiang, Y., Hou, J., Wu, Z., Yu, Z., 2018. Effects of applying Lactobacillus plantarum and Chinese gallnut tannin on the dynamics of protein degradation and proteases activity in alfalfa silage. Grass and Forage Science 73, 648-659.
Licitra, G., Hernandez, T.M., Van Soest, P.J., 1996. Standardization of procedures for nitrogen fractionation of ruminant feeds. Animal Feed Science and Technology 57, 347-358.
Mahanani, M.M.P., Kurniawati, A., Hanim, C., Anas, M.A., Yusiati, L.M., 2020. Effect of Leucaena leucocephala leaves as tannin source on rumen microbial enzyme activities and in vitro gas production kinetics. IOP Conference Series: Earth and Environmental Science 478, 012088.
Makkar, H.P.S., 2000. Quantification of tannins in tree foliage: A Laboratory Manual. Vienna: Joint FAO/IAEA.
Makkar, H.P.S., 2003. Effects and fate of tannins in ruminant animals, adaptation to tannins, and strategies to overcome detrimental effects of feeding tannin-rich feeds. Small Ruminant Research 49, 241-256.
Makkar, H.P.S., Blümmel, M., Becker, K., 1995. In vitro effects and interactions of tannins and saponins and fate of tannins in rumen. Journal of Science of Food and Agriculture 69, 481-493.
Menke, K.H., Steingass, H., 1988. Estimation of the energetic feed value obtained from chemical analysis and gas production using rumen fluid. Animal Research and Development 28, 7-55.
Min, B.R., Barry, T.N., Attwood, G.T., McNabb, W.C., 2003. The effect of condensed tannins on the nutrition and health of ruminants fed fresh temperate forages: A review. Animal Feed Science and Technology 106, 3-19.
Mhlongo, L.C., Kenyon, P., Nsahlai, I.V., 2025. Effect of Acacia mearnsii forage or tannin extract on rumen dry matter and crude protein degradation. Journal of Animal Physiology and Animal Nutrition 109, 22-29.
Mirzaei-Aghsaghali, A., Maheri-Sis, N., Mansouri, H., Razeghi, M.E., Mirza-Aghazadeh, A., Cheraghi, H., Aghajanzadeh-Golshani, A., 2011. Evaluating potential nutritive value of pomegranate processing by-products for ruminants using in vitro gas production technique. ARPN Journal of Agricultural and Biological Science 6, 45-51.
Mohamaden, W.I., Hegab, I.M., Shang-li, S., 2020. In situ ruminal degradation kinetics and blood metabolites as affected by feeding different sources of tannin and flavonoids to small-tailed Han rams. Livestock Science 239, 104029.
Muck, R.E., Moser, L.E., Pitt, R.E., 2003. Postharvest factors affecting ensiling. In: Buxton, D., Muck, R., Harrison, J. (Eds.) Silage Science and Technology. Vol. 42. Madison, WI: ASA-CSSA-SSSA, pp. 251-304.
Mueller-Harvey, I., 2006. Unravelling the conundrum of tannins in animal nutrition and health. Journal of the Science of Food and Agriculture 86, 2010-2037.
Ørskov, E.R., McDonald, I., 1979. The estimation of protein degradability in the rumen from incubation measurements weighted according to rate of passage. Journal of Agricultural Science 92, 499-503.
Patra, A.K., Saxena, J., 2011. Exploitation of dietary tannins to improve rumen metabolism and ruminant nutrition. Journal of Animal Physiology and Animal Nutrition 95, 215-234.
Perez-Maldonado, R.A., Norton, B.W., 1996. Digestion of 14C-labelled condensed tannins from Desmodium intortum in sheep and goats. British Journal of Nutrition 76, 501-513.
Rira, M., Morgavi, D.P., Popova, M., Maxin, G., Doreau, M., 2022. Microbial colonisation of tannin-rich tropical plants: Interplay between degradability, methane production and tannin disappearance in the rumen. Animal, 100589.
Santos, G.T., Oliveira, R.L., Petit, H.V., Cecato, U., Zeoula, L.M., Rigolon, L.P., Damasceno, J.C., Branco, A.F., Bett, V., 2000. Effect of tannic acid on composition and ruminal degradability of bermudagrass and alfalfa silages. Journal of Dairy Science 83, 2016-2020.
SAS, 2001. SAS/STAT User's Guide: Version 9.1. SAS Institute Inc., Cary, NC.
Tabacco, E., Borreani, G., Crovetto, G.M., Galassi, G., Colombo, D., Cavallarin, L., 2006. Effect of chestnut tannin on fermentation quality, proteolysis, and protein rumen degradability of alfalfa silage. Journal of Dairy Science 89, 4736-4746.
Tavendale, M.H., Meagher, L.P., Pacheco, D., Walker, N., Attwood, G.T., Sivakumaran, S., 2005. Methane production from in vitro rumen incubations with Lotus pedunculatus and Medicago sativa, and effects of extractable condensed tannin fractions on methanogenesis. Animal Feed Science and Technology 123, 403-419.
Theodorou, M.K., Williams, B.A., Dhanoa, M.S., McAllan, A., France, J., 1994. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology 48, 185-197.
Van den Bossche, T., Goossens, K., Haesaert, G., Wambacq, E., Vandaele, L., De Boever, J.L., 2024. Autumn grass treated with a hydrolyzable tannin extract versus lactic acid bacteria inoculant: Effects on silage fermentation characteristics and nutritional value and on performance of lactating dairy cows. Journal of Animal Physiology and Animal Nutrition 108, 111-125.
Van Soest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74, 3583-3597.
Wang, C., Pian, R., Chen, X., Lv, H., Zhou, W., Zhang, Q., 2020. Beneficial effects of tannic acid on the quality of bacterial communities present in high-moisture mulberry leaf and stylo silage. Frontiers in Microbiology 11, 586412.
Weatherburn, M., 1967. Phenol-hypochlorite reaction for determination of ammonia. Analytical Chemistry 39, 971-974.
Winters, A.L., Cockburn, J.E., Dhanoa, M.S., Merry, R.J., 2000. Effects of lactic acid bacteria in inoculants on changes in amino acid composition during ensilage of sterile and non-sterile ryegrass. Journal of Applied Microbiology 89, 442-451.
Xu, H., Wu, N., Na, N., Sun, L., Zhao, Y., Ding, H., Fang, Y., Wang, T., Xue, Y., Zhong, J., 2022. Fermentation weight loss, fermentation quality, and bacterial community of ensiling of sweet sorghum with lactic acid bacteria at different silo densities. Frontiers in Microbiology 13, 1013913.
Yanza, Y.R., Fitri, A., Suwignyo, B., Hidayatik, N., Kumalasari, N.R., Irawan, A., Jayanegara, A., 2021. The utilisation of tannin extract as a dietary additive in ruminant nutrition: A meta-analysis. Animals 11, 3317.
Zamiri, M.J., Rajaei Sharifabadi, H., Bagheri, A.S., Solhjoo, A., 2015. Effects of inclusion of licorice (Glycyrrhiza glabra L.) leaves, a tannin-containing plant, in a low-protein diet on feedlot performance and carcass characteristics of fat-tailed lambs. Tropical Animal Health and Production 47, 597-602.
Zhang, L.L., Wang, Y.M., Xu, M., 2014. Preparation and antimicrobial activity of tannin polymers from Platycarya strobilacea infructescence. Materials Research Innovations 18 (Suppl. 2), 1046-1049.
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Available Online from 14 February 2025

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