Abstract
Biochar, which is the product of biomass pyrolysis, has been suggested as a feed supplement to improve understanding of the rumen microbial ecosystem. The study aims to investigate the potential of activated charcoal to favorably modify rumen fermentation and reduce methane emission by measuring ruminal parameters, enzymes, and microbial communities. Thirty goats (weighing 14.35 ± 1.40 kg) were randomly put into three groups: a control diet group and groups with diet supplemented with activated charcoal at 50 mg/kg and 150 mg/kg. Each group comprised ten replicate pens. The basal diet contained 40% barley and 60% alfalfa hay, and diets were similar to isocaloric and isonitrogenous. The trial lasted 60 days following a 14-day adaptation. The biochar was obtained from pyrolysis of a river plant (Ceratophyllum demersum), impregnate with zinc chloride (ZnCl2), and went through the carbonization process at 700 ℃. Rumen fluids were sampled, filtered, and stored at -80 ℃ in liquid nitrogen to analyze eukaryotic diversity and enzyme activity; the rest was stored at -20 °C to measure fermentation parameters. The results showed that supplementation with activated biochar did not affect pH values and acetate/propionate molar ratio (P = 0.021). Ruminal ammonia (NH3-N), Total Volatile Fatty Acids, Acetate, and Butyrate (%) were significantly (p < 0.05) higher at 50 mg and 150 mg activated charcoal groups, compared with control group. Supplementation with activated biochar increased the activities of CMCase, Xylanase, Pectinase, and α-amylase in the rumen, but β-glucosidase and protease were not affected. These changes were associated with improved growth performance in the goats. Overall, the interplay of biochar’s physical and chemical characteristics plays a major role in different living systems by influencing the different metabolic enzymes and processes, availability of different nutrients, and environmental conditions. Dietary supplementation with activated biochar increased total bacteria, Fibrobacter succinogenes, Rumonococcus albus, and Ruminococcus flavefaciens) and methanogenic, but did not affect protozoa. In conclusion, this study would further promote biochar applications in ruminal fermentation processes.
References
Al-Azzawi M., Bowtell l., Hancock K. Preston S. 2012.Addition of activated carbon into a cattle diet to mitigate GHG emissions and improve production. Sustainability.13(15): 8254. https://doi.org/10.3390/su13158254
Antonopoulos D.A., Aminov R.I., Duncan P.A., White B.A., Mackie R.I. 2003. Characterization of the gene encoding glutamate dehydrogenase (gdhA) from the ruminal bacterium R. flavefaciens FD-1. Archives of microbiology. 179 (3) :184–190. https://doi.org/10.1007/s00203-002-0513-4
AOAC. 2006. Official Methods of Analysis. 18th Edition, Association of Official Analytical Chemists, Gaithersburgs, MD.
Bagherpoor Z., Rezaei J., Rouzbehan Y. 2023. Potential of biochar in enhancing the effectiveness of probiotics Bacilli and Lactobacilli on in vitro microbial populations, hydrolytic enzymes, and ruminal fermentation in sheep.Animal Production Research, 12(3): 29-47. https://doi.org/10.22124/ar.2023.25526.1790
Barboza S.P., Parker K.L., Hume I.D. 2009. Integrative wildlife nutrition Berlin, Heidelberg, Germany: Springer-Verlag, 342. https:// doi.org/10.1007/978-3-540-87885-8
Cabeza I., Waterhouse T., Sohi S., Rookej A. 2018. Effect of biochar produced from different biomass sources and at different process temperatures on methane production and ammonia concentrations in vitro. Animal Feed Science and Technology, 237: 1-7. https://doi.org/10.1016/j.anifeedsci.2018.01.003
Doreau M., Ferlay A. 1995. Effect of dietary lipids on nitrogen metabolism in the rumen:a review. Livestock Production Science, 43(2): 97–110. https://doi.org/10.1016/0301-6226(95)00041-I
European Centre for Disease Prevention and Control (ECDC). Historical data by year - West Nile fever seasonal surveillance. Available at: https://ecdc.europa.eu/en/west-nile-fever/surveillance-and-disease-data/historical Accessed: 31.07.2019.
Garillo E.P., Pradhan R., Tobioko H. 1995.Effect of activated carbon on growth, ruminal characteristics, blood profiles and feed digestibility in sheep. Asian-Australasian Journal of Animal Sciences, 8(1): 43-50. https://doi.org/10.5713/ajas.1995.43
Garillo E.P., Pradhan R., Tobioko H. 1994. Effects of activated charcoal on ruminal characteristics and blood profiles in mature goats. The West Japan Journal of Animal Science, 37:85-89.
Lammirato C., Miltner A., Kaestner M. 2011. Effects of Wood Char and Activated Carbon on the Hydrolysis of Cellobiose by β-Glucosidase from Aspergillus Niger. Soil Biology and Biochemistry, 43(9): 1936–1942. https://doi.org/10.1016/j.soilbio.2011.05.021
Lehmann J., Joseph S. 2015. Biochar for Environmental Management: Science and Technology and Implementation. Routledge, London.
Leng R.A., Inthapanya S., Preston T.R. 2012. Biochar lowers net methane production from rumen fluid in vitro. Livestock Research for Rural Development, 24(6): 103.
Lü F., Yang L., Liming S., Pinjing H. 2019. Powdered biochar doubled microbial growth in anaerobic digestion of oil. Applied Energy, 247(2): 605-614. https://doi.org/10.1016/j.apenergy.2019.04.052
Machmüller A., Soliva C.R., Kreuzer M. 2003. Methane-suppressing effect of myristic acid in sheep as affected by dietary calcium and forage proportion. The British journal of nutrition, 90(3):529–540. https://doi.org/10.1079/bjn2003932
Martin C., Michalet-Doreau B. 1995. Variations in mass and enzyme activity of rumen micro- organisms: Effect of barley and buffer supplements . The Journal of the Science of Food and Agriculture, 67:407-413. https://doi.org/10.1002/jsfa.2740670319
Martínez M.E., Ranilla M.J., Tejido M.L., Ramos S., Carro M.D. 2010. Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. I. Digestibility, fermentation parameters, and microbial growth. Journal of dairy science, 93(8): 3684-3698. https://doi.org/10.3168/jds.2009-2933
McFarlane Z.D., Myer P.R., Cope E.R., Evans N.D., Bone T.C., Biss B.E., Mulliniks J.T. 2017. Effect of biochar type and size on in vitro rumen fermentation of orchard grass hay. Agricultural Sciences, 8(4):316–325. https://doi.org/10.4236/as.2017.84023
Mitsumori M., Shinkai T., Takenaka A. 2012. Responses in digestion, rumen fermentation and microbial populations to inhibition of methane formation by a halogenated methane analogue. The British journal of nutrition, 108(3):482–491. https://doi.org/10.1017/S0007114511005794
Nguyen L.N., Vu M.T., Johir M.A., Pernice M., Ngo H.H, Zdarta J., Jesionowski T., Nghiem L. 2021. Promotion of direct interspecies electron transfer and potential impact of conductive materials in anaerobic digestion and its downstream processing-a critical review. Bioresource Technology, 341:125847. https://doi.org/10.1016/j.biortech.2021.125847
O’Toole A., Andersson D., Gerlach A., Glaser B., Kammann C., Kern J., Kuoppamäki K., Mumme J., Schmidt H.-P., Schulze M., Srocke F., Stenrød M. and Stenström J. 2016. Current and future applications for biochar. In Biochar in European Soils and Agriculture: Science and Practice, Eds. S. Shackley, G. Ruysschaert, K. Zwart, B. Glaser Routledge, London, UK , 253-280.
Pereira C., Muetzel R., Camps S., Arbestain M., Bishop P., Hina K., Hedley M. 2014. Assessment of the influence of biochar on rumen and silage fermentation: a laboratory-scale experiment. Animal Feed Science and Technology, 196:22–31. https://doi.org/10.1016/j.anifeedsci.2014.06.019
Qomariyah N., Retnani Y., Jayanegara A., Wina E., Permana IG. 2021. Influence of biochar and liquid smoke additives from cacao-pod husks on in vitroruminal fermentation characteristics. Advances in Animal and Veterinary Sciences, 9(4): 533-543. http://dx.doi.org/10.17582/journal.aavs/2021/9.4.533.543
Romanos, J., Beckner, M., Rash, T., Firlej, L., Kuchta, B., Yu, P. 2012. Nanospace Engineering of KOH Activated Carbon. Nanotechnology, 23(1): 15401. https://doi.org/10.1088/0957-4484/23/1/015401
Saleem A., Ribeiro G., Yang W., Ran T., Beauchemin K., McGeough E., Ominski K.H., Okine E.K., McAllister T.A. 2018. Effect of engi-neered biocarbon on rumen fermentation, microbial protein synthesis,and methane production in an artificial rumen (RUSITEC) fed a high for-age diet. Journal of Animal Science, 96(8):3121-3130. https://doi.org/10.1093/jas/sky204
Schmidt H.P., Hagemann N., Draper K., Kammann C. 2019. The use of biochar in animal feeding. Peer J, 7: 7373. https://doi.org/10.7717/peerj.7373
Schmidt H.P., Wilson K., Kammann C. 2017.Using biochar in animal farming to recycle nutrients and reduce greenhouse gas emissions. Geophysical Research Abstracts, 19: 5719.
Sirjani M.H., Rezaei J., Zahedifar M., Rouzbehan Y. 2022. Effect of adding biochar in diets containing probiotics on in vitro fermentation variables, health indicators, rectum bacteria, and blood enzymes of Holstein calves. Animal Production Research, 11(4): 11-19. https://doi.org/10.22124/ar.2023.23067.1727
Soo J., Malik B.A., Turner J.M., Persad R., Wine E., Siminoski K., Huynh H.Q. 2013. Use of exclusive enteral nutrition is just as effective as corticosteroids in newly diagnosed pediatric Crohn's disease. Digestive diseases and sciences, 58(12): 3584−3591. https://doi.org/10.1007/s10620-013-2855-y
Sun Y., Wu Q., Wang Y., Li C., Liang X. 2021. Protein-derived 3D amorphous carbon with N, O doping as high rate and long lifespan anode for potassium ion batteries. Journal of Power Sources, 512:230530. https://doi.org/10.1016/j.jpowsour.2021.230530
Teoh R., Caro E., Holman D.B., Joseph S., Meale S.J., Chaves A.V. 2019. Effects of hardwood biochar on methane production, fermentation characteristics, and the rumen microbiota using rumen simulation. Frontiers in Microbiology, 10: 1–13. https://doi.org/10.3389/fmicb.2019.01534
Terry S.A., Ribeiro G.O., Gruninger R.J., Chaves A.V., Beauchemin K.A., Okine E., McAllister T.A. 2019. A Pine Enhanced Biochar Does Not Decrease Enteric CH4 141.Emissions, but Alters the Rumen Microbiota. Frontiers in veterinary science, 6: 308. https://doi.org/10.3389/fvets.2019.00308
Toth J. D., Dou Z. 2016. Use and impact of biochar and charcoal in animal production systems. In Agricultural and environmental applications of biochar: advances and barriers. Eds. M. Guo, Z. He, S.M. Uchimiya, Soil Science Society of America. Inc, Madison: 199–224. https://doi.org/10.2136/sssaspecpub63.2014.0043.5
Van D.T.T., Mui N.T., Ledin I. 2006. Effect of method of processing foliage of Acacia mangium and inclusion of bamboo charcoal in the diet on performance of growing goats. Animal Feed Science and Technology, 130(3), 242–256. https://doi.org/10.1016/J.ANIFEEDSCI.2006.01.00.8
Van Soest P.J., Robertson J.B., Lewis B.A. 1991. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science ,74(10):3583–3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2
Villarreal J., Kahn C.A., Dunford J.V., Patel E., Clark R.F. 2015. A retrospective review of the prehospital use of activated charcoal. The American journal of emergency medicine, 33(1): 56-59. https://doi.org/10.1016/j.ajem.2014.10.019
Wang M., Wang J.J., Taft N.D., Hollier C.A., Myers G., Wang X. 2019. Effect of alkali-enhanced biochar on silicon uptake and suppression of gray leaf spot development in perennial ryegrass.Crop Protection, 119: 9–16. https://doi.org/10.1016/j.cropro.2019.01.013
Yang M., Zhan X., Wang K., Zhu S., Ye Z., Sheng K., Zhang G.X. 2022. Investigation of cascade valorization of Pistia stratiotes L. by hydrothermal treatment. Fuel. 324(1-3): 124473. https://doi.org/10.1016/j.fuel.2022.124473 OIE: Animal Diseases. Available at: http://www.oie.int/en/animal-health-in-the-world/information-on-aquatic-and-terrestrial-animal-diseases/. Accessed 07.08.2019.

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