In vitro evaluation of nano zinc oxide (nZnO) on mitigation of gaseous emissions

  • Sarker, Niloy Chandra (Agricultural and Biosystems Engineering Department, North Dakota State University) ;
  • Keomanivong, Faithe (Animal Sciences Department, North Dakota State University) ;
  • Borhan, Md. (Agricultural and Biosystems Engineering Department, North Dakota State University) ;
  • Rahman, Shafiqur (Agricultural and Biosystems Engineering Department, North Dakota State University) ;
  • Swanson, Kendall (Animal Sciences Department, North Dakota State University)
  • Received : 2018.09.21
  • Accepted : 2018.10.29
  • Published : 2018.11.30


Background: Enteric methane ($CH_4$) accounts for about 70% of total $CH_4$ emissions from the ruminant animals. Researchers are exploring ways to mitigate enteric $CH_4$ emissions from ruminants. Recently, nano zinc oxide (nZnO) has shown potential in reducing $CH_4$ and hydrogen sulfide ($H_2S$) production from the liquid manure under anaerobic storage conditions. Four different levels of nZnO and two types of feed were mixed with rumen fluid to investigate the efficacy of nZnO in mitigating gaseous production. Methods: All experiments with four replicates were conducted in batches in 250 mL glass bottles paired with the ANKOM$^{RF}$ wireless gas production monitoring system. Gas production was monitored continuously for 72 h at a constant temperature of $39{\pm}1^{\circ}C$ in a water bath. Headspace gas samples were collected using gas-tight syringes from the Tedlar bags connected to the glass bottles and analyzed for greenhouse gases ($CH_4$ and carbon dioxide-$CO_2$) and $H_2S$ concentrations. $CH_4$ and $CO_2$ gas concentrations were analyzed using an SRI-8610 Gas Chromatograph and $H_2S$ concentrations were measured using a Jerome 631X meter. At the same time, substrate (i.e. mixed rumen fluid+ NP treatment+ feed composite) samples were collected from the glass bottles at the beginning and at the end of an experiment for bacterial counts, and volatile fatty acids (VFAs) analysis. Results: Compared to the control treatment the $H_2S$ and GHGs concentration reduction after 72 h of the tested nZnO levels varied between 4.89 to 53.65%. Additionally, 0.47 to 22.21% microbial population reduction was observed from the applied nZnO treatments. Application of nZnO at a rate of $1000{\mu}g\;g^{-1}$ have exhibited the highest amount of concentration reductions for all three gases and microbial population. Conclusion: Results suggest that both 500 and $1000{\mu}g\;g^{-1}$ nZnO application levels have the potential to reduce GHG and $H_2S$ concentrations.


  1. Moss AR, Jouany JP, Newbold J, editors. Methane production by ruminants: its contribution to global warming. Ann Zootech EDP Sciences. 2000;231-253. doi:
  2. EPA 430-P-18-001. Draft inventory of us greenhouse gas emissions and sinks: 1990-2016. 2009. documents/2018_complete_report.pdf. Accessed: February 8, 2018.
  3. EIA. Emissions of greenhouse gases in the U. S. 2009. Report number:doe/eia-0573(2009). Report number: doe/eia-0573(2009). Accessed: January 11, 2018.
  4. Hughes MN, Centelles MN, Moore KP. Making and working with hydrogen sulfide: the chemistry and generation of hydrogen sulfide in vitro and its measurement in vivo: a review. Free Radic Biol Med. 2009;47(10):1346-53.
  5. Johnson KA, Johnson DE. Methane emissions from cattle. J Anim Sci. 1995; 73(8):2483-92.
  6. Hogan KB. Anthropogenic methane emissions in the United States, estimates for 1990. 1993. Accessed 2 Nov 2018.
  7. Wolin M, Miller T. Microbe interactions in the rumen microbial ecosystem. The rumen ecosystem (ed PN Hobson). 1988;343-59.
  8. Bauchop T, Mountfort DO. Cellulose fermentation by a rumen anaerobic fungus in both the absence and the presence of rumen methanogens. Appl Environ Microbiol. 1981;42(6):1103-10.
  9. Ushida K, Jouany J. Methane production associated with rumen-ciliated protozoa and its effect on protozoan activity. Lett Appl Microbiol. 1996; 23(2):129-32.
  10. Hristov A, Oh J, Lee C, Meinen R, Montes F, Ott T, et al. Mitigation of greenhouse gas emissions in livestock production: A review of technical options for non-CO2 emissions. FAO Animal Production and Health Paper No. 2013;177:1-206. doi:
  11. Dehority BA. Rumen microbiology. Nottingham: Nottingham University Press; 2003.
  12. Drewnoski M, Beitz DC, Loy DD, Hansen SL, Ensley SM. Factors affecting ruminal hydrogen sulfide concentration of cattle. Anim Ind Rep. 2011;657(1):11.
  13. Morine S, Drewnoski M, Hansen S. Increasing dietary neutral detergent fiber concentration decreases ruminal hydrogen sulfide concentrations in steers fed high-sulfur diets based on ethanol coproducts. J Anim Sci. 2014;92(7): 3035-41.
  14. Martin C, Morgavi D, Doreau M. Methane mitigation in ruminants: from microbe to the farm scale. Animal. 2010;4(03):351-65.
  15. Boadi D, Benchaar C, Chiquette J, Masse D. Mitigation strategies to reduce enteric methane emissions from dairy cows: update review. Can J Anim Sci. 2004;84(3):319-35.
  16. Benchaar C, Pomar C, Chiquette J. Evaluation of dietary strategies to reduce methane production in ruminants: a modelling approach. Can J Anim Sci. 2001;81(4):563-74.
  17. Robertson L, Waghorn G, editors. Dairy industry perspectives o methane emissions and production from cattle fed pasture or total mixed rations in New Zealand. Proceedings-new zealand society of animal production; 2002.
  18. Dong Y, Bae H, McAllister T, Mathison G, Cheng K. Lipid-induced depression of methane production and digestibility in the artificial rumen system (rusitec). Can J Anim Sci. 1997;77(2):269-78.
  19. Dohme F, Machmuller A, Wasserfallen A, Kreuzer M. Comparative efficiency of various fats rich in medium-chain fatty acids to suppress ruminal methanogenesis as measured with rusitec. Can J Anim Sci. 2000;80(3):473-84.
  20. Machmuller A, Kreuzer M. Methane suppression by coconut oil and associated effects on nutrient and energy balance in sheep. Can J Anim Sci. 1999;79(1):65-72.
  21. Wright A, Kennedy P, O'Neill C, Toovey A, Popovski S, Rea S, et al. Reducing methane emissions in sheep by immunization against rumen methanogens. Vaccine. 2004;22(29):3976-85.
  22. Kuzma J, VerHage P. Nanotechnology in agriculture and food production: anticipated applications: project on emerging nanotechnologies; 2006.
  23. Bollo E. Nanotechnologies applied to veterinary diagnostics. Vet Res Commun. 2007;31:145-7.
  24. Scott N. Nanotechnology and animal health. Revue Scientifique Et Technique-Office International Des Epizooties. 2005;24(1):425.
  25. Narducci D. An introduction to nanotechnologies: What's in it for us? Vet Res Commun. 2007;31:131-7.
  26. Swain PS, Rao SB, Rajendran D, Dominic G, Selvaraju S. Nano zinc, an alternative to conventional zinc as animal feed supplement: A review. Anim Nutri. 2016;2(3):134-41.
  27. Mu H, Chen Y, Xiao N. Effects of metal oxide nanoparticles (TiO2, Al2O3, SiO2 and ZnO) on waste activated sludge anaerobic digestion. Bioresour Technol. 2011;102(22):10305-11.
  28. Luna-delRisco M, Orupold K, Dubourguier H-C. Particle-size effect of CuO and ZnO on biogas and methane production during anaerobic digestion. J Hazard Mater. 2011;189(1):603-8.
  29. National Academies of Sciences, Engineering, and Medicine. Nutrient requirements of beef cattle. Washington DC: National Academies Press; 2016.
  30. McDougall E. Studies on ruminant saliva. The composition and output of sheep's saliva. Biochem J. 1948;43(1):99.
  31. Borhan MS, Capareda SC, Mukhtar S, Faulkner WB, McGee R, Parnell CB. Greenhouse gas emissions from ground level area sources in dairy and cattle feedyard operations. Atmosphere. 2011;2(3):303-29.
  32. Rahman S, Lin D, Zhu J. Greenhouse gas (GHG) emissions from mechanically ventilated deep pit swine gestation operation. J Civil Environ Eng. 2012;2:104.
  33. Sarker, N. C., Rahman, S., Borhan, M. S., Rajasekaran, P., Santra, S., & Ozcan, A. (2018). Nanoparticles in mitigating gaseous emissions from liquid dairy manure stored under anaerobic condition. J Envron Sci. (In Press) doi:
  34. Goetsch A, Galyean M. Influence of feeding frequency on passage of fluid and particulate markers in steers fed a concentrate diet. Can J Anim Sci. 1983;63(3):727-30.
  35. Sigg L. Redox potential measurements in natural waters: significance, concepts and problems. Redox: Springer; 2000. p. 1-12.
  36. Nutrition L. A. Target pH levels in silage. Dairy Herd Management 2016 Accessed 12 Jan 2018.
  37. Bhandari S, Ominski K, Wittenberg K, Plaizier J. Effects of chop length of alfalfa and corn silage on milk production and rumen fermentation of dairy cows. J Dairy Sci. 2007;90(5):2355-66.
  38. Grant R, Mertens D. Influence of buffer pH and raw corn starch addition on in vitro fiber digestion kinetics. J Dairy Sci. 1992;75(10):2762-8.
  39. Wu H, Yang D, Zhou Q, Song Z. The effect of pH on anaerobic fermentation of primary sludge at room temperature. J Hazard Mater. 2009;172(1):196-201.
  40. Shete S, Tomar S. Ruminating Over Methane Emissions. NISCAIR-CSIR. 2010; 31-32.
  41. Colmenarejo M, Sanchez E, Bustos A, Garcia G, Borja R. A pilot-scale study of total volatile fatty acids production by anaerobic fermentation of sewage in fixed-bed and suspended biomass reactors. Proc Biochem. 2004;39(10): 1257-67.
  42. Lee SJ. Relationship between oxidation reduction potential (ORP) and volatile fatty acid (VFA) production in the acid-phase anaerobic digestion process. 2008. doi:
  43. Blanc FC, Molof AH. Electrode potential monitoring and electrolytic control in anaerobic digestion. J Water Pollut Control Fed. 1973;45(4):655-67.
  44. Environmental Y. ORP Management in wastewater as an indicator of process efficiency. YSI, Yellow Springs, OH. 2008. Accessed 2 Nov 2018.
  45. Moran J. Tropical dairy farming: feeding management for small holder dairy farmers in the humid tropics: Csiro publishing; 2005.
  46. Zhisheng C. Effect of nano-zinc oxide supplementation on rumen fermentation in vitro. Chinese J Anim Nutr. 2011;8:023.
  47. Hook SE, Wright A-DG, McBride BW. Methanogens: methane producers of the rumen and mitigation strategies. Archaea. 2010;2010.
  48. Pouliquen F, Blanc C, Arretz E, Labat I, Tournier-Lasserve J, Ladousse A, et al. Ullmann's encyclopedia of industrial chemistry. 1985.