Investigation the buffering capacity of some native alkalizer and buffer compounds and their effect on fermentation and rumen digestion parameters in vitro

Document Type : Ruminant Nutrition


1 Department of Animal Sciences, Faculty of Agriculture, Birjand University, Birjand, Iran

2 Department of Animal Science, Birjand Faculty of Agriculture, Birjand, Iran


 Introduction: Buffering and alkalizing supplements have been used in dairy cows in order to increase production and prevention of ruminal acidosis for many years. Magnesium carbonate is typically obtained from magnesite mines and magnesium oxide is produced by the calcification of magnesium carbonate (reduction, oxidation and burning or heating with extreme heat). Some minerals such as bentonite and zeolite have buffering properties and a special role in the buffering capacity of rumen cation exchange. Buffers raise acetate to propionate ratio and improve fiber digestion. The purpose of this experiment was to evaluate the buffering capacity (BC) and buffering value index (BVI) of some native buffer and alkalizer agents including sodium bentonite, magnesium carbonate, magnesium oxide and sodium bicarbonate and their effects on ruminal fermentation and digestion parameters in vitro.
Materials and Methods: The first experiment was performed with different types (11 samples) of buffer and alkalizer agents to determine the buffering Capacity (BC) and buffer Value Index (BVI). The experimental samples were including magnesium carbonate A (MgCA), magnesium carbonate B (MgCB), magnesium carbonate C (MgC C), magnesium carbonate D (MgC D), magnesium oxide A (MgOA), magnesium oxide B (MgOB), magnesium oxide C (MgOC), magnesium oxide D (MgOD), sodium bentonite A (SBeA), sodium bentonite B (SBeB) and sodium bicarbonate (SBi). The buffering capacity was defined as the resistance to change in pH from 7 to 5. In order to determine BC, individual samples were dried and ground to pass through 1-mm screen. Buffering capacity was determined by titrating the 30-ml solution under continuous stirring from its initial pH to 5 with 1 N HCl and by titrating a similar prepared solution of samples from its initial pH to 7 with 1 N NaOH. In the second experiment in vitro batch culture technique used. The basal diet contains 80 percent of the concentrate and 20 percent of the forage. half-gram (DM) of base ration with magnesium carbonate (MgC), magnesium oxide (MgO), sodium bentonite (SBe) and sodium bicarbonate (SBi) (1 and 3% DM of ration) were added into the culture vials, then incubated with 50 ml of buffered rumen fluid. Rumen fluid was collected from two ruminally fistulated cows. The gas production was estimated. In vitro dry matter disappearance (IVDMD), pH and ammonia nitrogen (NH3-N) was measured after 8 and 24 h incubation times.
Results and Discussion: The results showed that the highest pH, buffering capacity and buffering value index were in MgOA and the least of these parameters indicated at MgCD, respectively. Compared to the control lower values were obtained for total gas production (b) of MgO 3%, MgC 3%, MgO 1% and SBe 3% additives. In the present study, addition of MgC and SBi (3% DM diet) also caused a remarkably increase in total gas production. Although the exact mechanisms are not known for the general responses in gas production to supplementation of MgC and SBi, one of the mechanisms of   increasing total gas production is considered to be the higher ruminal microbial numbers caused by the SBi and MgC addition. It has been demonstrated that SBi addition increased the number of ruminal total bacteria, cellulolytic and amylolytic bacteria in buffalo. 3% MgO supplementation increased (P < 0.05) the final pH and it decreased (P < 0.05) the IVDMD compared with the control (after 24h incubation). Significant increase in pH by additive MgO and bentonite can be due to the exchange of cations with hydrogen ion as a modifier of hydrogen ion. Also, in this experiment use of SBe decreased digestibility of dry matter. Some researchers reported that 0.5% MgO (DM diet) does not change the digestibility of dry matter compared to sodium bicarbonate. On the other hand, results of this section of experiment was in agreement with results of gas production, MgC and MgO treatments which had the most and least amount of produced gas (respectively), these also affected digestibility of dry matter. Generally, buffers and alkalizing compounds have no effect on digestibility of dry matter, and probably this reduction in magnesium oxide treatment may indicated a change in the microbial population of ruminal fluid because of the effects of these treatments. In addition, Concentration of NH3-N did not differ (P = 0.2443) between the treatments. Some compounds, such as bentonites, are believed to be able to absorb and release proteins and other nitrogen substrates, so absorbing it when ammonia is high in the rumen, and when the ammonia concentration is reduced, a large amount of it released and thus increases the efficiency of ammonia and protein in the rumen.
Conclusion: In general, the results of this experiment show that magnesium oxide, while increasing the pH of the rumen fluid, also has negative effects on the parameters of gas production, fermentation and possibly microbial activity, and also reduces the in vitro digestibility of dry matter. However, magnesium carbonate and sodium bentonite as a true buffer and due to the preservation and stability of rumen fluid acidity can improve fermentation conditions without having a negative effect on digestibility, so using 1% magnesium carbonate and sodium bentonite as a true buffer is suggested in the livestock diet.


1- Aghashahi, A., A. Nikkhah., S. A. Mirhadi., M. Zahedifar, and H. Mansouri. 2005. Effects of different level of unprocessed bentonite, processed bentonite, and clinoptilolite at different rumen degradable protein level, on ammonia concentration, soluble and digestible protein (in-vitro). Pajouhesh and Sazandegi, 70: 80-90. (In Persian).
2- Agnote, M. 2004. Drought feeding and managing sheep. 4th. pp: 36. Victorian Government. Victoria, Australia.
3- Ammerman, C. B., C. F. Chicco., J. E. Moore., P. A. Van Wallenghem, and L. R. Arrington. 1971. Effect of dietary magnesium on voluntary feed intake and rumen fermentation. Journal of Dairy Science, 54(9), 1288-1292.
4- AOAC International. 2012. Official Methods of Analysis. 19th ed. AOAC International, Gaithersburg, MD.
5- ASTM D2974-14. 2014. Standard Test Methods for Moisture, Ash, and Organic Matter of Peat and Other Organic Soils, ASTM International, West Conshohocken, PA.
6- ASTM D4972-13. 2013. Standard Test Method for pH of Soils, ASTM International, West Conshohocken, PA.
7- Beede D. K. 2017. Can we differentiate supplemental magnesium sources nutritionally? Tri-State Dairy Nutrition Conference. April. 17-19.
8- Bennink, M., R. T. R. Tyler., G.M. Ward, and D.E. Johnson. 1978. Ionic milieu of bovine and ovine rumen as affected by diets. Journal of Dairy Science, 61: 315–323.
9- Blümmel, M., H. P. S. Makkar, and K. Becker. 1997. In vitro gas production: a technique revisited. Journal of Animal Physiology and Animal Nutrition, 77: 24-34.
10- Broderick, G. A, and J.H. Kang. 1980. Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. Journal of Dairy Science, 63: 64-75.
11- Chalupa, W., D. T. Galligan, and J. D. Ferguson. 1996. Animal nutrition and management in the 21st century: dairy cattle. Animal Feed Science and Technology, 58: 1-18.
12- Cruywagen C. W., S. Taylor., M. M. Beya, and T. Calitz. 2015. The effect of buffering dairy cow diets with limestone, calcareous marine algae, or sodium bicarbonate on ruminal pH profiles, production responses, and rumen fermentation Journal of dairy Science, 98: 5506-5514.
13- Danesh Mesgaran, M., J. Amini, and M. Paktinat. 2013. In vitro usage of various non-organic compounds to subdue acidogenic value and enhance the fermentation of alfalfa hay-based diets by mixed rumen microbiota. Journal of Livestock Production, 4(10): 165-170.
14- Erdman, R. A. 1988. Dietary buffering requirements of the lactating dairy cow: A review. Journal of Dairy Science, 71: 3246–3266
15- Goff, J.P., 2004. Macromineral disorders of the transition cow. VETERINARY Clinics of NORTH America, 20: 471–494.
16- Goff, J.P., 2006. Macromineral physiology and application to the feeding of the dairy cow for prevention of milk fever and other periparturient mineral disorders. Animal Feed Science Technology, 126: 237–257.
17- Horn, G. W., J. L. Gordon., E. C. Prigge, and F. N. Owens. 1979. Dietary buffers and ruminal and blood parameters of subclinical lactic acidosis in steers. Journal of Animal Science, 48(3): 683-691.
18- Jahani, A. H., M. Danesh Mesgaran., A. Vakili., K. Rezayazdi, and M. Hashemi. 2011. Effect of various medicinal plant essential oils obtained from semi-arid climate on rumen fermentation characteristics of a high forage diet using in vitro batch culture. African Journal Microbial Research, 5: 4812-4819.
19- Jacques, K.A., D. E. Axe., T. R. Harris, and D. L. Harmon. 1986. Effect of sodium bicarbonate and sodium bentonite on digestion, solid and liquid flow, and ruminal fermentation characteristics of forage sorghum silage-based diets fed to steers. Journal of Animal Science, 63: 923-932.
20- Komisarczuk-Bony, S, and M. Durand. 1991. Effects of minerals on microbial metabolism. In: Jouany, J.P. (Ed.), Rumen Microbial Metabolism and Ruminant Digestion. INRA Editions, Paris, 179–198.
21- Koul, Y., U. Kumar., K. Sareen, and S. Singh. 1998. Effect of sodium bicarbonate supplementation on ruminal microbial populations and metabolism in buffalo calves. Indian Journal Animal Science, 68(7): 629-631.
22- Le Ruyet, P, and W.B. Tucker. 1992. Ruminal Buffers: Temporal Effects on Buffering Capacity and pH of Ruminal Fluid from Cows Fed a High Concentrate Diet. Journal of Dairy Science, 75: 1069-1077
23- Mahdavi Rad, N., M. Chaji., M. Bojarpour, and M. Dehghanbanadaki. 2018. Investigation the buffering capacity of several conventional buffer compounds in feeding of ruminant animals by acid titration method and their effect on gas production parameters. Iranian Journal of Animal Science, 48 (4): 559-571. (in Farsi)
24- Mao, S., W, Huo., J. Liu., R. Zhang, and W. Zhu. 2017. In vitro effects of sodium bicarbonate buffer on rumen fermentation, levels of lipopolysaccharide and biogenic amine, and composition of rumen microbiota. Journal of the Science of Food and Agriculture, 97(4): 1276-1285.
25- Moharrery, A. 2007. The determination of buffering capacity of some ruminant’s feedstuffs and their cumulative effects on TMR ration. American Journal of Animal and Veterinary Sciences, 2 (4):72-78.
26- Mojtahedi, M. 2013. Identification of nanostructure and nanoporous bentonite adsorbents and their efficiency on aflatoxin b1 detoxification in vitro and in vivo. Ph.D. Dissertation. Ferdowsi University of Mashhad, Iran. (In Persian)
27- Mojtahedi, M., M. Danesh Mesgaran., S. A. Vakili, and E. Abdi Ghezeljeh. 2013. Effect of esterified glucomannan on carryover of aflatoxin from feed to milk in lactating holstein dairy cows. Annual Review and Research in Biology, 3: 76-82.
28- Ørskov, E.R, and I. McDonald. 1979. The estimation of protein degradation in the rumen from incubation measurements weighted according to rate of passage. Journal of Agriculture Science, 92: 499–503.
29- Peirce, S.B., L. D. Muller, and H.W. Harpster. 1983. Influence of sodium bicarbonate and magnesium oxide on digestion and metabolism in yearling beef steers abruptly changed from high forage to high energy diets. Journal of Animal Science, 57(6): 1561-1567.
30- Perez-Ruchel, A., J. L. Repetto, and C. Cajarville. 2014. Use of NaHCO3 and MgO as additives for sheep fed only pasture for a restricted period of time per day: effects on intake, digestion and the rumen environment. Journal of Animal Physiology and Animal Nutrition, 98(6): 1068-1074.
31- Rayssiguier, Y, and C. Poncet. 1980. Effect of lactose supplement on digestion of Lucerne hay by sheep. II. Absorption of magnesium and calcium in the stomach. Journal of Animal Science, 51: 186–192.
32- Saleh, M. S, and A. B. Bonf. 2000. Bentonite supplementation to concentrate for lactating buffaloes. Egypt Journal of Nutrient Feeds, 6: 67-78.
33- SAS Institute Inc. 2009. Statistical Analysis System (SAS) User’s Guide, SAS Institute, Cary, NC, USA.
34- Shaw, F. D, and W. J. Pryor. 1972. Feeding wheat to cattle. 2. The effect of level of wheat and sodium bicarbonate on ruminal characteristics. Australian Veterinary Journal, 48: 504-507.
35- Theodorou, M. K., B. A. Williams., M. S. Dhanoa., A. B. McAllan, and J. France. 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.
36- Tucker, W. B., J. F. Hogue., M. Aslam., M. Lema., M. Martin., F. N. Owens., I. S. Shin., P. Le Ruyet, and G. D. Adams. 1992. A buffer value index to evaluate effects of buffers on ruminal milieu in cows fed high or low concentrate, silage, or hay diets. Journal of Dairy Science, 75: 1069-1077.
37- Urdaz, J.H., J.E.P. Santos, P. Jardon, and M.W. Overton. 2003. Importance of appropriate amounts of magnesium in rations for dairy cows. Journal of the American Veterinary Medical Association, 222: 1518-1523.
38- Varadyova, Z., M. Baran., W. Zawadzki, and P. Siroka. 2003. Effect of dolomite, magnesium oxide (MgO) and chalk (CaCO3) on in vitro fermentation of amorphous and crystalline cellulose and meadow hay using inoculum from sheep. Berl Munch Tierarztl Wochenschr, 116(1-2): 50-54.
39- Varadyova, Z., S.K. Kisidayov., K. Mihalikov, and M. Baran. 2006. Influence of natural magnesium sources on the in vitro fermentation and protozoan population in the rumen fluid collected from sheep. Small Ruminant Research, 61: 63–71.
40- Wilson, G.F. 1980. Effects of magnesium supplements on the digestion of forages and milk production of cows with hypomagnesaemia. Animal Production, 31: 153–157.