Use of acid-consuming bacteria and various buffers to improve digestion and fermentation of highly concentrated diets

Document Type : Research Paper


1 Former MSc Student of Animal Nutrition, Department of Animal Science, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Ahvaz, Iran

2 Professor, Department of Animal Science, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Ahvaz, Iran


Introduction: The use of highly concentrated rations in the fattening of ruminant animals is done to improve the performance per unit of feed consumption. Increasing the amount of concentrate in the ration improves the food conversion ratio and the weight gain of fattening lambs, and the production of volatile fatty acids. However, some in vivo experiments have reported that long-term use of highly concentrated diets is not always associated with increased performance. Therefore, feeding diets containing high amounts of concentrate (with a limited amount of effective fiber) often leads to metabolic disorders, the most important of which is subacute ruminal acidosis which causes a decrease in dry matter consumption and reduces the fiber digestibility, milk fat, lameness, and liver abscess, and even leads to the death of the animal. Moreover, sub-acute acidosis leads to significant economic losses. Therefore, preventive measures to prevent acidosis and improve starch digestion, such as the use of antibiotics, probiotics, and buffers, have been taken into consideration. Lactate-consuming bacteria metabolize lactic acid and control its accumulation in the rumen. This process is important when the animal diet contains a large amount of grain. Megasphaera elsdenii and Selenomonas ruminantium are the dominant strains that consume lactic acid in the rumen, and among these two strains, Megasphaer alsdeni consumes 65 to 95% of the lactate in the rumen. The use of biological methods in controlling pH changes can also reduce the dependence of livestock on chemical additives to some extent. Therefore, the present experiment was conducted to compare the effect of different chemical and biological buffers (acid-consuming bacteria) on the digestion and fermentation of highly concentrated diets.
Materials and methods: The experimental treatments included: 1. The control diet (without additives and containing 70% concentrate + 30% fodder or basal diet), 2. Basal diet + 3 mL of bacteria Megasphaera elsdenii (bacteria- 4.5 × 108 cfu/3mL), 3. Basal diet + 1% sodium bentonite, 4. Basal diet + 1% sodium bentonite + 3 mL of bacteria, 5. Basal diet + 1% sodium bicarbonate, 6. Basal diet + 1% sodium bicarbonate + 3 mL bacteria, 7. Basal diet + 1% magnesium oxide, 8. Basal diet + 1% magnesium oxide + 3 mL bacteria, 9. Basal diet + 1% zeolite, 10. Basal diet + 1% zeolite + 3 mL bacteria, 11. Basal diet + 1% sodium sesquicarbonate, and 12. Basal diet + 1% sodium-sesquicarbonate + 3 mL bacteria. Megasphaera elsdenii bacteria were isolated and prepared from Najdi goats at the Agricultural Sciences and Natural Resources University of Khuzestan (Ahvaz, Iran) and their activity was investigated in experiments. In gas production experiments, the experimental samples were ground with a mill containing a 1 mL sieve. About 200 mg of the dry matter of the desired sample was weighed and poured into 100 mL vials, and eight replicates were considered for each treatment. The produced gas of the samples was recorded at 0, 2, 4, 6, 8, 12, 24, 48, 72, and 96 hours of incubation using a digital barometer.
Results and discussion: The effect of experimental treatments on potential and rate of gas production, partitioning factor, microbial biomass production, microbial biomass production efficiency, pH, ammonia nitrogen concentration, apparent dry matter digestibility, and protozoa population was significant (P<0.05), and except for ammonia nitrogen, all parameters in buffer treatments were higher than the control. The highest gas production potential (68.26 mg), microbial biomass production (1212.31 mg), and microbial biomass production efficiency (79%) were observed in the treatment containing sodium bicarbonate + bacteria (P<0.05). The highest pH and ammonia nitrogen were for the treatment containing bacteria (6.60) and control (27.30 mg/100 mL). The total protozoa population was the highest in the treatment of sodium bentonite + bacteria.
Conclusions: In general, the results of the present experiment showed that the use of buffers improved digestion and fermentation conditions, and each of the buffers had a greater effect on one or more parameters than the others. In addition, acid-consuming bacteria as a pH regulator had competitive effects with chemical buffers, especially bicarbonate buffer, and even better in some cases. The effect of some chemical buffers used in the present experiment, such as bicarbonate, has been further investigated, but the effect of other chemical and biological buffers mentioned in the present experiment on the digestive and fermentation properties of ruminant animals has been investigated less. Therefore, it is recommended to investigate the effect of chemical buffers used in the present experiment alone or together with acid-consuming bacteria in feeding ruminant animals.


Main Subjects

Abdl-Rahim M. A. 2010. In vitro manipulation of rumen fermentation efficiency by fumaric acid –bentonitecoupled addition as an alternative to antibiotics. Journal of Agricultural Science, 2(2): 174-180.
Aderinboye R. Y., Akinlolu A. O., Adeleke M. A., Najeem G. O., Ojo V. O. A., Isah O. A. and Babayemi O. J. 2016. In vitro gas production and dry matter degradation of four browse leaves using cattle, sheep and goat inocula. Slovak Journal of Animal Science, 49(1): 32-43.
Aikman P. C., Henning P. H., Humphries D. J. and Horn C. H. 2011. Rumen pH and fermentation characteristics in dairy cows supplemented with Megasphaera elsdenii NCIMB 41125 in early lactation. Journal of Dairy Science, 94: 2840-2849.
Asadi A., Kiani A., Azarfar A. and Valipour A. 2016. Effects of Metafix with or without Monensin on performance and blood metabolites in Farahani lambs. Iranian Journal of Animal Science, 47: 421-428. (In Persian).
Baah J. M., Ivan A. N., Hristov K. M., Koenig L. M., Rode T. and McAllister A. 2007. Effects of potential dietary antiprotozoal supplements on rumen fermentation and digestibility in heifers. Animal Feed Science and Technology, 137: 126-137.
Beauchemin K. A. and Yang W. Z. 2005. Effects of physically effective fiber on intake, chewing activity, and ruminal acidosis for dairy cows fed diets based on corn silage. Journal of Dairy Science, 88: 2117-2129.
Bach A., Guasch I., Elcoso G., Duclos J. and Khelil-Arfa H. 2018. Modulation of rumen pH by sodium bicarbonate and a blend of different sources of magnesium oxide in lactating dairy cows submitted to a concentrate challenge. Journal of Dairy Science, 101(11): 9777-9788. 
Blümmel ‎M., ‎Steingaβ ‎H. and Becker K. 1997. The relationship between in vitro gas production, in vitro microbial biomass yield and 15 N incorporation and its implications for the prediction of voluntary feed intake of roughages. British Journal of Nutrition, 77(6): 911-921.
Brigatti M. F., Galan E. and Theng B. K. J. 2006. Strauctures and mineralogy of clay minerals. Handbook Clay Science, Elsevier Ltd.
Broderick G. A. and Kang J. H. 1980. Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. Journal of Dairy Science, 63: 64-75.
Calsimiglia S., Cardozo P. W., Ferrer A. and Bach A. 2008. Changes in rumen microbial fermentation are due to combined effect of type of diet and pH. Journal of Animal Science, 86: 702-711.
Carro M. D., Valdés C., Ranilla M. J. and González J. S. 2000.  Effect of forage to concentrate ratio in the diet on ruminal fermentation and digesta flow kinetics in sheep. Journal of Animal Science, 70: 127-134.
Danesh Mesgaran M., Amini J., Paktinat M.  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.
Dehority B. A. 2003. Rumen microbiology (2nd ed.). London: Academic Press. Pp. 11-151.
Der Bedrosian M. 2009. The effect of sodium bicarbonate or live yeast culture Saccharomyces cerevisiae on the metabolism and production of lactating dairy cows. Ph.D. Dissertation, University of Delaware, Newark, Delaware.
Direkvandi E., Mohammadabadi T. and Salem A. Z. 2020. Oral administration of lactate producing bacteria alone or combined with Saccharomyces cerevisiae and Megasphaera elsdenii on performance of fattening lambs.‏ Journal of Applied Animal Research, 48(1): 235-243.
Dixon J. B., Kannewischer I., TenorioArvide M. G. and Barrientos Velazquez A. L.  2008. Aflatoxin sequestration in animal feeds by quality-labelledsmectite clays: an introductory plan. Applied Clay Science, 40: 201-208.
Dschaak C., Eun M., Young J. S., Stott A. J. and Peterson S. 2010. Effects of supplementation of natural zeolite on intake, digestion, ruminal fermentation, and lactational performance of dairy cows. The Professional Animal Scientist, 26: 647-654.
Ekrami S. H. S. 2009. Utilization of growth promoters and bentonite in sheep rations. Ph.D. Dissertation, University of Al-Azhar, Egypt.
Faichney G. J., Teleki E. and Brown G. H. 2004. Effect of physical form of lucene hay on digestion and rate of passage in sheep. Australian Journal of Agricultural Research, 55: 1253-1262.
Fenn P. D. and Leng R. A. 1989. Wool growth and sulfur amino acid entry rate in sheep fed roughage based diets supplemented with bentonite and sulfur amino acids. Australian Journal of Agricultural Research, 40: 889-896.
Gasmi Boubaker A., Kayouli C. and Buldgen A. 2005. In viro gas produvtion and its relationship to in situ disappearance and chemical composition of some Mediterrnean browse species. Animal Feed Science and Technology, 123-124: 303-311.
Ghoniem A. H., El–Bltagy E. A. and Abdou A. A. 2018. Effect of supplementation dry yeast or bentonite and their combination as feed additives on productive performance of lactating buffalos. Journal of Animal and Poultry Production, 9(11): 423-431. ‏
Harrison J. R., White R., Kincaid E., Block T., Jenkins N. and Pierre S. T. 2012. Effectiveness of potassium carbonate sesquihydrate to increase dietary cation-anion difference in early lactation cows. Journal of Dairy Science, 95(7): 3919-3925.
Hu W. and Murphy M. R. 2005. Statistical evaluation of early- and mid-lactation dairy cow responses to dietary sodium bicarbonate addition. Animal Feed Science and Technology, 119: 43-54.
Khalifeh M. J., Mohammadabadi T., Chaji M., Salari S. and Khalil M. 2012. The effect of different levels of sodium bentonite on in vitro fermentation and digestibility of soybean meal. In: Proceedings of the 15th AAAP Animal Science Congress, 26-30 November. Thailand. Pp. 3133-3135.
Khoujeh B., Gharehbash A. M., Bayat Kouhsar J. and Moslemipour F. 2016. Effect of using different sources of buffer on digestibility and fermentation parameters in In vitro situation. Proceedings of the 7th Iranian Congress on Animal Science, Karaj, Iran. (In Persian).
Koul V., Kumar U., Sareen V. K. and Singh S. 1998. Effect of sodium bicarbonate supplementation on ruminal microbial populations and metabolism in buffalo calves. Indian Journal of Animal Science, 68: 629-631. 
Krause K. M. and Combs D. K. 2003. Effects of forage particle size, forage source and grain fermentability on performance and ruminal pH in midlactation cows. Journal of Dairy Science, 86: 1382-1397.
Kung L. and Hession A. O.1995. Preventing in vitro lactate accumulation in ruminal fermentations by inoculation with (Megasphaera elsdenii). Journal of Animal Science, 73(1): 250-256.
Mao S., Huo W., Liu J., Zhang R. and Zhu W. 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.
McDaniel M. R. 2009. The effects of dosing feedlot cattle with Megasphaera elsdenii strain NCIMB 41125 prior to the introduction of a grain-rich diet. Ph.D. Dissertation, Kansas State University, Kansas.
McDonald P., Edwards R. A., Greenhalgh J. F. D., Morgan C. A., Sinclair L. A. and Wilkinson R. G. 2011. Animal Nutrition (7th ed.). Harlow United Kingdom: Longman Group. Pp. 30-693.
Meissner H. H., Henning P. H., Leeuw K. J., Hagg F. M., Horn C. H., Kettunen A. and Apajalahti J. H. A. 2014. Efficacy and mode of action of selected non-ionophore antibiotics and direct-fed microbials in relation to Megasphaera elsdenii NCIMB 41125 during in vitro fermentation of an acidosis-causing substrate. Livestock Science, 162: 115-125.
Menke K. H, and Steingass H. 1988. Estimation of the energetic feed value obtained from chemical analysis and in vitro gas production using rumen fluid Animal. Research and Development, 28: 7-55.
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, Mashhad, Iran. (In Persian).
Mohammadabadi T., Bakhtiari M. A. and Alimirzaei P. 2018. Isolation and identification of lactate-producing and utilizing bacteria from the rumen of Najdi goats. Indian Journal of Small Ruminant, 24(2): 276-280.
Norollahi H. 2007. Effect of fattening period on growth and carcass characteristics of male Turkey-Ghashghaii lambs. Pajohesh and Sazandegi, 75: 132-137. (In Persian).
Orskov E. R. and 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.
Papi N. and Mostafa Tehrani A. 2017. Effects of dietary concentrate levels on growth performance, feed intake and carcass characteristics of fattening shall male lambs. Journal of Ruminant Research, 5(2): 59-70. (In Persian).
Philippeau C., Lettat A., Martin C., Silberberg M., Morgavi D. P., Ferlay A. and Nozière P. 2017. Effects of bacterial direct-fed microbials on ruminal characteristics, methane emission, and milk fatty acid composition in cows fed high-or low-starch diets. Journal of Dairy Science, 100(4): 2637-2650.
Plaizier J. C., Danesh Mesgaran D., Derakhshani H., Golder H., Khafipour E., Kleen J. L., Lean I., Loor J., Penner G. and Zebeli Q. 2018. Enhancing gastrointestinal health in dairy cows: A review. Animal, 12(2): 399-418. 
Prabhu R., Altman E. and Eiteman M. A. 2012. Lactate and acrylate metabolism by Megasphaera elsdenii under batch and steady-state conditions. Applied and Environmental Microbiology, 78: 8564-8570.
Silanikove N., Landau S., Kababya D., Bruckental I. and Nitsan Z. 2006. Analytical approach and effects of condensed tannins in carob pods (Ceratonia siliqua) on feed intake, digestive and metabolic responses of kids. Livestock Science, 99: 29-38.
Stevenson A. E. and Glare N. T. 1963. Measurement of feed intake by grazing cattle and sheep. New Zealand Journal of Agricultural Research, 6(1-2): 121-126.
Sulzberger S. A., Kalebich C. C., Melnichenko S. and Cardoso F. C. 2016. Effects of clay after a grain challenge on milk composition and on ruminal, blood, and fecal pH in Holstein cows. Journal of Dairy Science, 99(10): 8028-8040.
Theodorou M. K., Williams B. A., Dhanoa M. S., McAllan A. B. and France J. A. 1994. Simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology, 48: 185-197.
Vaghar Seyedi, S.M., Mojtahedi M., Ghiasi S.A. and Fathi Nasri M.H. 2018. Buffering capacity of some native alkalizer and buffer compounds and their effect on in vitro gas production and digestibility. Iranian Journal of Animal Science Research, 11 (40): 425-436. (In Persian).
Wallace R. J. and Newbold C. J. 1991. Effect of bentonite on fermentation in the rumen simulation technique (Rusitec) and rumen ciliate protozoa. Journal of Agricultural Science, 116: 163-175.
Wang Y. H., Xua M., Wang F. N., Yz Z. P., Yao J. H., Zan L. S. and Yang F. X. 2009. Effect of dietary starch on rumen and small intestine morphology and digesta pH in goats. Livestock Science, 122: 48-52.
Zebeli Q., Terrill S. J., Mazzolari A., Dunn S. M., Yang W. Z. and Ametaj B. N. 2012. Intraruminal administration of Megasphaera elsdenii modulated rumen fermentation profile in mid-lactation dairy cows. Journal of Dairy Research, 79(01): 16-25.