Fitting of pH conditions for the study of concentrate feeds fermentation by the in vitro gas-production technique
Z. Amanzougarene A and M. Fondevila A B
+ Author Affiliations
- Author Affiliations
A Departamento de Producción Animal y Ciencia de los Alimentos, Instituto Agroalimentario de Aragón (IA2), Universidad de Zaragoza-CITA. Miguel Servet 177, 50013, Zaragoza, Spain.
B Corresponding author. Email: mfonde@unizar.es
Animal Production Science 58(9) 1751-1757 https://doi.org/10.1071/AN16097
Submitted: 16 February 2016 Accepted: 16 March 2017 Published: 3 May 2017
Abstract
Two experiments were conducted to simulate in vitro the fermentation conditions under high-concentrate feeding. The concentration of bicarbonate ion in the buffer of the incubation solution was assayed in Experiment 1, by adjusting medium pH to 6.50, 6.25, 6.00, 5.75 and 5.50, in two incubation series of 12 h, using barley as the reference substrate. The pH diminished linearly (P < 0001) by lowering the buffer, and remained constant throughout 12 h, except for treatments 5.75 and 5.50, where pH dropped to 5.51 and 5.31 at 12 h. Gas production decreased linearly with a decreasing medium pH (P < 0.001), with the total volume of gas produced after 12 h being highly dependent (P < 0.01) on pH at 12 h (R2 = 0.629), thus demonstrating the importance of the incubation pH for estimation of fermentation of concentrate feeds. In Experiment 2, the effect of pH on direct and indirect proportion of gas was studied by adding 0.0, 0.1, 0.2, 0.3, 0.4 and 0.5 mmol of acetic acid, either with or without (water added instead) rumen inoculum, to the media. Linear multiple regressions established between the volume of gas produced and the addition of acetic acid, and the bicarbonate ion concentration showed high determination coefficients for water (R2 = 0.929) and rumen inoculum (R2 = 0.851). Without inoculum, indirect gas production ranged from 9.4 to 12.4 mL/mmol of acid for medium pH of 5.50–6.50. With rumen inoculum, indirect gas was 20.8 mL/mmol acid, although this may have been biased by the contribution of inoculum itself to direct fermentation.
Additional keywords: barley, bicarbonate buffer, indirect gas, in vitro, rumen fluid.
References
Analytical Software (2010) ‘Statistix 10 for Windows.’ (Analytical Software: Tallahassee, FL)AOAC (2005) ‘Official methods of analysis.’ 18th edn. (Association of Official Analytical Chemists: Gaithersburg, MD)
Bertipaglia LMA, Fondevila M, van Laar H, Castrillo C (2010) Effect of pelleting and pellet size of a concentrate for intensively reared beef cattle on in vitro fermentation by two different approaches. Animal Feed Science and Technology 159, 88–95.
| Effect of pelleting and pellet size of a concentrate for intensively reared beef cattle on in vitro fermentation by two different approaches.Crossref | GoogleScholarGoogle Scholar |
Beuvink JM, Spoelstra SF (1992) Interactions between substrate, fermentation end-products, buffering systems and gas production upon fermentation of different carbohydrates by mixed rumen microorganisms in vitro. Applied Microbiology Technology 37, 505–509.
| Interactions between substrate, fermentation end-products, buffering systems and gas production upon fermentation of different carbohydrates by mixed rumen microorganisms in vitro.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38XlsVKqtbw%3D&md5=c80a0e554d9cc94db61530ff6797696dCAS |
Broudiscou LP, Offner A, Sauvant D (2014) Effects of inoculum source, pH, redox potential and headspace di-hidrogen on rumen in vitro fermentation yields. Animal 8, 931–937.
| Effects of inoculum source, pH, redox potential and headspace di-hidrogen on rumen in vitro fermentation yields.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXotFCrsr8%3D&md5=59ae49b8138a5c4f4aaad3a9af46d8e6CAS |
Counotte GHM, Van’t Klooster AT, Van der Kuilen J, Prins RA (1979) An analysis of the buffer system in the rumen of dairy cattle. Journal of Animal Science 49, 1536–1544.
| An analysis of the buffer system in the rumen of dairy cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL3cXptVChtg%3D%3D&md5=a04e053ad7566d818e426e0cd2656ab2CAS |
Czerkawski JW, Breckenridge G (1977) Design and development of a long-term rumen simulation technique (RUSITEC). British Journal of Nutrition 38, 371–384.
| Design and development of a long-term rumen simulation technique (RUSITEC).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE1cXhs1Kiu7o%3D&md5=51abcd46e7e211644639176006b005c6CAS |
Erdman RA (1988) Dietary buffering requirements of the lactating dairy cow: a review. Journal of Dairy Science 71, 3246–3266.
| Dietary buffering requirements of the lactating dairy cow: a review.Crossref | GoogleScholarGoogle Scholar |
Fondevila M, Pérez-Espés B (2008) A new in vitro system to study the effect of liquid phase turnover and pH on microbial fermentation of concentrate diets for ruminants. Animal Feed Science and Technology 144, 196–211.
| A new in vitro system to study the effect of liquid phase turnover and pH on microbial fermentation of concentrate diets for ruminants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXnvVKjsrc%3D&md5=cbd3cb29e41df35319432e79ec0e9badCAS |
Getachew G, Crovetto GM, Fondevila M, Krishnamoorthy U, Singh B, Spanghero M, Steingass H, Robinson PH, Kailas MM (2002) Laboratory variation of 24 h in vitro gas production and estimated metabolizable energy values of ruminant feeds. Animal Feed Science and Technology 102, 169–180.
| Laboratory variation of 24 h in vitro gas production and estimated metabolizable energy values of ruminant feeds.Crossref | GoogleScholarGoogle Scholar |
Goering HK, van Soest PJ (1970) ‘Forage fiber analyses (apparatus, reagents, procedures and some applications).’ USDA agriculture handbook no. 379. (USDA-ARS: Washington, DC)
Grant RJ, Mertens DR (1992) Influence of buffer pH and raw corn starch addition on in vitro fibre digestion kinetics. Journal of Dairy Science 75, 2762–2768.
| Influence of buffer pH and raw corn starch addition on in vitro fibre digestion kinetics.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK3s%2FlsFOksw%3D%3D&md5=59d6d8a7f6cafd13d2d2f938f5518c9cCAS |
Hiltner P, Dehority BA (1983) Effect of soluble carbohydrates on digestion of cellulose by pure cultures of rumen bacteria. Applied and Environmental Microbiology 46, 642–648.
Hoover WH, Crooker BA, Sniffen CJ (1976) Effects of differential solid-liquid removal rates on protozoa numbers in continuous cultures of rumen contents. Journal of Animal Science 43, 528–534.
| Effects of differential solid-liquid removal rates on protozoa numbers in continuous cultures of rumen contents.Crossref | GoogleScholarGoogle Scholar |
Hungate RE (1966) ‘The rumen and its microbes.’ (Academic Press: New York)
Kohn RA, Dunlap TF (1998) Calculation of the buffering capacity of bicarbonate in the rumen and in vitro. Journal of Animal Science 76, 1702–1709.
| Calculation of the buffering capacity of bicarbonate in the rumen and in vitro.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXktFSgu7k%3D&md5=372bf81040f652a4116150c816cfbddaCAS |
Lanzas C, Fox DG, Pell AN (2007) Digestion kinetics of dried cereal grains. Animal Feed Science and Technology 136, 265–280.
| Digestion kinetics of dried cereal grains.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXnslGhtLg%3D&md5=82690c6bc33f72cae902946116294f12CAS |
Martínez ME, Ranilla MJ, Tejido ML, Saro C, Carro MD (2010) The effect of the diet fed to donor sheep on in vitro methane production and ruminal fermentation of diets of variable composition. Animal Feed Science and Technology 158, 126–135.
| The effect of the diet fed to donor sheep on in vitro methane production and ruminal fermentation of diets of variable composition.Crossref | GoogleScholarGoogle Scholar |
Menke KH, Raab I, Salewski A, Steingass H, Fritz D, Schneider W (1979) The estimation of the digestibility and metabolizable energy content of ruminant feeding stuffs from the gas production when they are incubated with ruminal liquor. Journal of Agricultural Science, Cambridge 93, 217–222.
| The estimation of the digestibility and metabolizable energy content of ruminant feeding stuffs from the gas production when they are incubated with ruminal liquor.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE1MXlsFKitbc%3D&md5=dcd36121528f99acf81a2ece481b0014CAS |
Mould FL, Morgan R, Kliem KE, Krystallidou E (2005) A review and simplification of the in vitro incubation medium. Animal Feed Science and Technology 123–124, 155–172.
| A review and simplification of the in vitro incubation medium.Crossref | GoogleScholarGoogle Scholar |
Opatpatanakit Y, Kellaway RC, Lean IJ, Annison G, Kirby A (1994) Microbial fermentation of cereal grains in vitro. Australian Journal of Agricultural Research 45, 1247–1263.
| Microbial fermentation of cereal grains in vitro.Crossref | GoogleScholarGoogle Scholar |
Russell JB, Dombrowski DB (1980) Effect of pH on the efficiency of growth by pure cultures of rumen bacteria in continuous culture. Applied and Environmental Microbiology 39, 604–610.
Rymer C, Givens DI (1998) Relationship between the rates of production of gas and VFA during in vitro incubations of high temperature dried grass. In ‘Proceedings of the British Society of Animal Science annual meeting 1998’, p. 55.
Rymer C, Moss AR, Deaville ER, Givens DI (1998) Factors affecting the amount of indirect gas produced by the in vitro gas production technique. In ‘In vitro techniques for measuring nutrient supply to ruminants’. (Eds ER Deaville, E Owen, AT Adesogan, C Rymer, JA Huntington, TLJ Lawrence) pp. 89–91. BSAS Occasional Publication No. 22.
Theodorou MK, Williams BA, Dhanoa MS, McAllan AB, 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.
| A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds.Crossref | GoogleScholarGoogle Scholar |
Theodorou MK, Lowman RS, Davies ZS, Cuddeford D, Owen E (1998) Principles of techniques that rely on gas measurements in ruminant nutrition. In ‘In vitro techniques for measuring nutrient supply to ruminants’. (Eds ER Deaville, E Owen, AT Adesogan, C Rymer, JA Huntington, TLJ Lawrence), pp. 55–63. BSAS Occasional Publication No. 22.
Tilley JMA, Terry RA (1963) A two stage technique for the in vitro digestion of forage crops. Journal of the British Grassland Society 18, 104–111.
| A two stage technique for the in vitro digestion of forage crops.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF3sXks1Cmurk%3D&md5=7da67ce44b99ed3fbfbbd86fde6190f1CAS |
