Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.43

  • SJR .263

  • Impact Factor .427 (2022)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Full Research Article

Indian Journal of Animal Research, volume 56 issue 2 (february 2022) : 140-144

Development of Protocol for Lyophilization of Goat Rumen Fluid

A. Ruba Nanthini, C. Valli, L. Radhakrishnan, D. Balasubramanyam, A. Mangalagowri
1Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 051, Tamil Nadu, India.
Cite article:- Nanthini Ruba A., Valli C., Radhakrishnan L., Balasubramanyam D., Mangalagowri A. (2022). Development of Protocol for Lyophilization of Goat Rumen Fluid. Indian Journal of Animal Research. 56(2): 140-144. doi: 10.18805/IJAR.B-4730.

ABSTRACT

Background: Rumen fluid has been used as microbial inoculum to treat indigestion in ruminant animals and to conduct in vitro rumen fermentation experiments. Lyophilization of the goat rumen fluid will provide continuous supply of rumen inoculums either for laboratory studies or for transfaunation in treating digestive disorders sequelae to high grain rations. However, no standard protocol is available for lyophilizing goat rumen fluid. Hence, this study was designed to develop a protocol to lyophilize goat rumen fluid as an alternate source for fresh goat rumen fluid.
Methods: The study was conducted using 5 × 3 × 3 factorial design with four different cryoprotectants viz., 10% skim milk powder, 10% skim milk powder + 5% sodium glutamate, 5% glycerol, 5% DMSO and no cryoprotectant, at three pre freezing (2, 24 and 48 hours) and three freeze drying (8, 24 and 32 hours) time intervals to standardize protocol for lyophilizing goat rumen fluid. The viability of rumen microbes in the “lyophilized goat ruminal inoculum”, was determined via in vitro gas production study.
Result: Pre freezing (-80°C deep freezer) duration of 48 hours with 32 hours of time duration in lyophilizer (-45°C) was ideal for lyophilizing goat rumen fluid with or without the addition of various cryoprotectants. Glycerol used at 5% as cryoprotectant resulted in significantly (P<0.05) highest gas production at all (12, 24 and 48) incubation hours studied indicating better viability of rumen microbes.

REFERENCES


  1. Abadias, M., Benabarre, A., Teixido, N., Usall, J., Vinas, I. (2001). Effect of freeze drying and protectants on viability of the biocontrol yeast Candida sake. International Journal of Food Microbiology. 65: 173-182.

  2. Belanche, A., Palma-Hidalgo, J.M., Nejjam, I., Serrano, R., Jiménez, E., Martín-García, I., Yanez-Ruiz, D.R. (2018). In vitro assessment of the factors that determine the activity of the rumen microbiota for further applications as inoculum. Journal of the Science of Food and Agriculture. doi: 10.1002 /jsfa.9157.

  3. Bircher, L., Geirnaert, A., Hammes, F., Lacroix, C., Schwab, C. (2018). Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes. Microbial biotechnology. 11(4): 721-733.

  4. Chaudhry, A.S., Mohamed, R.A.I. (2012). Fresh or frozen rumen contents from slaughtered cattle to estimate in vitro degradation of two contrasting feeds. Czech Journal of Animal Science. 57: 265-273.

  5. Denek, N., Can, A., Avci, M. (2010). Frozen rumen fluid as microbial inoculum in the two-stage in vitro digestibilityassay of ruminant feeds. South African Journal of Animal Science. 40: 251-256.

  6. DePeters, E.J. and George, L.W. (2014). Rumen transfaunation. Immunology letters. 162(2): 69-76.

  7. Fowler, A. and Toner, M. (2005). Cryo-injury and biopreservation. Annals of the New York Academy of Sciences. 1066: 119- 135.

  8. Hristov, A.N., Lee, C., Hristova, R., Huhtanen, P., Firkins, J.L. (2012). A meta-analysis of variability in continuous-culture ruminal fermentation and digestibility data. Journal of Dairy Science. 95(9): 5299-5307.

  9. Laflin, S.L. and Gnad, D.P. (2008). Rumen cannulation: procedure and use of a cannulated bovine. Veterinary Clinics of North America: Food Animal Practice. 24(2): 335-340.

  10. Lopez, S. (2005). In vitro and in situ techniques for estimating digestibility. In Quantitative Aspects of Ruminant Digestion and Metabolism, 2nd edition, CABI Publishing: Wallingford, UK, pp. 87-121

  11. Lovelock, J.E. (1953). The mechanism of the protective action of glycerol against haemolysis by freezing and thawing. Biochim Biophys Acta. 11: 28-36.

  12. McDougall, E.I. (1948). The composition and output of sheep’s saliva. The Biochemical Journal. 43(1): 99-109.

  13. Menke, K.H., Raab, L., Salewski, A., Steingass, H., Fritz, D., Schneider, W. (1979). The estimation of the digestibility and metabolizable energy content of ruminant feedingstuffs from the gas production when they are incubated with rumen liquor in vitro. The Journal of Agricultural Science. 93(1): 217-222.

  14. Morgan, C.A., Herman, N., White, P.A., Vesey, G. (2006). Preservation of micro-organisms by drying; A review. Journal of microbiological methods. 66(2): 183-193.

  15. Prates, A., De Oliveira, J.A., Abecia, L. and Fondevila, M. (2010). Effects of preservation procedures of rumen inoculum on in vitro microbial diversity and fermentation. Animal Feed Science and Technology. 155(2-4): 186-193.

  16. Snedecor. G.W. and Cochran, W.G. (1989). Statistical methods applied to experiments in agriculture and biology. 5th ed. Ames, Iowa: Iowa State University Press.

  17. Spanghero, M., Chiaravalli, M., Colombini, S., Fabro, C., Froldi, F., Mason, F., Moschini, M., Sarnataro, C., Schiavon, S., Tagliapietra, F. (2019). Rumen inoculum collected from cows at slaughter or from a continuous fermenter and preserved in warm, refrigerated, chilled or freeze-dried environments for in vitro tests. Animals. 9(10): 815.

  18. Yanez-Ruiz, D.R., Bannink, A., Dijkstra, J., Kebreab, E., Morgavi, D.P., O’Kiely, P., Reynolds, C.K.,Schwarm, A., Shingfield, K.J., Yu, Z. (2016). Design, implementation and interpretation of in vitro batch culture experiments to assess enteric methane mitigation in ruminants-A review. Animal Feed Science and Technology. 216: 1-18.
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)