Effect of free thymol and sustained release thymol on rumen fermentation and plasma metabolites in sheep

Document Type : Research Paper


1 PhD graduated student, Department of Animal Science, Faculty of Agriculture, Bu-Ali Sina University

2 Associate Professor, Department of Animal Science, Faculty of Agriculture, Bu-Ali Sina University

3 Professor of Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences

4 Assistant Professor, Department of Clinical Science, Faculty of Paraveterinary Science, Bu-Ali Sina University


This experiment was conducted to compare the effect of free and sustained release thymol on rumen fermentation characteristics, microbial protein and some plasma metabolites. Six ruminally cannulated sheep were used in a 3×3 merged Latin square design. Treatments were (a) basal diet+ 250 mg free Thymol, (b) basal diet+ 250 mg slow relrase Thymol and (c) control (basal diet without Thymol). The experiment consisted of three 14-d periods with 10 d adaptation and 4 d sampling. At sampling period, the feces were collected and at the end of each period blood sampling and rumen fluid collection were made at different times. The measured traits were: Feed digestability, N-NH3, plasma and glucose concentration, ruminal pH, microbial protein synthesis and efficiency of microbial protein synthesis. Dry matter intake (985.5 g and 917.5 g per day, respectively), crude protein digestibility (82.7% and 85.6%, respectively) and plasma glucose (114.8 mg/dL and 108 mg/dL, respectively) were increased by free and sustained release thymol consumption (P<0.05). Plasma urea concentration, microbial protein synthesis and N-NH3 were decreased in 5 and 7 hours after feeding (P<0.05). Number of diplodinia was 0.38×105 in diet with free thymol and this number was lower than those in other two treatments (P<0.05). Thymol consumption improved the efficiency of dietary protein utilization by decreasing N-NH3 and increased level of plasma glucose showed a positive effect on energy utilization of animal.


Adesogan A.T. 2009. Using Dietary Additives to Manipulate Rumen Fermentation and Improve Nutrient Utilization and Animal Performance. dairy.ifas.ufl.edu. Florida ruminant nutrition symposium, 20th symposiu. 13-38.
Ando T., Nishida T., Ishhida M., Hosoda K and Bayaru E. 2003. Effect of peppermint feeding on the digestibility, ruminal fermentation and protozoa. Livestock Production Science, 82: 245-248.
AOAC. 1990. "Official methods of analysis" 15th edition. Association of official analytical chemist, Arlington, U.S.A.
Bach A., Calsamiglia S. and Stern M. D. 2005. Nitrogen metabolism in the rumen. Journal of Dairy Science, 88: 9–21.
Bampidis V. A., Christodoulou V., Christaki E., Florou-Paneri P. and Spais A. B. 2005. Effect of dietary garlic bulb and garlic husk supplementation on performance and carcass characteristics of growing lambs. Animal Feed Science and Technology, 121: 273-283.
Benchaar C., Calsamiglia S., Chaves A. V., Fraser G. R., Colombatto D., McAllister T. A. and Beauchemin K. A. 2008. A review of plant-derived essential oils in in ruminant nutrition and production. Animal Feed Science and Technology, 145: 209–228.
Benchaar C., McAllister T. A. and Chouinard P. Y. 2005. Effects of cinnamaldehyde, yucca saponins extract and condensed tannins on fermentation characteristics, and ciliate protozoal populations in the rumen of lactating dairy cows. Journal of Dairy Science. 83 (Suppl. 1): 304.
Benchaar C., Petit H. V., Berthiaume R., Ouellet D. R., Chiquette J. and Chouinard P. Y. 2007. Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage. Journal of Dairy Science, 90: 886–897.
Benchaar C., Petit H. V., Berthiaume R., Whyte T. D. and Chouinard P. Y. 2006. Effects of addition of essential oils and monensin premix on digestion, ruminal fermentation, milk production, and milk composition in dairy cows. Journal of Dairy Science, 89: 4352–4364.
Borchers R. 1965. Proteolytic activity of rumen fluid in vitro. Journal of Animal Science, 24: 1033–1038.
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.
Budak D., Yılmaz A .2013. Effects of aromatic plants on rumen fermentation. Macedonian . Journal of Animal Science. 3(1): 75–80.
Burt S. 2004. Essential oils: Their antibacterial properties and potential applications in foods—A review. International Journal of Food Microbiology, 94: 223–253.
Busquet M., Calsamiglia S., Ferret A. and Kamel C. 2006. Plant extracts affect in vitro rumen microbial fermentation. Journal of Dairy Science, 89: 761–771.
Cardozo P. W., Calsamiglia S., Ferret A. and Kamel C. 2006. Effects of alfalfa extract, anise, capsicum, and a mixture of cinnamaldehyde and eugenol on ruminal fermentation and protein degradation in beef heifers fed a high-concentrate diet. Journal of Dairy Science, 84: 2801–2808.
Castillo C., Benedito J. L., Vazquez P., Pereira V., Mendez J., Juan Sotillo J. and Hernandez J. 2012. Effects of supplementation with plant extract product containing carvacrol, cinnamaldehyde and capsaicin on serum metabolites and enzymes during the finishing phase of feedlot-fed bull calves. Animal Feed Science and Technology, 171: 246– 250.
Chaturvedi S., Sharma P. K, Visht S. and Tyagi S. 2012. "Comparison of emulsification and ionic gelation method of preparation of mucoadhesive microsphere". The Pharma Innovation, 1: 1-10.
Dehority B. A. 1993. Laboratory manual for classification and morphology of rumen ciliate protozoa. CRC press, Inc, Boca Raton. FL.
Dryden G. M. 2008. Animal nutrition science, CABI, 302 pages.
Evans J. D. and Martin S. A. 2000. Effects of thymol on ruminal microorganisms. Curr. Microbiol. 41:336–340.
Hart K.J., Yanez-Ruiz D.R., Duval S.M., McEwan N.R and Newbold C.J. 2007. Plant extracts to manipulate rumen fermentation. Animal Feed Science and Technology, 1-28
Helander I. M., Alakomi H. L., Latva-Kala K., Mattila-Sandholm T., Pol L., Smid E. J., Gorris L. G. M. and Wright A. 1998. Characterization of the action of selected essential oil components on Gram negative bacteria. Journal of Agricultural and Food Chemistry, 46: 3590–3595.
Jamroz D., Wertelecki T., Houszka M. and Kamel C. 2006. Influence of diet type on the inclusion of plant origin active substances on morphological and histochemical characteristics of the stomach and jejunum walls in chicken. Journal ofAnimal PhysiologyandAnimal Nutrition, 90: 255–268.
Kholif  S. M., Morsy T. A., Abdo M. M., Matloup O. H and Abu El-Ella A. A. 2012. Effect of supplementing lactating goats rations with garlic, cinnamon or ginger oils on milk yield, milk composition and milk fatty acids profile. Journal of Life Sciences, 4(1): 27-34.
Kohlert C., Van Rensen I., Marz R., Schindler G., Graefe E. U. and Veit M. 2000. "Bioavailability and pharmacokinetics of natural volatile terpenes in animals and humans". Planta Medica, 66: 495–505.
Lin B., Lu Y., Salem A. Z. M., Wang J. H., Liang Q. and Liu J. X. 2013. Effects of essential oil combinations on sheep ruminal fermentation and digestibility of a diet with fumarate included. Animal Feed Science and Technology, 184: 24-32.
Lin B., Tan Z., Xiao G., Wang M., Cong Z., Wang S., Tang S., Zhou C., Sun Z. and Wang W. 2009. Evaluation of compositional and nutritional equivalence of genetically modified rice to conventional rice using in situ and in vitro techniques. Journal of the Science of Food and Agriculture, 89: 1490–1497.
Makkar H. P. S. and Becker K. 1999. Purine quantification in digesta from ruminants by spectrophotometric and HPLC methods. British Journal of Nutrition, 81: 107-113.
Mathis C. P. and Sawyer J. E. 2003. Urea in range cattle supplements. New Mexico state university. Cooperative extension service. Circular 583 college of agriculture and home economics.
McIntosh F. M., Williams P., Losa R., Wallace R. J., Beever D. A. and Newbold C. J. 2003. Effects of essential oils on ruminal microorganisms and their protein metabolism. Applied and Environmental Microbiology, 69: 5011–5014.
Meunier J. P., Cardot J. M., Gauthier P., Beyssac E. and Alric M. 2006. "Use of rotary fluidized-bed technology for development of sustained-release plant extracts pellets: Potential application for feed additive delivery". Journal of Animal Science, 84: 1850-1859.
Min B. R., Barry T. N., Attwood G. T. and McNabb W. C. 2003. The effect of condensed tannins on the nutrition and health of ruminants fed fresh temperature forages: A review. Animal Feed Science and Technology, 106: 3–19.
Molero R., Ibars M., Calsamiglia S., Ferret A. and Losa R. 2004. Effects of a specific blend of essential oil compounds on dry matter and crude protein degradability in heifers fed diets with different forage to concentrate ratios. Animal Feed Science and Technology, 114: 91–104.
Ozdogan M., Onenc S. S and Onenc A. 2011. Fattening performance, blood parameters and slaughter traits of Karya lambs consuming blend of essential oil compounds. African Journal of Biotechnology, 10(34): 6663-6669.
Patra A. K. 2011. Effect of essential oil on rumen fermentation, microbial ecology and ruminal production. Asian Journal of Animal and Veterinary Advances, 6(5): 416-428.
Patra A. K. and Saxena J. 2010. A new perspective on the use of plant secondary metabolites to inhibit methanogenesis in the rumen. Phytochemistry, 71: 1198–1222.
Patra A. K. and Yu Z. 2012. Effects of essential oils on methane production, fermentation, abundance and diversity of rumen microbial populations. Applied and Environmental Microbiology, 78: 4271–4280.
Perdok H., Langhout P. and Vugt P. V. 2003. Stimulating appetite. Feed mixture, 11: 10-13.
Piva A., Pizzamigli V., Morlacchini M., Tedeschi M. and Piva G. 2007. Lipid microencapsulation allows slow release of organic acids and natural identical flavors along swine intestine. Journal of Animal Science, 85: 486-493.
Pulina G Avondo M., Molle G., Francesconi A. H. D., Atzori A. S. and Cannas A. 2013. Invited Review: Models for estimating feed intake in small ruminants. Brasileira de Zootecnia, 42(9): 675-690.
Rowe R. C., Sheskey P. J. and Quinn M. E. 2009. Handbook of Pharmaceutical Excipients. Sixth edition. RPS Publishing. UK. ruminant nutrition and production. Animal Feed Science and Technology, 145: 209–228.
SAS. 2004. SAS/STAT software: changes and enhancements though release. 9.1 Statistical Analysis System Institute, Cary, NC.
Soltan M. A. E., Shewita R. S. and Al-Sultan S. I. 2009. Influence of essential oils supplementation on digestion, rumen fermentation, rumen microbial populations and productive performance of dairy cows. Asian-Australasian Journal of Animal Science, 3: 1–12.
Taghavi-Nezhad A. M., Alipour D., Flythe M. D., Zamani P. and Khodakaramian G. 2014. The effect of essential oils of Zataria multiflora and Mentha spicata on the in vitro rumen fermentation, and growth and deaminative activity of amino acid-fermenting bacteria isolated from Mehraban sheep. Animal Production Science, 54: 299–307.
Talebzadeh R., Alipour D., Saharkhiz M. J., Azarfar A. and Malecky M. 2012. Effect of essential oils of Zataria multiflora on in vitro rumen fermentation, protozoa population, growth and enzyme activity of anaerobic fungus isolated from Mehraban sheep. Animal Feed Science and Technology,172: 115–124.
Tekeli A., Çelik L. and Kutlu H. R. 2007. Plant extracts; a new rumen moderator in ruminant diets. Journal of Tekirdag Agricultural Faculty, 4(1).
Tshuma T. 2013. The effect of blood urea nitrogen on reproductive performance of beef heifers on different levels of nitrogen supplementation. PhD thesis. Faculty of Veterinary Science, Universiy of Pretoria. Africa.
Wallace R. J. 2004. Antimicrobial properties of plant secondary metabolites. Proceedings of Nutrition Society, 63: 621–629.
Wallace R. J., McEwan N. R., McIntosh F. M., Teferedegn B. and. Newbold C. J. 2002. Natural products as manipulators of rumen fermentation. Asian-Australasian Journal of Animal Science, 10: 1458–1468.
Wanapat M., Pichad K., Pawadee P. and Sadudee W. 2008 Effect of supplementation of garlic powder on rumen ecology and digestibility of nutrients in ruminants. Journal of the Scienceof Food and Agriculture, 88(13): 2231–2237.
Wang Y., Douglas G. B., Waghorn G. C., Barry T. N., Foote A. G. and Purchas R. W. 1996. Effects of condensed tannins upon the performance of lambs grazing Lotus corniculatus and lucerne (Medicago sativa). The Journal of Agricultural Science, 126: 87–98.
Wang Y., McAllister T. A., Yanke L. J., Xu Z. J., Cheeke P. R. and Cheng K. J. 2000. In vitro effects of steroidal saponins from Yucca schidigera extract on rumen microbial protein synthesis and rumen fermentation. Journal of the Scienceof Food and Agriculture, 80: 2114–2122.
Yang W. Z., Ametaj B. N., Benchaar C., He M. L. and Beauchemin K. A. 2010. Cinnamaldehyde in feedlot cattle diets: Intake, growth performance, carcass characteristics, and blood metabolites. Journal of Animal Science, 88: 1082-1092.
Yang W. Z., Benchaar C., Ametaj B. N., Chaves A. V. and McAllister T. A. 2007. Effects of garlic and juniper berry essential oils on ruminal fermentation and on the site and extent of digestion in lactating cows. Journal of Dairy Science, 90: 5671–5681.