The effect of Nepeta glomerulosa Boiss. (Lamiaceae) essential oil (NGEO) on in vitro gas production and ruminal fermentation

Document Type : Original Research Articles (Regular Papers)


1 Assistant Professor, Department of Animal Science, Faculty of Agriculture and Animal Science, University of Torbat-e Jam, Torbat-e Jam, Iran.

2 Assistant Professor, Research Center of Special Domestic Animals, University of Zabol, Zabol, Iran.



The study aimed to evaluate the chemical composition and the effects of Nepeta glomerulosa Boiss. (Lamiaceae) essential oil (NGEO) on in vitro gas production and ruminal fermentation. The essential oil (EO), obtained by steam distillation from Nepeta glomerulosa Boiss. (0, 150, 300, and 450 mg/L), was investigated in an in vitro culture medium using sheep rumen fluid and artificial saliva. A fattening diet was used as the substrate in the culture medium and gas production was measured. The profile of NGEO was determined by GC-mass analysis. The 1, 8-cineole (23.2%), α-pinene (15.3%), limonene (9.1%), and β-pinene (3.5%) were the major components in NGEO. Ammonia nitrogen and total volatile fatty acids (TVFA) concentrations did not change when NGEO was added to the culture medium, whereas TVFA tended to increase at the higher concentration of NGEO (p <0.1). The pH value of the culture medium linearly and quadratically decreased with increasing NGEO (p <0.05). The potential of gas production (bgas; linear, and quadratic, p <0.05) increased with increasing NGEO, however, the constant rate decreased linearly and quadratically (p <0.05). Dry matter (DMD) and organic matter degradability (OMD) were increased (linear and quadratic) with increasing NGEO in the culture medium. The partitioning factor (PF), microbial mass yield (MMY), and efficiency of microbial mass synthesis (EMMS) linearly and quadratically decreased when the concentration of NGEO increased. It seems that NGEO affected the fermentation process in vitro partly via improving TVFA production or by increasing DMD and OMD. Further in vitro and in vivo studies are needed to confirm that NGEO in the diet has no adverse effects on the health and production in ruminants.


  • Alekish, M.O., Ismail, Z.B., Awawdeh, M.S., Shatnawi, S., 2017. Effects of intramammary infusion of sage (Salvia officinalis) essential oil on milk somatic cell count, milk composition parameters and selected hematology and serum biochemical parameters in Awassi sheep with subclinical mastitis. Veterinary Word 10, 895-900.
  • Aronen, T.S., 1997. Interactions between Agrobacterium tumefaciens and coniferous defence compounds α-pinene and trans-stilbene. European Journal of Plant Pathology 27, 55-67.
  • Barnett, A.J.G., Reid, R., 1957. Studies on the production of volatile fatty acids from grass in artificial rumen. 1. Volatile fatty acids production from fresh grasses. Journal of Agricultural Science 48, 315-321.
  • Benchaar, C., Greathead, H., 2011. Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Animal Feed Science and Technology 166-167, 338-355.
  • Benchaar, C., Calsamiglia, S., Chaves, A.V., Fraser, G.R., Colombatto, D., McAllister, T.A., Beauchemin, K.A., 2008. A review of plant-derived essential oils in ruminant nutrition and production. Animal Feed Science and Technology 145, 209-228.
  • Benchaar, C., Chaves, A.V., Fraser, G.R., Beauchemin, K.A., McAllister, T.A., 2007. Effects of essential oils and their components on in vitro rumen microbial fermentation. Canadian Journal of Animal Science 87, 413-419.
  • Besharati, M., Palangi, V., Moaddab, M., Nemati, Z., Pliego, A.B., Salem, A.Z., 2020. Influence of cinnamon essential oil and monensin on ruminal biogas kinetics of waste pomegranate seeds as a biofriendly agriculture environment. Waste and Biomass Valorization (in press, doi: 10.1007/s12649-020-01167-2).
  • Blümmel, M., Makkar, H.P.S., Becker, K., 1997. In vitro gas production: a technique revisited. Journal of Animal Physiology and Animal Nutrition 77, 24-34.
  • Bodas, R., Prietoa, N., García-González, R., Andrésa, S., Giráldeza, F.J., Lopeza, S., 2012. Manipulation of rumen fermentation and methane production with plant secondary metabolites. Animal Feed Science and Technology 176, 78-93.
  • Calsamiglia, S., Busquet, M., Cardozo, P.W., Castillejos, L., Ferret, A., 2007. Invited review: Essential oils as modifiers of rumen microbial fermentation. Journal of Dairy Science 90, 2580-2595.
  • Cardozo, P., Calsamiglia, S., Ferret, A., Kamel, C., 2005. Screening for the effects of natural plant extracts at different pH on in vitro rumen microbial fermentation of a high-concentrate diet for beef cattle. Journal of Animal Science 83, 2572-2579.
  • Castillejos, L., Calsamiglia, S., Ferret, A., 2006. Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems. Journal of Dairy Science 89, 2649-2658.
  • Castillejos, L., Calsamiglia, S., Ferret, A., Losa, R., 2007. Effects of dose and adaptation time of a specific blend of essential oil compounds on rumen fermentation. Animal Feed Science and Technology 132, 186-201.
  • Castillejos, L., Calsamiglia, S., Martın-Tereso, J., Ter Wijlen, H., 2008. In vitro evaluation of the effects of ten essential oils at three doses on ruminal fermentation of high concentrate feedlot-type diets. Animal Feed Science and Technology 145, 259-270.
  • Cui, H., Bai, M., Rashed, M.M.A., Lin, L., 2018. The antibacterial activity of clove oil/chitosan nanoparticles embedded gelatin nanofibers against Escherichia ColiO157:H7 biofilms on cucumber. International Journal of Food Microbiology 266, 69-78.
  • Demirtaş, A., Öztürk, H., Pişkin, İ., 2018. Overview of plant extracts and plant secondary metabolites as alternatives to antibiotics for modification of ruminal fermentation. Ankara Üniversitesi Veteriner Fakültesi Dergisi 65, 213-217.
  • Fraser, G.R., Chaves, A.V., Wang, Y., McAllister, T.A., Beauchemin, K.A., Benchaar, C., 2007. Assessment of the effects of Cinnamon leaf oil on rumen microbial fermentation using two continuous culture systems. Journal of Dairy Science 90, 2315-2328.
  • Goel, G., Makkar, H.P.S., Becker, K., 2008. Effects of Sesbania sesban and Carduus pycnocephalus leaves and Fenugreek (Trigonella foenum-graecum L.) seeds and their extracts on partitioning of nutrients from roughage- and concentrate-based feeds to methane. Animal Feed Science and Technology 147, 72-89.
  • Helander, I.M., Alakomi, H.L., Latva-Kala, K., Mattila-Sandholm, T., Pol, I., Smid, E.J., Gorris, L.G.M., Von 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.
  • Jahani-Azizabadi, H., Danesh Mesgaran, M., Vakili, A.R., Rezayazdi, K., Hashemi, M., 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 of Microbiology Research 5, 4812-4819.
  • Karting, T., Still, F., Reinthaler, F., 1991. Antimicrobial activity of the essential oil of young pine shoots (Picea abies L.). Journal of Ethnopharmacology 35, 155-157.
  • Komolong, M.K., Barber, D.G., McNeill, D.M., 2001. Post-ruminal protein supply and N retention of weaner sheep fed on a basal diet of lucerne hay (Medicago sativa) with increasing levels of quebracho tannins. Animal Feed Science and Technology 92, 59-72.
  • Makkar, H.P., 2010. In vitro screening of feed resources for efficiency of microbial protein synthesis. In: Vercoe P., Makkar H., Schlink A. (Eds.), In Vitro Screening of Plant Resources for Extra-nutritional Attributes in Ruminants: Nuclear and Related Methodologies. Springer, Dordrecht. doi:10.1007/978-90-481-3297-3_7.
  • McGuffey, R.K., Richardson, L.F., Wilkinson, J.I.D., 2001. Ionophores for dairy cattle: current status and future outlook. Journal of Dairy Science 84 (Supplement), E194-E203.
  • McIntosh, F.M., Newbold, C.J., Losa, R., Williams, P., Wallace, R.J., 2000. Effects of essential oils on rumen fermentation. Reproduction Nutrition Development 40 (Suppl. 2), 221-222. (Abstract).
  • Menke, K.H., Steingass, H., 1988. Estimation of the energetic feed value obtained from chemical analysis and in vitro gas production using rumen fluid. Animal Research and Develop 28, 7-55.
  • Narimani, R., Moghaddam, M., Ghasemi Pirbalouti, A., Mojarab, S., 2017. Essential oil composition of seven populations belonging to two Nepeta species from Northwestern Iran. International Journal of Food Properties 20, 2272-2279.
  • Newbold, C.J., McIntosh, F. M., Williams, P., Losa, R., Wallace, R.J., 2004. Effects of specific blend of essential oil compounds on rumen fermentation. Animal Feed Science and Technology 114, 105-112.
  • Nooriyan Soroor, E., Rouzbehan, Y., 2014. The influence of golpar (Heracleum persicum) on in vitro rumen fermentation parameter, sand on methane production. Iranian Journal of Applied Animal Science 44, 385-395.
  • NRC, 2007. Nutrient Requirements of Small Ruminants: Sheep, goats, cervids, and new world camelids. 6th ed. National Research Council. National Academy Press, Washington, DC, USA.
  • Ørskov, E.R., 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.
  • Sajjadi, S.E., Ghassemi, N., 1999. Volatile constituents of Nepeta glomerulosa Boiss. subsp. carmanica. Flavour and Fragrance Journal 14, 265-267.
  • Salem, A.Z.M., Olivares, M., Lopez, S., Gonzalez-Ronquillo, M., Rojo, R., Camacho, L.M., Cerrillo, S.M.A., Mejia, H.P., 2011. Effect of natural extracts of Salix babylonica and Leucaena leucocephala on nutrient digestibility and growth performance of lambs. Animal Feed Science and Technology 170, 27-34.
  • Sallam, S.M.A., Bueno, I.C.S., Brigide, P., Godoy, P.B., Vittii, D.M.S.S., Abdalla, A.L., 2009. Efficiency of eucalyptus oil on in vitro ruminal fermentation and methane production. Nutritional and Foraging Ecology of Sheep and Goats 85, 267-272.
  • SAS, 2002. SAS User’s Guide: Statistics. Statistical Analysis Systems Institute Inc., Cary, North Carolina. USA.
  • Spanghero, M., Zanfi, C., Fabbro, E., Scicutella, N., Camellini, C., 2008. Effects of a blend of essential oils on some end products of in vitro rumen fermentation. Animal Feed Science and Technology 145, 364-374.
  • Wallace, R.J., 2004. Antimicrobial properties of plant secondary metabolites. Proceedings of the Nutrition Society, Edinburgh, Scotland.
  •  Xing-dong, L., Hua-li, X., 2014. Antifungal activity of the essential oil of Zanthoxylum bungeanum and its major constituent on Fusarium sulphureum and dry rot of potato tubers. Phytoparasitica 42, 509-517.