Effects of different levels of the savory essential oil on in vitro ruminal fermentation parameters, microbial protein synthesis and protozoal populations in two diets supplemented with fish and soybean oil

Authors

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

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

3 Professor at Department of Biotechnology, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran

Abstract

Background and objectives: Despite the central importance of ruminants in providing the human nutritional requirements, their relatively low production efficiency leads often to increased production costs and pollut the environment. In this connection, using supplements and feed additives have considered as efficient strategies for improving the efficiency of livestock production. The plant-based feed additives, including essential oils and plant extracts, are the products that improve the efficiency of the ruminant production, through ameliorating the rumen digestion and fermentation. On the other hand, using the supplements, such as the different fat sources as the energetic supplement in animal diet, has had several positive impacts such as reducing ruminal acidosis, reducing methane production and and consequently declining environmental pollution, and improving feed energy efficiency and the performance of animals. Regarding the growing importance of high-quality products and special interest of consumers for the safety and quality of animal products in recent years, the feed additives of natural and green origin have been of great interest for animal nutritionists in recent decades. In this context, the current research aimed at investigating in vitro the savory essential oil effects on rumen fermentation, microbial protein synthesis and protozoal population in two diets supplemented with fish and soybean oils.
Materials and methods: The essential oil was extracted by steam distillation using a clevenger apparatus, and was subsequently analysed qualitatively and quantitatively by Gas Chromatography-Mass Spectrometry (GC-MS). The essential oil effects on fermentation parameters were tested in vitro at 0, 150, 300, 450, and 600 mg/ in two diets supplemented with fish and soybean oils using the incubations of 24 h.
Results: None of the rumen fermentation parameters was affected by the diet type. The gas produced over 24 h of incubation, in vitro true dry matter and organic matter degradability decreased linearly with increasing doses of the essential oil (P < 0.01) in both experimental diets. However, the partitioning factor as well as the microbial protein, and the efficiency of microbial protein synthesis increased none-linearly (linearly and quadratically) and linearly, respectively, with the essential oil dosage (P < 0.01). The ruminal ammonia concentration decreased none-linearly with the essential oil dosage (P < 0.01). Total protozoa numbers as well as most of the rumen common protozoal genera were reduced by increasing doses of the essential oil (P < 0.01). Total VFA concentration increased at the doses up to 450 and 300 mg/L of the essential oil in the fish oil and soybean oil containing diets, respectively (P < 0.05), and decreased thereafter at higher doses. The acetate molar proportions decreased none-linearly and that of propionate increased quadratically in both diets with essential oil dosage (P < 0.05).
Conclusion: In total, these findings indicated that using the savory essential oil, especially at low and medium doses, improved rumen fermentation through enhancing microbial protein synthesis, reducing ammonia and protozoa population, and increasing TVFA. Hence, the above-mentioned doses of the this essential oil can improve rumen fermtaion thereby enhancing the farm animals productivity. However, the savory essential oil seems to have negative impact on some rumen microorganisms at 450 mg/L and higher doses.

Keywords


AOAC, 2000. Official methods of analysis‎,17th ed. Association of official analytical chemists.‎, VA, USA‎.
Abbasi, A., Maddah, S.M., Mahboubi, A., Khaledi, A., Vazini, H. and Esmaeili, D. 2017. Investigate the inhibitory effects of Satureja khuzestanica essential oil against housekeeping fabD and exoA genes of Pseudomonas aeruginosa from hospital isolates using RT-PCR technique. Annals of Medical and Health Sciences Research, 7: 246-250.
AbuGhazaleh, A., Schingoethe, D., Hippen, A., Kalscheur, K. and Whitlock, L. 2002. Fatty acid profiles of milk and rumen digesta from cows fed fish oil, extruded soybeans or their blend. Journal of Dairy Science, 85: 2266-2276.
Alizadeh, A. R., Alikhani, M., Ghorbani, G.R., Rahmani, H.R., Rashidi, L. and Loor, J. J. 2012. Effects of feeding roasted safflower seeds (variety IL-111) and fish oil on dry matter intake, performance and milk fatty acid profiles in dairy cattle. Journal of Animal Physiology and Animal Nutrition, 96: 466-73.
Bach, A., Calsamiglia, S. and Stern, M. D. 2005. Nitrogen metabolism in the rumen. Journal of Dairy Science, 88: E9-E21.
Belanche, A., Abecia, L., Holtrop, G., Guada, J., Castrillo, C., De La Fuente, G. and Balcells, J. 2011. Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme. Journal of Animal Science, 89: 4163-4174.
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 ruminant nutrition and production. Animal Feed Science and Technology, 145: 209-228.
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: 886-97.
Blummel, M., Makkar, H.P.S. and Becker, K. 1997. In vitro gas production: a technique revisited. Journal of Animal Physiology and Animal Nutrition, 77: 24-34.
Bodkowski, R., Czyż, K., Sokoła-Wysoczańska, E., Janczak, M., Cholewińska, P., Wyrostek, A. 2020. The Effect of Low-Temperature Crystallization of Fish Oil on the Chemical Composition, Fatty Acid Profile, and Functional Properties of Cow’s Milk. Animals 10, 1834.
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.
Burt, S. 2004. Essential oils: their antibacterial properties and potential applications in foods—a review. International Journal of Food Microbiology, 94: 223-253.
Cardozo, P., Calsamiglia, S., Ferret, A. and 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.
Carreño, D., Hervás, G., Toral, P. G., Belenguer, A. and Frutos, P. 2015. Ability of different types and doses of tannin extracts to modulate in vitro ruminal biohydrogenation in sheep. Animal Feed Science and Technology, 202: 42-51.
Castillejos, L., Calsamiglia, S., Martín-Tereso, J. and Ter Wijlen, H. 2008. In vitro evaluation of 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
Dehority, B. A. 2003. Rumen microbiology, Nottingham University Press Nottingham. 372p.
Donovan, D., Schingoethe, D., Baer, R., Ryali, J., Hippen, A. and Franklin, S. 2000. Influence of dietary fish oil on conjugated linoleic acid and other fatty acids in milk fat from lactating dairy cows. Journal of Dairy Science, 83: 2620-2628.
Dorman, H. and Deans, S. G. 2000. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88: 308-316.
Farsam, H., Amanlou, M., Radpour, M., Salehinia, A. and Shafiee, A. 2004. Composition of the essential oils of wild and cultivated Satureja khuzistanica Jamzad from Iran. Flavour and Fragrance Journal, 19: 308-310.
Fievez, V., Dohme, F., Danneels, M., Raes, K. and Demeyer, D. 2003. Fish oils as potent rumen methane inhibitors and associated effects on rumen fermentation in vitro and in vivo. Animal Feed Science and Technology, 104: 41-58.
Garcia, F., Colombatto, D., Brunetti, M. A., Martínez, M. J., Moreno, M. V., Scorcione Turcato, M., Lucini, E., Frossasco, G. and Martínez Ferrer, J. 2020. The reduction of methane production in the in vitro ruminal fermentation of different substrates is linked with the chemical composition of the essential oil. Animals, 10: 786.
Garcia, V., Catala-Gregori, P., Madrid, J., Hernandez, F., Megias, M. and Andrade-Montemayor, H. 2007. Potential of carvacrol to modify in vitro rumen fermentation as compared with monensin. Animal, 1: 675-680.
Gardinal, R., Calomeni, G., Zanferari, F., Vendramini, T., Takiya, C., Bertagnon, H., Batista, C., Della Libera, A. and Renno, F. 2018a. Different durations of whole raw soybean supplementation during the prepartum period: Measures of cellular immune function in transition cows. Journal of Dairy Science, 101: 661-674.
Gardinal, R., Calomeni, G., Zanferari, F., Vendramini, T., Takiya, C., Del Valle, T. and Renno, F. 2018b. Different durations of whole raw soybean supplementation during the prepartum period: Milk fatty acid profile and oocyte and embryo quality of early-lactating Holstein cows. Journal of Dairy Science, 101: 675-689.
Getachew, G., Makkar, H. P. and Becker, K. 2000. Tannins in tropical browses: effects on in vitro microbial fermentation and microbial protein synthesis in media containing different amounts of nitrogen. Journal of Agricultural Food Chemistry, 48: 3581-3588.
Hadian, J., Hossein Mirjalili, M., Reza Kanani, M., Salehnia, A. and Ganjipoor, P. 2011. Phytochemical and morphological characterization of Satureja khuzistanica Jamzad populations from Iran. Chemistry and Biodiversity, 8: 902-915.
Hart, K., Yáñez-Ruiz, D. R., Duval, S., McEwan, N. and Newbold, C. 2008. Plant extracts to manipulate rumen fermentation. Animal Feed Science and Technology, 147: 8-35.
Hegarty, R. and Klieve, A. 1999. Opportunities for biological control of ruminal methanogenesis. Australian Journal of Agricultural Research, 50: 1315-1320.
Honan, M., Feng, X., Tricarico, J. M. and Kebreab, E. 2021. Feed additives as a strategic approach to reduce enteric methane production in cattle: modes of action, effectiveness and safety. Animal Production Science. Doi:10.1071/AN20295
Hristov, A. N., Ivan, M., Rode, L. M. and McAllister, T. A. 2001. Fermentation characteristics and ruminal ciliate protozoal populations in cattle fed medium- or high-concentrate barley-based diets. Journal of Animal Science, 79: 515-524.
Hundal, J., Wadhwa, M. and Bakshi, M. 2019. Herbal feed additives containing essential oil: 1. Impact on the nutritional worth of complete feed in vitro. Tropical Animal Health and Production, 51: 1909-1917.
Kahvand, M. and Malecky, M. 2018. Dose-response effects of sage (Salvia officinalis) and yarrow (Achillea millefolium) essential oils on rumen fermentation in vitro. Annals of Animal Science, 18: 125-142.
Kholif, A. E. and Olafadehan, O. A. 2021. Essential oils and phytogenic feed additives in ruminant diet: chemistry, ruminal microbiota and fermentation, feed utilization and productive performance. Phytochemistry Reviews, Doi:10.1007/s11101-021-09739-3.
Lovett, D., Lovell, S., Stack, L., Callan, J., Finlay, M., Conolly, J. and O'Mara, F. 2003. Effect of forage.concentrate ratio and dietary coconut oil level on methane output and performance of finishing beef heifers. Livestock Production Science, 84: 135-146.
Lowry, O., Rosebrough, N., Farr, A. and Randall, R. 1951. Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193: 265–275.
Macheboeuf, D., Morgavi, D. P., Papon, Y., Mousset, J. L. and Arturo-Schaan, M. 2008. Dose–response effects of essential oils on in vitro fermentation activity of the rumen microbial population. Animal Feed Science and Technology, 145: 335-350.
Machmüller, A., Soliva, C. R. and Kreuzer, M. 2003. Effect of coconut oil and defaunation treatment on methanogenesis in sheep. Reproduction Nutrition Development, 43: 41-55.
Makkar, H., Blümmel, M. and Becker, K. 1995. Formation of complexes between polyvinyl pyrrolidones or polyethylene glycols and tannins, and their implication in gas production and true digestibility in in vitro techniques. British Journal of Nutrition, 73: 897-913.
Makkar, H., Sharma, O., Dawra, R. and Negi, S. 1982. Simple determination of microbial protein in rumen liquor. Journal of Dairy Science, 65: 2170-2173.
Makkar, H. P. S. 2003. Effects and fate of tannins in ruminant animals, adaptation to tannins, and strategies to overcome detrimental effects of feeding tannin-rich feeds. Small Ruminant Research, 49: 241-256.
Manso, T., Castro, T., Mantecón, A. and Jimeno, V. 2006. Effects of palm oil and calcium soaps of palm oil fatty acids in fattening diets on digestibility, performance and chemical body composition of lambs. Animal Feed Science and Technology, 127: 175-186.
Menke, K. 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.
Minson, D. J. 1997. Ruminants: the protein producers. Biologist, 44: 463-464.
Moss, A. R., Jouany, J.-P. and Newbold, J. 2000. Methane production by ruminants: its contribution to global warming. pp. 231-253. In Annales de zootechnie, Vol. 49, EDP Sciences. Paris.
Nolan, J. and Dobos, R. 2005. Nitrogen transactions in ruminants. pp. 177–206. In J. Dijkstra, J.M. Forbes, J. France (eds). Quantitative aspects of ruminant digestion and metabolism. CABI Publishing: Wallingford, UK.
Onodera, R., Nakagawa, Y. and Kandtsu, M. 1977. Ureolytic activity of the washed cell suspension of rumen ciliate protozoa. Agricultural and Biological Chemistry, 41: 2177-2182.
Ottenstein, D. and Bartley, D. 1971. Separation of free acids C2–C5 in dilute aqueous solution column technology. Journal of Chromatographic Science, 9: 673-681.
Patel, S. and Ambalam, P. 2018. Role of rumen protozoa: metabolic and fibrolytic. Advances in Biotechnology and Microbiology, 10:79-83.
Purba, R. A. P., Paengkoum, P. and Yuangklang, C. 2019. In vitro ruminal fermentation and methane production of PUFA containing rations as treated by flavonoid and essential oil from Piper betle L. Chemistry. Doi: 10.20944/preprints201904.0156.v1
Reyes-Becerril, M., Gijón, D., Angulo, M., Vázquez-Martínez, J., López, M. G., Junco, E., Armenta, J., Guerra, K. and Angulo, C. 2021. Composition, antioxidant capacity, intestinal, and immunobiological effects of oregano (Lippia palmeri Watts) in goats: preliminary in vitro and in vivo studies. Tropical Animal Health and Production, 53: 101. Doi: 10.1007/s11250-020-02450-z.
Santos, N. W., Yoshimura, E. H., Mareze-Costa, C. E., Machado, E., Agustinho, B. C., Pereira, L. M., Brito, M. N., Brito, N. A., and Zeoula, L. M. 2017. Supplementation of cow milk naturally enriched in polyunsaturated fatty acids and polyphenols to growing rats. PloS one 12, Doi:10.1371/journal.pone.0172909.
Santra, A. and Karim, S. 2002. Influence of ciliate protozoa on biochemical changes and hydrolytic enzyme profile in the rumen ecosystem. Journal of Applied Microbiology, 92: 801-811.
Shingfield, K., Ahvenjärvi, S., Toivonen, V., Ärölä, A., Nurmela, K., Huhtanen, P. and Griinari, J. M. 2003. Effect of dietary fish oil on biohydrogenation of fatty acids and milk fatty acid content in cows. Animal Science, 77: 165-179.
Shokryazdan, P., Rajion, M. A., Meng, G. Y., Boo, L. J., Ebrahimi, M., Royan, M., Sahebi, M., Azizi, P., Abiri, R. and Jahromi, M. F. 2017. Conjugated linoleic acid: a potent fatty acid linked to animal and human health. Critical Reviews in Food Science and Nutrition, 57: 2737-2748.
Silva, S. N. S. E., Chabrillat, T., Kerros, S., Guillaume, S., Gandra, J. R., de Carvalho, G. G. P., Silva, F. F. D., Mesquita, L. G., Gordiano, L. A., Camargo, G. M. F., Ribeiro, C. V. D. M., de Araújo, M. L. G. M. L., Alba, H. D. R., Silva, R. D. G. and de Freitas, J. E., Jr. 2021. Effects of plant extract supplementations or monensin on nutrient intake, digestibility, ruminal fermentation and metabolism in dairy cows. Animal Feed Science and Technology, 275: 114886.
Sylvester, J., Karnati, S., Yu, Z., Newbold, C. J. and Firkins, J. 2005. Evaluation of a real-time PCR assay quantifying the ruminal pool size and duodenal flow of protozoal nitrogen. Journal of Dairy Science, 88: 2083-2095.
Takenaka, A., Tajima, K., Mitsumori, M. and Kajikawa, H. 2004. Fiber digestion by rumen ciliate protozoa. Microbes and Environments, 19: 203-210.
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, protozoal population, growth and enzyme activity of anaerobic fungus isolated from Mehraban sheep. Animal Feed Science and Technology, 172: 115-124.
Torres, R. N. S., Moura, D. C., Ghedini, C. P., Ezequiel, J. M. B. and Almeida, M. T. C. 2020. Meta-analysis of the effects of essential oils on ruminal fermentation and performance of sheep. Small Ruminant Research, 189: 106148.
Van Soest, P. J., Robertson, J. B. and Lewis, B. A. 1991. Methods for dietary fiber, neutral ‎detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science, 74: 3583-3597.
Wanapat, M., Kang, S., Khejornsart, P. and Wanapat, S. 2013. Effects of plant herb combination supplementation on rumen fermentation and nutrient digestibility in beef cattle. Asian-Australasian Journal of Animal Sciences, 26: 1127-1136.
Williams, A.G., Coleman, G.S., 1992. Role of protozoa in the rumen, The Rumen Protozoa, Springer, pp. 317-347.
Zhou, R., Wu, J., Lang, X., Liu, L., Casper, D. P., Wang, C., Zhang, L. and Wei, S. 2020. Effects of oregano essential oil on in vitro ruminal fermentation, methane production, and ruminal microbial community. Journal of Dairy Science, 103: 2303-2314.