Effects of adding different levels of thyme powder and garlic cabbage powder in alfalfa, sainfoin and their mixture on digestibility and the amount of in vitro gas production

Document Type : Research Paper

Authors

Abstract

Introduction: Between 2 to 12% of a ruminant’s energy intake is typically lost through the enteric fermentation process. Ruminant production is associated with nitrogen loss especially in the form of ammonia from urine and manure management during composting. These contribute to greenhouse emission and environmental pollution in general. In the past few decades, feed additives such as antibiotics, ionophores, methane inhibitors, and anti-protozoa agents have been successfully used to reduce these rumen energy, nitrogen releases and metabolic disorders (Calsamiglia et al. 2007), though   increase production efficiency. Therefore, ruminants’ nutritionists try to use compounds that increase the efficiency of energy and protein intake by changing the population and activity of ruminal microorganisms (Fereydounpour et al. 2016).   These alternative compounds are probiotics, organic acids, and medicinal plants (Calsamiglia et al. 2007). The essential oils are secondary plant metabolites, which have antimicrobial properties and can be suitable substitutes for antibiotics to alter rumen microbial activity. Also, a research has shown that the essential oils and their constituents have the potential to improve nitrogen and energy utilization in ruminants (Patra et al. 2006). Talebzadeh et al (2012) reported that the essential oil of thyme at 150, 300, 450, and 600 micrograms per ml of culture medium was used only at concentrations above 450 mg decreased the real digestibility of organic matter. Adding powdered herbs such as thyme and garlic cabbage can affect the in vitro digestibility and the amount of in vitro gas production in sainfoin and alfalfa and their mixture. Thymol is a main phenolic component in thyme and carvacrol is also a minor component (Leung and Foster 1996). These compounds are involved in oxidation and reduction reactions. Thyroid has a large antimicrobial activity by preventing the bacteria growth. Researchers reported that active compounds in garlic containing allycine, diallylsulfide, and di-allylene sulfide, which can affect ruminal harmful bacteria. Considering the various compounds and effects of thyme and garlic and the need for further studies on medicinal plants, this study carried out to investigate the effects of adding levels of thyme powder and garlic on digestibility and fermentation parameters of sainfoin and alfalfa forage in laboratory conditions.
Material and methods: A study was conducted to investigate the effect of adding different levels   of thyme powder and garlic cabbage powder on digestibility and the amount of in vitro gas production at three levels of 0, 1.5, and 3% for alfalfa, sainfoin and mixed alfalfa and sainfoin based on a completely randomized design. Measurement of digestibility of alfalfa forage, sainfoin and mixture of 50% alfalfa and 50% sainfoin were carried out using thyme powder and garlic cabbage powder by Holden method (2000). In this method, the nylon bag was used instead of filtration and a daisy (rumen simulator) incubator was used instead of a hot water bath. Menke et al (1979) method was used to measure the amount of gas production. The amounts of short chain fatty acids, digestibility of dry matter, organic matter in dry matter, and metabolizable energy were estimated using related equations (Menke and Steinggass 1988). The data obtained from the method of gas production analyzed in a completely randomized design with repeated measurements. Data digestibility by Holden was investigated in a completely randomized design using SAS (2003) software.
Results and discussion: The results showed that adding 1.5% thyme and garlic to sainfoin forage improved digestibility of dry matter, organic matter digestibility, digestibility of organic matter in dry matter, and metabolizable energy compared with control group (P <0.05). Also, adding 3% garlic powder showed a significant increase in digestibility of organic matter in dry matter and metabolizable energy compared with the control group (P <0.05). But adding 1.5% and 3% thyme and garlic powder to alfalfa and mixed alfalfa and sainfoin had not a significant effect on dry matter digestibility, organic matter digestibility, digestibility of organic matter in dry matter and metabolizable energy compared with the control group. The results showed that adding 1.5 percent of garlic powder to sainfoin was reduced the amount of produced gas compared with the control group for 6, 24, 48 hours after incubation and also adding 3 percent of garlic powder to sainfoin was decreased the amount of produced gas compared with the control group for 6, 24, 48, 72, 96 and 120 hours after incubation (P <0.05). This trend was followed by the reduction of the amount of gas for treatment of 3% garlic + alfalfa at 3, 6, 12, 24, 48, 72, 120 times compared with the control group (P <0.05). Feeding 1.5% of garlic powder was increased the amount of produced gas at 3, 48, 72, and 120 hours of incubation compared with the control group (P <0.05). The results showed that adding 1.5 and 3% garlic powder to sainfoin forage had the lowest amount of digestible organic matter, short chain fatty acid, metabolizable energy, gas production potential and gas production rate compared with other treatments (P <0.05). Also, results showed that 3% of garlic powder in alfalfa had the lowest amount of digestible organic matter, short chain fatty acid, metabolizable energy, gas production potential compared with the control group (P <0.05). Adding plant additives had no significant effect on mixed of alfalfa and sainfoin compared with the control group. The essential oils that contain high levels of effective components (Fraser et al. 2007) or even low ones (Castillejos et al. 2006) can affect digestibility. This is due to the sensitivity of fibrolytic bacteria to the active components of all essential oils (Benchaar et al. 2007).
Conclusion: The effect of different levels of thyme and garlic on the nutritional value and digestibility of forage and sainfoin showed that addition of 1.5% of garlic and thyme improved the digestibility of forage by Holden method. Generally, adding thyme to forage at most of the times did not significantly improve the amount of gas produced compared with the control group, but at some times there was a numerical increase in the amount of produced gas. Adding 3% garlic at most of the times and 1.5% garlic at some of the times caused a significant reduction in the amount of produced gas compared with the control group. It can also be concluded that the use of 3% garlic reduced the fermentation parameters obtained from the gas test (total volatile fatty acid, metabolizable energy, and digestibility) in sainfoin and alfalfa. The results of this study suggest that the addition of thyme and garlic can change the fermentation parameters and the amount of gas production, which is different depending on their usage amount.
 

Aharoni Y, Gilboa N and Silanikove N, 1998. Analysis of the suppressive effect of tannins on ruminal degradation by compartmental models. Animal Feed Science Technology 71: 251-267.
Ahmadiar R and Hojibari F, 2010. The effect of different garlic products on ruminal microbial fermentation under laboratory conditions. Master's Thesis. Razi University of Kermanshah. (In Persian).
Anassori E, Dalir-Naghadeh B, Pirmohammadi R, Taghizadeh A, Asri-Rezaei S, Maham M, Farahmand-Azar S and Farhoomand P, 2011. Garlic: A potential alternative for monensin as a rumen modifier. Livestock Science 142: 276-287.
AOAC, 2005. Official method of Analysis. 18th Edition, Association of Officiating Analytical Chemists, Washington DC, Method 935.14 and 992.24.
Benchaar C and 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, Petit V, Berthiaume RH, Ouellet DR, Chiquette J and Chouinard PY, 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.
Beuvink JMW and 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 and Biotechnology 37:505-509.
Borchers R, 1965. Proteolytic activity of rumen fluid in vitro. Journal of Animal Science 24:1033–1038.
Burt S, 2004. Essential oils: Their antibacterial properties and potential applications in foods: areview. International Journal of Food Microbiology 94: 223–253.
Busquet M, Calsamiglia S, Ferret A, Carro MD and Kamel C, 2005. Effect of garlic oil and four of its compounds on rumen microbial fermentation. Journal of Dairy Science 88: 4393–4404.
Calsamiglia S, Busquet M, Cardozo PW and Castillejos L and Ferret A, 2007. Invited review: Essential oils as modifiers of rumen microbial fermentation. Journal Dairy Science 90: 2580-2595.
Castillejos L, Calsamiglia S and Ferret A, 2006. Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems. Journal Dairy Science 89: 2649–2658.
Demeyer DI and Van Nevel CJ, 1975. Methanogenesis an integrated part of carbohydrate fermentation and its control. University of New England Publishing Unit Armidale pp. 366–382.
DeRosa M, Gambacorta A and Gliozzi A, 1986. Structure, biosynthesis, and physicochemical properties of archaebacterial lipids. Microbiology Reviews 50:70-80.
Devant M, Gambacorta A and Bach A, 2007. Effects plant extracts supplementation on rumen fermentation and metabolism young Holstein bulls consuming high levels of concentrate. Journal of Animal Feed Science Technology 137: 46–57.
Fan M and Chen J, 2001. Studies on antimicrobial activity of extracts from thyme. Wei Sheng Wu Xue Bao 41: 499-504
Fattahnia F, Mohammadizadeh T, Azarfar A, Khatib Joe A and Tasley G, 2015. Effect of Zataria multiflora essential oil on rumen fermentation processes in diets containing different sources of starch and fat by gas production method. Journal of Ruminant Research3: 37-58. (In Persian).
Fereydounpour, M, Bayat Koohsar, J, Pooralmadari E G, Ebrahimi P, 2016. Effect of essential oils of different species of thyme on gas production, digestibility and fermentation parameters in laboratory conditions. Animal Production Research 714:109-117. (In Persian).
Fraser GR, Chaves AV, Wang Y, McAllister TA, Beauchemin KA and Benchaar C, 2007. Assessment of the effects of cinnamon leaf oil on rumen microbial fermentation using two continuous culture systems. Journal Dairy Science 90: 2315-2328.
Hess HD, Machmüller A, Diaz TE and Kreuzer M, 2001. Rusitec evaluation of the potential of saponin–rich tropical fruits to manipulate rumen fermentation and to reduce methanogenesis. Proceedings of the Society of Nutrition Physiology 10: 123.
Hodjatpanah-montazeri A, Danesh Mesgaran M, Vakili A, Ghorbani B and Tabatabaie F, 2014. In vitro Effect of Garlic Oil and Turmeric Extract on Methane Production from Gas Test Medium. Annual Research Review in Biology 4: 1439:1447.
Holden LA, 2000. Comparison of methods of in vitro dry matter digestibility for ten feeds. Journal Dairy Science 82: 1791-1797.
Ivan M, Neill, L Forster R, Alimon R, Rode LM and Entz T, 2000. Effects of Isotricha, Dasytricha, Entodinium and total fauna on ruminal fermentation and duodenal flow in wethers fed different diets. Journal of Dairy Science 83: 776–787.
Jani A, Danesh Mesgaran M and Vakili A, 2010. Effect of peppermint oil and NFC on in vitro gas production parameters. Fourth Iranian Animal Science Congress, Tehran, Iran. (In Persian).
Kamel C, Greathead HMR, Tejido ML, Ranilla MJ and Carro MD, 2007. Effects of allicin and diallyl disulfide on in vitro rumen fermentation of a mixed diet. Journal of Animal Feed Science Technology 145: 351-363.
Klevenhusen F, ZeitzaDuvalb JO, Kreuzera SM and Solivaa CR, 2011. Garlic oil and its principal component di-allyl disulfide fail to mitigate methane, but improve digestibility in sheep. Animal Feed Science Technology 166: 356–363.
Kongmun P, Wanapat M, Pakdee P and Navanukraw C, 2010. Effect of coconut oil and garlic powder on in vitro fermentation using gas production technique. Livestock Science 127: 38-44.
Leung A and Foster S, 1996. Encyclopedia of common natural ingredients drugs and cosmetics. 2nd edition, John Willey, Editor, New York: John Willey and Sons Inc Press.
Martin SA and Macy JM, 1985. Effects of monensin, pyromellitic diimide and 2-bromoethanosulfonic acid on rumen fermentation in vitro. Journal of Animal Science 60: 544–550.
Martinez S, Madrid J, Hernandez F, Megias MD, Sotomayor JA and Jordan MJ, 2006. Effect of thyme essential oils (Thymus hyemalis and Thymus zygis) and monensin on in vitro ruminal degradation and volatile fatty acid production. Journal of Agricultural and Food Chemistry 54: 6598-6602.
McAllister TA, Bae HD, Jones GA and Cheng KJ, 1994. Microbial attachment and feed digestion in the rumen. Journal of Animal Science 72: 3004–3018.
Menke K and Steinggass H, 1988. Estimation of the energetic feed value from chemical analyses and in vitro gas production using rumen fluid. Animal Research and Development 28:7-55.
Menke K, Raa L, Steingass H, Fritz D and Scheider W, 1979. The estimation of the digestibility and metabolisable energy content of ruminant feeding stuffs from the gas production technique when they are incubated with rumen liquor in vitro. Journal of Agricultural Science Cambridge 93: 217-222.
Patra AK, 2011. Effects of essential oils on Rumen fermentation, microbial ecology and Ruminant production. Asian Journal of Animal and Veterinary Advances 6: 416- 428.
Patra AK, Kamra DN and Agarwal N, 2006. Effect of spices on rumen fermentation, methanogenesis and protozoa counts in in vitro gas production test. International Congress Series 1293: 176–179.
Sahli F, Darej C and Moujahed N, 2018. Potential of white garlic powder (Allium sativum L.) to modify in vitro ruminal fermentation. South African Journal of Animal Science 48: 30-41.
Sallam SMA, Bueno ICS, Brigide P, Godoy PB, Vitti, DMSS and Abdalla AL, 2009. Efficacy of eucalyptus oil on in vitro ruminal fermentation and methane production, Options Mediterraneennes. Serie A, Seminaires Mediterraneens 85: 267-272.
SAS / STAT User's Guide. Version 9.1 Edition. 2003. SAS Inst. Cary, NC.
Taherinia MH, Chaji M, Mohammadabadi T, Eslami M and Sari M, 2014. Effects of diets contain garlic powder on degradability and digestion of ruminal and intestinal of fibrous and protein feedstuffs in Arabian sheep. Journal of Ruminant Research2:20-31. (In Persian).
Talebzadeh R, Alipour D, Saharkhiz MJ, 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 Technology 172:115- 124.
Van Soest PJ, Robertson JB and Lewis BA, 1991. Methods for dietary neutral detergent fiber, and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 3583-3597.
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 Science of Food and Agriculture 88: 2231–2237.
Yang WZ, Benchaar C, Ametaj BN, Chaves AV, He ML and McAllister TA, 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.