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Effect of mixture of herbal plants on ruminal fermentation, degradability and gas production

ABSTRACT.

Reducing livestock negative environmental impacts get great interest in last years. So, present study was carried out to determine the effect of adding different levels of mixture of thyme and celery versus salinomycin on ruminal fermentation, gas production, dry, organic matter and fiber degradation. Four experimental treatments were used by in-vitro batch culture technique, as follow: 60% CFM, 40% clover hay (control), control diet + 2.5 gm thyme + 2.5 gm celery kg-1 DM (T1), control diet + 5 gm thyme + 5 gm celery kg-1 DM (T2), control diet + 10 gm thyme + 10 gm celery kg-1 DM (T3), control diet + 0.4 gm Salinomycin kg-1 DM (T4). Ruminal pH value was significantly increased (p < 0.05) with T4 compared with other treatments. While, the T4 recorded the lowest value (p < 0.05) for microbial protein, short chain fatty acids concentrations (SCFA), total gas production, dry matter and organic matter degradability (DMd and OMd) compared with other treatments. Fiber fraction degradability (NDFd and ADFd) appeared no significant variance (p > 0.05) between control and other treatments except for T1 that recorded the lowest value (p < 0.05). It is concluded that mixture of thyme plus celery could be alternate for ionophores in the ruminant diets to enhance ruminal fermentation, reducing gas production without any negative effect on nutrients degradability.

Keywords:
thyme; celery; ionophores; gas production; rumen fermentation; nutrients digestibility

Introduction

A major goal of livestock production system today is restriction of using antibiotics and other synthetic medicinal chemistry as feed additives and growth promoters, versus natural growth promoters (NGPs) as effective alternatives to those products. Using antibiotic in ruminant nutrition as a supplement are widely applied to alter rumen fermentation and methane emission but due to the extreme of using antibiotic led to be resistance against different drugs by animals in addition antibiotics transfer to ruminant product (milk or meat) which could threaten human health (Zhan et al., 2017Zhan, J., Liu, M., Wu, C., Su, X., Zhan, K., & Zhao, G. Q. (2017). Effects of alfalfa flavonoids extract on the microbial flora of dairy cow rumen. Asian-Australasian Journal of Animal Sciences, 30(9), 1261-1269.; Bayat et al., 2018Bayat, A. R., Tapio, I., Vilkki, J., Shingfield, K. J., & Leskinen, H. (2018). Plant oil supplements reduce methane emissions and improve milk fatty acid composition in dairy cows fed grass silage-based diets without affecting milk yield. Journal of Dairy Science, 101(2), 1136-1151. ; Khattab, Abd El Tawab, Hadhoud, & Shaaban, 2020Khattab, M. S. A., Abd El Tawab, A. M., Hadhoud, F. A., & Shaaban, M. M. (2020). Utilizing of celery and thyme as ruminal fermentation and digestibility modifier and reducing gas production. International Journal of Dairy Scienc e, 15(1), 22-27.). So, European Union (EU) banned using antibiotics as feed additive, this radical change in laws resulted in an intensive development of research that relates to find effective natural compounds that could inhibit GHG and modulate the rumen fermentation (Matloup et al., 2017Matloup, O. H., Abd El Tawab, A. M., Hassan, A. A., Hadhoud, F. I., Khattab, M.S.A., Khalel, M. S., ... Kholif, A. E. (2017). Performance of lactating Friesian cows fed a diet supplemented with coriander oil: Feed intake, nutrient digestibility, ruminal fermentation, blood chemistry and milk production. Animal Feed Science and Technology, 226, 88-97. doi: 10.1016/j.anifeedsci.2017.02.012
https://doi.org/10.1016/j.anifeedsci.201...
)

Many studies have focused on the alternative strategies development to maintain animal productive performance and health (Khattab, Ebeid, Abd El Tawab, Abo El-Nor, & Aboamer, 2016Khattab, M. S. A., Ebeid, H. M., Abd El Tawab, A. M., Abo El-Nor, S. A. H., & Aboamer, A. A. (2016). Effect of supplementing diet with herbal plants on ruminal fiber digestibility and gas production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(6):1093-1097.; Khattab et al., 2017Khattab, M. S. A., El-Zaiat, H. M., Abd El Tawab, A. M., Matloup, O. H., Morsy, A. S., Abdou, M. M., ... Sallam, S. M. A. (2017). Impact of lemongrass and galangal as feed additives on performance of lactating Barki goats. International Journal of Dairy Science, 12(2), 184-189. doi: 10.3923/ijds.2017.184.189
https://doi.org/10.3923/ijds.2017.184.18...
; Khattab & Tawab, 2018Khattab, M. S. A., & Tawab, A. M. (2018). In vitro evaluation of palm fronds as feedstuff on ruminal digestibility and gas production. Acta Scientiarum Animal Sciences, 40, e39586.; Matloup et al., 2017Matloup, O. H., Abd El Tawab, A. M., Hassan, A. A., Hadhoud, F. I., Khattab, M.S.A., Khalel, M. S., ... Kholif, A. E. (2017). Performance of lactating Friesian cows fed a diet supplemented with coriander oil: Feed intake, nutrient digestibility, ruminal fermentation, blood chemistry and milk production. Animal Feed Science and Technology, 226, 88-97. doi: 10.1016/j.anifeedsci.2017.02.012
https://doi.org/10.1016/j.anifeedsci.201...
). Phytogenic feed additives commonly known as phytobiotic, or herbal plants can be defined as source of plant derived products supplemented to animal feeds in order to increase production and enhance performance. They originate from tubers, roots, fruits or leaves of herbs, spices and other plants (Alam et al., 2013Alam, M. J., Mamuad, L. L., Kim, S. H., Jeong, C. D., Sung, H. G., Cho, S. B., ... Lee, S. S. (2013). Effect of Phytogenic Feed Additives in Soybean Meal on In vitro Swine Fermentation for Odor Reduction and Bacterial Community Comparison. Asian-Australasian Journal of Animal Sciences, 26(2), 266-274. doi: 10.5713/ajas.2012.12511
https://doi.org/10.5713/ajas.2012.12511...
).

Phytogenic feed additives mainly meant plant secondary metabolites such as tannins, saponins and essential oils which investigated as a natural rumen modifier (Knapp, Laur, Vadas, Weiss, & Tricarico, 2014Knapp, J. R., Laur, G. L., Vadas, P. A., Weiss, W. P., & Tricarico, J. M. (2014). Invited review: Enteric methane in dairy cattle production: Quantifying the opportunities and impact of reducing emissions. Journal of Dairy Science, 97(6), 3231-3261. doi: 10.3168/jds.2013-7234
https://doi.org/10.3168/jds.2013-7234...
; Ishlak et al., 2015Ishlak, A., Gunal, M., & AbuGhazaleh, A. A. (2015). The effects of cinnamaldehyde, monensin and quebracho condensed tannin on rumen fermentation, biohydrogenation and bacteria in continuous culture system. Animal Feed Science and Technology, 207, 31-40. doi: 10.1016/j.anifeedsci.2015.05.023
https://doi.org/10.1016/j.anifeedsci.201...
; Cobellis, Trabalza-Marinucci, Marcotullio, & Yu, 2016Cobellis, G., Trabalza-Marinucci, M., Marcotullio, M. C., & Yu, Z., (2016). Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro. Animal Feed Science and Technology, 215, 25-36. doi: 10.1016/j.anifeedsci.2016.02.008
https://doi.org/10.1016/j.anifeedsci.201...
; Ali, Mohamed, Sameeh, Darwesh & Abd El-Razik, 2016Ali, S. I., Mohamed, A. A., Sameeh, M. Y., Darwesh, O. M., & Abd El-Razik, T. M. (2016). Gamma-Irradiation Affects Volatile Oil Constituents, Fatty Acid Composition and Antimicrobial Activity of Fennel (Foeniculum vulgare) Seeds Extract. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(1), 524-532. ). It is used in animal nutrition due to their effects of rumen microbial populations by improving ruminal fermentation efficiency and mitigate methane emissions (Khiaosa-ard & Zebeli, 2013Khiaosa-ard, R., & Zebeli, Q. (2013). Meta-analysis of the effects of essential oils and their bioactive compounds on rumen fermentation characteristics and feed efficiency in ruminants. Journal of Animal Science, 91(4), 1819-1830. doi: 10.2527/jas.2012-5691
https://doi.org/10.2527/jas.2012-5691...
). Methane is one of ruminal digestion secondary metabolites which released due to inability of the animal to benefit from H2 and CO2 production during fermentation, methanogens bacteria can be utilized hydrogen and their carbon dioxide to produce methane (Hobson & Stewart, 1997Hobson, P. N., & Stewart, C. S. (1997). The Rumen Microbial Ecosystem (2nd ed.). London, UK: Blackie.; Martin, Morgavi, & Doreau, 2009Martin, C., Morgavi, D. P., & Doreau, M. (2009). Methane mitigation in ruminants: from microbe to the farm scale. Animal, 4(3), 351-365. doi: 10.1017/S1751731109990620
https://doi.org/10.1017/S175173110999062...
). It was estimated that dairy farms contribute more than 3% of the total greenhouse gasses (Knapp et al., 2014Knapp, J. R., Laur, G. L., Vadas, P. A., Weiss, W. P., & Tricarico, J. M. (2014). Invited review: Enteric methane in dairy cattle production: Quantifying the opportunities and impact of reducing emissions. Journal of Dairy Science, 97(6), 3231-3261. doi: 10.3168/jds.2013-7234
https://doi.org/10.3168/jds.2013-7234...
) that is in addition to be considered a loss in feed energy by up to 12% in the form of emitted methane (Arndt, Powell, Aguerre, Crump, & Wattiaux, 2015Arndt, C., Powell, J. M., Aguerre, M. J., Crump, P. M., & Wattiaux, M. A. (2015). Feed conversion efficiency in dairy cows: Repeatability, variation in digestion and metabolism of energy and nitrogen, and ruminal methanogens. Journal of Dairy Science, 98(6), 3938-3950. doi: 10.3168/jds.2014-8449
https://doi.org/10.3168/jds.2014-8449...
; Li et al., 2018Li, Z. J., Ren, H., Liu, S. M., Cai, C. J., Han, J. T., Li, F., & Yao, J. H. (2018). Dynamics of methanogenesis, ruminal fermentation, and alfalfa degradation during adaptation to monensin supplementation in goats. Journal of Dairy Science, 101(2), 1048-1059. doi: 10.3168/jds.2017-13254
https://doi.org/10.3168/jds.2017-13254...
).

Also, several researchers found that phytobiotic have anti-inflammatory activities and strong antioxidant. it has been reported that many of herbal plants contain essential oils and / or active compounds such as lemongrass, turmeric, galangal, rosemary, clove and cinnamomum, etc. on modifying rumen fermentation and positively affect methane emission, volatile fatty acids (VFA's), protein, carbohydrates degradation and reduce ruminal bio-hydrogenation (Khattab et al., 2016Khattab, M. S. A., Ebeid, H. M., Abd El Tawab, A. M., Abo El-Nor, S. A. H., & Aboamer, A. A. (2016). Effect of supplementing diet with herbal plants on ruminal fiber digestibility and gas production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(6):1093-1097.; Khattab et al., 2017Khattab, M. S. A., El-Zaiat, H. M., Abd El Tawab, A. M., Matloup, O. H., Morsy, A. S., Abdou, M. M., ... Sallam, S. M. A. (2017). Impact of lemongrass and galangal as feed additives on performance of lactating Barki goats. International Journal of Dairy Science, 12(2), 184-189. doi: 10.3923/ijds.2017.184.189
https://doi.org/10.3923/ijds.2017.184.18...
). While, these positive effect in the ruminal fermentation accompanied with negative effect on fiber degradation (Khattab et al., 2017Khattab, M. S. A., El-Zaiat, H. M., Abd El Tawab, A. M., Matloup, O. H., Morsy, A. S., Abdou, M. M., ... Sallam, S. M. A. (2017). Impact of lemongrass and galangal as feed additives on performance of lactating Barki goats. International Journal of Dairy Science, 12(2), 184-189. doi: 10.3923/ijds.2017.184.189
https://doi.org/10.3923/ijds.2017.184.18...
). Supplementing diets with celery or thyme showed no significant pH, microbial protein, ammonia-N concentrations and improved short chain fatty acids concentrations and significantly lowered total gas production (Khattab et al., 2020Khattab, M. S. A., Abd El Tawab, A. M., Hadhoud, F. A., & Shaaban, M. M. (2020). Utilizing of celery and thyme as ruminal fermentation and digestibility modifier and reducing gas production. International Journal of Dairy Scienc e, 15(1), 22-27.).

Ionophores are feed additives inhibiting growth of gram-positive bacteria that produce hydrogen gas for alter rumen microbial populations through ion transfer cross cell membranes which used by archaea bacteria to produce methane (Ishlak et al., 2015Ishlak, A., Gunal, M., & AbuGhazaleh, A. A. (2015). The effects of cinnamaldehyde, monensin and quebracho condensed tannin on rumen fermentation, biohydrogenation and bacteria in continuous culture system. Animal Feed Science and Technology, 207, 31-40. doi: 10.1016/j.anifeedsci.2015.05.023
https://doi.org/10.1016/j.anifeedsci.201...
). Therefore, the objective of the present in vitro study to evaluate the effect of using different levels of thyme plus celery mix (natural feed additive) as an alternative to antibiotic feed additives on ruminal fermentation, ammonia nitrogen concentration and gas emission, dry matter and organic matter and fiber degradation.

Material and methods

Experimental treatments

In-vitro incubation procedures were carried out as described by Khattab, Azzaz, Abd El Tawab, and Murad (2019Khattab, M. S. A., Azzaz, H. H., Abd El Tawab, A. M., & Murad, H. A. (2019). Production Optimization of Fungal Cellulase and its Impact on Ruminal Degradability and Fermentation of Diet. International Journal of Dairy Science, 14(2), 61-68. doi: 10.3923/ijds.2019.61.68
https://doi.org/10.3923/ijds.2019.61.68...
), rumen fluid was collected before morning feeding from 3 ruminally cannulated Holstein dairy cows (mean weight 680 ± 30 kg), mixed and squeezed through 4-layers cheesecloth under continuous flushing with CO2 and immediately transported to laboratory at 39(C (used as a source of inoculum). Treatments were: 60% CFM, 40% clover hay (control), control diet + mix of (2.5 gm thyme and 2.5 gm celery kg-1 DM) (T1), control diet + mix of (5 gm thyme and 5 gm celery kg-1 DM) (T2), control diet + mix of (10 gm thyme and 10 gm celery kg-1 DM) (T3), control diet + 0.4 gm Salinomycin kg-1 DM (T4) (Table 1). Each treatment was tested in eight replicates accompanied by blank bottles (no substrate). the experiment run were repelicated twice in different weeks. Substrate (400 mg) was added to the incubation bottles of 100 mL capacity. Each bottle was filled with 40 mL of the incubation medium (292 mg K2HPO4, 240 mg KH2PO4, 480 mg (NH4)2SO4, 480 mg NaCl, 100 mg MgSO4.7H2O, 64 mg CaCl2.2H2O, 4 mg Na2CO3 and 600 mg cysteine hydrochloride) per 1 liter of double distilled water (ddH2O) and dispensed anaerobically in the 1:4 (v/v) ratio. Then the bootles were incubated at 39°C for 48h.

Table 1
chemical composition of feed ingredients (%).

Substrates sampling and gas production recording

After 48h of incubation, gas production (GP) was recorded using the pressure reading technique according to Khattab and Tawab (2018Khattab, M. S. A., & Tawab, A. M. (2018). In vitro evaluation of palm fronds as feedstuff on ruminal digestibility and gas production. Acta Scientiarum Animal Sciences, 40, e39586.) bottles were uncapped, pH was determined using Hanna digital pH meter, 0.8 mL of strained ruminal fluid was mixed with 0.2 mL of a solution containing 250 g of metaphosphoric acid L-1 for SCFA analysis by titration, after steam distillation of a 4 mL sample, by the method of Annison (1954Annison, E. F. (1954). Some observations on volatile fatty acids in the sheep's rumen. Biochemical Journal, 57(3), 400-405. doi: 10.1042/bj0570400
https://doi.org/10.1042/bj0570400...
). Contents were filtrated of each bottle to obtain the non-digested residue for determination of degradation percent.

Chemical analysis

The non-fermented residues were dried, weighed and digestibility calculated using the equations as described by (Khattab et al., 2016Khattab, M. S. A., Ebeid, H. M., Abd El Tawab, A. M., Abo El-Nor, S. A. H., & Aboamer, A. A. (2016). Effect of supplementing diet with herbal plants on ruminal fiber digestibility and gas production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(6):1093-1097.) as follows:

I n - v i t r o d r y m a t t e r d i s a p p e a r a n c e % = ( i n t i a l D M i n p u t - D M r e s i d u e - B l a n k ) i n t i a l D M i n p u t X 100

I n - v i t r o o r g a n i c m a t t e r d i s a p p e a r a n c e % = ( i n t i a l O M i n p u t - O M r e s i d u e - B l a n k ) i n t i a l O M i n p u t X 100

Dry matter of TMR was determined by drying at 105ºC for 48h (Association of Official Analytical Chemists [AOAC], 1990Association of Official Analytical Chemists [AOAC]. (1990). Official Methods of Analysis (15th ed.). Arlington, VA: AOAC International.; method 930.15). Samples were analyzed for CP (method 976.05), ether extract (method 920.39), and ash (method 942.05) according to AOAC (2000Association of Official Analytical Chemists [AOAC]. (2000). Official Methods of Analysis (17th ed.). Gaithersburg, MD: AOAC International.). The neutral detergent fiber (NDF) was determined using the (Van Soest, Robertson, & Lewis, 1991Van Soest, P. J., Robertson, J. B., & 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. doi: 10.3168/jds.S0022-0302(91)78551-2
https://doi.org/10.3168/jds.S0022-0302(9...
) procedure. The heat stable amylase and sodium sulphite were used to determine NDF. The acid detergent fiber (ADF) content was determined according to AOAC (1990Association of Official Analytical Chemists [AOAC]. (1990). Official Methods of Analysis (15th ed.). Arlington, VA: AOAC International.) method 973.18. The neutral detergent fiber and ADF values were measured on organic matter (OM) basis. Microbial protein production was calculated as 19.3 g microbial nitrogen per kg OMD according to Khattab el al. (2016Khattab, M. S. A., Ebeid, H. M., Abd El Tawab, A. M., Abo El-Nor, S. A. H., & Aboamer, A. A. (2016). Effect of supplementing diet with herbal plants on ruminal fiber digestibility and gas production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(6):1093-1097.). The NH3-N concentration was determined as described by Khattab, Abd-El-Gawad, Abo El-Nor, and El-Sherbiny (2015).

Statistical analysis

Data were statistically analysed using GLM procedure of Statistical Analysis System (SAS, 2009Statistical Analysis System [SAS]. (2009). SAS/STAT User’s guide, Version 9.2. Cary, NC: SAS Institute Inc.), version 9.2. Significant differences between means of treatments were carried out by the Duncan’s test, and the significance threshold was set at p < 0.05.

Results and discussion

Ruminal fermentation parameters:

No differences (p > 0.05) were observed between herbal plant (Mix of Thyme plus Celery) at all different levels compared with control treatments (Table 2). while, it was significantly decreased (p < 0.05) versus the salinomycin treatment in pH values. Also, Ammonia-N concentrations showed no significant (p > 0.05) changes between control and salinomycin compared with T2 that recorded the lowest value (p < 0.05) (31.00 mg 100 mL-1). Many studies stated the effect of herbal plants, spices or essential oils on ruminal pH. The data of the current study for ruminal pH were in line with that of Cardozo, Calsamiglia, Ferret, and Kamel, (2006Cardozo, P., Calsamiglia, S., Ferret, A., & 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 Animal Science, 84(10), 2801-2808. doi: 10.2527/jas.2005-593
https://doi.org/10.2527/jas.2005-593...
); Yang et al. (2007Yang, W. Z., Benchaar, C., Ametaj, B. N., Chaves, A. V., He, M. L., & 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(12), 5671-5681. doi: 10.3168/jds.2007-0369
https://doi.org/10.3168/jds.2007-0369...
); Kung Junior, Williams, Schmidt, and Hu (2008Kung Júnior, L., Williams, P., Schmidt, R. J., & Hu, W. (2008). A blend of essential plant oils used as an additive to alter silage fermentation or used as a feed additive for lactating dairy cows. Journal of Dairy Science, 91(12), 4793-4800. doi: 10.3168/jds.2008-1402
https://doi.org/10.3168/jds.2008-1402...
); Chaves et al. (2008Chaves, A. V., Stanford, K., Dugan, M. E. R., Gibson, L. L., McAllister, T. A., Van Herk, F., & Benchaar, C. (2008). Effects of cinnamaldehyde, garlic and juniper berry essential oils on rumen fermentation, blood metabolites, growth performance, and carcass characteristics of growing lambs. Livestock Science, 117(2-3), 215-224. doi: 10.1016/j.livsci.2007.12.013
https://doi.org/10.1016/j.livsci.2007.12...
); Chaves, Stanford, Gibson, McAllister, and Benchaar (2008Chaves, A. V., Stanford, K., Gibson, L. L., McAllister, T. A., & Benchaar, C. (2008). Effects of carvacrol and cinnamaldehyde on intake, rumen fermentation, growth performance, and carcass characteristics of growing lambs. Animal Feed Science and Technology, 145, 396-408. doi: 10.1016/j.anifeedsci.2007.04.016
https://doi.org/10.1016/j.anifeedsci.200...
); Fandino, Calsamiglia, Ferret, and Blanch (2008Fandino, I., Calsamiglia, S., Ferret, A., & Blanch, M. (2008). Anise and capsicum as alternatives to monensin to modify rumen fermentation in beef heifers fed a high concentrate diet. Animal Feed Science and Technology, 145(1-4), 409-417. doi: 10.1016/j.anifeedsci.2007.04.018
https://doi.org/10.1016/j.anifeedsci.200...
); Meyer et al. (2009Meyer, N. F., Erickson, G. E., Klopfenstein, T. J., Greenquist, M. A., Luebbe, M. K., Williams, P., & Engstrom, M. A. (2009). Effect of essential oils, tylosin and monensin on finishing steer performance, carcass characteristics, liver abscesses, ruminal fermentation and digestibility. Journal of Animal Science, 87(7), 2346-2354. doi: 10.2527/jas.2008-1493
https://doi.org/10.2527/jas.2008-1493...
); Yang, Benchaar, Ametaj, and Beauchemin (2010Yang, W. Z., Benchaar, C., Ametaj, B. N., & Beauchemin, K. A. (2010). Dose response to eugenol supplementation in growing beef cattle: ruminal fermentation and intestinal digestion. Animal Feed Science and Technology, 158(1-2), 57-64. doi: 10.1016/j.anifeedsci.2010.03.019
https://doi.org/10.1016/j.anifeedsci.201...
); Tager and Krause (2011Tager, L. R., & Krause, K. M. (2011). Effects of essential oils on rumen fermentation, milk production, and feeding behavior in lactating dairy cows. Journal of Dairy Science, 94(5), 2455-2464. doi: 10.3168/jds.2010-3505
https://doi.org/10.3168/jds.2010-3505...
), who found no effect of essential oils on ruminal pH. Levels used of essential oils or herbal plants could be influenced on ruminal pH (Evans & Martin, 2000Evans, J. D., & Martin, S. A. (2000). Effects of Thymol on Ruminal Microorganisms. Current Microbiology, 41(5), 336-340. doi: 10.1007/s002840010145
https://doi.org/10.1007/s002840010145...
) who suggested that supplementation of 400 mg L-1 of thymol oil increased ruminal pH while, it was not affected at lower dose (50, 100, 200) (in vitro). Increased in pH at a high level of Thyme could be due to inhibition of rumen microbial fermentation, and a reduction in acetate, lactate, propionate and methane concentrations. Also, Castillejos, Calsamiglia, and Ferret (2006Castillejos, L., Calsamiglia, S., & Ferret, A. (2006). Effect of essential oils active compounds on rumen microbial fermentation and nutrient flow in in vitro systems. Journal of Dairy Science, 89(7), 2649-2658. doi: 10.3168/jds.S0022-0302(06)72341-4.
https://doi.org/10.3168/jds.S0022-0302(0...
) observed the same results when using different levels of some EO, because it has including thymol, limonene, eugenol, guaiacol, vanillin and ϒ-terpinene and the increased ruminal pH consisted with a decrease in rumen fermentation and depression in total VFAs concentration.

Table 2
ruminal parameters of experimental diets supplemented with different levels of mix of Thyme plus Celery powder or Salinomycin.

Herbal plants powder or essential oils had variable influence of ruminal ammonia nitrogen concentration in the many studies. Reduction in NH3-N concentration is suggesting the potentiality of mix of celery plus thymol for inhibiting deamination. These results are in agreement with Wanapat, Cherdthong, Pakdee, and Wanapat (2008Wanapat, M., Cherdthong, A., Pakdee, P., & Wanapat, S. (2008). Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citrates Stapf.) powder supplementation. Journal of Animal Science, 86(12), 3497-3503. doi: 10.2527/jas.2008-0885
https://doi.org/10.2527/jas.2008-0885...
); Macheboeuf, Morgavi, Papon, Mousset, and Arturo-Schaan (2008Macheboeuf, D., Morgavi, D. P., Papon, Y., Mousset, J. L., & 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(1-4), 335-350. doi: 10.1016/j.anifeedsci.2007.05.044
https://doi.org/10.1016/j.anifeedsci.200...
); Cobellis et al. (2016Cobellis, G., Trabalza-Marinucci, M., Marcotullio, M. C., & Yu, Z., (2016). Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro. Animal Feed Science and Technology, 215, 25-36. doi: 10.1016/j.anifeedsci.2016.02.008
https://doi.org/10.1016/j.anifeedsci.201...
) who found that EOs especially cinnamaldehyde and cinnamon reduced NH3-N concentrations in the rumen. Also, McIntosh, Newbold, Losa, Williams, and Wallace (2000McIntosh, F. M., Newbold, C. J., Losa, R., Williams, P., & Wallace, R. J. (2000). Effects of Essential Oil on Rumen Fermentation. Reproduction Nutrition Development, 40, 221-222.) observed that EO inhibited deamination of amino acids measured in vitro by 25%, these potential related to inhibiting bacterial attachment to feed particles (Wallace, McEwan, McIntosh, Teferedegne, & Newbold, 2002Wallace, R. J., McEwan, N. R., McIntosh, F. M., Teferedegne, B., & Newbold, C. J. (2002). Natural products as manipulators of rumen fermentation. Asian-Australasian Journal of Animal Sciences, 15(10), 10-21. doi: 10.5713/ajas.2002.1458
https://doi.org/10.5713/ajas.2002.1458...
). Castillejos, Calsamiglia, Ferret, and Losa (2005Castillejos, L., Calsamiglia, S., Ferret, A., & Losa, R. (2005). Effects of a specific blend of essential oil compounds and the type of diet on rumen microbial fermentation and nutrient flow from a continuous culture system. Animal Feed Science and Technology, 119(1-2), 29-41. doi: 10.1016/j.anifeedsci.2004.12.008
https://doi.org/10.1016/j.anifeedsci.200...
) notified that the microbial species affected by antibiotic were the same as those affected by essential oils. The hyper-ammonia producing bacteria (Clostridium sticklandii and Peptostreptecoccus anaerobius) were associated with a reduction of ammonia production which inhibited by EO added. While the other hyper-ammonia producing species (Clostridium aminophilum) were not affected (McIntosh et al., 2003McIntosh, F. M., Williams, P., Losa, R., Wallace, R. J., Beever, D. A., & Newbold, C. J. (2003). Effects of Essential Oils on Ruminant Microorganisms and Their Protein Metabolism. Applied and Environmental Microbiology, 69(8), 5011-5014. doi: 10.1128/AEM.69.8.5011-5014.2003
https://doi.org/10.1128/AEM.69.8.5011-50...
). In addition, the differences in effects of Herbal plants powder or essential oils supplementation to diets on ammonia nitrogen between in vitro or in vivo studies might be qualified in part by the capacity of rumen microbial populations to adapt and/or degrade EO or powder components (Benchaar & Greathead, 2011Benchaar, C., & Greathead, H. (2011). Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Animal Feed Science and Technology, 166, 338-355. doi: 10.1016/j.anifeedsci.2011.04.024
https://doi.org/10.1016/j.anifeedsci.201...
).

While, microbial protein and short chain fatty acids concentrations (SCFA) showed no significantly decreased (p > 0.05) between herbal plant treatments compared with control treatments while, the salinomycin treatment recorded the lowest value (p < 0.05) (6.07 mmol and 191.48 mg gm-1 DM) respectively. Reduction of short chain fatty acids (SCFAs) in herbal plants treatments could be a good indicator of simultaneous with methane emission reduction in the rumen tract (Busquet, Calsamiglia, Ferret, & Kamel, 2006Busquet, M., Calsamiglia, S., Ferret, A. & Kamel, C. (2006) Plant extracts affect in vitro rumen microbial fermentation Journal of Dairy Science, 89(2), 761-771. doi: 10.3168/jds.S0022-0302(06)72137-3
https://doi.org/10.3168/jds.S0022-0302(0...
), these results are in agreement with Evans and Martin (2000Evans, J. D., & Martin, S. A. (2000). Effects of Thymol on Ruminal Microorganisms. Current Microbiology, 41(5), 336-340. doi: 10.1007/s002840010145
https://doi.org/10.1007/s002840010145...
) who observed that at high level of thymol (400 mg L-1) reduced the short chain fatty acids concentration and the proportion of propionate and acetate, while acetate to propionate ratio was increased. Similar results findings were noted by Castillejos et al. (2006Castillejos, L., Calsamiglia, S., & Ferret, A. (2006). Effect of essential oils active compounds on rumen microbial fermentation and nutrient flow in in vitro systems. Journal of Dairy Science, 89(7), 2649-2658. doi: 10.3168/jds.S0022-0302(06)72341-4.
https://doi.org/10.3168/jds.S0022-0302(0...
) for thymol and eugenol. While, Castillejos, Calsamiglia, Martín-Ereso, and Ter Wijlen (2008)Castillejos, L., Calsamiglia, S., Martín-Ereso, J., & 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(1-4), 259-270. doi: 10.1016/j.anifeedsci.2007.05.037
https://doi.org/10.1016/j.anifeedsci.200...
found that at all level of thymol (i.e., 5, 50 and 500 mg L-1) increased short chain fatty acids concentration, but did not affect the proportions of propionate, acetate, valerate, acetate to propionate ratio and branched-chain fatty acids concentration.

Gas production

Ruminal gas production of experimental diets supplemented with different levels of mix of Thyme plus Celery powder or Salinomycin are listed in Table 3. The values cleared that herbal plant addition to diet (T1 to T3) insignificantly lowered (p > 0.05) in total gas production compared with control diet (153.5, 153.5, 154 vs. 15725 mL, respectively). While, the salinomycin treatment recorded the lowest value (p < 0.05) being (137 ml) for total gas production compared with other treatments. Also, these results showed that gas production per each gram of DM, NDF or ADF recorded the same trend of total gas production.

Table 3
Ruminal gas production of experimental diets supplemented with different levels of mix of Thyme plus Celery powder or Salinomycin.

Addition of medical plants or EO have caused either a modifying rumen fermentation and positively affected gas emission in several of in vivo or in vitro studies (Macheboeuf et al., 2008Macheboeuf, D., Morgavi, D. P., Papon, Y., Mousset, J. L., & 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(1-4), 335-350. doi: 10.1016/j.anifeedsci.2007.05.044
https://doi.org/10.1016/j.anifeedsci.200...
; Patra & Yu, 2012Patra, A. K., & Yu, Z. (2012). Effects of Essential Oils on Methane Production and Fermentation by, and Abundance and Diversity of, Rumen Microbial Populations. Applied and Environmental Microbiology, 78(12), 4271-4280. doi: 10.1128/AEM.00309-12
https://doi.org/10.1128/AEM.00309-12...
; Lin, Lu, Wang, Liang, & Liu, 2012Lin, B., Lu, Y., Wang, J. H., Liang, Q., & Liu, J. X. J. (2012). The effects of combined essential oils along with fumarate on rumen fermentation and methane production in-vitro. Journal of Animal and Feed Sciences, 21, 198-210. doi: 10.22358/jafs/66064/2012
https://doi.org/10.22358/jafs/66064/2012...
; Cobellis et al., 2016Cobellis, G., Trabalza-Marinucci, M., Marcotullio, M. C., & Yu, Z., (2016). Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro. Animal Feed Science and Technology, 215, 25-36. doi: 10.1016/j.anifeedsci.2016.02.008
https://doi.org/10.1016/j.anifeedsci.201...
). The same results were found by Rezaei and Pour (2012Rezaei, N., & Pour, H. A. (2012). Evaluation effect of thyme extract on degradability of soybean meal with gas product technique. Advances in Bioresearch, 3, 45-47.) who noted that the gas emission was reduced on addition thyme methanolic extracts also, Chaudhry and Khan (2012Chaudhry, A. S. & Khan, M. M. H. (2012). Impacts of different spices on in vitro rumen dry matter disappearance, fermentation and methane of wheat or ryegrass hay based substrates. Livestock Science, 146, 84-90.) using in vitro gas production technique, five curry spices such as turmeric, cinnamon, clove, cumin and coriander as a natural antibiotics killing methanogenic bacteria which lead to reduction in methane production by 40%. While, Mariam, El-Zarkouny, El-Shazly, and Sallam (2014Mariam, A. G., El-Zarkouny, S. Z., El-Shazly, K. A. & Sallam, S. M. A. (2014). Impact of essential oils blend on methane emission, rumen fermentation characteristics and nutrient digestibility in barki sheep. Journal of Agricultural Science, 6(7), 144. doi: 10.5539/jas.v6n7p144
https://doi.org/10.5539/jas.v6n7p144...
) reported that the potential impacts of different levels of EOs mix such as thyme, eucalyptus, peppermint, cinnamon and lemon on ruminal fermentation and nutrient digestibility in Barki sheep, they found that there were no significant differences among treatments of essential oil mix on in vitro gas production and methane production, short chain fatty acids (SCFA), NH3-N concentration and protozoa count. While, the result of the study showed that the combination of the five EOs had an additive effect on lower response on rumen microbial fermentation and methane emission. Kim, Adesogan, and Shin (2012Kim, S. C., Adesogan, A. T. & Shin, J. H. (2012). Effects of dietary addition of wormwood (Artemisia montana Pampan) silage on growth performance, carcass characteristics, and muscle fatty acid profiles of beef cattle. Animal Feed Science and Technology, 177, 15-22. ) suggested that the herbal plant extracts (Wormwood, Allium sativum for. Pekinense; Artemisia princeps var. Orientalis; Garlic, Allium cepa; Ginger, Citrus unshiu; Onion, Zingiber officinale; Honeysuckle; Mandarin orange, Lonicera japonica) were shown to have properties to reduce acetate to propionate ratio and methane production, increase fibrolytic bacteria species and decrease methanogen population. It well-known that the cell wall contents (NDF and ADF) have negatively affected on gas production which tends to reduce the microbial activity (Khattab et al., 2015Khattab, M. S. A., Abd-El-Gawad, A., Abo El-Nor, S. A. H., & El-Sherbiny, M. (2015). The effect of diet supplemented with vegetable oils and/or monensin on the vaccenic acid production in continuous culture fermenters. Animal Nutrition, 1(4) 320-323.).

Nutrients digestibility.

Nutrients digestibility are listed in Table 4. The results of dry matter and organic matter digestibilities (DMd and OMd) cleared that there were no significant differences (p > 0.05) between herbal plant groups and control but, salinomycin group actually recorded the lowest value (p < 0.05) being 42.18 and 49.61 % respectively.

The data of fiber fraction degradability (NDFd and ADFd) appeared that there were no significant variance (p > 0.05) between control and other treatments except T1 there was recorded the lowest value (p < 0.05) being 28.81 and 19.57 % respectively.

Table 4
Ruminal DM, OM, NDF and ADF degradability (%) of experimental diets supplemented with different levels of mix of Thyme plus Celery powder or Salinomycin.

Dry matter and organic matter digestibilities were not affected by addition of herbal plants these results are in agreement with previous studies (Castillejos et al., 2005Castillejos, L., Calsamiglia, S., Ferret, A., & Losa, R. (2005). Effects of a specific blend of essential oil compounds and the type of diet on rumen microbial fermentation and nutrient flow from a continuous culture system. Animal Feed Science and Technology, 119(1-2), 29-41. doi: 10.1016/j.anifeedsci.2004.12.008
https://doi.org/10.1016/j.anifeedsci.200...
, Wanapat et al., 2008Wanapat, M., Cherdthong, A., Pakdee, P., & Wanapat, S. (2008). Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citrates Stapf.) powder supplementation. Journal of Animal Science, 86(12), 3497-3503. doi: 10.2527/jas.2008-0885
https://doi.org/10.2527/jas.2008-0885...
, Wanapat, Kang, Khejornsart, & Wanapat, 2013Wanapat, M., Kang, S., Khejornsart, P., & 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.; Nanon, Suksombat, Beauchemin, & Yang, 2014Nanon, A., Suksombat, W., Beauchemin, K. A., & Yang, W. Z. (2014). Short Communication: Assessment of lemongrass oil supplementation in a dairy diet on in vitro ruminal fermentation characteristics using the rumen simulation technique. Canadian Journal of Animal Science, 94, 731-736.; Patra and Yu (2014Patra, A. K., & Yu, Z. (2014). Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen. Applied Microbiology and Biotechnology, 98(2), 897-905. doi: 10.1007/s00253-013-4930-x
https://doi.org/10.1007/s00253-013-4930-...
); Ishlak et al., 2015Ishlak, A., Gunal, M., & AbuGhazaleh, A. A. (2015). The effects of cinnamaldehyde, monensin and quebracho condensed tannin on rumen fermentation, biohydrogenation and bacteria in continuous culture system. Animal Feed Science and Technology, 207, 31-40. doi: 10.1016/j.anifeedsci.2015.05.023
https://doi.org/10.1016/j.anifeedsci.201...
and Khattab et al., 2016Khattab, M. S. A., Ebeid, H. M., Abd El Tawab, A. M., Abo El-Nor, S. A. H., & Aboamer, A. A. (2016). Effect of supplementing diet with herbal plants on ruminal fiber digestibility and gas production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(6):1093-1097.). Also, Patra (2011Patra, A. K. (2011). Effects of essential oils on rumen fermentation, microbial ecology and ruminant production. Asian Journal of Animal and Veterinary Advances, 6, 416-428.) found that EOs have no negative effect on fiber degradation. Using medical plants in animal diets had no negative effect on activity and growth of the major cellulolytic bacterial population (Cobellis et al., 2016Cobellis, G., Trabalza-Marinucci, M., Marcotullio, M. C., & Yu, Z., (2016). Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro. Animal Feed Science and Technology, 215, 25-36. doi: 10.1016/j.anifeedsci.2016.02.008
https://doi.org/10.1016/j.anifeedsci.201...
). While, Fraser et al. (2007Fraser, 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(5), 2315-2328. doi: 10.3168/jds.2006-688
https://doi.org/10.3168/jds.2006-688...
) using in vitro two continuous culture systems were evaluated for its study the effect of cinnamon leaf oils on ruminal fermentation and observed significant reduction on microbial activity and decreased in digestibility. Rezaei and Pour (2012Rezaei, N., & Pour, H. A. (2012). Evaluation effect of thyme extract on degradability of soybean meal with gas product technique. Advances in Bioresearch, 3, 45-47.) how observed that the degradability of soybean meal using in vitro gas production technique was decreased on addition thyme methanolic extracts.

Conclusion

Under the conditions of the present study, it could be concluded that adding Thyme plus Celery mix (a natural feed additive) to ruminant diets had insignificant effect on ruminal fermentation parameters (reduction total gas production, SCFA and microbial protein), Also, there were no negative effect on DM, OM, NDF and ADF degradability at 2.5 or 5 g celery and thyme kg-1 DMd. Recently, the in vivo study is carrying out to illustrate more topics such as the effect of those herbal plants on feed intake, rumen fermentation, productive performance, and milk production and compassion.

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Publication Dates

  • Publication in this collection
    25 Sept 2020
  • Date of issue
    2021

History

  • Received
    01 July 2019
  • Accepted
    15 Apr 2020
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