In vitro methane production from silages based on Cenchrus purpureus mixed with Tithonia diversifolia in different proportions

María Alexandra Huertas González Olga Lucía Mayorga Mogollón Yuri Marcela García Saavedra Vilma Amparo Holguín Castaño Jairo Mora-Delgado About the authors

Abstract

ABSTRACT. Climate change (CC) affects food production, mainly those based on livestock systems. Producers must identify adaptation strategies to ensure the production, during periods of drought, and lack of forage. Besides contributing to CC, high emissions of ruminal methane (CH4) are energy loss potentially usable for livestock production. The objective was to estimate in vitro ruminal gas production (RGP) and determine the CH4 emissions from silages. Treatments were made with forage of Cenchrus purpureus mixed with Tithonia diversifolia T1= C.purpureus at 100%; T2= C.purpureus/ T.diversifolia in 33/67 percent ratio; T3= C.purpureus/ T.diversifolia 67/33; and T4= T.diversifolia at 100%. Samples of silages were analyzed, and they were inoculated with strains of Lactobacillus paracasei (T735); then they were fermented in vacuum-sealed bags for 67 days. RGP and CH4 were measured at 2, 4, 8, 12, 18, 24, 30, 36, and 48 hours. Additionally, modeling of CH4 production kinetics was conducted, using different equations. The results indicate that the highest cumulative CH4 production was for T1. This kinetics was represented using the Gompertz model. In conclusion, the inclusion of T.diversifolia to C.purpureus silages contributes to the decrease of methane at the ruminal level, which constitutes an adaptation practice at climate change.

Keywords:
digestibility; greenhouse gasses; small ruminants; silvi-pastures


Introduction

Currently, livestock has become an activity of great importance within the agricultural sector of tropical areas and especially in the Colombian territory (Bettencourt, Tilman, Narciso, Carvalho, & Henriques, 2015Bettencourt, E. M. V., Tilman, M., Narciso, V., Carvalho, M. L. D. S., & Henriques, P. D. D. S. (2015). The livestock roles in the wellbeing of rural communities of Timor-Leste. Revista de Economia e Sociologia Rural, 53(1), 63-80. doi: 10.1590/1234-56781806-94790053s01005
https://doi.org/10.1590/1234-56781806-94...
). Within livestock activity, it is known that ruminants have a digestive system that can use fibrous material with a high content of structural carbohydrates and convert them into foods of high nutritional quality, such as meat and milk (Friedrich, 2014Friedrich, T. (2014). Producción de alimentos de origen animal. Actualidad y perspectivas. Revista Cubana de Ciencia Agrícola, 48(1), 5-6. Recovered from https://www.redalyc.org/pdf/1930/193030122003.pdf
https://www.redalyc.org/pdf/1930/1930301...
). However, different studies have suggested that this digestive system also produces methane (CH4), a potent greenhouse gas (GHG) that contributes significantly to global warming (Cárdenas & Flores, 2012Cárdenas, J. A. B., & Flores, C. L. (2012). Emisión de metano entérico por rumiantes y su contribución al calentamiento global y al cambio climático. Revisión. Revista Mexicana de Ciencias Pecuarias, 3(2), 215-246. Recovered from https://www.researchgate.net/publication/236158591_Emision_de_metano_enterico_por_rumiantes_y_su_contribucion_al_calentamiento_global_y_al_cambio_climatico_Revision
https://www.researchgate.net/publication...
).

With increasing pressure from the global community to reduce methane emissions and the inverse correlation between energy utilization and CH4 production (Olivo & Soto-Olivo, 2010Olivo, M. L, & Soto-Olivo, A. (2010). Comportamiento de los gases de efecto invernadero y las temperaturas atmosféricas con sus escenarios de incremento potencial. Universidad, Ciencia y Tecnología, 14(57), 221-230. Recovered from http://ve.scielo.org/scielo.php?script=sci_arttext&pid=S1316-48212010000400002
http://ve.scielo.org/scielo.php?script=s...
), especially with ruminants, methane emissions studies have acquired great importance due to its negative effects on the environment (Ribeiro et al., 2016Ribeiro, R. S., Terry, S. A., Sacramento, J. P., Silveira, S. R., Bento, C. B., Silva, E. F., … Chaves, A. V. (2016). Tithonia diversifolia as a Supplementary Feed for Dairy Cows. PloS one, 11(12), e0165751. doi: 10.1371/journal.pone.0165751
https://doi.org/10.1371/journal.pone.016...
). It should be noted that the CH4 production at the enteric level is associated with the quality and quantity of the food ingested; diets with lower digestibility and higher content of structural carbohydrates, translate into increased gas production (Kulivand & Kafilzadeh, 2015Kulivand, M., & Kafilzadeh, F. (2015). Correlation between chemical composition, kinetics of fermentation and methane production of eight pasture grasses. Acta Scientiarum. Animal Sciences,37(1), 9-14. doi: 10.4025/actascianimsci.v37i1.24336
https://doi.org/10.4025/actascianimsci.v...
). More gas production from enteric fermentation means a lower energy efficiency of animals and the emission of CH4 through belching, that is, the energy that the animal underutilizes, because it is not converted into livestock products.

Livestock products constitute some important high protein sources for global food security because they provide 17% of global energy consumption and 33% of global protein consumption (Rojas-Downing, Nejadhashemi, Harrigan, & Woznicki, 2017Rojas-Downing, M., Nejadhashemi, A. P., Harrigan, T., & Woznicki, S. A. (2017). Climate change and livestock: impacts, adaptation, and mitigation. Climate Risk Management, 16, 145-163. doi: 10.1016/j.crm.2017.02.001
https://doi.org/10.1016/j.crm.2017.02.00...
; Rosegrant, Fernandez, & Sinha, 2009Rosegrant, M. W., Fernandez, M., & Sinha, A. (2009). Looking into the future for agriculture and AKST. In B. D. McIntyre, H. R. Herren, J. Wakhungu, R. T. Watson (Eds.), International Assessment of Agricultural Knowledge, Science and Technology for Development (IAASTD). Agriculture at a crossroads (p. 307-376). Washington, DC: Island Press.). Thus, animal production systems contribute to the livelihoods of one billion of the countryside population in the world (Hurst, Termine & Karl, 2007Hurst, P., Termine, P., & Karl, M. (2007). Agricultural workers and their contribution to sustainable agriculture and rural development. Geneve, CH: FAO. Recovered from http://www.fao.org/3/a-bp976e.pdf
http://www.fao.org/3/a-bp976e.pdf...
); mainly, the small ruminant production systems have been managed by small households from indigenous people and rural societies, some of them under extreme poverty conditions (Forero-Álvarez, 2013Forero-Álvarez, J. (2013). The economy of family farming production. Cuadernos de Desarrollo Rural, 10(spe70), 27-45. Recovered from http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0122-14502013000100002
http://www.scielo.org.co/scielo.php?scri...
; Maluf, Burlandy, Santarelli, Schottz, & Speranza, 2016Maluf, R. S., Burlandy, L., Santarelli, M., Schottz, V., & Speranza, J. S. (2015). Nutrition-sensitive agriculture and the promotion of food and nutrition sovereignty and security in Brazil. Ciência & Saúde Coletiva, 20(8), 2303-2312. doi: 10.1590/1413-81232015208.14032014
https://doi.org/10.1590/1413-81232015208...
). In these production systems, the traditional grazing practices have been the most common cause of pasture degradation and consequently, environmental impacts by increasing greenhouse emissions (Mahecha-Ledesma, Angulo-Arizala, & Barragán-Hernández, 2017Mahecha-Ledesma, L., Angulo-Arizala, J., & Barragán-Hernández, W. (2017). Calidad nutricional, dinámica fermentativa y producción de metano de arreglos silvopastoriles. Agronomía Mesoamericana, 28(2), 371-387. doi: 10.15517/ma.v28i2.22750
https://doi.org/10.15517/ma.v28i2.22750...
). Therefore, having livestock as a source, mainly in animal feeding practices (Verdecia et al., 2011Verdecia, D. M., Ramírez, J. L., Leonard, I., Álvarez, Y., Bazán, Y., Bodas, R., & López, S. (2011). Calidad de la Tithonia diversifolia en una zona del Valle del Cauto. REDVET. Revista electrónica de Veterinaria, 12(5), 1-13. (ID 63622168004)).

Thus, feeding practices and strategic supplementation based on local forage resources available in tropical areas constitutes an eco-friendly technological option (Castaño, Grisalez, Navia, & Delgado, 2015Castaño, V. H., Grisalez, S. O., Navia, A. V., & Delgado, J. R. M. (2015). Evaluación multicriterio de 44 introducciones de Tithonia diversifolia (hemsl.) Gray en Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
https://doi.org/10.15446/rfmvz.v62n2.519...
). In this sense, the use of forage bushes constitutes a viable alternative, Tithonia diversifolia is a shrub whose forage is a good source of protein in the diets and other primary and secondary metabolites (Rivera, Naranjo, Cuartas, & Arenas, 2013Rivera, J. E., Naranjo, J. F., Cuartas, C. A., & Arenas, F. A. (2013). Fermentación in vitro y composición química de algunos forrajes y dietas ofrecidas bajo un Sistema Silvopastoril en el trópico de altura. Livestock Research for Rural Development, 25(10), 1-12. Recovered from https://www.researchgate.net/profile/Julian_Rivera2/publication/287056357_In_vitro_fermentation_and_chemical_composition_of_some_forages_and_diets_offered_to_cattle_in_a_tropical_Silvopastoral_System/links/56caed1308ae96cdd06f6a61.pdf
https://www.researchgate.net/profile/Jul...
); some of them have shown a tendency to decrease rumen methane synthesis. In fact, the supplementation of ruminant diets with T. diversifoliahas been suggested as a promising dietary strategy (Ribeiro et al., 2016Ribeiro, R. S., Terry, S. A., Sacramento, J. P., Silveira, S. R., Bento, C. B., Silva, E. F., … Chaves, A. V. (2016). Tithonia diversifolia as a Supplementary Feed for Dairy Cows. PloS one, 11(12), e0165751. doi: 10.1371/journal.pone.0165751
https://doi.org/10.1371/journal.pone.016...
). Currently, T. diversifolia has been reported because it reduces methane output by 6-fold when compared to control due to the presence of secondary metabolites in the plant. These chemical characteristics of the T. diversifolia associated with its high consumption, high digestibility and high passage rate, are expressed in a significant decrease in methane production per unit of digested fodder (Meza et al., 2014Meza, G. A., Loor, N. J., Sánchez, A. R., Avellaneda, J. H., Meza, C. J., Vera, D. F., ... López, F. X. (2014). Inclusión de harinas de follajes arbóreos y arbustivos tropicales (Morus alba, Erythrina poeppigiana, Tithonia diversifolia e Hibiscus rosa-sinensis) en la alimentación de cuyes (Cavia porcellus Linnaeus). Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 61(3), 258-269. Recovered from http://www.scielo.org.co/pdf/rfmvz/v61n3/v61n3a05.pdf
http://www.scielo.org.co/pdf/rfmvz/v61n...
). This means that the implementation of this fodder species in the feeding of ruminants has been shown to maintain an optimal ruminal balance that improves productivity and decreases methane production (Galindo et al., 2011Galindo, J., Gonzáluez, N., Sosa, A., Ruíz, T., Torres, V., Aldana, A. I., ... Noda, A. C. (2011). Efecto de Tithonia diversifolia (Hemsl.) Gray (Botón de oro) en la población de protozoos y metanógenos ruminales en condiciones in vitro. Revista Cubana de Ciencia Agrícola, 45(1), 33-37. Recovered from https://www.semanticscholar.org/paper/Efecto-de-Tithonia-diversifolia-%28Hemsl.%29-Gray-de-en-Galindo-Gonz%C3%A1lez/b147f9ba2a5edc5566028c62b89bbac97fb7fdf9?p2df
https://www.semanticscholar.org/paper/Ef...
).

Despite the high protein content of T. diversifolia (10.3-25.6%) (La et al., 2009La, O., Valenciaga, D., Gonzalez, H., Orozco, A., Castillo, Y., Ruiz, O., ... Arzola, C. (2009). Effect of the combination of Tithonia diversifolia with Pennisetum purpureum cv. Cuba CT-115 on the in vitro gas kinetics and production. Cuban Journal of Agricultural Science, 43(2), 143-146. Recovered from https://www.researchgate.net/publication/292840960_Effect_of_the_combination_of_Tithonia_diversifolia_with_Pennisetum_purpureum_cv_Cuba_CT-115_on_the_in_vitro_gas_kinetics_and_production
https://www.researchgate.net/publication...
), just a few silage production studies have been conducted (Holguín, Ortíz Grisalez, Velasco Navia, & Mora-Delgado 2015Holguín, V. A., Ortíz Grisalez, S., Velasco Navia, A., & Mora-Delgado, J. (2015). Multi-criteria evaluation of 44 introductions of Tithonia diversifolia (Hemsl.) A. Gray in Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
https://doi.org/10.15446/rfmvz.v62n2.519...
); therefore, it is necessary to determine the extent of incorporation of this plant and the possibility of blending it with grass to obtain the maximum benefit for animal nutrition and farmer’s households.

On the other hand, in tropical areas, seasonal forage production has been a problem due to food shortage at certain times of the year associated with drought, and in other times by an overproduction caused to high rainfall (Miguel, Delagarde, & Ribeiro-Filho, 2019Miguel, M. F., Delagarde, R., & Ribeiro-Filho, H. M. N. (2019). Corn silage supplementation for dairy cows grazing annual ryegrass at two pasture allowances. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 71(3), 1037-1046. doi: 10.1590/1678-4162-9795
https://doi.org/10.1590/1678-4162-9795...
). Seen this way, the challenge is how to conserve fodder to defer its use between times of high production and shortages without losing quality; silage is an alternative, different studies (Florez Delgado, Capacho Mogollon, Quintero Muino, & Gamboa Vera, 2018Florez Delgado, D. F., Capacho Mogollon, A. E., Quintero Muino, S. M., & Gamboa Vera, K. Y. (2018). Effect of supplementation with orange silage on the caprine milk quality. Revista U.D.C.A Actualidad & Divulgación Científica, 21(2), 501-506. doi: 10.31910/rudca.v21.n2.2018.982
https://doi.org/10.31910/rudca.v21.n2.20...
; Galina, Ortiz-Rubio, Mondragón, Delgado-Pertíñez, & Elías, 2009Galina, M. A., Ortiz-Rubio, M. A., Mondragón, F., Delgado-Pertíñez, M., & Elías, A. (2009). Rendimiento de terneros alimentados con silo de maíz o láctico con un promotor de la fermentación ruminal. Archivos de Zootecnia, 58(223), 383-393. Recovered from https://www.redalyc.org/pdf/495/49515090007.pdf
https://www.redalyc.org/pdf/495/49515090...
) have shown silage of other non-legumes plants as protein sources. Therefore, it is necessary to investigate how silages with different proportions of grasses and protein forages could be an efficient alternative for production, and environmentally friendly (Steinfed et al., 2006Steinfed, H., Gerber, P., Wassenaar, T., Castel, V., Rosales, M., & De Haan, C. (2006). Livestock´s long shadow: environmental issues and options. Rome, IT: FAO.). The present study aims to estimate in vitro ruminal gas production (RGP) and determine the CH4 emissions from silages used as diets for lambs. This was done through an experiment, based on the evaluation of four diets, prepared with a grass mixed with a high protein plant. Consequently, our goal is to produce a strategic knowledge for the small-ruminant farmers. The main result suggested that the lower cumulative CH4 production was for those diets in which the T. diversifolia was included.

Material and methods

Silage preparation

The forage used for silage production was harvested at the Experimental Center of the National University of Colombia - Palmira Headquarters (CEUNP) in the Department of Valle del Cauca. This area is located at an altitude of 1,000 meters above the sea level; 02°06’ N and 65°03’ W; it presents an annual average rainfall of 1,000 mm and an average temperature of 24°C. This area has been classified as a tropical dry forest (Holdridge, 1987Holdridge, L. R. (1987). Ecología basada en zonas de vida (n. 83). San José, CR: Instituto Interamericano de Ciencias Agricoles.). The forage of Cenchrus purpureus and Tithonia diversifolia was harvested at 60 and 90 days, respectively. The forage was pre-dried for 24 hours; then, it was chopped to reduce the particle size to 2 cm, in a three-blade mill, 7.5 HP, 1400 rpm, and 4.5 Amps Gaitan Brand. Once the forage was chopped separately for each species, a manual mixture was made. Simultaneously, using a hand pump, the forage was sprayed in layers with an inoculum based on Lactobacillus paracasei bacteria (T735), it was obtained from macerated leaves of T. diversifolia, following the protocol explained by Holguín, Grisalez, Huertas, Fandiño and Delgado (2018Holguín, V., Grisalez, S. O., Huertas, A., Fandiño, L. C., & Delgado, J. R. M. (2018). Ganancia de peso en ovinos alimentados con un ensilaje de Pennisetum purpureun y Tithonia diversifolia. RIAA, 9(2), 1. Recovered from https://dialnet.unirioja.es/servlet/articulo?codigo=6535138
https://dialnet.unirioja.es/servlet/arti...
), developed in the Diagnostic Laboratory Veterinarian of the University of Tolima. The concentration of the inoculum used was 30 x 107 CFU mL-1. The silages were prepared using a silage bag packing machine and then they were stored in vacuum-sealed bags, as follows: Treatment 1 = 100% Cenchrus purpureus silage, Treatment 2 = Cenchrus purpureus silage in mixture with Tithonia diversifolia, with a proportion 33 and 67%, Treatment 3 = Cenchrus purpureus silage in mixture with Tithonia diversifolia, with a proportion 67 and 33%, respectively, and Treatment 4 = 100% Tithonia diversifolia silage. The silages were stored for 67 days.

Analytical phase

The bromatological analysis of the silages was carried out in the laboratory of Animal Ecophysiology at the University of Tolima, where proximal chemical analyses were performed. We followed the methods established by the Association of Official Analytical Chemists (AOAC, 1990Association of Official Analytical Chemists [AOAC]. (1990). Official Methods of Analysis (15th ed.). Arlington, VA: AOAC.) (Horwitz, Latimer, & AOAC International, 2010Horwitz, W., Latimer, G. W., & AOAC International . (2010). Official methods of analysis of AOAC International. Gaithersburg, MD: AOAC International.) for dry matter (DM), organic matter (OM), crude protein CP), ether extract (EE) and ash content (AC). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined by the protocol of Van Soest, Robertson and Lewis (1991Van Soest, P. V., 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(10), 3583-3597. doi: 10.3168/jds.S0022-0302(91)78551-2
https://doi.org/10.3168/jds.S0022-0302(9...
). After that, the in vitro laboratory phase was conducted at the Agrosavia Nutrition Laboratory in Mosquera-Cundinamarca.

In vitro incubation

The in vitro incubation procedure described by Schofield, Pell and Pitt (1994Schofield, P., Pitt, R. E., & Pell, A. N. (1994). Kinetics of fiber digestion from in vitro gas production. Journal of Animal Science, 72(11), 2980-2991. doi: 10.2527/1994.72112980x
https://doi.org/10.2527/1994.72112980x...
) was used. The ruminal liquor was obtained on an empty stomach of a fistulated sheep. The sheep were fed with kikuyo grass. The ruminal liquor was filtered using four layers of gauze and it was CO2-gassing constantly. Then, 0.6 g of silage were weighed, and they were introduced into 60 mL-bottles, provided with butyl rubber stoppers and staples; then, 8 mL of buffer solution (pH 6.5) (Van Soest et al., 1991Van Soest, P. V., 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(10), 3583-3597. doi: 10.3168/jds.S0022-0302(91)78551-2
https://doi.org/10.3168/jds.S0022-0302(9...
) and 2 mL of ruminal fluid were added to each bottle, maintaining a continuous gassing with CO2. The four treatments and a control group, each with four repetitions, were incubated for 48 hours under a temperature of 37°C. Thus, a total amount of 20 bottles were incubated, 16 containing substrate and inoculum (4 treatments* 4 repetitions) and 4 corresponding to the control group, whose function was to correct the production of gas generated by the microorganisms.

In vitro digestibility of dry matter

The in vitro dry matter digestibility (INDDM) was determined at 48 hours of incubation. To do this, the silage content of each bottle was poured into a 50 mL tubes-Falcon brand, previously identified, and weighed. The not digestible (NDM) was determined by drying the filtered material at 60°C for 48 hours; consequently, digestible (DDM) was determined by the difference.

Gas production

Following the methodology of Theodorou, Williams, Dhanoa, McAllan and France (1994Theodorou, M. K., Williams, B. A., Dhanoa, M. S., McAllan, A. B., & France, J. (1994). A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology, 48(3-4), 185-197. doi: 10.1016/0377-8401(94)90171-6
https://doi.org/10.1016/0377-8401(94)901...
), the gas production generated by enteric fermentation was quantified for each treatment at 2, 4, 8, 12, 18, 24, 30, 36, and 48 hours. The gas quantification was made using a digital transducer (ASHCROFT®) which measures the amount of gas according to the pressure accumulated in the bottle. Thus, the total gas production was determined as the sum of the partial productions in the sampling hours. A sample of gas obtained at each hour of measurement was stored in vacutainer tubes (BD of 7 mL) under vacuum for subsequent determination of methane concentration. To convert the pressure (PSI) data into gas volume (mL), we use the equation:

Y = -0.1375+ (5.385 * X) + (0.0777 * X2),

where: X is the pressure in PSI and Y is the gas volume in mL (Posada, Solano, & Vergara, 2006Posada, S. L., Solano, R. N., & Vergara, D. M. B. (2006). Relación entre presión y volumen para la implementación de la técnica in vitro de producción de gases en Medellín, Colombia. Revista Colombiana de Ciencias Pecuarias, 19(4), 407-414. Recovered from https://dialnet.unirioja.es/servlet/articulo?codigo=3239461
https://dialnet.unirioja.es/servlet/arti...
); gas production was expressed per gram of incubated dry matter (mL g-1 of DM).

Methane production

A sample of gas was taken from each treatment at each hour of measurement, which were stored in vacutainers (BD of 7 mL), under vacuum; subsequently, methane concentration was determined using a methane gas laser sight (CROWCON®).

Modeling methane production

To estimate the parameters of the fermentation kinetics, the cumulative methane gas production of the best treatment was adjusted to the models in Table 1. The adjustment of the data to each model and the parameter estimation was performed using non-linear models through INFOSTAT® software version 2018 (Di Rienzo et al., 2008Di Rienzo, J. A., Casanoves, F., Balzarini, M. G., Gonzalez, L., Tablada, M., & Robledo, C. W. (2008). InfoStat, versión 2008. Córdoba, AG: Grupo InfoStat, FCA, Universidad Nacional de Córdoba.). Thus, the model that best represents the methane production kinetics was selected,the best values in the goodness-of-fit criteria BIC and AIC.

Statistical analysis

The statistical analysis for the dependent variables was done using a completely randomized design in each case. The dependent variables were: Cumulative gas production (mL); methane production per gram of dry matter (ppm); parameters obtained from the most adjusted model, and in vitro digestibility of DM (%). The first two variables were analyzed using repeated measures over time, using mixed models and heterogeneous variances. The linear model for the observations of this experiment is as follows:

Y i j k = μ + τ i + β j + ( τ β ) i j + s k + ε i j k

Yijk: represents the production of methane observed in the ith level of factor Silage and jth level of the factor Time for the kth subject; (: represents the general average of the observed variable; (i: represents the effect of the ith level of silage factor; (j: represents the effect of the jth level of the Time factor; (((ij ): represents the interaction effect corresponding to the Silage and Time factor; sk: represents the random effect corresponding to the k-th subject, where sk(N(0, (s 2); ( ijk: represents the random error where ( ijk (N(0, (( 2). It is also assumed that the two random terms s k and ( ijk are independent. The comparison of means was made using the Fisher LSD test. For the statistical analyzes, the INFOSTAT® software, version 2018 (Di Rienzo et al., 2008Di Rienzo, J. A., Casanoves, F., Balzarini, M. G., Gonzalez, L., Tablada, M., & Robledo, C. W. (2008). InfoStat, versión 2008. Córdoba, AG: Grupo InfoStat, FCA, Universidad Nacional de Córdoba.) was used.

Table 1
Models used to model the in vitro gas kinetics of Cenchrus purpureus silages in admixture with Tithonia diversifolia.

Results

Bromatological analyses

The bromatological analysis (Table 2) shows higher CP and DM contents in the treatments in which the T. diversifolia was included in silage. On the contrary, ashes, NDF, and ADF were higher as the proportion of C. purpureus increases in the silage, the above related to a lower digestibility, and greater emission of methane (Barbosa et al., 2018Barbosa, A. L., Voltolini, T. V., Menezes, D. R., Moraes, S. A., Nascimento, J. C. S., & Rodrigues, R. T. S. (2018). Intake, digestibility, growth performance, and enteric methane emission of Brazilian semiarid non-descript breed goats fed diets with different forage to concentrate ratios. Tropical Animal Health and Production, 50(2), 283-289. doi: 10.1007/s11250-017-1427-0
https://doi.org/10.1007/s11250-017-1427-...
).

Table 2
Bromatological analysis of Cenchrus purpureus silages in mixture with Tithonia diversifolia.

In vitro digestibility of dry matter

In Table 3 it is possible to notice the in vitro digestibility of dry matter (IVDDM) at 48 hours of incubation. No statistical significance was found among the treatments studied. The positive interaction effect was confirmed in the treatment with 100% Tithonia diversifolia inoculated with L. paracasei which resulted in the best IVDDM (p < 0.0001).

Table 3
In vitro digestibility of dry matter from the vegetable silages of Cenchrus purpureus in mixture with Tithonia diversifolia.

Gas production

The silages showed increases in gas production over time, with a significant effect for the interaction treatments and incubation time (p = 0.0008). The gas accumulation for the T1 samples is greater from hour 2 (12.44 mL g DM-1), with no significant effect with T4 (Table 4). However, as of hour 8, a significant effect (p = 0.043) was observed in the gas production for T1 to the other treatments. Treatments 2, 3, and 4 show a similar behavior from hour 2 to hour 24, with a decrease in the gas production of T4 without significant differences between these treatments, but with T1 (p = 0.032). T4 treatment obviously maintains a decrease for the other treatments until 48 hours.

Methane production

Silages showed increases in methane production over time; a significant effect was observed for the interaction between treatments and hours (p = 0.0001). Thus, the accumulated methane production for T1 is greater since hour 2, showing a significant effect concerning the other treatments (p = 0.0005). This trend remains constant until hour 36; at that time, T3 also shows an increase concerning T1, with significant effects (p = 0.008) with respect to T2 and T4. For T2, methane production begins to decrease after 24 hours; T4 maintained a low methane production compared to T1 and T3, experiencing a slight increase at the hour 24. It was determined that the best treatments were T4 and T2, given the lower cumulative methane production. Therefore, we proceeded to perform the modeling of the kinetics of methane production with T2 treatment.

Modeling of methane production kinetics

The Mitscherlich and Gompertz models had the best goodness of fit, but between these two the second is the one that best predicts biologically the production of methane gas. That best prediction availability was verified by the CME, AIC, and BIC. In Table 6, the equations that represent the potential of in vitro gas production of T. diversifolia for the different models are shown. The fermentation potential of the substrate under the incubation conditions (asymptote of the curve) in the Gompertz model corresponds to 311.97 mL gr-1 of DM (parameter α), with a latency or fermentation delay phase of 2, 85 h (parameter β) and a fermentation rate of 008 mL h-1 (parameter γ).

Also, in Figure 1 the equation of greater adjustment is described, showing an increase over time in gas production, a trend that is interpreted by France, Dijkstra, Dhanoa, Lopez and Bannink (2000France, J., Dijkstra, J., Dhanoa, M. S., Lopez, S., & Bannink, A. (2000). Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations. British Journal of Nutrition, 83(2), 143-150. doi : 10.1017/S0007114500000180
https://doi.org/10.1017/S000711450000018...
) as an increase in microbial activity, although this does not imply any assumption about the constancy of microbial growth performance.

The gas higher production was observed in the treatment 4 (100% T. diversifolia), compared to silages with lower inclusion of this species (T2 and T3) or in all-grass silage (T1).

Table 4
Production of in vitro gas accumulated (mL g-1 DM) in the silage mixture of T. diversifolia (T) / C.purpureum (P).
Table 5
Production of in vitro gas methane accumulated (ppm g-1 DM) in the silage mixture of T. diversifolia (T) / C.purpureum (P).
Table 6
Statistical equations and estimators of the modeling of methane gas production kinetics for Cenchrus purpureus silages in admixture with Tithonia diversifolia.

Figure 1.
Modeling the in vitro production of methane accumulated (mL g-1 DM) in the silage mixture of T. diversifolia (T) / C. purpureus (C).

Discussion

The present study provides information about the biochemical characteristics of silages prepared with T. diversifolia, which allows us to appreciate the use of this species in the feeding of ruminant animals.

In short, it was identified that the treatments in which the T. diversifolia was included to silage, showed better nutritional characteristics compared with treatments without the inclusion of the species. Similar results to those found in this study were described by Donney’s et al. (2015Donney’s, G., Molina, I. C., Rivera, J. E., Villegas, G., Chará, J., & Barahona, R. (2015). Producción in vitro de metano de dietas ofrecidas en sistemas silvopastoriles intensivos con Tithonia diversifolia y sistemas tradicionales. In 3 Congreso Nacional de Sistemas Silvopastoriles y VIII Congreso Internacional de Sistemas Agroforestales INTA (p. 7-9). Puerto Iguazú, Argentina.). Also, La O et al. (2012La O, O., González, H., Orozco, A., Castillo, Y., Ruiz, O., Estrada, A., & Castro, B. I. (2012). Composición química, degradabilidad ruminal in situ y digestibilidad in vitro de ecotipos de Tithonia diversifolia de interés para la alimentación de rumiantes. Revista Cubana de Ciencia Agrícola, 46(1), 47-53. ) reported a protein content between 18.26 and 26.40% in different ecotypes of this species, and Mahecha, Escobar, Suárez, and Restrepo (2007Mahecha, L., Escobar, J. P., Suárez, J. F., & Restrepo, L. F. (2007). Tithonia diversifolia (hemsl.) Gray (botón de oro) como suplemento forrajero de vacas F1 (Holstein por Cebú). Livestock Research for Rural Development, 19(2), 1-6. Recovered from http://www.lrrd.org/lrrd19/2/mahe19016.htm
http://www.lrrd.org/lrrd19/2/mahe19016.h...
), found contents of 16.73% protein.

Despite not finding statistical significance among the treatments for the IDIVDM variable, we were able to verify that the treatments in which the species T. diversifolia was included show higher values of digestibility. However, Naranjo and Cuartas (2011Naranjo, J. F., & Cuartas, C. A. (2011). Caracterización nutricional y de la cinética de degradación ruminal de algunos de los recursos forrajeros con potencial para la suplementación de rumiantes en el trópico alto de Colombia. Revista CES Medicina Veterinaria y Zootecnia, 6(1), 9-19. Recovered from https://dialnet.unirioja.es/servlet/articulo;jsessionid=ECA2D78337903C8AD48A2D75E69BBA2E.dialnet01?codigo=3697941
https://dialnet.unirioja.es/servlet/arti...
) reported similar values but with significant differences depending on the ADF contents since as it is known, the digestibility of the forages is inversely related to the content of these fibers, which also depends on the internal composition and its structure (Moreira, Leonel, Vieira, & Pereira, 2013Moreira, L. M., Leonel, F. D. P., Vieira, R. A. M., & Pereira, J. C. (2013). A new approach about the digestion of fibers by ruminants. Revista Brasileira de Saúde e Produção Animal,14(2), 382-395. doi: 10.1590/S1519-99402013000200008
https://doi.org/10.1590/S1519-9940201300...
). In this regard, Holguín et al. (2015Holguín, V. A., Ortíz Grisalez, S., Velasco Navia, A., & Mora-Delgado, J. (2015). Multi-criteria evaluation of 44 introductions of Tithonia diversifolia (Hemsl.) A. Gray in Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
https://doi.org/10.15446/rfmvz.v62n2.519...
) found that the application of an inoculum based on Lactobacillus paracasei resulted in better digestibility compared to non-inoculated silage but was similar to silage inoculated with SilAll (p = 0.0060).

Also, in the present study (Table 4), we observed that the treatment 4 (100% T. diversifolia) maintains a decrease in gas production concerning other treatments until the last measurement hour, the above is directly related to the nutritional composition of the silage, it means that forages with a lower content of fibrous carbohydrates and a greater amount of soluble carbohydrates produce less gas (Rivera et at., 2015Rivera, J. E., Molina, I. C., Donneys, G., Villegas, G., Chará, J., & Barahona, R. (2015). Dinámicas de fermentación y producción in vitro de metano en dietas de sistemas silvopastoriles intensivos con L. leucocephala y sistemas convencionales orientados a la producción de leche. Livestock Research for Rural Development, 27(4), 1-15. Recovered from https://www.researchgate.net/publication/274314837_Dinamica_de_fermentacion_y_produccion_de_metano_en_dietas_de_sistemas_silvopastoriles_intensivos_con_L_leucocephala_y_sistemas_convencionales_orientados_a_la_produccion_de_leche
https://www.researchgate.net/publication...
; Kulivand & Kafilzadeh, 2015Kulivand, M., & Kafilzadeh, F. (2015). Correlation between chemical composition, kinetics of fermentation and methane production of eight pasture grasses. Acta Scientiarum. Animal Sciences,37(1), 9-14. doi: 10.4025/actascianimsci.v37i1.24336
https://doi.org/10.4025/actascianimsci.v...
). The study conducted by Molina Botero, Cantet, Montoya, Correa Londoño and Barahona Rosales (2013Molina Botero, I. C., Cantet, J. M., Montoya, S., Correa Londoño, G. A., & Barahona Rosales, R. (2013). In vitro methane production from two tropical grasses alone or in combination with Leucaena leucocephala or Gliricidia sepium. CES Medicina Veterinaria y Zootecnia, 8(2), 15-31. Recovered from http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S1900-96072013000200002
http://www.scielo.org.co/scielo.php?scri...
) with the species Leucaena leucocephala and Gliricidia sepium blended with Megathyrsus maximus and Dichantium aristatum grasses, reached 48 hours of maximum gas production (around 115 mL of gas g-1 DM); similar values, although higher, were found in our experiment.

On the other hand, we found that the accumulated methane production for T1 is greater since hour 2, showing a significant effect concerning the other treatments, a trend that remains constant until hour 36. For treatment 2, the methane production begins to decrease after 24 hours, which is because the gas and methane production of soluble fractions of high-quality forages may be higher during the first hours of fermentation (Freire et al., 2017Freire, J. M., Silva, A. M. D. A., Carneiro, H., Pereira Filho, J. M., Rocha, L. B., & Bidler, D. C. (2017). In vitro degradation and gas production of brachiaria grass with levels of biodiesel byproducts. Acta Scientiarum. Animal Sciences, 39(2), 175-179. doi: 10.4025/actascianimsci.v39i2.32832
https://doi.org/10.4025/actascianimsci.v...
); while silages prepared with greater inclusion of the species T. diversifolia maintained a low methane production. However, from the animal production point of view, it is not convenient to produce silage based on a 100% protein source, given the problems in the low acidification of the medium in the silo and the metabolic problems that could be caused in the animal at the rumen level. Therefore, it is suggested that silage based on a mixture of T. diversifolia and grass is the one that could be recommended for use as a supplement, especially in times of scarcity due to the climatic seasonality of tropical areas.

The gas higher production was observed in the treatment 4 (100% T. diversifolia), compared to silages with lower inclusion of this species. The authors suggest that this was due to the presence of secondary metabolites in the species T. diversifolia such as condensed tannins and saponins (Noguera, Saliba, & Mauricio, 2004Noguera, R. R., Saliba, E. O., & Mauricio, R. M. (2004). Comparación de modelos matemáticos para estimar los parámetros de degradación obtenidos a través de la técnica de producción de gas. Livestock Research for Rural Development, 16(11). Recovered from https://www.lrrd.org/lrrd16/11/nogu16086.htm
https://www.lrrd.org/lrrd16/11/nogu16086...
). This follows a greater digestibility of the silage with a greater proportion of T. diversifolia (Holguín et al., 2015Holguín, V. A., Ortíz Grisalez, S., Velasco Navia, A., & Mora-Delgado, J. (2015). Multi-criteria evaluation of 44 introductions of Tithonia diversifolia (Hemsl.) A. Gray in Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
https://doi.org/10.15446/rfmvz.v62n2.519...
). On the other hand, La et al. (2009La, O., Valenciaga, D., Gonzalez, H., Orozco, A., Castillo, Y., Ruiz, O., ... Arzola, C. (2009). Effect of the combination of Tithonia diversifolia with Pennisetum purpureum cv. Cuba CT-115 on the in vitro gas kinetics and production. Cuban Journal of Agricultural Science, 43(2), 143-146. Recovered from https://www.researchgate.net/publication/292840960_Effect_of_the_combination_of_Tithonia_diversifolia_with_Pennisetum_purpureum_cv_Cuba_CT-115_on_the_in_vitro_gas_kinetics_and_production
https://www.researchgate.net/publication...
) explain that these high values in the gas production in silage with the greater inclusion of T. diversifolia may be due to the concentration of easily fermentable carbohydrates. It also becomes clear that the optimization of microbial fermentation occurs in the presence of this protein forage in the incubation medium. In a recent experiment, Terry et al. (2016Terry, S. A., Ribeiro, R. S., Freitas, D. S., Delarota, G. D., Pereira, L. G. R., & Tomich, T. R., … Chaves, A. V. (2016). Effects of Tithonia diversifolia on in vitro methane production and ruminal fermentation characteristics. Animal Production Science, 56(3), 437-441. doi: 10.1071/AN15560
https://doi.org/10.1071/AN15560...
) demonstrated that in vitro VFA concentration increased when T. diversifolia was supplemented at 15.2% DM replacing fresh sugarcane and concentrates; also, they reported an increase in CH4 production with increasing concentrations of T. diversifolia. However, in the in vivo experiment, there was no effect (p= 0.82) of the inclusion of T. diversifolia on total VFA and as such, no increase in production parameters. Delgado et al. (2012Delgado, D. C., Galindo, J., González, R., González, N., Scull, I., & Dihigo, L. (2012) Feeding of tropical trees and shrub foliages as a strategy to reduce ruminal methanogenesis: studies conducted in Cuba. Tropical Animal Health and Production, 44, 1097-1104. doi: 10.1007/s11250-011-0045-5
https://doi.org/10.1007/s11250-011-0045-...
) found that T. diversifolia had methane reducing properties when supplemented at 30% into a star grass (Cynodon nlemfuensis) based diet.

Conclusion

The inclusion of T. diversifolia in the silage improves the quality of a grass-based diet, due to its high protein content, high ruminal degradability, and low fiber content. Besides, the lower production of methane in silage mixtures containing T. diversifolia, represents a lower energy loss and therefore greater production of volatile fatty acids.

Considering the fact that in the current study, the presence of secondary metabolites, which presents as a difficulty when assessing their effects on CH4 production, was not evaluated, we recommend doing this study to better understand that component of T. diversifolia in the CH4 production at the ruminal level.

Acknowledgements

We would like to thank the Colombian Agricultural Research Corporation AGROSAVIA Tibaitatá Research Center, for facilitating the spaces for the development of this experiment; to the Ecophysiology Laboratory and the Animal Nutrition Laboratory of the University of Tolima for allowing the use of laboratory equipment and analysis and to the Central Office of Research and Scientific Development of the University of Tolima for the financial support of this study

References

  • Association of Official Analytical Chemists [AOAC]. (1990). Official Methods of Analysis (15th ed.). Arlington, VA: AOAC.
  • Barbosa, A. L., Voltolini, T. V., Menezes, D. R., Moraes, S. A., Nascimento, J. C. S., & Rodrigues, R. T. S. (2018). Intake, digestibility, growth performance, and enteric methane emission of Brazilian semiarid non-descript breed goats fed diets with different forage to concentrate ratios. Tropical Animal Health and Production, 50(2), 283-289. doi: 10.1007/s11250-017-1427-0
    » https://doi.org/10.1007/s11250-017-1427-0
  • Bettencourt, E. M. V., Tilman, M., Narciso, V., Carvalho, M. L. D. S., & Henriques, P. D. D. S. (2015). The livestock roles in the wellbeing of rural communities of Timor-Leste. Revista de Economia e Sociologia Rural, 53(1), 63-80. doi: 10.1590/1234-56781806-94790053s01005
    » https://doi.org/10.1590/1234-56781806-94790053s01005
  • Cárdenas, J. A. B., & Flores, C. L. (2012). Emisión de metano entérico por rumiantes y su contribución al calentamiento global y al cambio climático. Revisión. Revista Mexicana de Ciencias Pecuarias, 3(2), 215-246. Recovered from https://www.researchgate.net/publication/236158591_Emision_de_metano_enterico_por_rumiantes_y_su_contribucion_al_calentamiento_global_y_al_cambio_climatico_Revision
    » https://www.researchgate.net/publication/236158591_Emision_de_metano_enterico_por_rumiantes_y_su_contribucion_al_calentamiento_global_y_al_cambio_climatico_Revision
  • Castaño, V. H., Grisalez, S. O., Navia, A. V., & Delgado, J. R. M. (2015). Evaluación multicriterio de 44 introducciones de Tithonia diversifolia (hemsl.) Gray en Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
    » https://doi.org/10.15446/rfmvz.v62n2.5199
  • Delgado, D. C., Galindo, J., González, R., González, N., Scull, I., & Dihigo, L. (2012) Feeding of tropical trees and shrub foliages as a strategy to reduce ruminal methanogenesis: studies conducted in Cuba. Tropical Animal Health and Production, 44, 1097-1104. doi: 10.1007/s11250-011-0045-5
    » https://doi.org/10.1007/s11250-011-0045-5
  • Di Rienzo, J. A., Casanoves, F., Balzarini, M. G., Gonzalez, L., Tablada, M., & Robledo, C. W. (2008). InfoStat, versión 2008. Córdoba, AG: Grupo InfoStat, FCA, Universidad Nacional de Córdoba.
  • Donney’s, G., Molina, I. C., Rivera, J. E., Villegas, G., Chará, J., & Barahona, R. (2015). Producción in vitro de metano de dietas ofrecidas en sistemas silvopastoriles intensivos con Tithonia diversifolia y sistemas tradicionales. In 3 Congreso Nacional de Sistemas Silvopastoriles y VIII Congreso Internacional de Sistemas Agroforestales INTA (p. 7-9). Puerto Iguazú, Argentina.
  • Florez Delgado, D. F., Capacho Mogollon, A. E., Quintero Muino, S. M., & Gamboa Vera, K. Y. (2018). Effect of supplementation with orange silage on the caprine milk quality. Revista U.D.C.A Actualidad & Divulgación Científica, 21(2), 501-506. doi: 10.31910/rudca.v21.n2.2018.982
    » https://doi.org/10.31910/rudca.v21.n2.2018.982
  • Forero-Álvarez, J. (2013). The economy of family farming production. Cuadernos de Desarrollo Rural, 10(spe70), 27-45. Recovered from http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0122-14502013000100002
    » http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0122-14502013000100002
  • France, J., Dijkstra, J., Dhanoa, M. S., Lopez, S., & Bannink, A. (2000). Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations. British Journal of Nutrition, 83(2), 143-150. doi : 10.1017/S0007114500000180
    » https://doi.org/10.1017/S0007114500000180
  • Freire, J. M., Silva, A. M. D. A., Carneiro, H., Pereira Filho, J. M., Rocha, L. B., & Bidler, D. C. (2017). In vitro degradation and gas production of brachiaria grass with levels of biodiesel byproducts. Acta Scientiarum. Animal Sciences, 39(2), 175-179. doi: 10.4025/actascianimsci.v39i2.32832
    » https://doi.org/10.4025/actascianimsci.v39i2.32832
  • Friedrich, T. (2014). Producción de alimentos de origen animal. Actualidad y perspectivas. Revista Cubana de Ciencia Agrícola, 48(1), 5-6. Recovered from https://www.redalyc.org/pdf/1930/193030122003.pdf
    » https://www.redalyc.org/pdf/1930/193030122003.pdf
  • Galina, M. A., Ortiz-Rubio, M. A., Mondragón, F., Delgado-Pertíñez, M., & Elías, A. (2009). Rendimiento de terneros alimentados con silo de maíz o láctico con un promotor de la fermentación ruminal. Archivos de Zootecnia, 58(223), 383-393. Recovered from https://www.redalyc.org/pdf/495/49515090007.pdf
    » https://www.redalyc.org/pdf/495/49515090007.pdf
  • Galindo, J., Gonzáluez, N., Sosa, A., Ruíz, T., Torres, V., Aldana, A. I., ... Noda, A. C. (2011). Efecto de Tithonia diversifolia (Hemsl.) Gray (Botón de oro) en la población de protozoos y metanógenos ruminales en condiciones in vitro. Revista Cubana de Ciencia Agrícola, 45(1), 33-37. Recovered from https://www.semanticscholar.org/paper/Efecto-de-Tithonia-diversifolia-%28Hemsl.%29-Gray-de-en-Galindo-Gonz%C3%A1lez/b147f9ba2a5edc5566028c62b89bbac97fb7fdf9?p2df
    » https://www.semanticscholar.org/paper/Efecto-de-Tithonia-diversifolia-%28Hemsl.%29-Gray-de-en-Galindo-Gonz%C3%A1lez/b147f9ba2a5edc5566028c62b89bbac97fb7fdf9?p2df
  • Holdridge, L. R. (1987). Ecología basada en zonas de vida (n. 83). San José, CR: Instituto Interamericano de Ciencias Agricoles.
  • Holguín, V. A., Ortíz Grisalez, S., Velasco Navia, A., & Mora-Delgado, J. (2015). Multi-criteria evaluation of 44 introductions of Tithonia diversifolia (Hemsl.) A. Gray in Candelaria, Valle del Cauca. Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 62(2), 57-72. doi: 10.15446/rfmvz.v62n2.5199
    » https://doi.org/10.15446/rfmvz.v62n2.5199
  • Holguín, V., Grisalez, S. O., Huertas, A., Fandiño, L. C., & Delgado, J. R. M. (2018). Ganancia de peso en ovinos alimentados con un ensilaje de Pennisetum purpureun y Tithonia diversifolia RIAA, 9(2), 1. Recovered from https://dialnet.unirioja.es/servlet/articulo?codigo=6535138
    » https://dialnet.unirioja.es/servlet/articulo?codigo=6535138
  • Horwitz, W., Latimer, G. W., & AOAC International . (2010). Official methods of analysis of AOAC International Gaithersburg, MD: AOAC International.
  • Hurst, P., Termine, P., & Karl, M. (2007). Agricultural workers and their contribution to sustainable agriculture and rural development Geneve, CH: FAO. Recovered from http://www.fao.org/3/a-bp976e.pdf
    » http://www.fao.org/3/a-bp976e.pdf
  • Kulivand, M., & Kafilzadeh, F. (2015). Correlation between chemical composition, kinetics of fermentation and methane production of eight pasture grasses. Acta Scientiarum. Animal Sciences,37(1), 9-14. doi: 10.4025/actascianimsci.v37i1.24336
    » https://doi.org/10.4025/actascianimsci.v37i1.24336
  • La, O., Valenciaga, D., Gonzalez, H., Orozco, A., Castillo, Y., Ruiz, O., ... Arzola, C. (2009). Effect of the combination of Tithonia diversifolia with Pennisetum purpureum cv. Cuba CT-115 on the in vitro gas kinetics and production. Cuban Journal of Agricultural Science, 43(2), 143-146. Recovered from https://www.researchgate.net/publication/292840960_Effect_of_the_combination_of_Tithonia_diversifolia_with_Pennisetum_purpureum_cv_Cuba_CT-115_on_the_in_vitro_gas_kinetics_and_production
    » https://www.researchgate.net/publication/292840960_Effect_of_the_combination_of_Tithonia_diversifolia_with_Pennisetum_purpureum_cv_Cuba_CT-115_on_the_in_vitro_gas_kinetics_and_production
  • La O, O., González, H., Orozco, A., Castillo, Y., Ruiz, O., Estrada, A., & Castro, B. I. (2012). Composición química, degradabilidad ruminal in situ y digestibilidad in vitro de ecotipos de Tithonia diversifolia de interés para la alimentación de rumiantes. Revista Cubana de Ciencia Agrícola, 46(1), 47-53.
  • Mahecha, L., Escobar, J. P., Suárez, J. F., & Restrepo, L. F. (2007). Tithonia diversifolia (hemsl.) Gray (botón de oro) como suplemento forrajero de vacas F1 (Holstein por Cebú). Livestock Research for Rural Development, 19(2), 1-6. Recovered from http://www.lrrd.org/lrrd19/2/mahe19016.htm
    » http://www.lrrd.org/lrrd19/2/mahe19016.htm
  • Mahecha-Ledesma, L., Angulo-Arizala, J., & Barragán-Hernández, W. (2017). Calidad nutricional, dinámica fermentativa y producción de metano de arreglos silvopastoriles. Agronomía Mesoamericana, 28(2), 371-387. doi: 10.15517/ma.v28i2.22750
    » https://doi.org/10.15517/ma.v28i2.22750
  • Maluf, R. S., Burlandy, L., Santarelli, M., Schottz, V., & Speranza, J. S. (2015). Nutrition-sensitive agriculture and the promotion of food and nutrition sovereignty and security in Brazil. Ciência & Saúde Coletiva, 20(8), 2303-2312. doi: 10.1590/1413-81232015208.14032014
    » https://doi.org/10.1590/1413-81232015208.14032014
  • Meza, G. A., Loor, N. J., Sánchez, A. R., Avellaneda, J. H., Meza, C. J., Vera, D. F., ... López, F. X. (2014). Inclusión de harinas de follajes arbóreos y arbustivos tropicales (Morus alba, Erythrina poeppigiana, Tithonia diversifolia e Hibiscus rosa-sinensis) en la alimentación de cuyes (Cavia porcellus Linnaeus). Revista de la Facultad de Medicina Veterinaria y de Zootecnia, 61(3), 258-269. Recovered from http://www.scielo.org.co/pdf/rfmvz/v61n3/v61n3a05.pdf
    » http://www.scielo.org.co/pdf/rfmvz/v61n3/v61n3a05.pdf
  • Miguel, M. F., Delagarde, R., & Ribeiro-Filho, H. M. N. (2019). Corn silage supplementation for dairy cows grazing annual ryegrass at two pasture allowances. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 71(3), 1037-1046. doi: 10.1590/1678-4162-9795
    » https://doi.org/10.1590/1678-4162-9795
  • Molina Botero, I. C., Cantet, J. M., Montoya, S., Correa Londoño, G. A., & Barahona Rosales, R. (2013). In vitro methane production from two tropical grasses alone or in combination with Leucaena leucocephala or Gliricidia sepium CES Medicina Veterinaria y Zootecnia, 8(2), 15-31. Recovered from http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S1900-96072013000200002
    » http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S1900-96072013000200002
  • Moreira, L. M., Leonel, F. D. P., Vieira, R. A. M., & Pereira, J. C. (2013). A new approach about the digestion of fibers by ruminants. Revista Brasileira de Saúde e Produção Animal,14(2), 382-395. doi: 10.1590/S1519-99402013000200008
    » https://doi.org/10.1590/S1519-99402013000200008
  • Naranjo, J. F., & Cuartas, C. A. (2011). Caracterización nutricional y de la cinética de degradación ruminal de algunos de los recursos forrajeros con potencial para la suplementación de rumiantes en el trópico alto de Colombia. Revista CES Medicina Veterinaria y Zootecnia, 6(1), 9-19. Recovered from https://dialnet.unirioja.es/servlet/articulo;jsessionid=ECA2D78337903C8AD48A2D75E69BBA2E.dialnet01?codigo=3697941
    » https://dialnet.unirioja.es/servlet/articulo;jsessionid=ECA2D78337903C8AD48A2D75E69BBA2E.dialnet01?codigo=3697941
  • Noguera, R. R., Saliba, E. O., & Mauricio, R. M. (2004). Comparación de modelos matemáticos para estimar los parámetros de degradación obtenidos a través de la técnica de producción de gas. Livestock Research for Rural Development, 16(11). Recovered from https://www.lrrd.org/lrrd16/11/nogu16086.htm
    » https://www.lrrd.org/lrrd16/11/nogu16086.htm
  • Olivo, M. L, & Soto-Olivo, A. (2010). Comportamiento de los gases de efecto invernadero y las temperaturas atmosféricas con sus escenarios de incremento potencial. Universidad, Ciencia y Tecnología, 14(57), 221-230. Recovered from http://ve.scielo.org/scielo.php?script=sci_arttext&pid=S1316-48212010000400002
    » http://ve.scielo.org/scielo.php?script=sci_arttext&pid=S1316-48212010000400002
  • Plata Pérez, L., González Ramírez, P. I., & Calderón Sánchez, E. (2017). La función de crecimiento de Richard y los modelos de crecimiento neoclásicos. EconoQuantum, 14(1), 99-118. doi: 10.18381/eq.v14i1.6543
    » https://doi.org/10.18381/eq.v14i1.6543
  • Posada, S. L., Solano, R. N., & Vergara, D. M. B. (2006). Relación entre presión y volumen para la implementación de la técnica in vitro de producción de gases en Medellín, Colombia. Revista Colombiana de Ciencias Pecuarias, 19(4), 407-414. Recovered from https://dialnet.unirioja.es/servlet/articulo?codigo=3239461
    » https://dialnet.unirioja.es/servlet/articulo?codigo=3239461
  • Ribeiro, R. S., Terry, S. A., Sacramento, J. P., Silveira, S. R., Bento, C. B., Silva, E. F., … Chaves, A. V. (2016). Tithonia diversifolia as a Supplementary Feed for Dairy Cows. PloS one, 11(12), e0165751. doi: 10.1371/journal.pone.0165751
    » https://doi.org/10.1371/journal.pone.0165751
  • Rivera, J. E., Naranjo, J. F., Cuartas, C. A., & Arenas, F. A. (2013). Fermentación in vitro y composición química de algunos forrajes y dietas ofrecidas bajo un Sistema Silvopastoril en el trópico de altura. Livestock Research for Rural Development, 25(10), 1-12. Recovered from https://www.researchgate.net/profile/Julian_Rivera2/publication/287056357_In_vitro_fermentation_and_chemical_composition_of_some_forages_and_diets_offered_to_cattle_in_a_tropical_Silvopastoral_System/links/56caed1308ae96cdd06f6a61.pdf
    » https://www.researchgate.net/profile/Julian_Rivera2/publication/287056357_In_vitro_fermentation_and_chemical_composition_of_some_forages_and_diets_offered_to_cattle_in_a_tropical_Silvopastoral_System/links/56caed1308ae96cdd06f6a61.pdf
  • Rivera, J. E., Molina, I. C., Donneys, G., Villegas, G., Chará, J., & Barahona, R. (2015). Dinámicas de fermentación y producción in vitro de metano en dietas de sistemas silvopastoriles intensivos con L. leucocephala y sistemas convencionales orientados a la producción de leche. Livestock Research for Rural Development, 27(4), 1-15. Recovered from https://www.researchgate.net/publication/274314837_Dinamica_de_fermentacion_y_produccion_de_metano_en_dietas_de_sistemas_silvopastoriles_intensivos_con_L_leucocephala_y_sistemas_convencionales_orientados_a_la_produccion_de_leche
    » https://www.researchgate.net/publication/274314837_Dinamica_de_fermentacion_y_produccion_de_metano_en_dietas_de_sistemas_silvopastoriles_intensivos_con_L_leucocephala_y_sistemas_convencionales_orientados_a_la_produccion_de_leche
  • Rojas-Downing, M., Nejadhashemi, A. P., Harrigan, T., & Woznicki, S. A. (2017). Climate change and livestock: impacts, adaptation, and mitigation. Climate Risk Management, 16, 145-163. doi: 10.1016/j.crm.2017.02.001
    » https://doi.org/10.1016/j.crm.2017.02.001
  • Rosegrant, M. W., Fernandez, M., & Sinha, A. (2009). Looking into the future for agriculture and AKST. In B. D. McIntyre, H. R. Herren, J. Wakhungu, R. T. Watson (Eds.), International Assessment of Agricultural Knowledge, Science and Technology for Development (IAASTD). Agriculture at a crossroads (p. 307-376). Washington, DC: Island Press.
  • Schofield, P., Pitt, R. E., & Pell, A. N. (1994). Kinetics of fiber digestion from in vitro gas production. Journal of Animal Science, 72(11), 2980-2991. doi: 10.2527/1994.72112980x
    » https://doi.org/10.2527/1994.72112980x
  • Steinfed, H., Gerber, P., Wassenaar, T., Castel, V., Rosales, M., & De Haan, C. (2006). Livestock´s long shadow: environmental issues and options Rome, IT: FAO.
  • Terry, S. A., Ribeiro, R. S., Freitas, D. S., Delarota, G. D., Pereira, L. G. R., & Tomich, T. R., … Chaves, A. V. (2016). Effects of Tithonia diversifolia on in vitro methane production and ruminal fermentation characteristics. Animal Production Science, 56(3), 437-441. doi: 10.1071/AN15560
    » https://doi.org/10.1071/AN15560
  • Theodorou, M. K., Williams, B. A., Dhanoa, M. S., McAllan, A. B., & France, J. (1994). A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology, 48(3-4), 185-197. doi: 10.1016/0377-8401(94)90171-6
    » https://doi.org/10.1016/0377-8401(94)90171-6
  • Van Soest, P. V., 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(10), 3583-3597. doi: 10.3168/jds.S0022-0302(91)78551-2
    » https://doi.org/10.3168/jds.S0022-0302(91)78551-2
  • Verdecia, D. M., Ramírez, J. L., Leonard, I., Álvarez, Y., Bazán, Y., Bodas, R., & López, S. (2011). Calidad de la Tithonia diversifolia en una zona del Valle del Cauto. REDVET. Revista electrónica de Veterinaria, 12(5), 1-13. (ID 63622168004)

Publication Dates

  • Publication in this collection
    30 Nov 2020
  • Date of issue
    2021

History

  • Received
    06 Dec 2019
  • Accepted
    07 July 2020
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