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Article

Rumen Fermentation Profile and Greenhouse Gas Mitigation of Three Forage Species from Agroforestry Systems in Dry and Rainy Seasons

by
Gary Meza-Bone
1,2,
Carlos Meza-Bone
1,
Juan Avellaneda-Cevallos
1,3,
Ronald Cabezas-Congo
1,
Ronald Villamar-Torres
1,2,
María Cabanilla-Campos
1,
Walter Vivas-Arturo
4,
Frank Intriago-Flor
4,
Fabricio Meza-Bone
5,
Aurelio Zapatier-Santillan
1,2,
Ricardo Bastidas-Espinoza
2,
Tito Solís-Barros
1,
Luz Muñoz-Arboleda
6,
Walter Garcia-Cox
5 and
Marcos Barros-Rodríguez
7,*
1
Carrera Agropecuaria, Facultad de Ciencias Pecuarias y Biológicas, Universidad Técnica Estatal de Quevedo, km 7 ½ vía Quevedo-El Empalme, Quevedo CP 120301, Los Ríos, Ecuador
2
Instituto Superior Tecnológico Ciudad de Valencia, km 3 1/2 vía a Valencia Sector El Pital 1, Quevedo CP 120301, Los Ríos, Ecuador
3
Programa de Pastos y Ganadería, Estación Experimental Tropical Pichilingue, Instituto Nacional de Investigaciones Agropecuarias-INIAP, km 7 vía Quevedo-El Empalme, Quevedo CP 120301, Los Ríos, Ecuador
4
Facultad de Ciencias Zootécnica, Universidad Técnica de Manabí, Sitio Las Animas km 2.5 vía Boyaca, Portoviejo 130709, Manabí, Ecuador
5
Carrera Forestal, Facultad de Ciencias Agrarias y Forestal, Universidad Técnica Estatal de Quevedo, km 7 ½ vía Quevedo-El Empalme, Quevedo CP 120301, Los Ríos, Ecuador
6
Facultad de Ciencias Agropecuarias, Universidad Nacional de Colombia, Sede Palmira, Palmira AA 237, Valle del Cauca, Colombia
7
Facultad de Ciencias Agropecuarias, Universidad Técnica de Ambato, Sector el Tambo-La Universidad, Vía a Quero, Cevallos CP 1801334, Tungurahua, Ecuador
*
Author to whom correspondence should be addressed.
Fermentation 2022, 8(11), 630; https://doi.org/10.3390/fermentation8110630
Submission received: 30 September 2022 / Revised: 28 October 2022 / Accepted: 8 November 2022 / Published: 11 November 2022
(This article belongs to the Special Issue Rumen Fermentation)
Versions Notes

Abstract

:
The variability of climatic conditions throughout the year and the physiological maturity of plant material can have effects on the nutritional value of plants and the productive performance of animals. The aim was to determine the effect of three tropical forage species (Tithonia diversifolia, Erythrina poeppigiana and Cratylia argentea) harvested at four cutting ages (30, 45, 60 and 75 days), during the dry and rainy seasons on in vitro gas production, rumen pH, volatile fatty acids (VFA) and CH4 production. A completely randomized design in a factorial arrangement (three forage species, four cutting ages and two seasons) was used. Four Brahman bulls of 250.0 ± 20.5 kg of live weight were used, provided with a ruminal cannula for the extraction of ruminal fluid. The animals were kept separately in individual pens, where they were fed with P. maximum and water was provided ad libitum. Gas production among forage species was lower (p = 0.0001) in T. diversifolia and E. poeppigiana in the dry season. The cutting age had an influence on gas production, showing lower production (p = 0.0164) at 30 and 45 days in the dry season. The VFA production showed differences for the effect between forages in the rainy season, with higher values (p < 0.05) for acetic, propionic and butyric acid in the species E. poeppigiana, C. argéntea and T. diversifolia respectively. CH4 production showed differences between forage species, showing the lowest (p = 0.0001) production of CH4 in T. diversifolia for the dry and rainy seasons. It is concluded that the use of shrubby species (T. diversifolia, E. poeppigiana and C. argéntea) in ruminant production systems in the rainy and dry seasons, with an early regrowth age (30 and 45 days), present a promising alternative on the reduction of CH4.

1. Introduction

Ruminant production systems in tropical, subtropical, semi-arid and arid zones around the world are restricted by the limited availability and low quality of forage resources throughout the year [1,2]. Forage resources characterized mainly by presenting variability in their nutritional components [3,4], probably influenced by: (a) variability of climatic conditions during the year (spring, summer, autumn and winter), (b) geographical area, (c) physiological development of the plant and (d) soil fertility [5]. The supply of poor-quality forage sources affects the productive performance of animals, by generating considerable energy losses that fluctuate between 2–15% of the energy available for production, as a consequence of the generation of greenhouse gases (GHG): CH4, CO2 and N2O, mainly in response to the presence of structural carbohydrates (cellulose and hemicellulose) in the feed [6,7]. In this way, it is estimated that livestock activity is responsible for generating approximately 14.5% of global GHG emissions of anthropogenic origin. Of which, it is estimated that livestock contributes with about 47% of CH4, 27% of CO2 and 29% of N2O [8,9,10].
The emission of GHG into the environment exacerbates the present problem of global warming. Therefore, there is currently a growing interest in reducing GHG emissions, mainly CH4 [11], due to its warming potential 28 times more than CO2 [12]. GHG production is an inevitable consequence of the anaerobic fermentation of feed in ruminants, observed with greater emphasis in animals that feed on low-quality forage (high in fiber), typical of semi-arid and arid areas where rain is scarce and solar radiation and environmental temperature is high [13]. An alternative to this problem is the use of tree or shrub legumes with forage potential, due to their ability to reduce GHG generation [14], in response to the presence of secondary metabolites (tannins, saponins, oils essential) that can modify the ruminal fermentation profile and reduce the production of CH4 [15,16]. However, the quality of the forage can vary according to the time of year, even among plants of the same species, probably due to a greater deposition of structural carbohydrates in the cell wall of the plants, due to the high solar radiation and lower humidity in the dry season, compared to the rainy season [17]. Aspects that limit the productive performance of animals and increase susceptibility to diseases in response to the decrease in the nutritional value of ingested forage, evidenced mainly by the low availability of protein, highly soluble carbohydrates and the reduction in the proportion of leaf area in the plant (ratio; leaves/stems) [18].
Many plants with forage potential for ruminant feeding have been undervalued, probably due to ignorance of their benefits and the variability of their nutritional value at different times of the year (spring, summer, autumn and winter) [19]. The use of these resources could favor the yield of forage biomass per unit of land used, unlike herbaceous forages that have a shallow root system and are affected in times of little rain, with delayed growth and decreased productivity. its nutritional value [20,21]. In this context, a notable variability has been evidenced on the protein content in Ceirchrus ciliaris L. influenced by the time of year (17.1% in the summer and 10.5% in the winter) [22]. In addition, forage trees and shrubs are usually rich in secondary compounds, mainly condensed tannins (CT) which, when combined with traditional grasslands, have the ability to decrease CH4 production [23]. Effects that are probably attributed to the decrease in methanogenic archaea and protozoa in the rumen in response to: (1) inhibition of H2 exchange between species (methanogen-protozoa) due to the effect of CT on the protein adhesin and subsequent change in the permeability of the cell membrane that affects the growth and activity of methanogen microorganisms, (2) formation of tannin-protein complexes that decreases the availability of nutrients for ruminal microorganisms and compromises their development, and (3) decrease in available H2 for CO2 reduction to CH4 [24]. However, the effects of tannins fluctuate depending on the type, source, molecular weight, dose and the adaptation of animals to their consumption. To such an extent that negative effects on productive performance, feed consumption, digestion and animal health have been evidenced. Effects that are probably attributed to: (i) interaction of CT with salivary glycoproteins and subsequent decrease in feed palatability, (ii) greater permanence of feed in the rumen as a consequence of decreased substrate digestibility, (iii) intoxication by the excessive consumption of CT (>55 g CT/kg DM) and (iv) reduced intestinal enzymatic activity of trypsin, amylase and amino acid synthesis [25].
The use of forage shrubs is presented as a promising alternative in ruminant feeding. Tithonia diversifolia is a shrub whose forage contains between 19.3–22.9% crude protein (CP), 18.4–25.5% dry matter (DM), 84.0–86.6% organic matter (OM) and 18.1–19.4% crude fiber (CF) [26,27] and 37.9–48.1 g/kg MS of TC [28]. Erythrina poeppigiana is a legume characterized by a high concentration of PC (20.2–32.0%), DM (23.2%), MO (90.7%), neutral detergent fiber (NDF: 53.4–60.9%), acid detergent fiber (ADF: 31.5–37.3%) and CT (0.4%) [29,30]. Cratylia argentea is also a legume with high nutritional value (CP: 19.5–24.0%, OM: 88.5%, NDF: 56.4%, ADF: 32.3% and TC: 0.2%) [30,31]. Therefore, it is hypothesized that T. diversifolia, E. poeppigiana and C. argéntea are forages that can be useful as feed for ruminants throughout the year and probably reduce CH4 production. Based on this background, the aim of this research work was to determine the effect of three tropical forage species at four cutting ages, during the dry and rainy seasons, on in vitro gas production, rumen pH, volatile fatty acids and methane production.

2. Materials and Methods

2.1. Location

The research was carried out at the Experimental Farm “La María”, of the State Technical University of Quevedo, located in the Province of Los Ríos, in the Canton Mocache, located at km 7 ½ of the Quevedo-El Empalme road, in an area classified as tropical humid forest, at a height of 73 m above sea level.

2.2. Animals

Four Brahman bulls of 250.0 ± 20.5 kg of live weight were used, provided with a ruminal cannula (four inches of internal diameter, Bar Diamond, Parma, Idaho, USA) for the extraction of ruminal fluid. The animals were kept separately in individual pens, where they were fed with P. maximum and water was provided ad libitum

2.3. Forage Samples

The forage was harvested from an agroforestry system established in 2016, located at the following geographic coordinates (Latitude: 1°05′0.19″ South, Longitude: 79°29′59.86″ West). The entire plant (leaves and stems) of three shrub species (T. diversifolia, E. poeppigiana and C. argentea) was used and an equalization cut was made and subsequently experimental cuts were made 30, 45, 60 and 75 days for the dry and rainy season. The plant samples were harvested in the rainy season from January to March (with a monthly rainfall of 270–600 mm and a temperature of 22–30 °C) and for the dry season from September to November (with a monthly rainfall of 0 mm and a temperature of 20–30 °C), the samples were taken from the same plants in both seasons manually at 0.50 cm from the soil towards the apex of the plant. Each shrub species were identified by Dr. F Meza-Bone, professor at the Facultad de Ciencias Agrarias y Forestal, Universidad Técnica Estatal de Quevedo (in spanish). A voucher specimen of T. diversifolia, E. poeppigiana and C. argentea has been deposited in our laboratory (voucher No. UTEQ-1001, 1002 and 1003 respectively). The sampled plant material (5 kg per plant: 20 plants of each species) leaves and stems were dried at 60 °C (72 h) in a forced air oven until reaching constant weight to determine the dry matter and chemical composition, later it was ground in a hammer mill (THOMAS-Wiley, USA, Model 4) and the resulting plant material was passed through a 1 mm sieve to homogenize the particle size.

2.4. Gas, CH4 Production and Rumen pH

The rumen content (solid and liquid fraction) was obtained from 4 bulls provided with ruminal cannula. Rumen content was collected before feeding and kept at 39 °C in a sealed plastic container during transport to the laboratory. The nitrogen-rich medium (artificial saliva) was prepared as described by Menke and Steingass [32]. Gas production was determined according to the methodology described by Theodorou et al. [33] which consists of placing 0.400 g of dry matter from each treatment was placed in serum bottles of 100 mL nominal capacity, 60 mL of inoculum (70:30 medium; artificial saliva/inoculum; rumen content) was added under a constant CO2 flow. The bottles were sealed and incubated at 39–40 °C. Gas pressure and volume were measured manually at 3, 6, 9, 12, 24, 48, 72 and 96 h after incubation with a DELTA OHM model DO 9704 pressure transducer (Delta OHM, Padova, Italy) and plastic syringes. CH4 production was quantified with a GX-6000 gas monitor (RKI Instruments gas analyzer, GX-6000, Tokyo, Japan) following the methodology described by Elghandour et al. [34]. For each treatment, four bottles were used and four additional bottles as blank. At the end of 96 h, the gas and CH4 data (mLgas or CH4/g MS incubated) were fitted to the equation y = D (1 − e−k t) described by Krishnamoorthy et al. [35], where:
y = cumulative gas production at a given time (mL)
D = potential cumulative gas production (mL)
k = rate of gas production (h-I)
t = time of fermentation (h)
The rumen pH was measured from the same bottles with the help of a pH meter (BANTE-221 Portable pH/ORP Meter, London, UK) at 6, 12 and 24 h post incubation.

2.5. Fermentation Profile

Four additional bottles for each treatment were prepared following the same methodology indicated above for Gas and CH4 production. 6 h after incubation, 8 mL of sample was collected from each treatment and repetition and mixed with 2 mL of 25% metaphosphoric acid and stored at −4 °C until analysis, AGVs were analyzed by means of a gas chromatograph (Clarus 400, PerkinElmer, Shelton, CT, USA) following the methodology described by Ryan [36].

2.6. Experimental Design and Statistical Analysis

A completely randomized design was used in a factorial arrangement (three forage species—four cutting ages and two seasons) with four repetitions per treatment, as follows:
Yijk = µ + Ai + Bj + Ck + ABij + ACik +BCjk + ABCijk + ∑ijk
where:
Yijk = Observation of the response variable obtained from the treatment with the i-th level of A, the j-th level of B, the k-th level of C
µ = Overall mean
Ai = Effect of the i-th level of the forage species factor
Bj = Effect of the j-th factor level of the cutting ages
Ck = Effect of the k-th level of the epoch factor
ABij = Effect of the interaction of the i-th level of factor A and the j-th level of factor B
ACij = Effect of the interaction of the i-th level of factor A and the k-th level of factor C
BCjk = Effect of the interaction of the j-th level of factor B and the k-th level of factor C
All the variables were analyzed according to the proposed experimental design, using the PROC GML of the SAS. All means were compared using Tukey test.

3. Results

3.1. Gas, CH4 Production and Rumen pH

Gas production showed differences (p < 0.05) between forage species, cutting age, interaction between forage species and cutting age, and on the dry and rainy seasons (Table 1). The accumulated gas production between forage species was lower (p < 0.05) in T. diversifolia (108.0 mLgas/0.400 g incubated DM) and E. poeppigiana (114.01 mLgas/0.400 g incubated DM) in the dry season, and in the rainy season it was lower (p < 0.05) in T. diversifolia (96.99 gas/0.400 g incubated DM). On the other hand, the effect between seasons (dry and rainy) showed lower (p < 0.05) accumulated gas production in T. diversifolia (96.9 mLgas/0.400 g incubated DM) and C. argéntea (118.3 mLgas/0.400 g incubated DM) in the rainy season, however, in the dry season E. poeppigiana showed lower (p < 0.05) gas production (114.01 mL gas/0.400 g incubated DM) compared to the other forages.
Regarding the cutting age effect in the dry season, it showed lower (p = 0.0164) gas production at 30 and 45 days (108.20 and 114.31 mLgas/0.400 g incubated DM, respectively), however, in the dry season lower (p = 0.0001) gas production at 30 days (103.53 mLgas/0.400 g incubated DM) was evidenced. The effect between seasons (dry and rainy) in the cutting ages showed differences, obtaining 4.3% lower (p < 0.05) gas production at 30, 45 and 60 days in the rainy season compared to the dry season. Regarding the interaction between forage species and cutting ages, no differences were observed for the two seasons (dry and rainy: p = 0.8737 and p = 0.0552, respectively) (Table 1).
CH4 production showed an effect between forage species, showing the lowest CH4 production in T. diversifolia for both the dry and rainy seasons, 73.5 and 51.7 mL CH4/0.400 g incubated DM, respectively. On the other hand, the effect between seasons on the CH4 production was lower in E. poeppigiana and C. argéntea for the rainy season (64.0 and 73.5 mL CH4/0.400 g incubated DM, respectively). Regarding the production of CH4 influenced by the cutting age effect, it was less than 30 days for both the dry and rainy seasons (p = 0.0001 and p = 0.0001, respectively). However, the effect between seasons (dry and rainy) showed lower (p < 0.05) CH4 production at 45, 60 and 75 days of the rainy season. The interaction between forage species and cutting ages showed an effect (p = 0.0001) on CH4 production in the dry season (Table 2).
The rumen pH at 6, 12 and 24 h did not show differences (p > 0.05) in the variables evaluated (Table 1).

3.2. Fermentation Profile

Volatile Fatty Acids and total VFAs in the dry season did not show differences in the effect of forage, age and interaction between forage and age (p = 0.5218, p = 0.9306 and p = 0.9547, respectively). However, in the rainy season, the production of acetic acid, propionic acid and butyric acid among forage species was higher in E. poeppigiana (76.39 mol %), C. argéntea (16.31 mol %) and T. diversifolia (8.16 mol %) respectively. On the other hand, the effect between seasons (dry and rainy) showed differences, obtaining higher mol % of propionic acid and butyric acid in the dry season for T. diversifolia, E. poeppigiana and C. argéntea respectively (p < 0.05). Regarding the effect of cutting ages in both the dry and rainy seasons, no differences were observed for all the variables evaluated (p > 0.05). However, the effect between seasons (dry and rainy) the dry season showed higher (p < 0.05) mol % of butyric acid in all cutting ages, while in the rainy season the acetic acid was higher (p < 0.05) for all cutting ages. The interaction between forage species and cutting age did not show differences (p > 0.05) on VFA production in the dry season. However, a higher production of acetic acid was observed in the rainy season (p = 0.0007) (Table 3).

4. Discussion

4.1. Gas, CH4 Production and Rumen pH

The nutritional value of plants varies according to the geographical area, seasonality of the year, forage species and regrowth age. The results obtained in the present study on in vitro gas production were influenced by the forage species, cutting age, interaction between forage species and cutting age, and the dry and rainy seasons (Table 1). This variability of responses could probably be due to the nutritional quality of the forage, associated with: (1) time of year (spring, summer, autumn and winter), (2) physiological maturity of the plant, which exerts changes on the components of the cell wall and protein [16,37,38] and, (3) presence of bioactive compounds (CT), which influence the rate of degradation and digestibility of the feed [39].
Under these concepts, the lower gas production evidenced in the rainy season in T. diversifolia and C. argentea compared to the dry season can probably be derived from the lower content of structural carbohydrates, influenced by the limited amount of sunlight available in the rainy season [13]. Contrary to this, the dry season, when presenting a greater intensity of solar radiation and temperature, could generate changes on the fibrous components in the plant, showing a greater amount of structural carbohydrates (cellulose and hemicellulose) [17,19,40]. In the same way, the moisture deficit in the dry season probably negatively influences the growth of forages, decreasing the leaf area, increasing the amount of structural carbohydrates, decreasing the photosynthetic activity that affects the production and nutritional quality of the plants [41]. In addition, the gas production could be influenced by the low amount of N in the forages in the dry season, resulting in a decrease in microbial growth as an effect of the low availability of NH3 in the rumen [42]. Or by the decrease in the N balance in animals, by increasing the amount of lignin in the plant and generating losses of endogenous and microbial N in the feces [19]. On the other hand, the results evidenced regarding the cut-off age on in vitro gas production (Table 1) are probably due to the greater deposit of structural carbohydrates in the plant, as the physiological development is prolonged [37,38].
The differences between the cutting ages on CH4 production is probably associated with: (1) the highest fiber concentration, (2) variations in the passage rate and (3) the presence of secondary compounds (CT) (Table 2). According to Carmona et al. [43], the CH4 emission rate is related to the physical-chemical characteristics of the forage. It is evident that, with a greater presence of structural carbohydrates, the production of CH4 increases [44]. On the other hand, the lower production of CH4 at a lower regrowth age is probably associated with a greater amount of highly soluble carbohydrates, showing a close relationship between the physiological maturity of the plant and the fiber content [37,38]. In such a way that the forage species, cutting age and time of year influence the production of CH4, possibly due to the variability in its chemical composition. Previous studies have shown that the use of tropical tree or shrub plants containing secondary metabolites in ruminant feed can minimize CH4 emission [30,45]. low percentages of fibrous fractions in the plant are associated with a lower production of CH4 [46].
The ruminal pH evidenced in the present study (Table 1) is consistent with that reported by Vargas-Ortiz et al. [25], who consider positively benefit the activity of microorganisms that degrade cellulose.

4.2. Fermentation Profile

The results obtained in the production of VFA in this study are probably due to the interaction between the seasons studied (dry and rainy) and the nutritional composition of the plants, showing better results in the rainy season (Table 3). The higher production of acetic acid in the rainy season could probably be due to a rapid degradation of the forage species in relation to the cutting age, due to the contribution of nutrients of rapid fermentation in physiologically young plants [47]. In this context, Rodríguez et al. [48] mention that the VFA profile can be modified by the type of fermented substrate. The lack of effects on the VFA profile in the present study in the dry season is probably due to the moisture deficit, which negatively influences the growth, production and nutritional quality of the forage [13]. High temperature, evaporation and wind speed, and low humidity can also have detrimental effects on forage quality [41]. It has been shown that as the plant advances in its state of physiological maturity, the grasses and forages present a greater amount of structural carbohydrates, related to the control of the water balance in the plant, in such a way that, in conditions of moisture stress (water scarcity), especially in dry seasons, the stomata are closed and transpiration is reduced to prevent water loss, with subsequent decrease in dry matter content [49,50].

5. Conclusions

The use of shrub and tree species such as Tithonia diversifolia, Erythrina poeppigiana and Cratylia argéntea in ruminant production systems in the rainy and dry seasons, with an early regrowth age between 30 and 45 days, present a promising alternative to the reduction of CH4, possibly becoming a sustainable strategy in the tropics to reduce enteric GHG production in ruminants and improve their productive performance.

Author Contributions

Conceptualization, G.M.-B., C.M.-B., J.A.-C., R.C.-C. and M.B.-R.; data tabulation, M.B.-R., R.V.-T., M.C.-C., W.V.-A. and F.I.-F.; writing—review and editing, M.B.-R., F.M.-B., A.Z.-S., R.B.-E., T.S.-B., L.M.-A. and W.G.-C. All authors have read and agreed to the published version of the manuscript.

Funding

The research was funded by the State Technical University of Quevedo, Ecuador through the FOCICYT-2016 project.

Institutional Review Board Statement

All procedures followed the recommendations of the ARRIVE Guide [Kilkenny et al., 2010] and were approved by the Bioethics Committee of the Faculty of Livestock and Biological Sciences of the State Technical University of Quevedo (resolution 20/2019).

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this paper are available on request from the corresponding author.

Acknowledgments

All authors thank the State Technical University of Quevedo, Ecuador, for financing this research and the Technical University of Ambato, Ecuador, for the collaboration through the Ruminology Laboratory of the Faculty of Agricultural Sciences.

Conflicts of Interest

The authors declare no conflict of interest.

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Table
Table 1. In vitro gas production parameters (mL gas/0.400 g incubated DM) and rumen pH of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Table 1. In vitro gas production parameters (mL gas/0.400 g incubated DM) and rumen pH of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Dry SeasonsRainy Seasons
Gas ProductionRumen pHGas ProductionRumen pH
Dk6 h12 h24 hDk6 h12 h24 h
Forage
T. diversifolia108.0 bA0.045 bA7.62 aA7.62 aA7.70 aA96.9 bB0.047 bA7.62 aA7.63 aA7.67 aA
E. poeppigiana114.0 bB0.040 bA7.60 aA7.66 aA7.65 aA116.3 aA0.037 cB7.60 aA7.65 aA7.62 aA
C. argentea121.7 aA0.053 aA7.62 aA7.67 aA7.67 aA118.3 aB0.054 aA7.63 aA7.69 aA7.65 aA
Ages (day)
30108.2 bA0.037 cA7.60 aA7.67 aA7.67 aA103.5 cB0.037 cA7.61 aA7.68 aA7.64 aA
45114.3 abA0.037 cB7.61 aA7.66 aA7.65 aA108.9 bB0.044 bcA7.61 aA7.68 aA7.63 aA
60117.3 aA0.049 bA7.62 aA7.63 aA7.65 aA112.6 abB0.047 bA7.63 aA7.65 aA7.62 aA
75118.4 aA0.062 aA7.61 aA7.62 aA7.71 aA117.1 aA0.056 aB7.62 aA7.62 aA7.69 aA
Inteaction
T. diversifolia30100.8 bA0.034 bA7.65 aA7.70 aA7.73 aA88.8 dA0.035 dA7.65 aA7.70 aA7.71 aA
45105.4 abA0.033 bB7.61 aA7.67 aA7.70 aA95.0 dB0.039 cdA7.62 aA7.67 aA7.67 aA
60112.5 abA0.040 bB7.61 aA7.57 aA7.67 aA96.1 cdB0.045 bcdA7.62 aA7.61 aA7.64 aA
75113.1 abA0.073 aA7.61 aA7.53 aA7.69 aA107.8 bB0.068 aB7.60 aA7.53 aA7.67 aA
E. poeppigiana30105.2 abA0.036 bA7.57 aA7.66 aA7.62 aA107.4 bcA0.033 dB7.58 aA7.67 aA7.58 aA
45115.0 abA0.036 bB7.61 aA7.69 aA7.64 aA113.0 abA0.038 cdA7.61 aA7.65 aA7.61 aA
60116.8 abB0.064 aA7.59 aA7.67 aA7.62 aA122.1 aA0.038 cdB7.59 aA7.66 aA7.61 aA
75118.9 abB0.067 aA7.63 aA7.62 aA7.71 aA122.6 aA0.039 cdB7.62 aA7.62 aA7.69 aA
C. argentea30118.5 abA0.041 bA7.59 aA7.65 aA7.68 aA114.3 abB0.042 cdA7.60 aA7.66 aA7.64 aA
45122.4 aA0.040 bB7.61 aA7.64 aA7.62 aA118.6 abB0.054 abcA7.61 aA7.71 aA7.61 aA
60122.6 aA0.064 aA7.68 aA7.67 aA7.66 aA119.5 aB0.059 abB7.68 aA7.69 aA7.62 aA
75123.3 aA0.067 aA7.61 aA7.71 aA7.73 aA121.0 aB0.068 abA7.63 aA7.71 aA7.72 aA
SE 4.030.0030.020.060.041.350.0030.030.050.04
Forage0.00010.00010.27110.45660.14110.00010.00010.24750.22360.3252
Ages 0.01640.00010.77530.68940.17540.00010.00010.86620.43630.2363
Forage × Ages0.87370.00010.10730.47770.51020.05520.00160.33610.52810.6432
a–d The letters in lowercase superscript in the column make significant reference (p < 0.05) to the forage species and cutting ages.; AB The letters in uppercase superscript in the rows make a significant reference (p < 0.05) between dry and rainy seasons.; D = potential cumulative gas production (mL), k = rate of gas production (h-I).; SE = standard error.
Table
Table 2. CH4 production parameters (mLCH4/0.400 g incubated DM) of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Table 2. CH4 production parameters (mLCH4/0.400 g incubated DM) of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Dry Seasons Rainy Seasons
CH4 Production CH4 Production
DkDk
Forage
T. diversifolia73.5 cA1.23 aA51.7 cA1.21 aA
E. poeppigiana95.7 bA1.06 bA64.0 bB1.20 aA
C. argéntea132.1 aA1.04 bA73.5 aB0.91 bB
Ages (days)
3042.1 dA1.24 aA35.7 dA1.34 aA
4574.2 cA1.18 aA54.1 cB1.11 bA
60136.2 bA1.14 aA75.2 bB0.90 cA
75149.3 aA0.88 bA87.2 aB1.08 bA
Inteaction
T. diversifolia3021.3 fA1.39 aA19.9 dA1.39 bA
4550.9 eA0.92 aA38.0 cdA1.39 abA
60116.0 bcA0.92 deA73.7 abB0.88 eA
75105.7 cA1.22 abcA75.0 abB1.25 bcA
E. poeppigiana3052.8 eA1.26 abB33.8 cdB1.64 aA
4592.0 cdA1.10 bcdA54.1 bcB1.04 cdeA
60182.6 aA0.75 eB74.7 abB0.95 deA
75201.1 aA1.12 bcdA93.3 aB1.16 bcdA
C. argéntea3052.1 eA1.05 bcdA53.3 bcA1.05 cdeA
4579.5 dA1.04 bcdA70.1 abA0.89 eB
60110.2 cA0.97 cdeA77.2 abA0.87 eA
75141.2 bA1.08 bcdA93.3 aB0.83 eB
SE 5.220.055.010.05
Forage0.00010.00010.00010.0001
Ages0.00010.00010.00010.0001
Forage × Ages0.00010.00130.05270.0001
a–f The letters in lowercase superscript in the column make significant reference (p < 0.05) to the forage species and cutting ages.; AB The letters in uppercase superscript in the rows make a significant reference (p < 0.05) between dry and rainy seasons.; D = potential cumulative CH4 production (mL), k = rate of CH4 production (h-I).; SE = standard error.
Table
Table 3. In vitro proportion of volatile fatty acids in the rumen (mol %) of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Table 3. In vitro proportion of volatile fatty acids in the rumen (mol %) of shrub and tree species at different cutting ages, in the dry and rainy seasons.
Dry SeasonsRainy Seasons
Rumen VFA (mol %)Rumen VFA (mol %)
AceticPropionicButyricTotal FVA (mmol/L)AceticPropionicButyricTotal FVA (mmol/L)
Forage
T. diversifolia73.19 aB16.54 aA10.27 aA35.56 aA76.03 abA15.81 bB8.16 aB30.18 aB
E. poeppigiana73.36 aB16.69 aA9.94 aA34.28 aA76.39 aA15.92 bA7.70 bB29.34 aA
C. argéntea73.82 aB16.75 aA9.43 aA34.04 aA75.72 bA16.31 aA7.97 abB32.28 aA
Ages (days)
3073.53 aB16.49 aA9.99 aA34.70 aA76.31 aA15.83 aA7.86 aB29.94 aA
4573.29 aB16.66 aA10.06 aA33.84 aA75.88 aA16.11 aA8.01 aB28.02 aB
6073.34 aB16.65 aA10.01 aA34.65 aA76.02 aA16.00 aA7.98 aB32.56 aA
7573.67 aB16.86 aA9.47 aA35.32 aA75.96 aA16.10 aA7.93 aB31.88 aA
Inteaction
T. diversifolia3073.08 aB16.17 aA10.74 aA35.57 aA76.50 abcA15.53 aA7.97 aA29.07 aA
4573.51 aB16.34 aA10.16 aA36.51 aA75.65 bcdA15.90 aA8.45 aA29.36 aA
6073.18 aB16.58 aA10.25 aA36.19 aA75.89 abcdA15.98 aA8.13 aB28.59 aA
7572.98 aB7.09 aA9.93 aA33.99 aA76.07 abcdA15.84 aB8.10 aA33.72 aA
E. poeppigiana3073.61 aB16.50 aA9.89 aA35.17 aA76.10 abcdA16.01 aA7.90 aA27.57 aA
4572.65 aB17.20 aA10.16 aA32.12 aA75.99 abcdA16.17 aA7.85 aA28.31 aA
6073.39 aB16.66 aA9.95 aA35.03 aA76.75 aA15.65 aA7.61 aA32.88 aA
7573.81 aB16.42 aA9.77 aA34.81 aA76.73 abA15.84 aA7.44 aA33.33 aA
C. Argéntea3073.89 aB16.78 aA9.33 aA33.35 aA76.33 abcA15.96 aA7.71 aA33.17 aA
4573.70 aB16.44 aA9.86 aA32.88 aA76.02 abcdA16.26 aA7.72 aA26.41 aA
6073.46 aA16.71 aA9.83 aA32.74 aA75.42 cdA16.38 aA8.20 aA36.21 aA
7574.22 aA17.08 aA8.71 aA37.18 aA75.10 dA16.64 aA8.26 aA33.33 aA
SE 0.790.450.821.750.220.190.224.45
Forage0.52180.79210.35990.42550.00060.01040.02000.6326
Ages0.93060.78460.79600.77970.11550.24010.85820.5947
Forage × Ages0.95470.64940.98720.38870.00070.15180.16310.8688
a–d The letters in lowercase superscript in the column make significant reference (p < 0.05) to the forage species and cutting ages.; AB The letters in uppercase superscript in the rows make a significant reference (p < 0.05) between dry and rainy seasons.; SE = standard error.
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Meza-Bone, G.; Meza-Bone, C.; Avellaneda-Cevallos, J.; Cabezas-Congo, R.; Villamar-Torres, R.; Cabanilla-Campos, M.; Vivas-Arturo, W.; Intriago-Flor, F.; Meza-Bone, F.; Zapatier-Santillan, A.; et al. Rumen Fermentation Profile and Greenhouse Gas Mitigation of Three Forage Species from Agroforestry Systems in Dry and Rainy Seasons. Fermentation 2022, 8, 630. https://doi.org/10.3390/fermentation8110630

AMA Style

Meza-Bone G, Meza-Bone C, Avellaneda-Cevallos J, Cabezas-Congo R, Villamar-Torres R, Cabanilla-Campos M, Vivas-Arturo W, Intriago-Flor F, Meza-Bone F, Zapatier-Santillan A, et al. Rumen Fermentation Profile and Greenhouse Gas Mitigation of Three Forage Species from Agroforestry Systems in Dry and Rainy Seasons. Fermentation. 2022; 8(11):630. https://doi.org/10.3390/fermentation8110630

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Meza-Bone, Gary, Carlos Meza-Bone, Juan Avellaneda-Cevallos, Ronald Cabezas-Congo, Ronald Villamar-Torres, María Cabanilla-Campos, Walter Vivas-Arturo, Frank Intriago-Flor, Fabricio Meza-Bone, Aurelio Zapatier-Santillan, and et al. 2022. "Rumen Fermentation Profile and Greenhouse Gas Mitigation of Three Forage Species from Agroforestry Systems in Dry and Rainy Seasons" Fermentation 8, no. 11: 630. https://doi.org/10.3390/fermentation8110630

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