Skip to main content
PMC home page
Veterinary World logoLink to Veterinary World
. 2023 Jan 30;16(1):215–221. doi: 10.14202/vetworld.2023.215-221

Supplementation of Water Spinach (Ipomoea aquatica) on the utilization of Mimosa pigra and Leucaena leucocephala leaf for in vitro fermentation

Channy Sambo 1, Sreychou Heng 1, Pisey Vong 2, Kuyhor Te 1, Sath Keo 3, Mom Seng 4, Samnang Ven 1,✉
PMCID: PMC9967708  PMID: 36855361

Abstract

Background and Aim:

Ipomoea aquatica (Water Spinach) is the most potential for livestock growth performance, including chickens, pigs, cattle, and goats, especially in a tropical country like Cambodia. It is not only an alternative feed source but also one kind of supplemented feed for goat raising. Supplementation with Water Spinach in the utilization of low-quality tree foliage results in an increase in dry matter intake in goat production. This study aimed to identify the effectiveness of supplementation of Water Spinach in the utilization of Mimosa pigra and Leucaena leucocephala leaf in in vitro fermentation.

Materials and Methods:

The study was designed according to a 2 × 2 factorial arrangement in randomized design of seven treatments with different ratios consisted of different three types of dietary treatments, including M. pigra, L. leucocephala, and Water Spinach. The treatments were arranged according to a completely randomized design and were as follow: T1 = M. pigra leaf (100%); T2 = L. leucocephala leaf (100%); T3 = M. pigra leaf and L. leucocephala leaf (50% and 50%); T4 = M. pigra leaf and Water Spinach (99.5% and 0.5%); T5 = L. leucocephala leaf and Water Spinach (99.5% and 0.5%); T6 = M. pigra leaf and Water Spinach (99% and 1%); and T7 = L. leucocephala leaf and Water Spinach (99% and 1%). A total of 200 mg (dry matter) of dietary treatments were prepared in a 60 mL syringe. Each treatment was replicated 3 time. Gas recording of each treatment lasted for 3 days. In vitro was performed for 72 h, was followed by Makkar method. Gas production was recorded at 2, 4, 8, 12, 24, 36, 48, and 72 h of incubation by using strict anaerobic technique. A mixture of rumen fluid and dietary treatments were carried out under continuous flushing with CO2 in sharking incubator at 39°C. After incubating for 72 h, the ammonia concentration (NH3-N) was measured and recorded to identify pH, nutrient digestibility, and ammonia concentration (NH3-N).

Results:

Nutrient digestibility of the treatment with Water Spinach supplement in the utilization of L. leucocephala was obtained at a higher digestibility than treatment with M. pigra (p < 0.05). Gas production was different between groups (p < 0.05). Treatment with only M. pigra leaf had the highest gas production (A), while treatment with Water Spinach supplementation had the lowest gas production (A). At 0–24 h, the treatment with L. leucocephala leaf and Water Spinach 0.5% had the highest gas production, but after 24 h, M. pigra leaf and Water Spinach 1% and L. leucocephala leaf and Water Spinach 0.5% produced more gas compared to the other treatments (p < 0.05).

Conclusion:

The supplementation of Water Spinach 1% in treatment with M. pigra and L. leucocephala leaf resulted in increased degradability, gas production, and NH3–N concentration without a change in the pH value rumen condition. Based on these results, it is recommended that the level of Water Spinach supplementation should be 1% of dietary intake. Future studies should consider investigating the rumen ecology associated with Water Spinach supplementation. Feeding with Water Spinach remains a good supplement for ruminant performance; therefore, further studies should be conducted using Water Spinach in ruminant feeding in both metabolic and feeding trials.

Keywords: In vitro fermentation, Leucaena leucocephala leaf, Mimosa pigra, Nutrient digestibility, Supplementation, Utilization, Water Spinach (Ipomoea Aquatica)

Introduction

Most grass feeding ruminants contain a high fiber level of more than 40% and a low protein level of <10%, resulting in limited growth performance during the dry season [1]. Varieties of feeding programs are important to provide an appropriate nutrient balance to achieve the nutrient requirement for ruminant maintenance, growth, and production of animals [2]. In Cambodia, the source of ruminant feeding is based on natural grasses that are readily available in the rural area, especially legume trees and shrub foliage such as Mimosa and Leucaena.

However, Mimosa pigra is affected by biodiversity, fishing, and crops [3]. Using leguminous leaves in livestock feed would prevent this plant from becoming a serious concern through the overgrowth in the environment and plays an important role in sustainable development [4]. Mimosa pigra has been known as a high crude protein source that contained in the leaf and it was also generally used as a feed source in swines, rabbits, and goats [5]. Legume trees like; Leucaena leucocephala were introduced as an animal feed supplement [6]. It is a digestible protein and was often recommended as ruminant feed; however, feeding with only L. leucocephala may not affect intake of dry matter (DM) and other nutrients, resulting in digestive problems in the ruminant animals [7]. These two legume trees are considered to be a source of protein consumption in ruminant, but it contains high levels of tannin, which results in low digestion in the ruminant and contributes to low gas production [8]. Water Spinach is known as vegetative forage, which contains an adequate protein level for animal requirements when combined with legume trees [1]. It is, therefore, hypothesized that the growth performance of goats fed leguminous tree species as a basal diet would be improved by supplementing it with Water Spinach. Thus, the supplementation of Water Spinach would be considered as a quality additive to improve livestock production while utilizing M. pigra and L. leucocephala leaves. The increase in growth rate was consistent with the additional level of Water Spinach and it maintained great DM digestibility [9]. However, there are limited available studies on using Water Spinach supplementation with legume trees as a basal diet in ruminants. Therefore, utilizing M. pigra and L. leucocephala leaves in different proportions with different levels of Water Spinach supplement would be a promising concept to investigate a novel digestibility system in goats. To conduct this experiment, in vitro fermentation is a convenient technique compared to other digestibility methods in ruminant experiments [10].

Therefore, this study aimed to investigate supplementation of Water Spinach on the utilization of M. pigra and L. leucocephala leaf on in vitro fermentation using beef cattle rumen fluid as rumen inoculum to evaluate ammonia concentration, pH, and nutrient digestibility in in vitro gas production methods.

Materials and Methods

Ethical approval

In this study, we did not use live animals; therefore, ethical approval is not required. Rumen fluid was obtained from two beef cattle sacrificed in the slaughterhouse.

Study period and location

The study was conducted from July to August 2019 on beef cattle. Twenty days of adaptation period was performed in two animals with basal diet in individual pens of slaughterhouse area located in the north of Phnom Penh, before rumen fluid was collected. A total of 10 days included 1 day of rumen fluid collection in the slaughterhouse, 3 days of in vitro fermentation in laboratory, and 6 days of nutrient digestibility analysis. In vitro performance, and nutritional analysis was conducted in the Chemical Analysis Lab., Graduate school, Royal University of Agriculture, Dangkao district, Phnom Penh.

Dietary substrate, treatments, and experimental design

The study designed was a 2 × 2 factorial arrangement in a randomized design of 7 treatments performed in triplicate, while the first factor was different levels of Water Spinach supplementation (0, 0.5, and 1% DM of the dietary substrate), and the second factor was tree forage utilization (non-supplemented, 50% of Leucaena and Mimosa, and 100% of Leucaena and M. pigra utilization DM of dietary substrate). The treatments were arranged according to a completely randomized design and were as follows: T1 = M. pigra leaf (100%); T2 = L. leucocephala leaf (100%); T3 = M. pigra leaf and L. leucocephala leaf (50% and 50%); T4 = M. pigra leaf and Water Spinach (99.5% and 0.5%); T5 = L. leucocephala leaf and Water Spinach (99.5% and 0.5%); T6 = M. pigra leaf and Water Spinach (99% and 1%); and T7 = L. leucocephala leaf and Water Spinach (99% and 1%). A total of 200 mg (DM) dietary treatments were prepared in a 60 mL syringe.

Chemical composition analysis

To identify dietary chemical composition, substrates were milled to pass through a 1 mm screen (Cyclotec 1093 Sample mill, Tecator, Hoganas, Sweden), including 1–2 m lengths of L. leucocephala and M. pigra stems. The end of the stem was cut to measure about 30–50 cm long and was maintained for laboratory analysis [11].

Medium solution preparation

A medium solution of 1000 mL was prepared for 33 plastic syringes that were obtained from the study conducted by Menke and Steingass [12]. Distilled water (475 mL), 240 mL macromineral solution (5.7 g Na2HPO4, 6.2 g KH2PO4, and 0.6 g MgSO4 made up to 1 L with distilled water), 240 mL buffer solution (35 g NaHCO3 and 0.4 g NH4HCO3 made up to 1 L with distilled water), 0.12 mL micromineral solution (13.2 g CaCl2H2O, 10 g MnCl4H2O, 1 g CoCl26H2O, and 0.8 g FeCl26H2O made up to 1 L with distilled water), 100 mg of resazurin, 2 mL of reducing buffer (1M NaOH, 336 mg Na2S.9H2O made up to 1 L with distilled water), and adding 47.5 mL distilled water to finalize the volume.

Animals and preparation of rumen inoculums

Animals were placed on a routine feeding plan for at least 20 days and kept in individual pens with free access to clean, fresh water, and mineral blocks. On day 21, about 1500 mL of rumen fluid was obtained from the animals before feeding a morning mix of concentrate, grass, and rice straw while at the slaughterhouse. Both large and small ruminants have the same digestive system with a four-compartment stomach, The digestive systems have the same function that results from the same condition, such as pH level, ammonia concentrate, and nutrient digestibility in the rumen fluid [13]. The rumen fluid was transferred into prewarmed thermos-flasks and transported to the laboratory. The artificial saliva solution or buffer media preparation was prepared following that used in the study of Menke and Steingass [12]. A total of 1,500 mL rumen inoculum was prepared by the mixture of rumen fluid (500 mL) and artificial saliva 1,000 mL into 1.5 L thermos-flasks.

In vitro fermentation of a substrate

About 200 mg (DM) of dietary treatments were weighed into 60 mL plastic syringes. The method used for in vitro fermentation was based on the technique described by Makkar et al. [14]. Strict anaerobic techniques were used in all steps during the rumen fluid transfer and incubation periods. The syringes with a mixture of substrate treatments were prewarmed in a water bath at 39°C for 1 h before filling with 30 mL of the rumen inoculum mixture [12]. Three blank syringes without samples but containing 30 mL of the medium containing the buffer and rumen fluid at a ratio of 2:1 were used as the control group.

Gas production kinetics

Rumen fluid was collected from the slaughtered cattle at 4 am before feeding for the incubation solution and diluted at a (1:2) ratio (reducing medium: rumen fluid) in the collected artificial saliva in a CO2 tank. The 30 mL of rumen inoculum was added to each 60 mL plastic syringe with the dietary substrate (200 mg of DM). All syringes were incubated in a shaking water bath at 39°C for 72 h. During the incubation, the gas production kinetics were recorded at 0, 2, 4, 8, 12, 24, 36, 48, and 72 h. Cumulative gas production data were calculated based on the gas production at each time using the Fitcurve program fitted to the model of Orskov and Mcdonald [15] as follows: Y = A + B (1-e(-ct)), where A = The gas production from the immediately soluble fraction; B = The gas production from the insoluble fraction; C = The gas production rate constant for the insoluble fraction (B); t = Incubation time; (A + B) = The potential extent of gas production; and Y = Gas produced at a time “t”. After incubating for 72 h, the ammonia concentration (NH3–N) was measured and recorded to identify pH, nutrient digestibility, and ammonia concentration (NH3–N).

Nutrient digestibility

To determine nutrient digestibility, the residue of in vitro fermentation was used to analyze the nutrient dry matter degradability (DMD) and organic matter degradability (OMD) based on the Association of Official Analytical Chemists [16].

pH measurement

pH measurement was performed at 72 h of incubation using a pH meter (SM802, Romania). The pH meter is calibrated to pH buffers 7 and 4 before performing the pH measurement. Fermentation liquid was poured into a beaker and the probe was placed in a beaker to read and record the pH.

Ammonia concentration (NH3–N)

Fermentation liquid was sampled at 18 h postinoculation and filtered through four layers of cheesecloth. Samples were kept in plastic bottles to which 2.5 mL of 1 M H2SO4 was added to stop the fermentation process of microbe activity and then centrifuged at 3,000× g for 10 min. The supernatant was stored at −20°C before the analysis of ammonia nitrogen (NH3–N) using the micro-Kjeldahl method [11].

Statistical analysis

All obtained data were subjected to the General Linear Model (to analyze the interactions) procedures of the Statistical Analysis System Institute [17] through a 2 × 2 factorial arrangement in a randomized design of seven treatments performed in triplicate. The statistical model included L. leucocephala leaf: Water Spinach, and M. pigra leaf: Water Spinach interactions. Differences among the treatment means with p < 0.05 were contrasted using Tukey’s multiple comparison test [18].

Results

Chemical composition of dietary

The chemical compositions of the dietary feed are presented in Table-1. Mimosa pigra and L. leucocephala leaves contained CP at 15.6% and 24.5%, respectively. Comparing the CP contained in the three main dietary substrates, Water Spinach had a higher protein level than M. pigra and L. leucocephala leaves (Table-1).

Table-1.

Nutrient composition of dietary feed (% DM).

Nutrient composition Water Spinach M. pigra leaf L. leucocephala leaf
DM 10.0 39.6 24.9
OM 87.4 92.3 91.3
CP 30.0 15.6 24.5
NDF 26.2 32.9 25.1
ADF 24.8 27.9 15.9
CF 14.9 23.2 15.6
Ash 12.6 7.7 8.7
Open in a new tab

DM=Dry matter, OM=Organic matter, CP=Crude protein, NDF=Neutral detergent fiber, ADF=Acid detergent fiber, CF=Crude fiber, L. leucocephala=Leucaena leucocephala, M. pigra=Mimosa pigra, T1=Mimosa pigra leaf; T2=L. leucocephala leaf; T3=Mimosa pigra leaf + L. leucocephala leaf (50:50); T4=Mimosa pigra leaf + Water Spinach 0.5%; T5=L. leucocephala leaves + Water Spinach 0.5%; T6=Mimosa pigra leaf + Water Spinach 1%, and T7=L. leucocephala leaf + Water Spinach 1%

pH and ammonia concentrations in the rumen fluid

Ruminal pH and NH3–N concentrate are presented in Table-2, which was not affected by the supplementation of Water Spinach and the pH range was within 6.77–6.87. In this study, the treatment of L. leucocephala leaf contained 44.9 mg/dL of NH3–N compared to M. pigra leaf, which contained 32.3 mg/dL.

Table-2.

Nutritional digestibility and ammonia concentration in rumen.

Treatment pH NH3–N (mg/dL) Digestibility (%)
DM OM
T1 6.83 32.3b 41.5b 40.0b
T2 6.80 44.9a 57.8a 55.2a
T3 6.83 38.0a,b 54.8a,b 53.3a
T4 6.80 31.9b 53.1a,b 51.2a
T5 6.83 44.0a 57.5a,b 55.6a
T6 6.77 31.0b 53.9a 52.2a
T7 6.87 44.5a 59.3a,b 57.2a
SEM 0.016 1.557 1.623 1.257
p-value 0.815 0.003 0.034 0.000
Open in a new tab

T1=Mimosa pigra leaf; T2=L. leucocephala leaf; T3=Mimosa pigra leaf + L. leucocephala leaf (50:50); T4=Mimosa pigra leaf + Water Spinach 0.5%; T5=L. leucocephala leaves + Water Spinach 0.5%; T6=Mimosa pigra leaf + Water Spinach 1%, and T7=L. leucocephala leaf + Water Spinach 1%, SEM=Standard error of the mean, L. leucocephala=Leucaena leucocephala, M. pigra=Mimosa pigra. a,b,cSuperscripts differences in the same column show significant differences among. p < 0.05

Gas production

Table-3 illustrates the values of A, B, A + B, and C of each dietary group and the statistically significant differences are indicated by p < 0.05. After 72 h of incubation with Water Spinach on the utilization of M. pigra and L. leucocephala leaves, there was an increase in gas production from the immediately soluble fraction (A), gas production from the insoluble fraction (B), gas potential extent of gas production (A + B), and cumulative gas production at 24 h, (p < 0.05) and it remained stable until 72 h (Table-3). A high gas production (mL/0.2 g DM) was found in the supplementation of Water Spinach with the utilization of L. leucocephala leaf (Table-3), which indicated that the treatment with L. leucocephala leaf + Water Spinach 0.5% gas production was higher than the treatment with L. leucocephala leaf + Water Spinach 1%.

Table-3.

Supplementation of Water Spinach on the utilization of M. pigra leaf and L. leucocephala leaf on in vitro gas production.

Treatment Gas production kinetics (mL/0.2 g DM substrate) Gas 72 h (mL/0.2 g DM substrate)
A B C A+B
T1 0.86a 26.5d 0.06c 27.4c 27.2c
T2 −0.07a,b 35.1cd 0.11a,b 35.0b,c 34.9b,c
T3 0.16a,b 33.5c,d 0.09b 33.7b,c 33.6b,c
T4 0.25a,b 26.6b,c 0.09a,b 26.8b 26.7b
T5 −1.01b 38.9a,b 0.11a 37.5a,b 37.5a,b
T6 −1.37a,b 25.9a 0.09ab 26.9a 27.7a
T7 −2.84a,b 38.1a 0.013a,b 37.0a 37.0a
SEM 0.316 1.38 0.004 1.214 1.219
p-value 0.039 0.020 0.006 0.006 0.005
Open in a new tab

A=The gas production from the immediately soluble fraction, B=The gas production from the insoluble fraction, C=The gas production rate constant for the insoluble fraction (B), A + B=The gas potential extent of gas production, L. leucocephala=Leucaena leucocephala, M. pigra=Mimosa pigra, SEM=Standard error of the mean, T1=Mimosa pigra leaf; T2=L. leucocephala leaf; T3=Mimosa pigra leaf + L. leucocephala leaf (50:50); T4=Mimosa pigra leaf + Water Spinach 0.5%; T5=L. leucocephala leaves + Water Spinach 0.5%; T6=Mimosa pigra leaf + Water Spinach 1%, and T7=L. leucocephala leaf + Water Spinach 1%. a,b,cSuperscripts differences in the same column show significant differences among. p < 0.05 treatment

Nutrient digestibility

The effect of Water Spinach supplementation on pH value, ammonia concentration, and digestibility of each feed group are presented in Table-2. The addition of Water Spinach did not change the pH value (p = 0.815). The range of pH values is between 6.77 and 6.88. The group with M. pigra leaf had the lowest ammonia levels (p < 0.05), however, ammonia concentration significantly increased with Water Spinach supplementation (p = 0.003). Supplementation with 1% of Water Spinach using L. leucocephala and M. pigra as basal diet resulted in better digestion compared to the 0.5% supplemented and control groups. In addition, DMD and OMD from the treatment group with L. leucocephala leaf with Water Spinach were higher than the treatment group with M. pigra leaf. A high DM digestibility was present in the group supplemented with 1% of Water Spinach (T7) which accounted for 59.3% compared to T4 (53.1%) and T5 (57.5%).

Discussion

This study aimed to investigate the supplementation of Water Spinach on the utilization of M. pigra and L. leucocephala leaf on in vitro fermentation using beef cattle rumen fluid as rumen inoculum to evaluate ammonia concentration, pH, and nutrient digestibility in in vitro gas production methods. The findings of this study are similar to the study of Wittayakun et al. [19], who reported that DM, CP, and OM content of M. pigra was 40.22%, 18.11%, and 92.93%, respectively. However, the M. pigra leaf chemical composition in this study had a low neutral detergent fiber (NDF) (32.9%) and acid detergent fiber (ADF) (27.9%) compared to values obtained by Wittayakun et al. [19], who reported that the NDF and ADF content of M. pigra was 54.7% and 39.9%, respectively. The chemical composition of Water Spinach in this study found that CP and crude fiber (CF) were 30.0% and 14.9%. In contrast, a previous study demonstrated that CP and NDF content was 19.5% and 41.5%, respectively [19]. According to the research by Hasanah et al. [20] and Opene et al. [21], Water Spinach contained CP levels between 10.65%–24.6%, 13%–21.62% of CFs, 10.3% DM, and 89.42% OM. Some reports have varying results, which may be due to different varieties, periods of sample collection, and soil types. In this study, L. leucocephala leaf contained 24.5% of CP, while the study by Makmur et al. [22] reported that L. leucocephala contained 21%–25% of CP. This result is similar to this study. It may be because of the dietary substrate sampling method; the previous study by Makmur et al. [22] collected all parts of plants, such as the leaves, flowers, and soft stems to investigate the chemical composition of each sample. However, this experiment used only the leaves of L. leucocephala.

To respond to the microbial efficiency in ruminants, the pH measurements in this study were within the normal range (6.77–6.87) compared to those reported by Truong and Van Thu [23]. Therefore, this supplementation of Water Spinach did not affect the ruminal pH (p = 0.81) (Table-2). On the other hand, this pH result did not significantly decrease because the dietary substrates were not fermentable carbohydrates [24]. This is similar to the results reported by Viennasay and Wanapat [25], which showed that the pH value was high (6.80–6.92) when feeding with concentrated feed.

With regard to the report of Rigueira et al. [26], NH3–N ranged from 3.3 to 4.4 mg/dL can be changed by the type or quality of feeding. The findings of this study are in accordance with the study of Lanyasunya et al. [27], which demonstrated increased NH3–N levels, the increase in NH3–N was 66.3–104.7 mg/L in cows fed with a large variety of plant forage. In this study, the treatments with Water Spinach supplement presented a significantly higher NH3–N concentrate than treatments without the Water Spinach supplement (Table-2). This increase may be due to Water Spinach being easily broken down by microorganisms.

The results of gas production are in accordance with the study of Emmanuel et al. [28], which reported that Water Spinach produced more gas after utilization with M. pigra and L. leucocephala leaves. While comparing with or without Water Spinach supplementation, gas production from M. pigra and L. leucocephala was reported to be 27.2 mL/0.2 g DM and 34.9 mL/0.2 g DM, respectively. This result was higher compared to the study by Malik et al. [29], who indicated that gas production from M. pudica utilization was about 24.4 mL/0.2 g DM. This may be because of different varieties of Mimosa, which originate from different locations.

The total gas production increased considerably by 20% when Water Spinach was used as a substitute for forage source in ruminants [30]. On the other hand, a previous study confirmed that supplementation with Water Spinach feed resulted in high gas production in ruminants and an increase in the digestibility and N retention of foliage in small ruminants, which could contribute to the high gas production [9]. This may contribute to the negative impact on protein and energy metabolism when goats are fed with a high level of dietary Water Spinach [9]. Hasanah et al. [20] found that the increase in gas production was shown in the low percentage of supplementation compared to the high concentration of supplement, which is similar to this study results. This study found that there was a high gas production in the utilization of L. leucocephala supplement with 0.5% of Water Spinach. With regards to the result reported by Soto et al. [31], gas production ranged between 34.4 and 37.6 mL/0.2 g DM for 24 h of incubation with vegetable waste as the diet. This may be because the large forage diet digestibility of each substrate may result in increased gas production [32]. On the other hand, the low content of lignin in Water Spinach may result in increased digestion in ruminants [20]. Using only legume trees in ruminant feed may contribute to low gas production (Table-3); this may be because of the high lignin content in M. pigra and L. leucocephala. In contrast, the gas production shown in Table-3 was considerably increased when there was supplementation with Water Spinach. A high amount of gas production was shown in the utilization of L. leucocephala compared to M. pigra by supplementation of Water Spinach (Ipomoea Aquatica). This is because of the high CP and NDF content of L. leucocephala, which results in high gas production (Table-3).

Nutritional digestibility is shown in Table-2. The DMD and OMD from the treatment with L. leucocephala leaf with Water Spinach were higher than the treatment with M. pigra leaf. This may be due to the high level of NDF and ADF in forage diets which affect the ruminal pH and result in limited rumen microbial growth, which affects intestinal digestibility [20]. In addition, DM and OM digestibility is presented in Table-2, which illustrates that DM digestibility in the treatment with supplementation of 1% Water Spinach in L. leucocephala was 59.3% (T7) compared to the treatment (T6) with M. pigra (53.9%), and OM digestibility was 57.2% (T7) and 52.2% (T6). These can be contributed that the 1% supplementation of Water Spinach in L. leucocephala usage, which resulted in more than 5% DM digestibility compared to the utilization of M. pigra.

This result seems to be consistent with the study by Wittayakun et al. [19], which showed that DM digestibility of M. pigra (giant sensitive tree) leaves and leaves with rachis was only 46.74% and 40.81%, respectively, and the OM digestibility was about 42.17% and 40.63%, respectively. This previous result was low compared to this study; it is possibly because there was no supplementation in feeding method. Adding Water Spinach in the utilization of M. pigra and L. leucocephala may provide a protein source for cellulolytic bacteria that grow in the digestive system; they also play an important role in the breakdown of other nutrients [33]. This possibly contributes to the great nutrient digestibility in the rumen when there was a Water Spinach supplement.

Conclusion

This study on the supplementation of Water Spinach with the utilization of M. pigra and L. leucocephala leaves on in vitro fermentation has shown that M. pigra and L. leucocephala leaves are plants that are rich in protein, which is a microbial requirement for growth, digestion, and fermentation. These supplements have proven to be recyclable food supplements for ruminants to ensure a balance based on nutrient values and can increase ammonia levels without causing a decrease in pH value. Moreover, a 0.5% Water Spinach supplementation of the dietary substrate could be sufficient for ruminant growth and performance when legume forage is used as a basal diet. According to the results of this experiment, it may contribute data for further studies to investigate the growth rate and methane production from the supplementation of Water Spinach in rumen digestibility.

Authors’ Contributions

SK, KT, MS, PV, and SV: Supervised the study. CS: Conducted the experiment. SV: Drafted the manuscript. SK: Designed the study. KT: Collected the data. MS, PV, and SH: Data analysis All authors have read and approved the final manuscript.

Acknowledgments

The authors are thankful to the United States Agency for International Development (USAID) under Cooperative Agreement No. AID-OAA-L-14-00006 for funding this research. Special thanks are given to the Centre for Livestock Development Studied for allowing the research to be conducted at the forage station. Thanks are also due to the Department of Animal Sciences and Chemical Analysis of Graduated School, Royal University of Agriculture, for the use of the laboratory to analyze the samples.

Footnotes

The authors are thankful to the United States Agency for International Development (USAID) under Cooperative Agreement No. AID-OAA-L-14-00006 for funding this research

Competing Interests

The authors declare that they have no competing interests.

Publisher’s Note

Veterinary World remains neutral with regard to jurisdictional claims in published institutional affiliation.

References

  • 1.Valdivié-Navarro M, Martínez-Aguilar Y, Mesa-Fleitas O, Botello-León A, Hurtado C.B, Velázquez-Martí B. Review of Moringa oleifera as forage meal (leaves plus stems) intended for the feeding of non-ruminant animals. Anim. Feed Sci. Technol. 2020;260(2):114338. [Google Scholar]
  • 2.Moorby J.M, Fraser M.D. Review:New feeds and new feeding systems in intensive and semi-intensive forage-fed ruminant livestock systems. Animal. 2021;15(Suppl 1):100297. doi: 10.1016/j.animal.2021.100297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Witt A.B, Floyd K.S, Nunda W, Beale T, Shanungu G, Kriticos D.J. Mimosa pigra in eastern and southern Africa:Distribution and socio-ecological impacts. Austral Ecol. 2020;45(6):788–799. [Google Scholar]
  • 4.Schneider F, Tarawali S. Sustainable development goals and livestock systems. Rev. Sci. Tech. 2021;40(2):585–595. doi: 10.20506/rst.40.2.3247. [DOI] [PubMed] [Google Scholar]
  • 5.CABI. Synthesis Report of the UNEP/GEF. Removing Barriers to Invasive Alien Plants and Their Management in Africa (RBIPMA) Project, Implemented in Four African Countries (Ethiopia, Ghana, Uganda and Zambia) between 2005 and 2010. 2nd ed. Nairobi, Kenya: CABI; 2019. pp. 39–184. [Google Scholar]
  • 6.Saha H.M, Kahindi R.K, Muinga R.W. Evaluation of manure from goats fed Panicum basal diet and supplemented with Madras thorn, Leucaena or Gliricidia. Trop. Subtrop. Agroecosyst. 2008;8(3):251–257. [Google Scholar]
  • 7.Sarkar S, Mohini M, Sharma A, Tariq H, Pal R.P. Effect of supplementing Leucaena leucocephala leaves alone or in conjunction with malic acid on nutrient utilization, performance traits, and enteric methane emission in crossbred calves under tropical conditions. Trop. Anim. Health Prod. 2021;53(5):514. doi: 10.1007/s11250-021-02941-7. [DOI] [PubMed] [Google Scholar]
  • 8.Ku-Vera J.C, Ayala-Burgos A.J, Solorio-Sánchez F.J, Briceño-Poot E.G, Ruiz-González A, Piñeiro-Vázquez A.T, Barros-Rodríguez M, Soto-Aguilar A, Espinoza-Hernández J.C, Albores-Moreno S, Chay-Canul A.J. Nutritional Strategies of Animal Feed Additives. New York, United States of America: Nova Science Publishers; 2013. Tropical tree foliages and shrubs as feed additives in ruminant rations; pp. 59–76. [Google Scholar]
  • 9.Hasanah H, Achmadi J, Pangestu E, Agus A. Supply production of water spinach waste as a supplement and fermentability feed ruminant in Klaten district, Indonesia (case study at dry season 2018) J. Comput. Theor. Nanosci. 2018;17(2–3):1196–1202. [Google Scholar]
  • 10.Gatachew G, DePeters E.J, Robinson P.H, Fadel J.G. Use of an in vitro rumen gas production technique to evaluate microbial fermentation of ruminant feeds and its impact on fermentation products. J. Anim. Feed. Sci. Technol. 2005;123–124(1):547–559. [Google Scholar]
  • 11.AOAC. Official Methods of Analysis. 16th ed. Gaithersburg, MD, USA: Association of Official Analytical Chemists; 1997. [Google Scholar]
  • 12.Menke H.H, Steingass H. Estimation of the energetic feed value obtained from chemical analysis and in vitro gas production using rumen fluid. Anim. Res. Dev. 1988;28:7–55. [Google Scholar]
  • 13.Russell J.B. Rumen Microbiology and Its Role in Ruminant Nutrition. 2nd ed. Vol. 2. Ithaca, New York: Department of Microbiology. Cornell University; 2002. pp. 5–6. [Google Scholar]
  • 14.Makkar H.P, Blummel M, Becker K. Formation of complexes between polyvinyl pyrrolidones or polyethylene glycols and tannins, and their implication in gas production and true digestibility in in vitro techniques. Br. J. Nutr. 1995;73(6):897–913. doi: 10.1079/bjn19950095. [DOI] [PubMed] [Google Scholar]
  • 15.Orskov E.R, Mcdonald I. The estimation of protein degradability in the Rumen from incubation measurements weighted according to rate of passage. J. Agric. Sci. 1979;92(2):499–503. [Google Scholar]
  • 16.AOAC. Official Method of Analysis of the Association of Official Analytical Chemist W Harwitz. 18th ed. Maryland, United States of America: AOAC International; 2005. [Google Scholar]
  • 17.SAS. User's Guide:Statistic. 4th ed. Cary, North Carolina: Version. 9. SAS Institude Inc; 2013. [Google Scholar]
  • 18.Crichton N. Information point:Tukey multiple comparison test. J. Clin. Nurs. 1999;8(3):299–304. [Google Scholar]
  • 19.Wittayakun S, Innaree W, Pranamornkith P. Yield, nutrient content and rumen in vitro digestibility of giant sensitive tree (Mimosa pigra) as dairy feed. Asian J. Agric. Biol. 2017;5(4):346–351. [Google Scholar]
  • 20.Hasanah H, Achmadi J, Pangestu E, Agus A. Evaluation of water spinach (Ipomoea aquatica) as forage substitution on in vitro gas production, digestibility, and kinetic fermentation. IOP Conf. Ser. Environ. Earth Sci. 2019;346(1):012069. [Google Scholar]
  • 21.Opene A.O, Ogunka-Nnoka C.U, Uwakwe A.A. Comparative study on the nutrient composition and in vitro antioxidant properties of leaf and stems of Ipomoea involucrata. Int. J. Agric. Innov. Res. 2018;7(2):272–279. [Google Scholar]
  • 22.Makmur M, Zain M, Agustin F, Sriagtula R, Putri E.M. In vitro rumen biohydrogenation of unsaturated fatty acids in tropical grass-legume rations. Vet. World. 2020;13(4):661–668. doi: 10.14202/vetworld.2020.661-668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Truong N.B, Van Thu N. Effect of dietary neutral detergent fiber level on nutrient intake, digestibility, rumen parameters and nitrogen retention of crossbred wagyu cattle. J. Anim. Sci. Technol. 2022;10(2):316–322. [Google Scholar]
  • 24.Dijkstra J, Ellis J.L, Kebreab E, Strathe A.B, Lopez S, France J, Bannink A. Ruminal pH regulation and nutritional consequences of low pH. Anim. Feed Sci. Technol. 2012;172(1–2):22–33. [Google Scholar]
  • 25.Viennasay B, Wanapat M. Enhancing lactating dairy cow rumen fermentation and production with Flemingia silage containing phytonutrients. Livest. Sci. 2020;241:104201. [Google Scholar]
  • 26.Rigueira J.P, de Jesus N.G, Júnior V.R, Monção F.P, Costa N.M, David G.S, Silva F.V, da Cunha Siqueira Carvalho C. Effects of different banana crop wastes on nutrient intake and digestibility, microbial protein synthesis, feeding behavior, and animal performance of ¾Holstein ×Zebu heifers in a semi-arid rangeland. Trop. Anim. Health Prod. 2021;53(2):209. doi: 10.1007/s11250-021-02660-z. [DOI] [PubMed] [Google Scholar]
  • 27.Lanyasunya T.P, Wang H, Kariuki S.T, Mukisira E.A, Abdulrazak S.A, Kibitok N.K, Ondiek J.O. The potential of Commelina benghalensis as a forage for ruminants. J. Anim. Sci. Technol. 2008;144(3–4):185–195. [Google Scholar]
  • 28.Emmanuel A, Bhawat S.R, Pawar M.M, Chahuan H.D, Makwana R.B. In vitro gas production technique of evaluation of feed. Anim. Feed Sci. Technol. 2013;1:103. [Google Scholar]
  • 29.Malik P.K, Bhatta R, Ravi N, Saravanan M, Baruah L. International Grassland Congress Proceedings. India: University of Kentucky; 2020. Effect of Graded Levels of Condensed Tannin (CT) from Mimosa Pudica on in vitro Methane Production. [Google Scholar]
  • 30.Hasanah H, Achmadi J, Pangestu E, Agus A. Supply production of water spinach waste as a supplement and fermentability feed ruminant in Klaten district, Indonesia. J. Comput. Theor. Nanosci. 2020;17(2–3):1196–1202. [Google Scholar]
  • 31.Soto E.C, Yanez D.R.R, Khelil H, Molina A.E. Challenging Strategies to Promote the Sheep and Goat Sector in the Current Global Context. Zaragoza: CIHEAM/CSIC/Universidad de Leon/FAO; 2011. An in vitro evaluation of the potential use of greenhouse wastes to replace barley in goats'diets; pp. 285–291. [Google Scholar]
  • 32.Saleem A.M, Ribeiro G.O, Jr, Yang W.Z, Ran T, Beauchemin K.A, McGeough E.J, Ominski K.H, Okine E.K, McAllister T.A. Effect of engineered biocarbon on rumen fermentation, microbial protein synthesis, and methane production in an artificial rumen (RUSITEC) fed a high forage diet. J. Anim. Sci. 2018;96(8):3121–3130. doi: 10.1093/jas/sky204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Khafipour E, Li S, Tun H.M, Derakhshani H, Moossavi S, Plaizier J.C. Effects of grain feeding on microbiota in the digestive tract of cattle. Anim. Front. 2016;6(2):13–19. [Google Scholar]

Articles from Veterinary World are provided here courtesy of Veterinary World

RESOURCES