Evaluation of nutritive traits and in vitro production of gas for Alfalfa hays and Fenugreek (Trigonella foenum-graecum) on feeding ruminants

Introduction

Belonged to the family (Fabaceae), the annual legume Fenugreek (Trigonella foenum-graecum) originated in the Mediterranean region. Fenugreek, as an additive, has many nutritional and medicinal values for humans and animals, and this trigonelline is a valuable alkaloid with practical therapeutic effects, especially for diabetes. Iranian fenugreek of Zanjani or Borazjani type is the best type for cultivation and forage production. This plant has a high nutritional value and contains valuable substances such as calcium, phosphorus, iron, carotene, vitamin C, and crud protein. Some different species of fenugreek have been introduced as a forage plant with good nutritional quality for ruminants, which, unlike alfalfa, does not cause bloating in livestock. In addition to that, it has natural steroid compounds used to increase livestock growth (Mir et al., 1997; Mir et al., 1998) [11,12] . In some regions, traditional ranchers were aware of the fattening potential of fenugreek, and they added fenugreek to their fodder before selling it (Azad, 2000) [5] . Fenugreek is cultivated as a spice in India and most Mediterranean regions. The Latin name of this plant is "Foenum-graecum," which means "Greek forage," identifying the utilized of it as a forage (Acharya et al., 2006) [1] . As reported, the fenugreek reduces the growth of the rumen protozoal population by up to 50% (Goel et al., 2008a) [7] . Also, previous studies have shown that several saponin-containing plants inhibited the growth of ruminal protozoa (Diaz et al., 1994; Newbold et al., 1997) [6,13] . Fenugreek saponins can improve fermentation efficiency (Goel et al., 2008b; Klita, 1996) [8,10] . There is not enough scientific information about the effect of adding fenugreek range forage on ruminant nutrition. This forage is produced in summer more than human consumption and is discarded in large fenugreek cultivation farms as well as Fruits and Vegetable Markets. Considering the high nutritional value of this forage and the lack of scientific information about the nutritional value of this additive in Iran, this study was conducted .Therefore, this study aimed to do a nutritional evaluation of the Iranian fenugreek in the diet of ruminants. www.extensionjournal.com Protein (CP) was calculated as 6.25×N. The Natural Detergent Fiber (NDF), Acid Detergent Fiber (ADF), Acid Detergent Lignin(ADL), and Crude Fiber(CF) were determined by the Fibertec system with modifications according to Van Soest (1967) [22] ; sulfite was omitted from the NDF analysis. Ether Extract (EE) was measured using a Soxhlet extractor. -Non-Fiber Carbohydrate (NFC) and Organic Matter (OM) were calculated as (100-CP-EE-Ash-NDF) and (100-Ash), respectively. In an adiabatic bomb calorimeter, gross energy (GE) was determined.

Animals' management:

The rumen fluid was collected from four mature fistulated Taleshi male cows (Safaei et al., 2007) [20] of Iran (age= 4 years; weighted = 415 kg), and animals were fed daily twice, with (DMIntake= 8 kg/day) diet contained: [16] .

Production of gas -In vitro

In the lab of the Institute of Animal Science Research\Karaj-Iran, the measurements of the production of gas -In vitrowere carried out. Fermentation of FTG samples was carried out with rumen liquor collected from 4 fistulated Taleshi male cows of Iran (Menke & Steingass, 1988) [15] . The inoculum was formulated as outlined by Menk and Steingass (1988). It was made by combining rumen liquor with artificial saliva at a ratio of 1:4 (v/v). For a total volume of 1L, the artificial saliva included a solution of buffer (237 ml), solution of main element (237 ml), solution of trace element (0.12 ml), solution of resazurin (1. 22 ml) (which was resazurin "100 mg" dissolved in distilled water "100 ml"), and 49. 5 ml of a reduction solution (this was freshly prepared separately and comprised 2 ml of 1 N NaOH, Na2S-7H2O (285 mg), and distilled water (47. 5 ml) to make 1 L of saliva, topped up with distilled water (475 ml). The buffer solution contained NaHCO3 at 35 g and NH4HCO3 at 4 g. The solution of main element contained Na2HPO4 at 5. 70g, KH2PO4 at 6. 20 g, and Mg SO4-7H2O at 0. 60 g the solution of trace element included CaCl2-2H2O at 13. 20 g, MnCl2-4H2O at 10. 00 g, CoCl2-6H2O at 1. 00 g, and FeCl2-6H2O at 0. 80 g, diluted to 100 ml with distilled water. After the reduction process was completed (indicated by the decoloration of resazurin upon adding the reduction solution), the rumen liquor was added to the medium. All procedures were conducted with continuous CO2 reflux. Around 200 mg of ground TFG samples were measured into 100 ml glass syringes. A volume of 30 ml of the fluid buffer mixture was placed into the 100 ml glass syringes. The glass syringes, which contained the TFG samples and rumen fluid buffer mixture, were kept at an incubation temperature of 39°C. After 30 minutes of incubation, each syringe was shaken gently. The production of gas has been measured at the incubation times 2, 4, 6, 8, 12, 24, 48, 72, and 96 h. In triplicate, treatment samples were incubated, in addition to three blank syringes, which only contained a rumen liquor buffer mixture. By subtracting the volume of blanks' produced gas, the production of gas for FTG samples was determined (Menke & Steingass, 1988) [15] . The data of the production of gas was fitted to the model given by Ørskov and McDonald (1979) [17] by the software Fit Curve (newway).

Where is the gas production from the immediate soluble fraction (ml); b = the gas production from the immediately insoluble fraction (ml); c = the gas production rate constant for the insoluble fraction (ml.h -1 ); a + b = the potential gas production (ml); t = the incubation time (h), and Y is the gas production at time t. The contents of ME (MJ kg -1 DM) of TFG samples were calculated by using the following equation given by Menke and Steingass (1988) [15]: ME (MJ kg-1 DM) = 2.20 + 0.136 GP + 0.057 CP Where GP = the 24-hour production of gas (ml 200 -1 mg), and CP = the crude protein (%).

The digestibility of Dry Matter of TFG samples was calculated by using the following equation given by Menke and Steingass (1988) [15]: DMD (%) = 15 + 10.1 (a+b) + 623(c) + 0.51 CP Where a + b =potential gas production (ml); c = the constant gas production rate for the insoluble fraction (ml.h -1 ); CP =crude protein from the TFG sample (%). The Digestibility of organic matter of TFG samples was calculated by using the following equation given by Menke and Steingass (1988) [15]: OMD (%) = 14.88 + 0.889 GP + 0.45 CP + 0.0651 XA (4) Where GP = the 24-hour net gas production (ml 200-1 mg DM), CP = the crude protein from the TFG sample (%), and XA = the ash content from the TFG sample (%). Short-chain fatty acid (SCFA) was calculated using the equation of Makkar (1995) and Menke et al. (1997) [14] ; SCFA (mmol) = 0.0222GP -0.00425 (5) Where GP = the 24-h production of gas (ml g -1 DM). NE (MJ/kg DM) = 0.115*GP + 0.0054*CP + 0.014*EE -0.0054*CA _ 0.36 Where GP =the 24-hour production of gas (ml/200mg DM), CP=crude protein, EE=ether extract, and CA=crude ash (%DM).

Statistical analysis

The treatments were TFG and ALF, with three replications in randomization, which were designed entirely. By using the software Statistical Analysis Systems of Virgin 9.1proc, all data were analyzed. GLM and a, b, and c (parameters of the gas test) were obtained by Fit Curve (new-way) software (Ørskov & McDonald, 1979; R, 2015) [17] . www.extensionjournal.com

Results and Discussion

The chemical compositions of the two forage species (TFG, ALF) are presented in Table 1 and Fig. 1. The fenugreek hay was significantly (p<0.01) higher in CP, Ash, EE, NFC, and GE than ALF, n but NDF, ADF, ADL, and OM of TFG were less than ALF hay (p<0.01). [9] .

Table 2 and Fig. 2 show gas production data during the fermentation period. The cumulative volume of gas production increased with increasing incubation time. Even though the availability of several models for estimating the kinetic production of gas, but, the method of Ørskov and McDonald (1979) [17] has been chosen, because of the relationship of its parameters with the intake, the digestibility, and the degradation characteristics of the forages. With those reported by Kamlak et al. (2005) [9] , cumulative gas production and estimated parameters in ALF hay were comparable.

Fig 2:

Gas production of forages The estimated parameters (b and a + b) of TFG hay, therefore, were higher -significantly (p<0.01)-than those of ALF, but c of TFG was lower (p<0.01). The DMD, OMD, ME, and NE contents of TFG hay were higher -significantly (p<0.01)-than those of ALF hay. [7] tested the fenugreek seeds effect and their aqueous and alcoholic/aqueous extracts on the production of methane and other fermentation products in all-forage diets as well as forage and concentrate mixtures in the rumen. The researchers concluded that supplementing fenugreek seeds with all-forage diets or concentrate-forage mix diets, both induced the degradability of OM in the rumen and reduced methane production per decomposed material. Therefore, it has been suggested that this supplement may improve ruminal fermentation efficiency. In Mir et al. (1997) [11] study, total volatile fatty acids, ruminal pH, and ruminal ammonia concentration were not significantly different in castrated calves fed fenugreek silage and alfalfa silage. The in vitro gas production rate for fenugreek silage was significantly higher than that for alfalfa silage (17.4 vs. 12.6% per hour). However, the final gas production after 72 hours was no different for the two silages (22. . Estrogens of foreign origin generally increase the storage of lean tissue in castrated ruminants and reduce subcutaneous and intramuscular fat. Phytoestrogens have been shown to have beneficial effects in stimulating growth and increasing growth rate in the diet of ruminants and nonruminants (Trenkle & Borroughs, 1978) [21] . Petit et al. (1995) [18] reported that fenugreek seed steroid saponins increase feed intake and weight gain in mice. Mir et al. (1998) [12] investigated the effect of dietary feeding of fenugreek silage and alfalfa silage with barley supplement (at three levels of 0, 15, and 30%) balanced by adding soybean meal for the same nitrogen level on the growth performance of male calves. In this study, feed intake, mean daily weight gain, and feed intake in fenugreek treatment were similar to those in alfalfa treatment. The researchers suggested that the nutritional value of fenugreek silage in ruminant growth was comparable to that of alfalfa silage .

On the other hand, Al-Amer and Basiouni (2005) [3] reported that growth hormone levels in goats treated with 60 g of fenugreek seeds per day for 7 weeks were higher than the control treatment.

Conclusion

Finally, the value of TFG hay -nutritionally-was higher than that of ALF hay, because, it contained more CP, OMD, and ME that were used to feed ruminants and reduce lost cost feeding.

Acknowledgement

AcknowledgmentsThe Animal Science Research Institute carried out this research, and the authors are grateful to the institute's laboratories in Karaj, Iran.

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