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Animal Feed Science and
Technology
journal homepage:www.elsevier.com/locate/anifeedsci
Defatted
Moringa oleifera
seed meal as a feed additive
for sheep
H. Ben Salem
a,∗, H.P.S. Makkar
baInstitut National de la Recherche Agronomique de Tunisie (INRAT), Laboratoire des Productions Animales et Fourragères,
rue Hédi Karray, 2049 Ariana, Tunisia
bInstitute for Animal Production in the Tropics and Subtropics (480b), University of Hohenheim, D-70593 Stuttgart, Germany
a r t i c l e
i n f o
Article history:
Received 14 September 2007 Received in revised form 15 July 2008 Accepted 24 July 2008
The objective was to evaluate use of defattedMoringa oleiferaseed meal (DMM) as an additive in sheep diets. Effects of DMM on feed intake, diet digestibility, microbial N supply, blood metabolites and growth of lambs fed an oat-vetch hay based diet were evaluated using 24 Barbarine lambs in four groups. All lambs were fed hayad libitum,and 100 g of soyabean meal (SBM) mixed with 0 (Control), 2 (Low), 4 (Medium) or 6 (High) g DM of DMM per day. A 45-day feed-ing study followed by a 6-day digestion study was completed. The DMM was higher (P<0.001) in crude protein (592 g/kg DMversus 471 g/kg DM) and lower (P<0.001) in NDFom (105 g/kg DMversus 169 g/kg DM) than SBM. Feeding DMM had no effect on hay intake, diet digestibility or N balance. Lambs grew at rates of 63.8, 88.5, 97.0 and 76.6 g/day, respectively (Q;P=0.076), possibly due the trend to an increase in the microbial N supply (Q;P=0.109) and the trend (Q;P=0.086) to higher N retention with higher levels of DMM in the diet. The highest daily gains tended (Q;P=0.076) to be highest in the lambs fed the intermediate levels of DMM. Results suggest that DMM has the potential to improve rumen fermentation, the medium levels are recommended as an additive to improve the growth rate of lambs fed hay-SBM diets.
© 2008 Elsevier B.V. All rights reserved.
Abbreviations: CP, crude protein; DM, dry matter; DMM, defatted Moringa seed meal; DOMi, digestible OM intake; EMNS, efficiency of microbial N supply; LW, live weight; NDFom, ash-free neutral detergent fibre; OM, organic matter; SBM, soyabean meal.
∗Corresponding author. Tel.: +216 71 230 024; fax: +216 71 231 592. E-mail address:bensalem.hichem@iresa.agrinet.tn(H. Ben Salem).
1. Introduction
Moringa oleifera Lam. (syn. Moringa pterygosperma Gaert.) is native to the western and sub-Himalayan parts of northwest India, Pakistan and Afghanistan. It is now widely cultivated across Africa (e.g., Nigeria, Senegal, Tanzania), South America and South-east Asia (e.g., Malaysia, Indonesia). Almost every part of this plant has value as food or feed. For example Moringa leaves are exceptionally rich in pro-vitamin A, vitamins B and C, Fe and several amino acids, and their consumption by children is recommended to overcome malnutrition in some countries (Fuglie, 2001). The beneficial effects of Moringa leaves to human and ruminant diets are well documented (e.g.,Foidl et al., 2001; Makkar et al., 2007). The seeds are considered to be antipyretic, acrid and bitter. The extracted oil, known commercially as “Ben” or “Behen” oil, has been used for illumination, and is particularly suitable as a lubricant. The fatty acid composition of Moringa oil is quite similar to olive oil in its contents of C18:1 and C18:0 (Anwar and Bhanger, 2003), which explains its potential as an edible oil. The cake remaining after oil extraction has been shown to retain the active ingredients for coagulation of various undesir-able moieties from a solution, making it a marketundesir-able commodity (Folkard and Sutherland, 1996). For example, Moringa press cake is used for water purification instead of common chemical coagulants such as aluminium sulphate. Proteins in the cake have a high positive charge (Folkard et al., 2001) and an antibiotic effect (Makkar et al., 2007) and have the potential to modify rumen fermentation. These proteins have also been shown to decrease degradability of feed proteins in anin vitrorumen system (Hoffmann et al., 2003; Makkar et al., 2007), and thus could enhance the post ruminal protein supply. Defatted seed cake is free of most plant secondary metabolites such as tannins, saponins, alka-loids, inhibitors of trypsin and amylase, lectin and cyanogenic glucosides, but contains glucosinolates (Makkar and Becker, 1997).
This study was conducted to evaluate effects of feeding defatted Moringa seed meal (DMM) on nutrient utilization, microbial protein supply to the intestine and growth performance in sheep.
2. Materials and methods
2.1. Preparation of defatted Moringa meal
Seeds ofMoringa oleiferaLamarck were purchased from India (Veg India Exports, Erode, India). They were packaged in plastic bags (0.5 kg Moringa seeds/bag), then shipped to Tunisia in cartoons. On arrival, the seeds were stored at 4◦C until used. Three weeks prior to the commencement of the
current experiment, seeds were shelled by hand to obtain kernels and shells. The shells were discarded and the kernels from each bag were ground to pass a 1-mm screen, then extracted with petroleum ether at 40–60◦C in a Soxhlet apparatus. Oil content averaged 380 g/kg kernels. The fat-free Moringa
seed meal, referred to as DMM, was used as a feed. Four samples of DMM from seeds in four different bags were conserved at 4◦C until analysed.
2.2. Animal, diet and treatments
The study was conducted in March–May 2006 at the experimental sheepfold of the National Insti-tute of Agricultural Research of Tunisia (INRAT) in Tunis, at 36.50N and 10.11E and an altitude of 4 m above mean sea level.
determination, all feed refusals and faeces from each lamb were weighed and mixed thoroughly and 0.1 of each refusal and faeces were collected and conserved, or frozen, respectively. When the 6-day col-lection was completed, feed refusals and faecal samples from each lamb were thawed (for faeces) and then mixed thoroughly to create one homogenous mixture of feed refusals and faeces for each lamb. Composited samples of refusals, and faeces and representative samples of feeds (n= 4), were dried at 50◦C, ground to pass a 1-mm screen, and then preserved for laboratory analysis. Urine excreted daily
by each lamb was collected in plastic vessels containing 100 ml of 0.1 N sulphuric acid to keep the pH below 3. After recording individual urine volume, 0.1 of the urine voided by each lamb was frozen. At the end of the collection period, urine samples were pooled by lamb and 30 ml of urine from each were stored at−15◦C until analysed. Each lamb received, immediately after feeding the morning meal,
clean water in a 10 l plastic bucket. Volumes of drinking water provided (1.5 of the volume consumed the previous day), and that remaining in the bucket the following day before feeding, were measured to determine water consumed.
On day 30 of the growth phase, and the last day of total collection, blood was withdrawn from the jugular vein of the lambs 1 h before feeding the morning meal. Samples were collected in 10 ml vacutainer tubes containing EDTA to prevent clotting, and then centrifuged at 3000×gfor 20 min at 4◦C. Plasma was decanted and frozen (−20◦C) until analysed.
2.3. Laboratory analyses and calculations
Feed-offered and -refused, and faeces, were analysed for DM by drying at 103◦C for 16 h in a forced
air oven, and for ash, ether extract and N according to methods 942.05, 920.39 and 988.05, respectively, ofAOAC (1990). These samples were also analysed (Van Soest et al., 1991) for ash-free neutral detergent fibre (NDFom) using procedures modified for use in an Ankom 200 Fibre Analyzer (ANKOM Technology, Fairport, NY, USA). Sodium sulphite and␣-amylase were not used in the NDF analysis. Concentrations of
Ca, K, Na, Mg, Zn, Cu, Mn and Fe of feeds were quantified using an atomic absorption spectrophotometer (AOAC, 1990, method 968.08), and the P concentration of feeds was determined colorimetrically (AOAC, 1990, method 965.17).
Rumen fluid was obtained from two ruminally cannulated Barbarine rams (average live weight 45±1.5 kg) before the morning meal, filtered through two layers of cheese cloth and pooled to form one rumen fluid sample for the gas production method. Rams were housed indoors and received a diet consisting of oaten hayad libitumand 300 g of concentrate (barley, 0.8 kg/kg and soyabean meal, 0.2 kg/kg) to fulfil maintenance requirements. Rumen fluid was freshly sampled for the two runs of incubation. It was collected with a manual pump, and transferred into pre-warmed thermos flasks. Feed samples (n= 4) were weighed in triplicate (0.2 g) into glass syringes before buffered rumen fluid was added (30 ml), andin vitroOM digestibility (OMD) and metabolisable energy (ME) were estimated using equations ofMenke and Steingass (1988)as
OMD (g/kg DM)=14.88+0.889×Gp+0.45×CP+0.0651×XA
ME (MJ/kg DM)=2.20+0.136Gp+0.0057×CP+0.0002859×CP2
where CP is crude protein in g/100 g DM, XA is ash in g/100 g DM and Gp is the net gas production (ml) from 200 mg of substrate after 24 h of incubation.
Apparent digestibility of DM, OM, CP and NDFom was calculated from dietary intakes of each constituent and the amount recovered in faeces.
Table 1
Chemical composition and estimated metabolizable energy (ME) and organic matter digestibility (OMD) of feeds (n= 4)
Oat-vetch haya Protein seed meals
Soyabean Moringa S.E.b
Organic matter (g/kg DM) 930 (6) 904 942 4.2*** Crude protein (g/kg DM) 101 (3) 471 592 4.0*** Ether extract (g/kg DM) 30 (3) 24 3.5 1.2*** Neutral detergent fibre (NDFom, g/kg DM) 586 (10) 169 105 2.6*** Ca (g/kg DM) 13.7 (0.4) 6.7 3.8 0.33*** Fe (mg/kg DM) 408.3 (7.7) 231.6 66.5 1.02*** ME (MJ/kg DM) 7.02 (0.3) 9.11 7.61 0.37* OMD (g/kg DM) 546 (6.8) 681 598 3.84***
aValues in parentheses are S.D.
b For protein meals only. Standard error followed with the significance level. ***P<0.001; **P<0.01; *P<0.05; ns,P>0.05.
microbes and 1000 converts mg to g. The efficiency of microbial-N supply (EMNS) was calculated as
EMNS= microbial N (g)
DOM intake (kg)
Plasma samples were analysed for glucose, Ca and total protein using Biomaghreb kits (Soukra, Tunisia) and an UV–Vis spectrophotometer (Spectronic 601, Milton Roy Company, Syracuse, New York, USA).
Table 2
Feed and water intakes,in vivodigestibility of diets, and growth rate of lambs fed increasing levels of defatted Moringa seed meal
Diets S.E. Contrasts
Control Low DMM Medium DMM High DMM Linear Quadratic DM intake (g/day)
Hay 952 975 1093 1063 81.4 ns ns
Soyabean meal 96 96 96 96
DMM 0 2 4 6
Total 1048 1073 1192 1164 81.4 ns ns
ME intake (MJ/day) 7.55 7.73 8.57 8.38 0.572 ns ns
Water intake (l/day) 3.4 3.7 4.1 4.0 0.24 ns ns
Apparent digestibility of diet (g/kg)
DM 660 647 651 674 13.0 ns ns
OM 679 666 668 696 12.3 ns ns
CP 668 653 635 638 22.9 ns ns
NDFom 599 611 609 648 11.7 0.014 ns
Live weight (kg)
Initial LW 27.5 26.2 28.4 27.7 2.06 ns ns
Average daily gain (g/day) 64 88 97 77 11.9 ns 0.076
Gain/feed 0.082 0.100 0.101 0.084 0.0105 ns ns
Table 3
Nitrogen balance and microbial N supply in sheep fed increasing levels of defatted Moringa meal
Diets S.E. Contrasts
Control Low DMM Medium DMM High DMM Linear Quadratic
N intake (g/day) 24.2 25.9 27.3 26.7 1.10 ns ns
Faecal N (g/day) 8.1 8.9 10.0 9.6 0.74 ns ns
Urinary N (g/day) 6.7 5.3 4.7 5.8 0.49 ns 0.023
N retention (g/day) 9.5 11.6 12.5 11.3 0.84 ns 0.086
g/kg N intake 392 449 461 420 27.2 ns ns
Urinary allantoin (mg/kg LW0.75) 45 46 58 38 8.3 ns ns
EMNS supply (g/kg DOMi)a 4.57 4.84 6.82 3.74 0.989 ns ns
ns,P>0.05.
aEMNS, efficiency of microbial N supply.
2.4. Statistical analysis
Nutrient and ME contents and OMD of SBM and DMM were analysed by ANOVA method using the GLM procedures ofSAS (1991). Treatment means were compared by the least significant difference method (LSD).
Data from thein vivoexperiment were subjected to analysis of variance using the GLM procedure ofSAS (1991)according to the model:
Yi=+DMMi+ei
whereYi: dependent variable;: general mean; DMMi: level of DMM (i= 1–4);ei: residual error.
Significance of differences between dietary treatments was determined using single degree of free-dom linear and polynomic contrasts withinSAS (1991)to determine effects of DMM levels incorporated in the diets. Significance was accepted ifP<0.05, and a tendency to a difference was accepted ifP<0.10.
3. Results
The DMM was higher (P<0.001) in CP, P and Mg, but lower (P<0.001) in NDFom, ether extract, Ca, Na, Mn, Zn and Fe than soyabean meal (Table 1). The ME and OMD were higher (P<0.05 andP<0.001, respectively) in soyabean meal than in DMM.
Increasing DMM addition had no impact on hay, ME and water intakes, or apparent digestibilities of DM, OM and CP (Table 2). However, lambs had a higher digestibility of NDFom (L;P=0.014) as the feeding level of DMM increased. The average daily gain of lambs fed the diet containing medium level of DMM tended to be higher than those fed the other diets (Q;P=0.076), but the gain to feed ratio was not affected.
Nitrogen balance was positive with all dietary treatments (Table 3), and enrichment of diets with increasing levels of the DMM tended to increase N retention in lambs fed the diet containing medium level of DMM (Q;P=0.086), primarily due to an equivalent reduction in urinary N output. Similarly, urinary excretion of allantoin and the microbial N supply increased, albeit non-significantly (Q;P=0.236 andP=0.109, respectively).
Table 4
Plasma metabolites in sheep receiving increasing levels of defatted Moringa meal
Diets S.E. Contrasts
Control Low DMM Medium DMM High DMM Linear Quadratic Plasma metabolites
Ca (mmol/l) 7.7 9.9 8.7 8.6 0.54 ns 0.051
Glucose (g/l) 0.70 0.84 0.81 0. 80 0.039 ns 0.058
Total proteins (g/l) 73 73 73 73 1.06 ns ns
DMM addition had no effect on concentrations of plasma proteins (Table 4), but levels of calcium (Q;P=0.051) and glucose (Q;P=0.058) were higher at medium feeding levels of DMM.
4. Discussion
The main finding from this study was that progressive DMM addition to the diet caused an overall quadratic impact on average daily gain of lambs with a progressive increase up to 4 g/day of DMM, and thereafter decrease at 6 g/day. Maximum growth at 4 g/day is supported by N retained and efficiency of microbial-N production data, which were highest at this level. The small shift of N excretion from urine to faeces at 4 g/day of DMM incorporation could assist in making crop-livestock systems more efficient since soil and the crops benefit more from faecal N than N voided by ruminants through urine (Delve et al., 2001). Additionally, this will reduce the likelihood of N leechate entering ground water.
In addition to the improvement of the protein value of the diet at 4 g/day of DMM incorporation in the diet, lambs tended to have higher plasma glucose level.Annison et al. (2002)reported a linear relation between glucose entry rate and ME intake such that the entry rate increased by about 10 g glucose per day/MJ of ME. This suggests that 4 g/day level of DMM in the diet increased the energy value of the diet, which may be supported by the highest gain/feed ratio obtained using this diet. This may have been at least partly due to changed rumen microbial population, and more efficient rumen fermentation. These changes could also be due to protection of feed proteins from rumen degradation, possibly due to the interaction of cationic proteins in DMM to the rumen microbes, thereby making them available in the intestine (Hoffmann et al., 2003). Although DMM is high in protein, (Makkar and Becker, 1997), the decreased daily gain of lambs receiving 6 g of DMM could be due to the cationic nature of its proteins that appear to elicit more-than-required antibacterial response (Makkar et al., 2007), and/or due to the bitter taste resulting from glucosinolates in DMM (Makkar and Becker, 1997). Most bioactive moieties are known to produce quadratic responses similar to that obtained here for DMM (Makkar et al., 2007). Results suggest that future work should explore the positive effects of DMM use as an additive, possibly due to its cationic proteins having antibacterial properties. It is possible that DMM response is similar to that of monensin and other antibiotic additive and could be used to positively manipulate rumen fermentation.
5. Conclusions
This is the first study on ruminant response to feeding of DMM in lamb diets and our findings would orientate future investigations on the use of DMM in livestock feeding. Our results suggest that DMM, incorporated at levels up to 4 g/day, had positive effects on rumen fermentation, digestion and performance. The apparent reduction in overall performance of the lambs at a feeding level of only 6 g/day could possibly be due to the presence of glucosinolates. In addition, the concentration of the active moiety (cationic proteins) at 6 g/day of DMM incorporation in the feed could be higher than the concentration required for eliciting the optimal response.
Acknowledgements
Partial financial support by the International Atomic Energy Agency (IAEA) is acknowledged. This study was completed under the framework of the technical cooperation project funded by IAEA and the Ministry of High Education, Scientific Research and Technology of Tunisia (INRAT-IAEA, TUN 5/021).
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