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The Use of Faba Bean and Sweet Lupin Seeds Alone or in Combination for Growing Lambs. 1. Effects on Growth Performance, Carcass Traits, and Blood Parameters.

Byline: Antonia Lestingi, Francesco Toteda, Arcangelo Vicenti, Davide De Marzo and Anna Maria Facciolongo

Abstract

The study investigates faba bean and lupin seeds when used alone or in combination as protein supplements for growing lambs, and their effects on growth, carcass characteristics, and hematological traits. Twenty- four Gentile di Puglia male lambs were weaned at 38 2 days of age, and divided into three homogeneous groups, based on body weight (BW). For eight weeks before slaughter, the lambs were assigned to one of three isoenergetic and isonitrogenous dietary treatments, differing in the protein supplement: a faba bean diet (FB) containing 300 g/kg diet (on as fed basis) of faba bean seeds; a faba bean plus lupin diet (FB+L) containing 150 g/kg diet of faba bean seeds plus 150 g/kg diet of lupin seeds (both on as fed basis); a lupin diet (L) containing 250 g/kg diet of lupin seeds (on as fed basis). Individual body weights and feed intakes were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio.

In the 1st and 8th weeks of the trial, blood samples were taken in order to measure glucose, total cholesterol, NEFA, creatinine, urea and total protein concentration and respective electrophoresis fractions. Carcass traits and tissue composition of leg and loin were determined. Slaughter body weight was lower for lambs fed on diets containing lupin than for lambs fed on diets containing faba beans. In Group L, ADG was lower than FB and FB+L groups. Group L lambs had lower ADFI than the other two groups. For Group FB lambs, loin weight was a higher percentage of half carcass weight than for Groups L and FB+L lambs. Leg and loin dissection data showed Group L lambs to have a greater percentage of bone in the leg than Group FB+L lambs. Group L lambs had higher concentrations of urea than the other two groups at the end of the trial. The use of lupin alone was associated with lower growth performances whereas, when lupin is given with faba bean the results suggest positive effects.

Keywords: lamb, protein supplement, growth performance, carcass traits, blood parameters

INTRODUCTION

The advantage of using cereal-based diets in intensive feeding of growing lambs is that they give better animal production performances than forage-only diets, although cereal-based diets generally require additional protein supplementation to meet the animal's nutritional requirements (Jacques et al., 2011). The high protein content (42- 54%) of imported soybeans, together with their excellent balance of amino acids and high energy levels, make them Europe's predominant protein supplement in commercial livestock rations (Turner et al., 2012). However, consumers are beginning to distrust the widespread use of genetically modified soybeans, and moreover high demand, rapid price increases and recent environmental concerns have increased interest in using locally-produced legumes in animal rations (Vasta et al., 2008).

Homegrown legume seeds have been used primarily for their high protein and starch contents, despite the presence of some anti-nutritional factors (Masoero et al., 2005) that may limit their use.

In Mediterranean environments, faba beans and lupin seeds are potential alternative to soybean meal in organic breeding systems (Masoero et al., 2005; Facciolongo et al., 2014); moreover they are legumes suitable for rotation with winter wheat. Faba bean (Vicia faba L.) is primarily used in animal feeding as grain, and to a lesser degree as fodder. Faba grain is a good source of starch (450- 500 g kg-1 dry matter) and protein (230-300 g kg-1 dry matter) with a well balanced content of amino acids (Ortiz et al., 1993). The use of a lamb fattening diet largely based on faba bean gave growth performances and meat characteristics similar to the most frequently used diets using soybean meal as their main protein source (Antongiovanni et al., 2002; Lanza et al., 2011).

Lupin seed is a legume with high crude protein (CP, 35-40%) content, which has a high degradability (RDP 665 g/kg of CP), and contains both low starch content and high content of non- starch polysaccharides (van Barneveld, 1999). The seeds of traditional varieties contain substantial amounts of alkaloids (5-20 g kg-1 DM; Guillaume et al., 1987) that can reduce intakes (Kung et al., 1991), but newer varieties of sweet lupin have low alkaloid contents (less than 0.05% DM), especially sweet white lupin (Lupinus albus L.). They also appear to be free of other anti-nutritional factors such as lecithin, antitrypsin and hemaglutinin (Guillaume et al., 1987). However, the high ruminal degradability of protein make their use difficult as replacements for soybean meals in the ruminant rations (INRA, 1988), but this issue is controversial (Guillaume et al., 1987; Tracy et al., 1988; Kung et al., 1991; Murphy and McNiven, 1994; Stanford et al., 1996; Vicenti et al., 2009).

Sweet lupin and faba bean differ in their chemical composition, in rumen degradability (Masoero et al., 2005) and in the amino acid composition (Degussa, 2006) of their proteins, and this may be reflected in animal performances. It is possible that combined use of the two legumes in animal feed could achieve a better balance and thus improve animal performances. Moreover, we are not aware of reports on the effect of the use of faba beans or lupin on lamb blood parameters, some of which are important indicators of the animal's nutritional state and welfare and may be useful to identify metabolic imbalances or disorders. Therefore, this study aimed to use faba bean and lupin seeds as protein supplements, both alone and in combination, and to evaluate their effects on the growth performances, carcass characteristics, and hematological parameters of growing lambs.

MATERIALS AND METHODS

Animals and diets

All procedures involving animals were conducted according to the Italian government guidelines (Directive 91/629/EEC, received in Italy by D.L. 533/92 and modified by D.L. 331/98). The study was conducted in a farm in southern Italy (latitude: 415'54"24 N; longitude: 1646'43"68 E) at 50 m above sea level, during 8 weeks, from December 2011 to February 2012. Twenty-four Gentile di Puglia male lambs, weaned at 382 days of age and at an average initial body weight (BW) of 15.80.5 kg (mean standard error), were divided into three homogenous groups for BW and age. They were housed in individual pens (1 m2 per head) with continuous access to water, and the temperature in the pens ranged from 7C to 15C. Lambs were assigned to one of three dietary treatments: FB group received 300 g/kg diet (on as fed basis) of faba bean seeds; FB+L group received 150 g/kg diet of faba bean seeds plus 150 g/kg diet of lupin seeds; L group received 250 g/kg diet of lupin seeds.

The three pelleted total mixed rations (PTMR) (Table I) were formulated to be isocaloric and isonitrogenous to meet the nutritional requirements of lambs (INRA, 1988). Lambs were adapted to the ration during 10 days. Feed was offered daily at 08:00 h at a rate of 110% of ad libitum intake calculated by weighing-back refusal weekly. Feed samples were taken weekly and stored at -20C until analysis. Straw was offered on the rack as a source of roughage. Straw intake was very low and was not recorded. Individual body weights and feed intakes were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR).

Slaughtering and carcass traits

At 98 days of age the lambs were slaughtered by exsanguination according to veterinary police rules (D.P.R. 320/54) after fasting for 12 h and recording of slaughter body weights. Hot carcass, skin, fleece, head, pluck and full and empty gastrointestinal tract (GIT) were weighed. Carcasses were then hung by the Achilles tendon and chilled at 4C (80-82% relative humidity) for 24 h and reweighed. The weight of digestive content (full-empty GIT) was used to calculate the net dressing percentage (carcass weight after chilling/empty body weight). The cold carcasses were then divided into two symmetrical sides. The right side was separated into cuts (neck, steak, brisket, shoulder, abdominal region, loin, leg) according to ASPA methods (1996). Loins and legs were dissected into tissue components (lean, separable fat and bone).

Table I.- Ingredient and chemical composition of the experimental diets.

Item###Diets-1

###FB###FB+L###L

Ingredient composition, % (as-fed basis)

Barley###20.0###20.0###20.0

Corn###20.0###20.0###20.0

Corn gluten###2.2###-###-

Oats###12.0###14.0###18.0

Faba bean###30.0###15.0###-

Lupin###-###15.0###25.0

Straw###10.0###10.0###10.0

Wheat bran###2.0###2.2###3.2

Calcium carbonate###1.8###1.8###1.8

Dicalcium phosphate###1.0###1.0###1.0

NaCl###0.5###0.5###0.5

Vitamins###0.5###0.5###0.5

Chemical composition (dry matter basis)

Moisture, % (as fed)###13.6###13.7###13.7

CP, %###14.9###15.4###15.7

Ether extract, %###3.2###3.4###4.0

Crude fiber, %###9.2###9.5###9.9

Ash, %###4.6###4.4###4.0

NDF, %###21.9###22.7###23.8

ADF, %###10.9###11.7###12.4

ADL, %###2.0###2.2###2.4

PDIN2 (g/kg dry matter)###103.0###102.0###101.0

PDIE3 (g/kg dry matter)###101.0###100.2###99.0

Meat Forage Units (n/kg DM)###0.90###0.92###0.93

Feed chemical composition

Samples of each PTMR were ground in a hammer mill with a 1 mm screen and analysed using the following AOAC procedures (2004): dry matter (DM) (Method 934.01), ether extract (EE) (Method 920.39), ash (Method 942.05), crude protein (CP) (method 954.01), crude fibre (CF) (Method 945.18), acid detergent fibre (ADF) and acid detergent lignin (ADL) (Method 973.18), and amylase-treated neutral detergent fibre (NDF) (Method 2002.04). The protein value (PDIE: protein digested in the small intestine allowed by the energy, and PDIN: protein digested in the small intestine allowed by the nitrogen) was estimated using the equations proposed by INRA (1988).

Blood parameters

In the 1st and 8th weeks of the trial, blood samples taken from the jugular vein of each animal with a Vacutainer(R) were used to analyse the energy (glucose, total cholesterol, NEFA) and protein (creatinine, urea, total protein and respective electrophoresis fractions) metabolism parameters. The blood samples were collected and centrifuged, within 30' at 3000 rpm for 15'; the serum obtained was divided into aliquots, transported at 4C to the laboratory, and frozen at -20C until analyses were performed. Serum concentration of parameters was assessed using Assel reagents and a SEAC photometer with interferential filters. Serum protein electrophoresis was performed on agarose gel according to the Helena BioSciences method.

Statistical analyses

Statistical analysis was performed using the GLM procedure of the SAS application package (SAS, 2000) with the model including diet treatment effect and experimental error. The statistical analysis of blood parameters was carried out using the ANOVA model for repeated measurements, which considered the diet (D), the sampling time (S) and the interaction (D x S). When the diet and the sampling time effects were significant (P less than 0.05) means were compared using Student's t test.

RESULTS AND DISCUSSION

In vivo performance and slaughtering data Slaughter body weight was lower (Pless than 0.05) for lambs fed with lupin seeds than for lambs fed with faba beans (Table II). There were no differences in slaughter body weights between lambs fed with lupin plus faba beans and those fed with the two protein sources alone. On the other hand, ADG in Group L was lower (Pless than 0.05) than FB and FB+L (27.9% and 23.5% lower, respectively). There were also differences between dietary treatments in average daily feed intake (ADFI). Group L had lower (Pless than 0.01) ADFI than the other two groups, while there were no differences in FCR between dietary treatments.

Lupin protein is highly degradable in the rumen and this may partly explain the poor animal performance observed in our trial. Masoero et al. (2005) reported that lupin meal is more N-soluble than faba bean meal (87.18% vs. 71.12%), and the same authors observed that in vitro rumen degradability of lupin protein is always greater than the faba bean degradation value (70.82% vs. 58.80% during the first 8 hours of incubation). The degree of lupin protein degradation in the rumen varies from 42 to 95%, based on work by Orskov and McDonald (1979), using a synthetic-fiber bag technique. The lower levels of rumen protein degradation were associated with ground whole seed at high fractional outflow rates (high feeding levels), while the high degradation rates (low by-pass values) were associated with fine grinding and/or low fractional outflow rates (low level feeding).

Although the method of seed preparation can influence the proportion of lupin protein that "by-passes" rumen fermentation, in most instances only a low proportion of lupin protein and amino acids reaches the abomasum intact (van Barneveld, 1999).

Moreover, lupin seeds supply monogastric animals and ruminants with less methionine than other amino acids, and contribute excessive amounts of arginine (van Barneveld, 1999). The comparatively high levels of arginine are a problem when feeding lupin to livestock, because arginine and lysine are antagonists and compete for a common carrier at cell level (van Barneveld, 1999). The lower ADFI observed in lambs fed with lupin seeds may be attributed to the alkaloid content of lupin, although it has been reported that sheep are relatively more tolerant of alkaloids than monogastrics (Stanford et al., 1996). Although the variety of sweet lupine used in the present study is known to have such a low alkaloid content, ranging from 0.007 to 0.222% (Stanford et al., 1996) that we did not measure it, it is still possible that it negatively influenced ADFI and growth.

In fact, some of the inconsistencies in the performance reported for lupin-fed lambs (Tracy et al., 1988) and cattle (Guillaume et al., 1987) may have been caused by these potential toxins, and reduced feed intake and growth of pigs have also been reported to occur with dietary alkaloid levels of 0.03% or greater (Pearson and Carr, 1977).

The section data are reported in Table III. FB lambs had a higher (Pless than 0.05) percentage of loin weight in the half-carcass weight compared with the lambs fed with the other two experimental diets. The abdominal region was also a greater percentage of half-carcass weight in FB+L lambs than in FB and L lambs (Pless than 0.05). The leg and loin dissection data (Table IV) showed that Group L had a larger (Pless than 0.05) percentage of leg bone than FB+L lambs, while there were no significant differences between groups regarding loin dissection, or regarding bone, fat and lean ratios of leg and loin.

Blood parameters

The blood parameters shown in Table V are within the range reported for ovine species (El Barody et al., 2002; Abdel-Ghani et al., 2011). The protein source and timing of blood sampling did not affect glucose, total cholesterol and NEFA concentrations. However, at the end of the trial, the lambs in Group FB had tendentially higher plasma NEFA concentrations than the lambs fed on diets containing lupin alone. The lower energy requirements of the lambs with a lower weight gain could explain lower NEFA mobilizations from adipose tissue (Glukman et al., 1987). Blood sampling time and feeding treatment both had a significant (Pless than 0.01) effect on urea concentration. Group L lambs had higher (Pless than 0.05) concentrations of urea than the other two groups at the end of the trial. Urea is an end-product of nitrogen (N) metabolism in many species, and its concentration reflects the level of ammonia production in the rumen (Lewis, 1957).

The relatively high urea blood content observed in Group L could be due to the greater protein degradability of lupin compared with faba bean

Table II.- Effect of protein source on growth performances and carcass traits in growing lambs (% of empty body weight).

###Diets1

###Item###r.m.s.e.###P-value

###FB###FB+L###L

Initial BW (kg)###15.97###15.93###15.80###2.20###0.984

Slaughter BW (kg)###25.98a###25.43ab###23.04b###2.80###0.050

Average daily gain (kg/day)###0.18a###0.17a###0.13b###0.04###0.013

Average daily feed intake (kg/d)###0.86a###0.86a###0.73b###0.03###less than 0.001

Feed conversion ratio (kg/kg)###4.97###5.17###6.38###1.50###0.094

Empty BW (kg)###24.07###23.53###21.62###3.47###0.268

Skin###14.21###15.44###14.97###1.66###0.233

Omentum###0.97###0.96###1.00###0.39###0.987

Head###4.40###4.48###4.54###0.53###0.828

Pluck###5.64###5.51###5.32###0.92###0.746

Net dressing percentage###52.54###52.81###53.14###2.30###0.910

Table III.- Effect of protein source on percentage of carcass cuts (% of half carcass weight) in growing lambs.

###Diets1

###Item###r.m.s.e.###P-value

###FB###FB+L###L

Half carcass (kg)###6.08###6.02###5.80###1.03###0.500

Half carcass composition (%)

Neck###9.55###9.51###9.55###0.66###0.985

Steaks###14.40###14.68###14.82###0.94###0.586

Brisket###10.79###11.27###11.24###0.93###0.366

Shoulder###16.62###16.70###16.79###0.49###0.739

Loin###7.42a###6.92b###6.86b###0.49###0.033

Abdominal region###5.79b###6.17a###5.36b###0.45###0.002

Leg###31.55###30.99###31.26###0.93###0.465

(92.7 vs 86.3%; Masoero et al., 2005). In fact, lower blood urea levels were observed in lambs receiving protected protein (Abdel-Ghani et al., 2011). As reported by Otto et al. (2000), a lower urea concentration may be related to higher amino acid and protein synthesis allowing greater growth performances also confirmed by the higher growth rate of the lambs receiving faba bean. Moreover, the urea concentration reflects the changes in protein metabolism (Hammond, 1998) and other studies have also reported a rise in blood urea concentration during growth (Abdel-Ghani et al., 2011). This may be linked to increasing ability of the lamb rumen to degrade proteins and carbohydrates as the lambs grow (Grunwaldt et al.,2005).

The timing of blood sampling had a significant (Pless than 0.01) effect on creatinine concentration, and a significant interaction (Pless than 0.01) was found between dietary treatment and blood sampling times. Moreover, at the end of the trial, blood creatinine values were significantly higher (Pless than 0.01) in Group L lambs than in the other two groups. Other studies have reported an increase in blood creatinine levels during growth (Abdel-Ghani et al., 2011); this is probably due to growth and the consequent increase in muscle mass. Istasse et al. (1990) found a positive correlation between creatinine, carcass weight and lean tissue in cattle. The significant increase in creatinine of Group L lambs might also be due in this case to the higher rumen degradability of lupin proteins in comparison

Table IV.- Effect of protein source on leg and loin tissue composition in growing lambs.

###Diets1

###Item###r.m.s.e.###P-value

###FB###FB+L###L

Leg (kg)###1.92###1.82###1.74###0.32###0.462

Leg composition (%)

Lean###72.70###74.45###70.94###3.75###0.132

Fat###6.78###6.44###6.79###2.57###0.994

Bone###20.52ab###19.11b###22.28a###2.67###0.043

Lean/bone###3.55###4.42###3.20###1.55###0.213

Lean+fat /bone###3.88###4.77###3.51###1.60###0.211

Lean /fat###12.07###12.98###12.87###5.27###0.915

Loin (kg)###0.45###0.42###0.39###0.08###0.267

Loin composition (%)

Lean###62.60###62.71###62.63###2.41###0.944

Fat###14.40###15.03###13.81###3.26###0.723

Bone###23.00###22.26###23.56###1.93###0.365

Lean /bone###2.73###2.81###2.68###0.24###0.433

Lean + fat /bone###3.37###3.52###3.28###0.37###0.365

Lean /fat###4.52###4.35###4.87###1.20###0.633

Table V.- Effects of protein source and time sampling on blood parameters in growing lambs.

###Diets-1

###FB###FB+L###L

###Item###Sampling time###Sampling time###Sampling time

###r.m.s.e###P-value

###(week)###(week)###(week)

###1###8###1###8###1###8###D###T###DxT

Glucose (mg/dl)###68.10###68.90###65.50###65.40###66.10###72.90###7.22###0.191###0.154###0.325

Total cholesterol (mg/dl)###53.00###49.67###53.70###50.50###55.90###52.80###9.58###0.607###0.204###0.993

NEFA (mEq/l)###1.34###1.48###1.30###1.37###1.30###0.97###0.49###0.234###0.687###0.296

Urea (mg/dl)###21.24ab###25.58b###23.27ab###27.32b###23.72ab###32.37a###3.97###0.002###less than 0.001###0.132

Creatinine (mg/dl)###0.95ab###1.10b###0.99ab###1.08b###0.90ab###1.35a###0.18###0.153###less than 0.001###0.006

Total protein (g/dl)###6.14###6.34###6.06###6.27###6.15###6.58###0.63###0.605###0.072###0.845

Albumin (%)###55.42###48.22###55.93###47.40###55.81###50.60###4.79###0.562###less than 0.001###0.549

Albumin/Globulin###1.24###0.96###1.27###0.91###1.29###1.04###0.20###0.468###less than 0.001###0.694

1-globulin (%)###6.18###8.48###6.66###7.98###5.94###7.45###1.72###0.458###less than 0.001###0.655

2-globulin (%)###15.74ab###15.82ab###16.08ab###16.68a###14.30ab###14.25b###1.84###0.002###0.658###0.845

-globulin (%)###7.18ab###6.97b###7.47ab###6.67b###7.67ab###8.45a###1.21###0.017###0.812###0.128

-globulin (%)###15.73###20.56###14.14###21.22###16.35###19.24###3.83###0.069###0.001###0.230

with faba bean proteins. In fact, Abdel-Ghani et al. (2011) found lower concentrations of blood creatinine and urea when lambs were fed diets containing ruminally protected proteins. Nozad et al. (2012) reported a positive correlation between blood urea and creatinine in cattle.

Total protein values were not affected by dietary treatment and sampling times. Albumin concentration was affected by the sampling time and was significantly (Pless than 0.01) lower at the end of the trial. The A/G ratio was also affected by the sampling time and was also lower (Pless than 0.01) at the end of the experiment. The a1-globulin and - globulin percentages were higher (Pless than 0.01) at the end of the trial. Dietary treatment had a significant (Pless than 0.01) effect on the a2-globulin percentage, which was higher in the Group FB+L than in L at the end of the trial. Kaneko et al. (1989) also reported decreases in albumin concentrations and increases in globulin fractions in older animals. It has been reported that the protein sources used in this study have nutraceutical properties (Duranti, 2006) and can reduce blood cholesterol levels.

The anticholesterolemic action of these legume grains seems to depend on mechanisms which include their effect on the serum levels of HDL, VLDL and LDL lipoproteins, known to migrate respectively into the a1, a2 and AY fractions of serum protein electrophoresis (Kaneko, 1989). The results of the present study show that the protein sources did not affect blood total cholesterol concentrations despite significant increases in the a1-protein fractions. This a1-globulin increase could confirm the mechanism reported in rats by Sirtori et al. (2004) for lupin, and by Macarulla et al. (2001) for faba bean. Since El Barody et al. (2002) showed an increase in cholesterol levels of lambs (of 4 different genotypes) from weaning to four months of age, it could therefore be assumed that the two protein sources have acted to reduce cholesterol.

In conclusion, the use of lupine and faba bean seeds in association as the main protein source provides lamb growth performances and slaughtering data comparable to those obtained when faba bean seeds are used alone. However, when faba been seeds were the sole protein source in the diet, the half-carcasses presented a higher percentage of loin than the half-carcasses of lambs fed with the two protein supplements in association. Use of the two protein sources together improved lamb growth performances and decreased the leg bone percentage compared to lupin used alone. Blood parameters were little affected by dietary treatments. On the whole, our results suggest that association of the two protein sources

in lamb feed has a positive effect on lamb growth and slaughtering data.

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Author:Lestingi, Antonia; Toteda, Francesco; Vicenti, Arcangelo; Marzo, Davide De; Facciolongo, Anna Maria
Publication:Pakistan Journal of Zoology
Article Type:Report
Date:Aug 31, 2015
Words:5270
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