Animal (2012), 6:5, pp 824833 & The Animal Consortium 2011 doi:10.

1017/S1751731111002229

animal

Semen quality, testosterone, seminal plasma biochemical and antioxidant proles of rabbit bucks fed diets supplemented with different concentrations of soybean lecithin
Y. A. Attia1- and K. I. Kamel2
1 Animal and Poultry Production Department, Faculty of Agriculture, Damanhour University, Damanhour 22516, Egypt; 2Waterfowl and Rabbit Breeding Department, Animal production Research Institute, Agricultural Research Center, P. O. Box 12618, Dokki, Giza, Egypt

(Received 27 June 2011; Accepted 10 October 2011; First published online 23 November 2011)

A total of 28 adult V-line rabbits were fed ad libitum a control diet or a diet supplemented with 0.5%, 1.0% and 1.5% soybean lecithin (SL) for 12 weeks. Bucks that received 0.5%, 1.0% or 1.5% dietary SL had a higher ejaculate volume, mass motility, sperm concentration, total sperm output and total motile sperm. Dietary SL reduced the percentage of dead sperm and increased the normal sperm, and this concurred with an increase in blood testosterone concentration. Blood and seminal plasma total lipid, acid phosphatase and seminal plasma alkaline phosphatase were signicantly increased because of inclusion of SL. Interestingly, SL reduced blood and seminal plasma thiobarbituric acid-reactive substances while increasing blood and seminal plasma glutathione content, glutathione S-transferase, glutathione peroxidase and superoxide dismutase activity. Conception rate and litter size at birth and weaning were also signicantly improved. Practically, it could be suggested that SL is a suitable supplement for improving semen quality, antioxidant status, reproductive traits and the economic efciency of V-line rabbit bucks and 1% is an adequate concentration.
Keywords: lecithin, rabbits, semen quality, antioxidant enzymes

Implications Rabbits are a source of animal protein and improving rabbit production could contribute to increasing life standard worldwide practically in developing countries. The effect of soybean lecithin in rabbit nutrition is not completely known because of lack of data on semen quality and reproductive indices and antioxidant status. The use of lecithin could contribute toward an increase in reproductive efciency and it may be an important resource as a supplement in rabbit nutrition.

Introduction Soybean lecithin (SL) is a by-product from the processing of soybean oil that, apart from being a source of energy, also serves as an emulsier and has the potential to facilitate fat absorption (Lechowski et al., 1999). Lecithin is also a polyunsaturated phosphatidylcholine (PPC), which is high in energy functional and structural elements of all biological
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E-mail: yfat_alexu40@hotmail.com

membranes. Lecithin, as a PPC, is a naturally occurring mixture of diglycerides of stearic, palmitic, linoleic and oleic acids that links to the choline ester of phosphoric acid (Fiume, 2001). PPC plays a rate-limiting role in the activation of numerous membrane-located enzymes, including superoxide dismutase (SOD) and glutathione (GSH), which are important antioxidants and protect cell membranes from damage by reactive oxygen species (ROS). ROS-induced damage to mitochondrial DNA may lead to reduced mitochondrial function. Lecithin may protect mitochondrial DNA, preserving the age-related decline in mitochondrial membrane potentials and hence their activity (Ulkowski et al., 2005). Lipids are involved in the structural and functional functions of the sperm, and their prole may be modied according to physiologic events and/or diet (Mourvaki et al., 2010). Animals cannot synthesize polyunsaturated fatty acids (PUFA) n-6 or n-3 fatty acids de novo as they lack the appropriate fatty acid desaturase enzymes. The n-6 PUFA linoleic acid (LA) and the n-3 PUFA a-linolenic acid therefore need to be provided in the diet as they are absolutely necessary for numerous processes, including growth, reproduction, vision

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Effect of soybean lecithin on semen quality of buck rabbits and brain development (Gurr et al., 2002). There is considerable evidence that dietary PUFA supplementation can inuence biosynthetic pathways involved in both prostaglandin synthesis and steroidogenesis that play multiple roles in the regulation of reproductive function. Furthermore, the PUFA composition of the cell membranes of the sperm and oocyte is important during fertilization (Aitken and Baker, 1995). Lecithin has been widely used in animal diets, such as in sheep (Jenkins and Fotouhi, 1990), lambs (Lough et al., 1991), horses (Holland et al., 1998), sh (Liu et al., 2004), swine (Soares and Lopez-Bote, 2002) and laying hens (Attia et al., 2009a). On the other hand, the use of lecithin in rabbit diets is rare. Supplementation of animal diets with oils rich in PUFA, such as LA, has positively inuenced reproductive functions (Fellner et al., 1995; Rocha et al., 2009). SL is also a rich source of alpha, gamma and delta tocopherol; however, the contents of tocopherol vary considerably from 13 (Scholeld, 1981) to 25.5 mg/100 g (Wang and Wang, 2008). SL was also shown to decrease hepatic cellular damage and improve oxidative stability, and exerted neuroprotective activity through its antioxidant action (Aabdallah and Eid, 2004; Das and Vasudevan, 2006; Das et al., 2007; Wang and Wang, 2008). The effect of the antioxidant properties of lecithin could be attributed to the synergistic inuence between amino-alcohol phospholipids and g- and d-tocopherols (Judde et al., 2003). Therefore, the aim of this study was to investigate the effect of SL supplementation on semen quality, blood testosterone concentration, blood and seminal plasma biochemistry including antioxidant status and fertility traits. Material and methods The experimental work of this study was carried out at El-Sabahia Poultry Research Station (Alexandria), Animal Production Research Institute, Agricultural Research Center, Ministry of Agriculture, during 2010 to 2011.

Animals, experimental design and diets A total of 28 male V-line rabbits (aged between 7 and 8 months) were randomly distributed among four dietary treatments in a straight-run experimental design. The rst treatment served as a control (Con; 0% SL); the second, third and fourth treatments were supplemented with 0.5%, 1.0% and 1.5% SL in the diet, respectively (Table 1). The diets were formulated based on the National Research Council (NRC, 1977) to meet rabbits nutrient requirements. The rabbit bucks were individually housed in galvanized wire cages, offered free access to fresh water and feed and subjected to a 14 : 10 lightdark cycle. Feed intake and BW gain Daily feed intake (g/kg BW) and weekly BW (g/buck) were recorded. Semen quality Semen collection occurred weekly over the 12 weeks; thus, 84 ejaculates were obtained per treatment. Ejaculates were collected using an articial vagina maintained at 458C to 468C and a teaser doe. The volume of each ejaculate was recorded after removal of the gel mass. The reaction time (RT)

Table 1 Proximate analysis of pelleted basal diet (% on a dry matter basis) Soybean lecithin concentration (%) Ingredients proles and determined composition (%) Clover hay Yellow corn Barley Wheat bran Soybean meal Soybean lecithin Molasses Di-calcium phosphate Sodium chloride Vitamin 1 mineral mixDL-methionine Total Calculated analysisDigestible energy (MJ/kg) CP Crude ber Ether extract
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0 40.0 10.0 13.0 15.0 17.5 0.0 3.0 0.8 0.3 0.3 0.1 100.0 10.69 17.2 13.9 2.43

0.5 40.0 8.7 14.0 16.3 17.0 0.5 2.0 0.8 0.3 0.3 0.1 100.0 10.74 17.1 14.1 2.92

1.0 40.0 7.0 14.0 18.0 17.0 1.0 1.5 0.8 0.3 0.3 0.1 100.0 10.77 17.2 14.2 3.40

1.5 40.5 5.5 14.0 19.5 16.5 1.5 1.0 0.8 0.3 0.3 0.1 100.0 10.77 17.2 14.5 3.88

The vitamin and mineral premix/kg contained the following IU/g for vitamins or minerals: A 4 000 000; D3 5 000 000; E 16.7 g; K 0.67 g; B1 0.67 g; B2 2 g; B6 0.67 g; B12 0.004 g; B5 16.7 g; pantothinc acid 6.67 g; biotein 0.07 g; folic acid 1.67 g; choline chloride 400 g; Zn 23.3 g; Mn 10 g; Fe 25 g; Cu 1.67 g; I 0.25 g; Se 0.033 g and Mg 133.4 g (Rabbit premix produced by Holland Feed Inter. Co.). Based on National Research Council (1994).

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Attia and Kamel was the time interval from the introduction of the teaser doe into the males cage to ejaculation; it was measured in seconds using a stopwatch and was considered as an indication of libido. Immediately after collection, semen was maintained at 358C in a water bath for evaluation. Fresh semen (two drops) was placed on a warm slide and covered with a cover slip (20 3 20 mm) to determine mass motility. Mass motility from at least three elds was examined at 378C under a microscope with phase-contrast optics, at 403, and assessed from 0% to 100%. A weak eosin solution was used at a rate of 1 : 99 before counting the cells for the evaluation of sperm concentration (3106/ml) according to Smith and Mayer (1955) using the improved Neubauer hemocytometer slide (GmbH1Co., Brandstwiete 4, 2000 Hamburg 11, Germany). The total sperm output was calculated by multiplying semen ejaculate volume and semen concentration. Assessment of live and abnormal spermatozoa was performed using an eosinnigrosin blue-staining mixture (Blom, 1950). The percentage of live, dead and abnormal spermatozoa was determined using stains that penetrated cells with damaged membranes. Normal live sperm exclude the eosin stain and appear white in color, whereas dead sperm take up eosin and appear pinkish in color because of loss of membrane integrity. Normal sperm have an oval head with a long tail. Abnormal sperm have head, midpiece or tail defects, such as a large or a misshapen head or a crooked or a double tail. The total number of motile sperm was calculated by multiplying the percentage of motile sperm and the total sperm outputs. The total functional sperm fraction (TFSF) was also calculated by multiplying the total sperm output and sperm motility and normal-morphology sperm (Correa and Zavos, 1996).

Blood testosterone The testosterone concentration in plasma was measured using immunoassay commercial kits (Biosource-Europe S.A. 8, rue de Llndustrie.B-1400 Nivelles, Belgium). Reproductive traits and economic efciency Reproductive criteria such as conception rate and litter size at birth (total born and total born alive) and at weaning were recorded. In addition, litter weights at birth and at day 28 were recorded. Bucks of each group were mated with 30 nulliparous female rabbits and their parameters were recorded according to the International Rabbit Reproduction Group (IRRG, 2005). Conception rate was also estimated as the number of kindled does divided by the number of mated doe 3 100. Economic efciency was assessed following the guidelines of El-Deek et al. (2010) using inputoutput analysis. Statistical analysis Data were analyzed as a randomized design using the General Linear Model procedure of SAS (1996). The StudentNewmanKeuls test was used for testing the mean differences. P-values , 0.05 were accepted as signicant.
Results

Semen and blood plasma constituents Blood samples were collected from the ear vein of each buck every other week and placed immediately on ice in heparinized tubes. Blood and seminal plasma was collected from blood by centrifugation at 860 3 g for 20 min at 48C and stored at 2608C (Attia et al., 2009b). Total protein (TP), albumin (Alb), total lipid (TL), the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), alkaline phosphatase (AlP) and acid phosphatase (AcP) in blood and seminal plasma samples and globulin, urea, creatinine, triglycerides and cholesterol in blood plasma were determined by colorimetric enzymatic methods using commercial kits purchased from bio-diagnostic R company (Recycling Crusher-SBM ). Thiobarbituric acid-reactive substances (TBARS) were assayed in the seminal and blood plasma using the method of Tappel and Zalkin (1959). Seminal and blood plasma GSH was determined using commercial GSH reduced kits according to the method of Beutler et al. (1963). Glutathione peroxidase (GPx) activity was assayed using the method of Chiu et al. (1976). SOD activity was assayed according to Misra and Fridovich (1972). Glutathione S-transferase (GST) activity was determined according to Habig et al. (1974) using p-nitrobenzylchloride as a substrate.
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Dietary fatty acid proles Table 2 shows the fatty acid prole of the experimental diets. The results showed that SL-supplemented diets contained higher concentrations of unsaturated fatty acids (UFA) and PUFA, especially C18:2, than the unsupplemented group, and their contents increased with increasing concentration of SL. Both saturated fatty acids and monounsaturated fatty acids exhibited a progressive decrease with increasing SL above 0.5%. BW and feed intake Table 3 shows that SL signicantly increased BW of bucks; however, a stepwise increase was shown in BW up to a 1% concentration, although a further increase in the SL content did not affect BW over that of 1%. On the other hand, SL showed the opposite trend in feed intake. Semen characteristics Table 3 indicates that rabbit bucks that received SL up to 1% exhibited a stepwise signicant increase in the ejaculate volume, mass motility, sperm concentration, total sperm output and the total motile sperm (P , 0.05), while a further increase did not induce an improvement over that of 1% in SL. However, the opposite trend was shown in the RT and percent dead sperm, although the difference among the different concentrations of supplemented SL was not signicant for RT. Dead sperm decreased progressively up to 1% SL and stabilized thereafter. Normal sperm signicantly increased gradually as the SL supplementation increased compared with the Con group but the difference between 0.5% and 1.0% was not signicant.

Effect of soybean lecithin on semen quality of buck rabbits

Table 2 Fatty acid proles of the experimental diets Soybean lecithin as % of the whole fatty acids Fatty acid C10:0 (capric acid) C12:0 (lauric acid) C14:0 (myristic acid) C16:0 (palmitic acid) C16:1 (palmitoleic acid) C18:0 (stearic acid) C18:1 (oleic acid) C18:2 (linoleic acid) C20:0 (arachidic acid) SFA UFA PUFA MUFA Total fatty acids 0 6.27 6.10 4.78 22.48 0.00 8.17 30.01 18.01 4.18 51.99 48.01 18.01 30.01 100.0 0.50 3.61 5.10 1.04 19.18 2.59 4.25 29.12 28.32 6.79 39.97 60.03 28.32 31.71 100.0 1.0 7.07 0.36 4.71 13.06 3.02 3.52 26.35 38.87 3.04 31.76 68.24 38.87 29.37 100.0 1.5 4.62 0.47 5.66 9.59 3.42 2.89 24.65 45.82 2.89 26.08 73.89 45.82 28.07 100.0

SFA 5 saturated fatty acids; UFA 5 unsaturated fatty acids; PUFA 5 polyunsaturated fatty acid; MUFA 5 monounsaturated fatty acids.

Table 3 BW, feed intake and semen characteristics of V-line male rabbits as affected by supplementation with different SL concentrations SL (%) Criteria BW and feed intake of bucks BW (g) Feed intake (g/kg per day) Semen quality criteria Ejaculate volume (ml) Reaction time (s) Mass motility (%) Sperm concentration (3106/ml) Total sperm output (3106) Total motile sperm (3106) Dead sperm (%) Normal sperm (%) TFSF 0 3150c 68.1a 0.80c 10.9a 69.1c 333.9c 269.2c 186.5c 28.1a 77.4c 144.5d 0.5 3200b 58.2b 0.86b 6.5b 75.1b 394.8b 346.7b 263.6b 22.9b 80.1b 212.6c 1.0 3353a 54.2c 0.93a 6.1b 80.7a 419.5a 400.4a 327.6a 18.1c 80.3b 266.1b 1.5 3317a 54.7c 0.94a 5.9b 80.9a 427.4a 409.1a 334.7a 17.6c 83.4a 282.6a s.e.m.

P-value
0.001 0.001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001

13.5 0.44 0.011 0.17 0.61 3.35 5.39 4.59 0.63 0.31 3.82

SL 5 soybean lecithin; s.e.m. 5 standard error of mean; TFSF 5 total functional sperm fraction. n 5 7 animals per treatment (the value represents 84 ejaculates per treatment). a,b,c Means within a row with different superscripts are signicantly different (P , 0.05).

A stepwise signicant increase with increasing SL supplementation was shown in TFSF. Correlation analysis (Table 4) showed that the relationship between dietary lecithin content and semen quality was not signicant after 1 week of supplementation. The correlation between lecithin content and most of the semen quality traits was signicant in week 2, except for ejaculate volume and RT. Only the correlation between lecithin concentration and sperm concentration was not signicant in week 3, lecithin concentration and ejaculate volume or abnormal sperm in week 4 and lecithin concentration and ejaculate volume in weeks 6 and 8. By week 10, all correlations between lecithin concentration and semen quality traits were signicant. The r-value showed a progressive

increase over time and had a strong correlation from week 8 onward, indicating that 8 weeks was the minimum period to obtain a signicant effect of dietary treatments on semen quality. For the entire period, the correlation between lecithin concentration and semen quality criteria was signicant and of intermediate magnitude. It should be mentioned that the negative correlation obtained for RT, abnormal sperm and dead sperm indicates that increasing the lecithin concentration decreased RT and percentage of abnormal and dead sperm.

Blood and seminal plasma biochemical constituents Data in Table 5 show that blood and seminal plasma TP, Alb and globulin were signicantly increased with the inclusion
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Attia and Kamel

Table 4 Correlations between SLC and semen quality of V-line rabbits at different ages Statistical analyses of SLC Week 1 r-value P-value Week 2 r-value P-value Week 3 r-value P-value Week 4 r-value P-value Week 5 r-value P-value Week 6 r-value P-value Week 7 r-value P-value Week 8 r-value P-value Week 9 r-value P-value Week 10 r-value P-value Week 11 r-value P-value Week 12 r-value P-value Whole period r-value P-value EV RT MM SC TSO TMS AS LS TFSF NS DS

0.048 0.80 0.173 0.38 0.424 0.025 0.174 0.337 0.274 0.1585 0.092 0.640 0.492 0.0078 0.327 0.0889 0.644 0.0002 0.722 0.0001 0.74333 0.0001 0.733 0.0001 0.317 0.0001

0.093 0.63 0.007 0.970 20.463 0.013 20.557 0.002 20.701 0.0001 20.885 0.0001 20.829 0.0001 20.765 0.0001 20.775 0.0001 20.759 0.0001 20.748 0.0001 20.759 0.0001 20.552 0.0001

20.109 0.61 0.545 0.003 0.683 0.0001 0.667 0.0001 0.714 0.0001 0.724 0.0001 0.738 0.0001 0.675 0.0001 0.673 0.0001 0.642 0.0002 0.671 0.0001 0.651 0.0002 0.553 0.0001

0.310 0.110 0.452 0.016 0.305 0.115 0.816 0.0001 0.718 0.0001 0.741 0.0001 0.877 0.0001 0.738 0.0001 0.716 0.0001 0.873 0.0001 0.823 0.0001 0.793 0.0001 0.462 0.0001

0.255 0.190 0.391 0.039 0.466 0.012 0.639 0.0002 0.633 0.0003 0.528 0.0039 0.762 0.0001 0.723 0.0001 0.737 0.0001 0.877 0.0001 0.831 0.0001 0.843 0.0001 0.426 0.0001

0.213 0.28 0.531 0.004 0.619 0.0004 0.740 0.0001 0.735 0.0001 0.733 0.0001 0.808 0.0001 0.784 0.0001 0.823 0.0001 0.893 0.0001 0.842 0.0001 0.848 0.0001 0.493 0.0001

0.056 0.78 20.526 0.004 20.467 0.012 20.258 0.185 20.513 0.0053 20.597 0.0008 20.644 0.0002 20.813 0.0001 20.713 0.0001 20.795 0.0001 20.838 0.0001 20.869 0.0001 20.471 0.0001

20.147 0.45 0.343 0.074 0.460 0.0139 0.616 0.0005 0.667 0.0001 0.723 0.0001 0.568 0.0016 0.569 0.0016 0.650 0.0002 0.650 0.0002 0.740 0.0001 0.733 0.0001 0.511 0.0001

0.193 0.33 0.572 0.002 0.660 0.0001 0.784 0.0001 0.755 0.0001 0.757 0.0001 0.849 0.0001 0.834 0.0001 0.837 0.0001 0.915 0.0001 0.873 0.0001 0.890 0.0001 0.513 0.0001

20.056 0.78 0.526 0.004 0.467 0.0123 0.258 0.185 0.513 0.0053 0.597 0.0008 0.644 0.0002 0.814 0.0001 0.713 0.0001 0.795 0.0001 0.838 0.0001 0.869 0.0001 0.471 0.0001

0.147 0.45 20.343 0.074 20.460 0.014 20.616 0.001 20.667 0.0001 20.723 0.0001 20.568 0.0016 20.569 0.002 20.650 0.0002 20.650 0.0002 20.740 0.0001 20.733 0.0001 20.511 0.0001

SLC 5 soybean lecithin concentration; EV 5 ejaculate volume; RT 5 reaction time; MM 5 mass motility; SC 5 sperm concentration; TSO 5 total sperm output; TMS 5 total motile sperm; AS 5 abnormal sperm; LS 5 live sperm; TFSF 5 total functional sperm fraction; NS 5 normal sperm; DS 5 dead sperm.

of SL in buck diets. Blood plasma Alb to globulin ratio decreased signicantly due to SL supplementation, while the opposite trend was shown in seminal plasma when SL was supplemented at only 1.5%. Blood plasma urea and creatinine were signicantly decreased due to SL supplementation; however, the decrease was signicant in plasma urea only when 1.5% SL was supplemented. However, the decrease in blood plasma creatinine was maximized at 1.5% SL, with no difference between 0.5% and 1.0% SL. Blood and seminal plasma TL increased signicantly with SL supplementation and the increase was maximized at 1.5% concentrations. However, SL supplementation at 1% and 1.5% signicantly decreased blood plasma cholesterol
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compared with the Con group, whereas the effect of 0.5% SL was intermediate. On the other hand, supplementation of SL had no signicant effect on blood plasma triglycerides. Blood plasma and seminal plasma AST and ALT were signicantly decreased with increasing SL supplementation that was minimized at 1.5% SL for blood plasma ALT and seminal plasma AST. Rabbit bucks exhibited a signicant increase in blood plasma AcP and seminal plasma AlP and AcP that maximized at 1.5% SL.

Blood and seminal plasma antioxidant status Table 6 shows that blood and seminal plasma GSH, GPx, SOD and GST activities increased signicantly with

Effect of soybean lecithin on semen quality of buck rabbits

Table 5 Seminal plasma (TP, Alb, TL, ALT, AST, AlP and AcP) by supplementation with different SL concentrations SL (%) Criteria Blood plasma constituents TP (g/dl) Alb (g/dl) G (g/dl) A/G (%) Creatinine (mg/dl) Urea (mg/dl) TL (mg/dl) Trig (mg/dl) Cholesterol (mg/dl) AST (IU) ALT (IU) AcP (U/l) Seminal plasma constituents TP (g/dl) Alb (g/dl) G (g/dl) A/G (%) TL (mg/l) AST (IU) ALT (IU) AlP (U/l) AcP (U/l) 0 6.58b 3.31c 3.27b 1.10a 1.37a 45.0a 459c 43.4 125.1a 55.7a 29.4a 48.2c 5.35d 3.15d 2.20b 1.48b 394d 33.8a 24.1a 52.6c 32.8c 0.5 6.99a 3.45b 3.54a 1.02b 1.30b 44.9a 505b 44.3 119.6ab 50.0b 26.5b 52.2b 5.69c 3.42c 2.27b 1.52b 399c 28.6c 21.4b 59.2b 35.7b 1.0 7.10a 3.48b 3.63a 0.99b 1.30b 44.8 a 510b 44.0 115.4b 47.9b 24.6c 53.0b 6.15b 3.70b 2.45a 1.62b 407b 30.2b 18.3c 59.5b 36.0ab 1.5 7.20a 3.62a 3.58a 0.99b 1.28c 41.8b 539a 43.3 116.0b 46.5b 22.9d 55.8a 6.38a 4.10a 2.28b 1.95a 411a 26.5d 18.2c 68.3a 36.5a s.e.m.

P-value
0.001 0.001 0.009 0.006 0.0001 0.0001 0.0001 0.91 0.0033 0.0001 0.0001 0.0001 0.0001 0.0001 0.009 0.001 0.0001 0.0001 0.0001 0.0001 0.0001

0.072 0.040 0.081 0.028 0.007 0.33 9.4 0.78 1.71 1.10 0.47 0.90 0.07 0.06 0.04 0.05 1.2 0.34 0.17 0.19 0.22

TP 5 Total protein; alb 5 albumin; TL 5 total lipid; ALT 5 alanine aminotransferase; AST 5 aspertate aminotransferase; AlP 5 alkaline phosphatase; AcP 5 acid phophatase; SL 5 soybean lecithin; s.e.m. 5 standard error of mean; G 5 globulin; A/G 5 albumin/globulin ratio; Trig 5 triglycerlids. n 5 42 samples per each treatment. a,b,c,d Means within a row with different superscripts are signicantly different (P , 0.05).

increasing SL content while decreasing blood and seminal plasma TBARS. These improvements were maximized at 1.5% for TBARS and GSH in blood and seminal plasma and SOD and GST in blood plasma. The effect of SL on blood and seminal plasma GPx and seminal plasma GPx, SOD and GST was maximized at 1%. Blood plasma testosterone concentration increased signicantly due to SL additions, with no differences between various supplemented concentrations.

Discussion The results indicated that SL is a rich source of PUFA and n-6 was the dominant fatty acid, and this was in agreement with the results of Jenkins and Fotouhi (1990), who found that the addition of 5.2% SL led to an increase in the dietary concentrations of LA and linolenic fatty acid. The LA (n-6) is now regarded as a nutritionally essential fatty acid (Holman, 1998), and all the classic symptoms of essential fatty acid deciency (dermatitis, growth retardation, infertility) can be completely cured by the use of n-6 fatty acids alone. These symptoms are related to the biological functions of n-6 fatty acids: (1) 18:2; n-6 is a structural component in the ceramides of the water barrier of the skin (Hansen, 1989). (2) Arachidonic acid (20:4; n-6) is a precursor of eicosanoids, which are local hormones that participate in a number of physiological as well as pathophysiological conditions, for example, parturition initiation, platelet aggregation, renal electrolyte regulation, blastocyte implantation and activation of immune cells (Newberry et al., 1999). (3) n-6 fatty acids possibly also play a role as a second messenger in the process of signal transduction across cell membranes (Chen et al., 1999). The effect of SL on productive performance and semen quality of rabbit bucks is rare in the literature. It was found that SL increased BW and decreased feed intake compared
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Reproductive performance and economic efciency Table 7 indicates that the conception rates of females mated with rabbit bucks supplemented with different concentrations of SL were signicantly improved compared with the Con group. The total and live litter size at birth and litter size at weaning as well as economic efciency were also signicantly higher than those of the Con group but without a difference in the various SL contents. However, no signicant differences were found between SL groups in bunny weight at birth and at 28 days of age. The correlation between lecithin concentration and plasma testosterone was strong and signicant while it was intermediate and signicant for litter size at birth and weaning and weak but signicant for conception rate (Table 8). The relationship between litter weight at birth and weaning was not signicant.

Attia and Kamel

Table 6 The blood and seminal plasma TBARS, GSH, GPx, SOD, GST and blood plasma testosterone concentration by supplementation with different concentrations of SL SL (%) Criteria Blood plasma antioxidant constituents TBARS (nmol/ml) GSH (mg/dl) GPx (mg/l) SOD (IU) GST (IU) Testosterone (ng/dl) Seminal plasma antioxidant constituents TBARS (nmol/ml) GSH (mg/dl) GPx (mg/l) SOD (IU) GST (IU) 0 1.998a 31.2d 6.81c 2.32c 3.20d 265.7b 1.127a 15.1d 4.42c 7.37b 1.026c 0.5 1.930b 34.4c 7.47b 2.43c 3.48c 333.7a 0.989c 18.8c 4.78b 7.25b 1.351b 1.0 1.855c 44.8b 7.98a 2.65b 4.00b 336.3a 1.045b 22.1b 4.89ab 7.67a 1.497a 1.5 1.844c 49.2a 8.03a 2.82a 4.28a 349.0a 0.987c 23.7a 5.04a 7.60a 1.492a s.e.m.

P-value
0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001 0.0021 0.0001

0.015 0.54 0.065 0.016 0.054 2.73 0.007 0.22 0.069 0.080 0.003

TBARS 5 thiobarbituric acid reactive substances; GSH 5 glutathione content; GPx 5 glutathione peroxidase; SOD 5 superoxide dismutase; GST 5 glutathione S-transferase; SL 5 soybean lecithin; s.e.m. 5 standard error of mean. n 5 42 samples per each treatment. a,b,c,d Means within a row with different superscripts are signicantly different (P , 0.05).

Table 7 Reproductive indices of bucks supplemented with different concentrations of SL SL (%) Criteria Conception rate (%) Litter size (kits/litter) at birth Live (kits/litter) at birth Litter size (n) at weaning Economic efciency (%) Bunny weight (g) At birth At 28 days 0 63.3b 7.1b 6.1b 5.2b 38.8b 59.2 582 0.5 83.3a 9.1a 8.9a 7.8a 137.5a 58.6 589 1.0 86.7a 9.0a 8.3a 7.8a 140.5a 59.8 590 1.5 86.7a 9.9a 9.0a 8.2a 152.0a 60.3 587 s.e.m. 7.21 0.72 0.83 0.46 8.20 0.623 8.82

P-value
0.059 0.0001 0.0001 0.0001 0.0001 ns ns

s.e.m. 5 standard error of mean. Economic efciency 5 net revenue/feed cost. Net revenue 5 litter size at weaning by 12 litter per kit. Feed cost 5 feed intake by price per kg of feed with or with lecithin supplementation. a,b Means within a row with different superscripts are signicantly different (P , 0.05).

Table 8 Correlation between SLC and reproductive traits Test SLC; r-value P-value 0.655 0.0001 CR 0.20 0.02 LSB 0.42 0.0001 LSW 0.530 0.0001 LWB 0.071 0.49 LWW 0.024 0.81

SLC 5 soybean lecithin concentration; Test 5 testosterone; CR 5 conception rate; LSB 5 litter size at birth; LSW 5 litter size at weaning; LWB 5 litter weight at birth; LWW 5 litter weight at weaning.

with the Con group. The positive effect on BW of bucks were 2%, 7% and 5%, meanwhile, the decrease in feed intake was 14%, 20% and 20% for 0.5%, 1.0% and 1.5% concentrations of SL, respectively. This indicated that SL at
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1% was adequate for improving feed utilization and semen quality of bucks despite the decrease in feed intake. The decrease in feed intake could be attributed to the increase in the availability of energy, essential fatty acids and choline and stimulation of lipid absorption (Attia et al., 2009a). In the literature, a high content of PUFA in the chicken sperm membrane inuences sperm function and modication of dietary fatty acids can affect spermatozoa (Bongalhardo et al., 2009). In addition, Mourvaki et al. (2010) concluded that daily supplementation with n-3, n-6 and n-9 fatty acids signicantly increased semen volume and concentration, while decreasing abnormal spermatozoa. SL improved semen quality, biochemical constituents of blood and seminal plasma and the antioxidant status of both

Effect of soybean lecithin on semen quality of buck rabbits uids, which could be attributed to increasing dietary tocopherol intake, which was at least doubled when SL was added at 1%. Thus, the improvements in semen quality could be attributed to increasing oxidative stability (Scholeld, 1981; Judde et al., 2003; Ulkowski et al., 2005; Wang and Wang, 2008), a neuroprotective effect (Aabdallah and Eid, 2004), decreasing cellular damage (AST and ALT) and TBARS while increasing GSH, SOD, GST and GPx (Das and Vasudevan, 2006; Das et al., 2007). These enhancements were accompanied by better reproductive performance (conception rate) and litter size up to weaning. Blood and seminal plasma constituents exhibited positive signicant responses to SL, showing a constant effect of SL on both uids. The improvements found herein are in general agreement with those reported by Al-Daraji et al. (2010) and Das et al. (2007). The latter authors showed that lecithin decreased AST and ALT activities and TBARS, indicating a decrease in sperm damage while increasing spermatozoa livability and concentration. The increase in the AlP and the AcP observed in blood and seminal plasma is in agreement with the result of Al-Daraji et al. (2001), who reported that both AlP and AcP are involved in the metabolism of spermatozoa via the hydrolysis of carbohydrates. The increase in the antioxidant status of both blood and seminal plasma (GSH, GPx, SOD and GST) and the decrease in TBARS showed higher cell stability. In fact, vertebrate sperm included rabbit display high rates of metabolic activity and are rich in PUFA (Castellini et al., 2000 and 2006), rendering them particularly susceptible to ROS-induced oxidation. It is known that lipid peroxidation is one of the major reactions leading to phospholipid loss, membrane damage and the loss of motility in mammalian spermatozoa (Mann and Lutwak-Mann, 1981). Previous results showed that the sperm of rabbits supplied with dietary PUFA are more susceptible to oxidative damage and undergo an early acrosome reaction in vitro (Castellini et al., 2003). However, in this study, supplementation with SL induced the opposite trend due to its higher tocopherol content as an antioxidant agent of SL (Aabdallah and Eid, 2004; Wang and Wang, 2008; Attia et al., 2009a) and resulted in higher cell stability of higher antioxidant enzymes and lower TBARS (Table 6). The GPx enzyme plays a particularly important role in the antioxidant protection of the cell by converting hydrogen peroxides into less harmful components (Olafsdottir and Reed, 1988) and GPx plays an important role against lipid peroxidation in spermatozoa in vitro (Alvarez and Storey, 1989). In this study, SL not only increased the productive performance, semen quality and antioxidant proles but also improved blood plasma testosterone (Table 5). Testosterone is required for the maturation of male germ cells and sperm production and quality (Attia et al., 1993 and 1995; Walker, 2009). The PUFA or their derived eicosanoids with the hypothalamopituitarygonadal axis play an important role in the hormonal control of spermatogenesis (Etches, 1996) and modulate steroid synthesis in different steroidogenic tissues (Mohn et al., 2005). Gromadzka et al. (2002) found that dietary fat signicantly inuenced plasma testosterone concentration and the activity of 17b-hydroxysteroid dehydrogenase, the key enzyme in the testosterone synthesis pathway in male rat gonads (Korach, 1997). Animals received rapeseed oil diets as a UFA source, which stimulated the testicular function in rats, suggesting that UFA stimulates enzyme activity and androgen secretion into blood (Korach, 1997). Testosterone is metabolized to estrogens by aromatase (Simpson et al., 1994) and estrogens seem to regulate sperm motility (Carreau et al., 2007). SL not only improved semen quality of bucks (Table 3) but also reproductive indices as an increase in the conception rate and litter size at birth was observed (Table 6). In this regard, Cerolini et al. (1997) suggest that the PUFA may play a role in maintaining the survival of the spermatozoon in the female reproductive tract before interaction with the oocyte. In mammals, the lipid composition of sperm membrane was found to play a major role in the physicochemical modication leading to fertilization (Langlais and Roberts, 1985). Phospholipids are the major lipid components of spermatozoa; they contain large amounts of PUFAs; thus, a diet enriched with lecithin as a rich source of n-6 PUFAs and antioxidants may be utilized successfully to improve the reproductive capacity of animals (Maldjian et al., 2003). Lipid and fatty acid compositions of spermatozoa may be important predictors of fertility. Similar to the present ndings, Kelso et al. (1996) and Bongalhardo et al. (2009) also indicated that sh oil or corn oil increased fertility and this effect may be more pronounced and may persist for longer durations. In addition, Brun et al. (2002) found that the mass motility (number of motile sperm per ejaculate) signicantly inuenced the conception rate but volume, percentage of motile sperm and concentration had no effect. They also found that litter size (total born) was signicantly inuenced by concentration and all variables dependent on it, particularly the number of total and motile sperms. In conclusion, SL signicantly increased productive and reproductive performance and semen quality of rabbit bucks and 1% is an adequate concentration.

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