Effect Of Cottonseed Shell On Reproductive Function In Xinjiang Fine-wool Male Sheep

Cotton seed shell is the by-product of cotton, it is rich in crude fiber and has a certain amout of fat and protein which can be used as available feed resources. However, it contains toxic substances free gossypol and long-term animal feeding can produce poisoning hazards that affect the growth and reproduction. In this experiment, we selectd fine wool male sheep as the test subject to disscuss the influence of different amount of cotton seed shell in reproduction of fine wool male sheep. We also isolated and cultured Leydig cells of age ram march to study the effects of gossypol acetic acid on it.The purpose of the experiment is to provide the Scientific Basis for its application in Ruminant animal feed. The main contents of this paper and the results are as follows:1 Effects of the cotton seed shell on fine wool male sheep antioxidant capacity and the internal organsForty eight three months weaned fine wool male sheep at an average initial body weight of 19.22kg were randomly allocated into 3 groups (sixteen sheep in each group). The dietary treatments were as follows:1) Control group (fed basal diet),2) Group 1:(basal diet+500g cotton seed shell+500g reed,3) Group 2:(basal diet+1000g cotton seed shell). Experiment test period for 7 days, trial period for eight months.At the age of 5,7,9,11 mouth, Thirty six sheep with 3 pigs per group were randomly selected and humanely slaughtered.We selected different organ samples to determine the related indicators. Separately in May age, in July age, in September age and in November age takes the Main research results were as follows:1) The average daily gain(ADG) of Control group was 157g, Group 1 was 163g and 176g in group 2. No significant difference was found in every group(P>0.05).2) Results of gossypol residues in tissues and organs showed that with the time extended, gossypol residues in sheep tissues improved. The accumulation of gossypol residues in different organs had different tissue compatibility.Its residual quantity from high to low was as follows:Testicle> liver> kidney> spleen> longissimus dorsi muscle> gluteus.3) The blood serum oxidation index changed when sheep fed Cotton seed shell. At the age of 11 month, serum T-AOC, GSH, CAT and GSH-PX of Group 1 and 2showed to be less. Compared with control, serum T-AOC, GSH, CAT and GSH-PX of Group 1 decreased 3.12%(P<0.05),25.17%(P<0.05),20.45%(P<0.05) and 28.50%(P<0.05),respectively. Group 2 decreased 9.37%(P<0.05),55.82%(P<0.05),34.09%(P<0.05) and 42.27%(P<0.05) respectively. We also found that serum MDA of test experiment increased. Compared with control, serum MDA of Group 1 increased 29.50% and Group 2 increased 80.09%(P<0.05)。Results of serum enzyme activities indicated that with 8 mouths of experiment serm ALT and AST greatly improved(P<0.05). Compared with Group 1, they increased by 22.91% and 3.63% (P<0.05). Compared with control, serm LDH of Group 1 and 2 had decreased 5.21% and 46.40%(P<0.05), respectively. Compared with Group 1, serm LDH decreased 45.43%(P<0.05); serm Na, K-ATPase of Group 1 and 2 decreased 70.9% and77.41%(P<0.05).4) The results of liver oxidation resistance index analysis:At the age of 11 mounth, liver oxidation resistance index significantly decreased. Compared with control, liver T-AOC,GSH,CAT and GSH-PX of Group 1 decreased 24.84%(P<0.05),29.17%(P<0.05),18.35%(P<0.05),20.23%(P<0.05) and Group 2 decreased 31.84%(P<0.05),51.00%(P<0.05),20.37%(P<0.05) and 31.42%(P<0.05); Compared with control, Liver MDA improved 22.09%(P<0.05);Compared with Group 1, liver T-AOC,GSH,CAT and GSH-PX of Group 2 decreased 9.32%(P<0.05),30.82%(P<0.05),2.49%(P<0.05) and 14.02%(P<0.05),respectively. Liver MDA improved by 19.31%(P<0.05)。2 Effects of cotton seed shells on blood serum sex hormone, testicle microstructure and seminal fluid quality in male fine-wool sheep1) Through the experiment, we found that sperm motility, live sperm rate, pH and sperm concentration greatly reduced (P<0.05).2) Results of the blood serum sex hormone indicated that serum testosterone and blood serum insulin, T3 and T4 changed.Compared with control, There were higher serum testosterone in group 1 and 2.They had improved by 38.70% and 290.32% (P<0.05),separately.Besides, serum insulin, T3 and T4 greatly reduced (P<0.05).3) Results of light microscope were as follows:compared with the control group, testicle acorn tube distance of Group 1 increased and the performance haematoma changed, some tubulary acorn tube cell obvious reduced; Testicle tubular of Group 2 crackered, cell quantity of Group 2 obviously decreased, some cell necrosised.3 Effects of Gossypol on testicle intersititial cell in virto1) MTT results of gossypol on testicle intersititial cell were as followes:1. Laevo-rotatory gossipol function:after 48 hours of treatment, laevo-rotatory gossipol greatly inhibit the intersititial cell increment when the concentration of Laevo-rotatory gossipol was 64,128,256,512 mg/L.2. Racemization gossipol function:after 48 hours of treatment, racemization gossipol greatly inhibit the intersititial cell increment when the concentration of Laevo-rotatory gossipol was 64,128,256,512mg/L.With the increase of racemization gossypol, the inhibition ehchanced.2) After 24h of laevo-rotatory gossipol and racemization gossipol treatments, gossypol could affect the Na-K-ATP enzyme activity in intersititial cell.When the concentration attained 512 mg/L, Na-K-ATP enzyme activity in intersititial cell greatly decereased (P<0.05)3) After 48h of laevo-rotatory gossypol treatment,with the increasement of laevo-rotatory gossypol, the quantity of dead cells also increased.4) Results of apoptosis and cell cycle showed that S phase cells of intersititial cell reduced, G0/G1 phase cells increased, Cells into the proliferative phase reduced and DNA synthesis slowed down when using racemization gossypol. Gossypol acetic acid could inhibit the proliferation of intersititial cell and changed the cell cycle.