Study On The Biological Effect And Mechanism Of Bamboo Leaf Flavonoid In Broilers

Bamboo Leaf Flavone is a physiologically active compound extracted from bamboo leaves. It is now widely used in medical health and food production but rarely used in broiler production. This experiment studied at the optimum extraction process of bamboo leaf flavone, the antebacterial and antioxidative effect of it in vitro, the safety evaluation of broilers, the protective effect on acute liver injury in broilers, the effect on immunity in broilers, the effect on cecal microflora community diversity, the effect and mechanism of production performance in broilers under heat stress and routine breeding and provide theoretical foundation in production and.Trial 1:This expriment studied the effect of volume fraction of extractant, extraction time, temperature, liquid to solid ratio on four kinds of bamboo leaves such as Mao, Ci, Qinsi, Mu bamboo through the application of ultrasonic wave and tested their antebacterial and antioxidative activities. The extraction rate of Mao, Ci, Qinsi, Mu bamboo were 16.25%,12.35%,9.58% and 5.49%, respectively. The results suggested the effects of extraction methods were ordered as follows. Ethanol volume fraction B> Reaction time A > Extraction temprature C> Liquid to solid ratio D. The optimum extraction procedures were extracting in volume fraction of 76% ethanol with liquid-to-solid ratio 10:1(mL/g), the leaching rate of flavone reaches its highest point. There was a positive correlation between the effect of four bamboom leaf flavone on one bacteria and total content of bamboom leaf flavone. Tests on MIC and MBC via means of doubling dilution suggested that the minimum MIC of Mao, Ci, Qinsi, Mu bamboom towards Samonella are 153.19 μg /mL,81.51 μg/mL,139.00 μg/mL, and 219.95 μg/mL, respectively. The minimum MBC were 306.38μg/mL,163.03 μg/mL,278.00 μg/mL,219.05 μg/mL, respectively. The minimum MIC of Mao, Ci, Qinsi, Mu bamboom towards Escherichia Coli were 306.38 μg/mL,162.80 μg/mL,278.00 μg/mL,439.90 μg/mL, respectively. The minimum MBC were 612.75 μg/mL,325.60 μg/mL,556.00 μg/mL,439.90 μg/mL, respectively. The minimum MIC of Mao, Ci, Qinsi, Mu bamboom towards Staphylococcus aureus were 153.19 μg/mL,81.40 μg/mL,278.00 μg/mL,109.98 μg/mL respectively. The minimum MBC were 306.38 μg/mL,162.80 μg/mL,278.00 μg/mL,219.95 μg/mL, respectively. It suggested that the sensitivity of four kinds of bamboom leaf flavone towards Escherichia Coli and Salmonella from high to low was Ci, Qins, Mao, Mu bamboo and the sensitivity of the four kinds of bamboom leaf flavones toward Staphylococcus aureus from high to low was Ci, Mao, Mu, Qinsi bamboom. The variation of sensitivity toward different types of bacteria reflects different types of flavonoids. We tested the antioxidant activities of four kinds of flavonoids by means of oxygen free radical clearance method, hydroxyl free radical clearance method, DPPH free radical clearance method, respectively. The results suggested that the clearance rate of oxygen free radical from high to low was Ci> Qinsi> Mao> Mu bamboo, the clearance rate of hydroxyl free radical from high to low was Ci> Mao> Qinsi> Mu, the clearance rate of DPPH free radical from high to low was Ci> Mao> Qinsi> Mu. As a result, bamboom leaf flavone has a strong antioxidant effect in vitro with the best effect extracted from Ci bamboom.Trial 2:The safety evaluation of bamboom leaf flavone on broilers. We selected 200 healthy broilers with same weights and randomLy separated them into five groups. Each group had four repetitions and each repetition had 10 broilers. The broilers were given bamboo leaf flavone for continuous 7 days at recommended clinical dosage of one time, three times, five times and ten times. The results suggested that during experimental period, target broilers increased weight, the number of erythrocyte and the weight of leokocyte significantly at the dosage of three times. The antioxidant, spleen and bursa of Fabricius indexes were significantly increased at the dosage of three times and five times while decreased at excessive dosage. There is no significant difference in the contents of AST, ALT, BUN, and CRE, compared with the control group in serum of experimental groups. Compared with the control group, there were no abnormal changes in the structure of main organ tissues at the dosage of ten times. The results suggested that bamboom leaf flavone is safe with no toxic and side effect in clinical use.Trial 3:The protective effect of bamboom leaf flavone on acute liver injury in broilers. In this experiment, we adopted 36 on 37-day-old healthy male AA broilers and randomLy separated them into six groups. We established acute liver injury model with CC14 and classified them into the blank group, the model group, the positive group, low dosage group (20mg/kg.b.w), medium dosage group (40mg/kg.b.w) and high dosage group (100mg/kg.b.w) with administration of bamboom leaf flavone. The results suggested that the growth wate of the model group was significantly lower than other groups (P<0.05). The liver, spleen, kidney swelled significantly with darker color and organ indexes were higher than the blank group. The activities of ALT, AST, MDA in serum were significantly higher than other groups. The content of SOD in serum was lower than other groups. Compared with the blank group, the positive group and administration groups showed no significant discrepancy in main biochemical indexes in serum and TC, TG remain constant. Meanwhile, administration groups reduced damage to liver cells by CC14, remained proliferation and differentiation of liver cells and inhibited excessive apoptosis. Bamboom leaf flavone can improve body antioxidant ability and decreased damage of lipid superoxidation. It can protect liver through guaranteeing normal metabolise of fat. The best dosage is 40mg/kg.b.w.Trial 4:The effect of bamboom leaf flavone on immune organs and function in broilers. In this experiment, we adopted 300 one-day-old male AA broilers and randomLy separated them into four groups. Each group had three repetitions and each repetition had 25 broilers. Except for the control group, other three groups were fed with basal diets added bamboom leaf flavone (200 mg/kg,400 g/kg, and 800 mg/kg, respectively) for continous 42 days. The results suggested that bamboom leaf flavone could increase immune organ indexes, improved EtRFC%, promoted proliferation and differentiation of spleen, thymus and bursa of Fabricius and inhibited apoptosis. The antibody titers of ND and AI in experimental group were increased, promoting gene expression of IL-2 and INF-γ in spleen and improving the contents of IL-2 and INF-γ in serum. This experiment suggested that bamboom leaf flavone has an effect on improving immunity of broilers.Trial 5:The effect of bamboom leaf flavone on cecal microflora community diversity of broilers. The classification was the same as Trial four. With the use of Illumina MiSeq high-throughput sequencing, we analyzed the effect of bamboom leaf flavone on cecal microflora community diversity of broilers. The results suggested that we obtained 1,240,937 significant sequences with 34,370 sequences of each sample on average. We got 95,287 OUTs through 97% sequence similarity. The dominant microflora was Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria. Compared with the control group, there was a significant discrepancy in intestinal microflora community diversity at Phylum and Genus level at each time point. The results suggested that bamboom leaf flavone has a significant effect on the diversity of microflora in intestine of broilers in a dose-dependent manner.Trial 6:The effect of bamboom leaf flavone on broilers under heat stress. In the study, we adopted 150 seven-day-old AA broilers and randomLy separated them into five groups. Each group had three repetitions and each repetition had 10 broilers. Except for the control group, other four groups were fed with basal diets added bamboom leaf flavone (400 mg/kg,800mg/kg,1600 mg/kg, and 3200mg/kg, respectively) for continous five weeks. The results suggested that the ratio of food to gain and cooked loss rate were decreased, the slaughter performances and water holding capacity were increased, the flesh color and meat tenderness ware improved. The content of AST in serum, MDA in liver and MDA in leg muscle with the addition of bamboom leaf flavone decreased significantly while the content of SOD in liver, the activities of T-AOCB and SOD in breast muscle and SOD in leg muscle increased. The results suggested that through increasing antioxidant ability of broilers, bamboom leaf flavone can improve the production performance of broilers under heat stress, the slaughter performance and meat quality. The best dosage is 1.6g/kg.b.w.Trial 7:The effect of bamboom leaf flavone on production performance of broilers v ia genetic pathway. The classification was the same as Trial four. Bamboom leaf flavone c an improve growth performance, food conversion rate, slaughter performance and meat qu ality. Meanwhile, bamboom leaf flavone significantly increased the activities of SOD, T-AOC, CAT, and the content of HDL-C and decreased the content of MDA, TG and T C in blood. By this way, it can affect blood lipid level while it has no significant effect on t he content of ALB, AST and BUN. Through regulating gene expression of PPARy, ATG L. ADSL in liver and LPL, ADSL in breast muscle, it can affect fat metabolism of broiler s and improve inosinic acid in muscle so as to improve chicken flavor. In a word, bamb oom leaf flavone has antebacterial and antioxidative effect in vitro and is safe with no toxic at the dosage of ten times in clinical use. Through improving antioxidant ability, promoting proliferation and differentiation of liver cells and inhibiting exce ssive apoptosis, it can reduce damage to liver cells from CC14 Adding bamboom 1 eaf flavone in basal diets can promote gene expression of IL-2 and INF-γ in splee n and improve the content of IL-2 and INF-γ in serum to promote the developme nt of immune organs and immune function of broilers. Through affecting cecal mic roflora varieties, it can affect the weight of broilers. The ability of relieving produc tion performance under heat stress is related to the improvement of antioxidant lev el of broilers. It can improve production performance of broilers under normal feed ing and regulate PPAR-γ, ATGL, ADSL gene in liver and LPL, ADSL gene in brea st muscle to affect fat deposition and the content of inosinic acid. The dosage of 4 00-800mg/kg is highly recommended.