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Study on the Mechanism of Growth-Promoting and Bacter-Inhibiting of Pulsatilla Chinensis in AA Broilers

Author: HouXiang
Tutor: WangTian
School: Nanjing Agricultural College
Course: Animal Nutrition and Feed Science
Keywords: Pulsatilla chinensis Anemonin Bacteria-inhibiting Immunologic function Production performance
CLC: S831
Type: Master's thesis
Year: 2009
Downloads: 11
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Abstract

Recent studies suggest that addition Pulsatilla chinensis the original role of anti-bacte-rial and resistant, it has function of enhancing the body immunity, inhibiting macrophage of secretion, anti-cancer, anti-inflammatory, antitoxic, diarrhea, sedation, analgesia, killing coccidia inhibiting peristalsis, and anti-endotoxemia. The purpose of this study was just to confirm the main active ingredient of anemonin in antimicrobial, and studied the effect of Pulsatilla chinensis on immune function production performance, physiological function and discussed its growth mechanism to chickens. So that we can provide an scientific basis for Baitouweng as the further development of antimicrobial feed additives to poultry production.Test 1 The preparation of anemonin by the method of water distillation. we took appropriate of Pulsatilla chinensis, crushing and plused some amount of water. Then 40℃water bath pot Purchase for 24 h. Used rotary evaporator vacuum distillation to collect distilled liquid, cooling, removed the float in the above-insoluble impurities, plusd sodium chloride, chlorine vibration extraction, and combined extract. And then used the method of decompression steamed to solvents, and leaved the provocative legacy sallow oily liquid. Finally, we left the crude anemonin standing a few days, re-crystallization and got the crystallization anemonin. In the end, to analysis anemonin, we used high performan-ce liquid chromatography and nuclear magnetic resonance.Test 2 Effects of Pulsatilla chinensis extract on production performance, blood parameters of AA broilers.480 1-day-old healthy chickens were used and randomly divided into 5 treatment groups with 6 duplicates:control group, treatment group supplement with 0.5%,1% and 2% Pulsatilla chinensis extract respectively, and antibiotic treatment group at the 40 days. Broiler body weight, feed consumption, blood parameters and immunity index were determined respectively. The results showed Pulsatilla chinensis extract supplement improved broiler body weight and feed conversation ratio, and 2% supplemental group got the best compare to other supplemental group and control, which increased body weight gain by 5.42%(P<0.05) and improved feed conversion ratio by 5.50% (P<0.05) on 42 days. Spleen index, thymus index, bursa of Fabricius index were increased respectively by 10.52%,8.68%,47.62%, compared with the control group on 42 days. In addition, Pulsatilla chinensis extract supplement increased the content of serum total protein and albumin, the activity of SOD, GSH-PX, the levels of T3, T4. GOT and GPT were slightly enhanced (P>0.05)Test 3 Study on antibacterial activity of Pulsatilla chinensis. (1) bacteriostatic test in vitro:through the Oxford Cup method we tested the effect of vitro antibacterial of Pulsatilla chinensis decoction and anemonin. The results showed that 0.5%,1% decoction and 0.1% anemonin had bacteriostasis to E. coli, Proteus, Enterobacter cloacae, and 0.1 mg/mL anemonin of E. coli, Proteus inhibition zone diameter of greater than 0.4mg/L Enrofloxacin. (2) antimicrobial activity in vivo:Pulsatilla chinensis extract used as an additive for chicken feeding. The division and management just as test one. The number of cecal E. coli and lactic acid bacteria were counted by plate to study the impact of Pulsatilla chinensis on chickens intestinal flora. And then, determine the consistency of antibacterial role by the vivo and vitro test. The results showed that the number of E. coli reduced 2.73%、8.45% and 9.94%(P>0.05) of different concentrations of Pulsatilla chinensis extract in the early stage compared with blank group,8.45% and 9.94% (P>0.05); While in the late stage, the number of actic acid bacteria had upward trend for treatment group compared to blank group, but there was no significant difference (P>0.05).Test 4 Effect of Pulsatilla chinensis on immunologic function of chicken. (1) in vivo tests:we tested the impact of different concentrations(100、200、400、800 mg/mL) of Pulsatilla chinensis distilled water on chickens splenocyte by the MTT method. The results showed that all of the four concentrations can stimulate proliferation of splenocyte induced by ConA. While 400,800 mg/mL concentration had a significantly level (P<0.05); It also had an significantly enhanced effect to splenocyte induced by 4 LPS (P<0.05) of this four concentration Pulsatilla chinensis distilled water, and proliferated with the increase concentration Pulsatilla chinensis. (2) in vitro test:Pulsatilla chinensis extract used as an additive for chicken feeding. The division and management just as test one. The thymus, spleen, bursa of Fabricius were rmoved to calculated immune organ index at 21 and 42 d. The results showed that the spleen index increased by 12.84%,14.68% treated by 0.5%,1% Pulsatilla chinensis extract compared to control group in the early stage(P>0.05); While the thymus index increased 20.98%/1.95%、12.68% beside the Bursal index increased 2.94%, 12.41%,6.86% treaded by 0.5%,1%,2% concentration group (P>0.05); In the late stage, spleen index increased by 9.65%,14.04%,10.52%(P>0.05), thymus index increased by 7.31%,3.65%,8.68%(P>0.05) and bursal index increased by 47.62%,52.38% and 47.62% (P<0.05) by 0.5%,1% and 2% Pulsatilla chinensis extract compared with the control group.Test 5 The effect of Pulsatilla chinensis on antioxidative function of chicken. Pulsatilla chinensis extract used as an additive for chicken feeding. The division and management just as test two. The blood was picked respectively 21 and 42 d, and tested activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX). The results as follows:in the early stage, there was no significant changes for SOD activity treated by Pulsatilla chinensis compared to control group (P>0.05). While the activity of GSH-PX had a significant increase treated by Pulsatilla chinensis, and 2% value a significantly level (P<0.05); At later stage, SOD value a significant increase for the treated group compared to control group (P<0.05), and GSH-PX activity had an increase for treated group but did not significant (P>0.05).These results suggest that anemonin was the main active ingredient to Pulsatilla chinensis. Pulsatilla chinensis can inhibit the growth of E. coli, promote the growth of lactic acid bacteria and improve the bacteria environmentof chickens; It also can enhance the growth of immune organs and slow the degradation of lymphoid organ. Pulsatilla chinensis distilled liquid can stimulate proliferation of splenocyte induced by ConA or LPS, and proliferated with the concentration increase of Pulsatilla chinensis distilled liquid, which has been strengthen the immune function of chickens; Pulsatilla chinensis can improve activity of serum SOD and GSH-PX, and the activity tended to increase with the increased Pulsatilla chinensis. It can improve the antioxidant capacity and reduce damage of organizati by free radical to which conducive to the growth of chickens; Pulsatilla chinensis can also improve the metabolism and speed the aqqradation of protein. All of these results were supported by an increase of body weigh and weight ratio of chickens.

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