槲皮素抑菌作用的体内和体外研究

发布时间：2018-03-28 14:23

本文选题：槲皮素　切入点：蛋鸡　出处：《东北农业大学》2015年硕士论文

【摘要】：本试验选取240只健康、生产性能相近的39周龄海赛白壳蛋鸡随机分4组,每组6个重复,每个重复10只鸡,试验8周。对照组饲喂基础饲粮,3个试验组分别在基础饲粮中添加槲皮素(quercetin)0.02%,0.04%和0.06%,选取大肠杆菌、沙门氏菌巴氏杆菌、粪链球菌金黄色葡萄球菌变形杆菌、绿脓杆菌肠球菌作为体外试验受试菌株,采用牛津杯方法进行试验。研究槲皮素对8种禽类易感染细菌的体内和体外抑制作用及机制,为探求槲皮素作为改善蛋鸡肠道内环境和减少禽类易感染疾病发生的新型饲料添加剂提供理论依据,以便更好地指导生产实践。本研究包括以下4部分:⑴槲皮素对蛋鸡盲肠肠道菌群的影响与对照组相比,试验组2和3大肠杆菌菌落总数极显著降低(P0.01)。3个试验组金黄色葡萄球菌、沙门氏菌、绿脓杆菌、粪链球菌、肠球菌总数均显著降低(P0.01、P0.05、P0.05、P0.01、P0.05);3个试验组乳酸杆菌菌落总数有升高趋势(P=0.072);3个试验组双歧杆菌菌落总数显著提高(P0.05,P0.01,P0.05)。⑵槲皮素对8种禽类易感染细菌的体外抑制作用随着槲皮素浓度的增加,各受试菌株的抑菌圈直径极显著增加(P0.01)。大肠杆菌、沙门氏菌巴氏杆菌、变形杆菌、绿脓杆菌属革兰氏阴性菌(G-),金黄色葡萄球菌粪链球菌肠球菌属革兰氏阳性菌(G+),综合受试菌株的最小抑菌浓度(MIC)和抑菌圈直径的试验结果可知,槲皮素对G-杆菌的抑菌效果更好,G-杆菌对槲皮素的敏感度较高。⑶槲皮素和抗生素的抑菌效果比较将特异性抗生素作为阳性对照,针对槲皮素的不同浓度和抑菌圈做回归曲线,结果表明,各受试菌株达到MIC时抑菌圈直径所需的抗生素浓度均高于MIC,且抗生素的浓度不相同。⑷槲皮素对4种禽类易感染细菌的体外抑菌机制与对照组相比,槲皮素可影响受试菌株的超微结构,使受试菌株结构完整性和胞膜受损,细菌胞浆内容物渗漏,最终细菌空化死亡。槲皮素作用于受试菌株后,粪链球菌胞外渗出的碱性磷酸酶(AKP)量随着槲皮素添加量的增加呈上升趋势(P=0.069),其他三种受试菌株胞外渗出的AKP量极显著增加(P0.01)。随着槲皮素添加量的增加,受试菌株菌液中β-半乳糖苷酶含量可溶性蛋白浓度ATP酶活性(大肠杆菌除外)均极显著增加(P0.01)。综上所述,适宜剂量槲皮素可以通过提高蛋鸡肠道有益菌群(双歧杆菌)菌落总数,降低有害菌群(大肠杆菌、金黄色葡萄球菌、沙门氏菌、绿脓杆菌、粪链球菌、肠球菌)菌落总数发挥体内抑菌作用。在体外,适宜剂量槲皮素通过影响细胞壁和细胞膜发挥抑菌作用。
[Abstract]:In this study, 240 healthy and similar 39 week-old sea white hens were randomly divided into 4 groups, 6 replicates in each group and 10 in each replica. For 8 weeks, the control group was fed with basic diet, and the three groups were fed with quercetin, quercetinine 0.02% and 0.06% respectively in the basal diet. Escherichia coli and Salmonella were selected. Pasteurella, Streptococcus faecalis? Staphylococcus aureus? Proteus, Pseudomonas aeruginosa? Enterococcus spp., as a strain tested in vitro, was tested by Oxford Cup method. The inhibitory effect and mechanism of quercetin on 8 avian infectious bacteria in vivo and in vitro were studied. In order to provide theoretical basis for quercetin as a new feed additive to improve the intestinal environment and reduce the incidence of avian infectious diseases in laying hens. In order to better guide the production practice, this study included the following four parts: 1 quercetin effect on the intestinal flora of the caecum of laying hens, compared with the control group, the total number of Escherichia coli colonies in the test groups 2 and 3 was significantly lower than that in the control group, and the total number of Escherichia coli colonies in the three experimental groups was significantly lower than that in the control group. Salmonella, Pseudomonas aeruginosa, Streptococcus faecalis, The total number of Enterococcus significantly decreased P0.01P0.05P0.05P0.01P0.01P0.05; the total number of Lactobacillus in three experimental groups showed an increasing tendency to increase P0.072; the total number of bifidobacterium in three groups significantly increased the inhibitory effect of Quercetin on 8 avian infectious bacteria in vitro with the increase of Quercetin concentration, and the total number of Bifidobacterium in three groups was significantly higher than that of P0.05, P0.01 and P0.052.Quercetin inhibited the infection of 8 avian infectious bacteria in vitro with the increase of Quercetin concentration. The diameter of bacteriostasis circle of each tested strain was significantly increased P0.01. Escherichia coli, Salmonella? Pasteurella, Proteus, Pseudomonas aeruginosa, Gram-negative, Staphylococcus aureus? Fecal streptococcus? Enterococcus belongs to Gram-positive bacteria G, the minimum inhibitory concentration (MIC) of the tested strains and the diameter of the inhibition circle can be seen. The antimicrobial effect of quercetin on G- bacillus was better. The sensitivity of G- bacillus to quercetin was higher than that of antibiotics. The specific antibiotics were used as positive control, and the regression curves were made according to the different concentration and inhibition circle of quercetin. The results showed that the concentration of antibiotics needed for the diameter of bacteriostasis circle of each tested strain reached MIC was higher than that of MICM, and the antibacterial concentration of quercetin was different from that of control group, and the antibacterial mechanism of quercetin against four avian infectious bacteria in vitro was higher than that of control group. Quercetin could affect the ultrastructure of the tested strain, damage the structural integrity and cell membrane of the tested strain, leak the cytosolic contents of the bacteria, and eventually the bacteria died of cavitation. The amount of alkaline phosphatase (AKP) exudated by Streptococcus faecalis increased with the increase of quercetin addition. The amount of AKP exudated from the other three strains was significantly increased with the increase of quercetin content. The content of 尾-galactosidase in the liquid of the tested strain? Soluble protein concentration? The activity of ATP enzyme (except Escherichia coli) was significantly increased (P 0.01). In conclusion, the optimal dose of quercetin could decrease the number of harmful bacteria (E. coli, Staphylococcus aureus) by increasing the total number of beneficial bacteria (Bifidobacterium) in the intestinal tract of laying hens. Salmonella, Pseudomonas aeruginosa, Streptococcus faecalis, Enterococcus) colony total play an in vivo bacteriostatic effect. In vitro, the appropriate dose of quercetin by affecting the cell wall and cell membrane play a bacteriostatic effect.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831.5

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