本文選題：槲皮素　切入點(diǎn)：蛋雞　出處：《東北農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：本試驗(yàn)選取240只健康、生產(chǎn)性能相近的39周齡海賽白殼蛋雞隨機(jī)分4組,每組6個(gè)重復(fù),每個(gè)重復(fù)10只雞,試驗(yàn)8周。對照組飼喂基礎(chǔ)飼糧,3個(gè)試驗(yàn)組分別在基礎(chǔ)飼糧中添加槲皮素(quercetin)0.02%,0.04%和0.06%,選取大腸桿菌、沙門氏菌巴氏桿菌、糞鏈球菌金黃色葡萄球菌變形桿菌、綠膿桿菌腸球菌作為體外試驗(yàn)受試菌株,采用牛津杯方法進(jìn)行試驗(yàn)。研究槲皮素對8種禽類易感染細(xì)菌的體內(nèi)和體外抑制作用及機(jī)制,為探求槲皮素作為改善蛋雞腸道內(nèi)環(huán)境和減少禽類易感染疾病發(fā)生的新型飼料添加劑提供理論依據(jù),以便更好地指導(dǎo)生產(chǎn)實(shí)踐。本研究包括以下4部分:⑴槲皮素對蛋雞盲腸腸道菌群的影響與對照組相比,試驗(yàn)組2和3大腸桿菌菌落總數(shù)極顯著降低(P0.01)。3個(gè)試驗(yàn)組金黃色葡萄球菌、沙門氏菌、綠膿桿菌、糞鏈球菌、腸球菌總數(shù)均顯著降低(P0.01、P0.05、P0.05、P0.01、P0.05);3個(gè)試驗(yàn)組乳酸桿菌菌落總數(shù)有升高趨勢(P=0.072);3個(gè)試驗(yàn)組雙歧桿菌菌落總數(shù)顯著提高(P0.05,P0.01,P0.05)。⑵槲皮素對8種禽類易感染細(xì)菌的體外抑制作用隨著槲皮素濃度的增加,各受試菌株的抑菌圈直徑極顯著增加(P0.01)。大腸桿菌、沙門氏菌巴氏桿菌、變形桿菌、綠膿桿菌屬革蘭氏陰性菌(G-),金黃色葡萄球菌糞鏈球菌腸球菌屬革蘭氏陽性菌(G+),綜合受試菌株的最小抑菌濃度(MIC)和抑菌圈直徑的試驗(yàn)結(jié)果可知,槲皮素對G-桿菌的抑菌效果更好,G-桿菌對槲皮素的敏感度較高。⑶槲皮素和抗生素的抑菌效果比較將特異性抗生素作為陽性對照,針對槲皮素的不同濃度和抑菌圈做回歸曲線,結(jié)果表明,各受試菌株達(dá)到MIC時(shí)抑菌圈直徑所需的抗生素濃度均高于MIC,且抗生素的濃度不相同。⑷槲皮素對4種禽類易感染細(xì)菌的體外抑菌機(jī)制與對照組相比,槲皮素可影響受試菌株的超微結(jié)構(gòu),使受試菌株結(jié)構(gòu)完整性和胞膜受損,細(xì)菌胞漿內(nèi)容物滲漏,最終細(xì)菌空化死亡。槲皮素作用于受試菌株后,糞鏈球菌胞外滲出的堿性磷酸酶(AKP)量隨著槲皮素添加量的增加呈上升趨勢(P=0.069),其他三種受試菌株胞外滲出的AKP量極顯著增加(P0.01)。隨著槲皮素添加量的增加,受試菌株菌液中β-半乳糖苷酶含量可溶性蛋白濃度ATP酶活性(大腸桿菌除外)均極顯著增加(P0.01)。綜上所述,適宜劑量槲皮素可以通過提高蛋雞腸道有益菌群(雙歧桿菌)菌落總數(shù),降低有害菌群(大腸桿菌、金黃色葡萄球菌、沙門氏菌、綠膿桿菌、糞鏈球菌、腸球菌)菌落總數(shù)發(fā)揮體內(nèi)抑菌作用。在體外,適宜劑量槲皮素通過影響細(xì)胞壁和細(xì)胞膜發(fā)揮抑菌作用。
[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.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S831.5

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