本文關(guān)鍵詞： 雞 致病性E.coli 復(fù)方中草藥超微粉 防治試驗(yàn)　出處：《河北科技師范學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：為了篩選出有效的防治雞致病性大腸桿菌病復(fù)方中草藥超微粉,本試驗(yàn)以篩選的致病性最強(qiáng)優(yōu)勢血清型地方流行株QH1(O_(78))建立病理模型,進(jìn)行單味中草藥體外抑菌試驗(yàn),利用雞致病性大腸桿菌病的中獸醫(yī)辨證理論觀點(diǎn),選擇體外抑菌敏感單味中草藥作為主藥,根據(jù)各種單味中草藥配伍中君、臣、佐、使的藥理性質(zhì)組合成復(fù)方中草藥超微粉,通過體外抑菌試驗(yàn)和人工感染動(dòng)物試驗(yàn)篩選出抑菌效果好的復(fù)方中草藥超微粉,用人工感染14、49日齡雛雞致病性E.coli驗(yàn)證其復(fù)方中草藥超微粉防治效果,并檢測復(fù)方中草藥超微粉對(duì)人工感染14日齡雛雞致病性E.coli后的免疫機(jī)能指標(biāo)、血清生化指標(biāo),揭示其作用機(jī)理;通過對(duì)雛雞的急性、亞慢性毒性試驗(yàn)評(píng)價(jià)復(fù)方中草藥超微粉使用的安全性。本研究的試驗(yàn)結(jié)果如下:1.從秦皇島地區(qū)不同養(yǎng)雞場分離鑒定出56株E.coli,其中46株分離株為致病性E.coli,占分離菌株的82.1%;46株致病性E.coli屬于11種血清型,以O(shè)_(78),O89,O142及O1為優(yōu)勢血清型;46株致病性E.coli攜帶16種不同毒力基因。根據(jù)E.coli分離菌株的動(dòng)物致病性試驗(yàn)、血清型及毒力基因攜帶情況,篩選出了致病性最強(qiáng)優(yōu)勢血清型地方流行株QH1(O_(78))。對(duì)1,14,49日齡雛雞的LD50分別為3.16×106 CFU/mL,1.07×107 CFU/m L,5×108 CFU/mL。2.單味中草藥體外抑菌試驗(yàn),利用平板瓊脂打孔法和改良微量二倍稀釋-平板法,從26種中草藥中篩選出對(duì)雞致病性E.coli的地方株QH1(O_(78))、QH2(O89)、QH4(O1)均敏感的中草藥金銀花、黃連、烏梅、五味子,其抑菌圈直徑在20.3~22.7mm之間,MIC在15.65~31.25 mg/mL之間。3.復(fù)方中草藥超微粉的篩選,利用雞致病性大腸桿菌病的中獸醫(yī)理論辨證理論觀點(diǎn)與各種單味中草藥配伍中的藥理性質(zhì)組合5個(gè)復(fù)方中草藥,通過體外抑菌和人工感染動(dòng)物性試驗(yàn),篩選出了抑菌效果最好的復(fù)方Ⅲ。4.復(fù)方中草藥超微粉(復(fù)方Ⅲ)對(duì)人工感染14,49日齡雛雞致病性E.coli防治試驗(yàn)的結(jié)果顯示,C組(1%禽用頭孢噻呋鈉)和D~F組(復(fù)方Ⅲ添加量分別為1.5%、2.0%、2.5%)的死亡率明顯低于B組(陽性對(duì)照組),E組和F組的預(yù)防與治療效果均高于B組與C組,其中以E組對(duì)14,49日齡雛雞致病性大腸桿菌病防治效果最好,保護(hù)率分別為86.7%,90.0%;治愈率分別為92.9%,90.0%;有效率分別為89.7%,93.3%。5.復(fù)方中草藥超微粉(復(fù)方Ⅲ)對(duì)人工感染14日齡雛雞致病性E.coli血清中生化指標(biāo)、免疫機(jī)能的檢測,A~F組分別在試驗(yàn)后的第7,14,21天采血分離血清進(jìn)行相關(guān)指標(biāo)檢測。結(jié)果顯示,E組(2.0%復(fù)方Ⅲ添加)效果最好,能提高血清中免疫蛋白IgG,IgM,IgA水平;提高血清中LZM,TP,GLB的含量;降低血清中GOT,TG,TC,BUN,UA含量。6.急性性毒性試驗(yàn)結(jié)果顯示復(fù)方Ⅲ對(duì)雛雞是無急性毒副作用,最大耐受量超過25g/(kg·d)。亞慢性毒性試驗(yàn)結(jié)果顯示,復(fù)方Ⅲ對(duì)雛雞長期服用是安全的。
[Abstract]:In order to screen out the superfine powder of Chinese herbal medicine for the prevention and treatment of chicken pathogenic Escherichia coli, the pathological model was established by screening the most dominant serotype local epidemic strain QH1 / OF78, and the bacteriostasis test of single Chinese herbal medicine was carried out in vitro. Based on the theory of Chinese veterinarian syndrome differentiation of pathogenic Escherichia coli disease in chicken, the Chinese herbal medicine with single Chinese herbal medicine was selected as the main medicine in vitro. According to the compatibility of each kind of Chinese herbal medicine with Zhong Jun, Chen and Zuo, the pharmacological properties were combined into compound Chinese herbal medicine ultrafine powder. In vitro bacteriostasis test and artificial infection animal test were used to screen the superfine powder of compound Chinese herbal medicine with good bacteriostasis effect. The pathogenicity E.coli of 14 ~ 49 days old chicks infected artificially was used to verify the control effect of the compound Chinese herbal medicine ultrafine powder. The immune function index, serum biochemical index and mechanism of compound Chinese herbal medicine ultrafine powder on 14 day old chickens infected by artificial infection were detected, and the mechanism of their action was revealed. The results of this study are as follows: 1. 56 strains of E. coli were isolated from different chicken farms in Qinhuangdao area, of which 46 strains were pathogenicity E.coli, accounting for the isolation of bacteria. There are 11 serotypes in pathogenic E. coli. 46 strains of virulence E. coli carried 16 different virulence genes. According to the animal pathogenicity test, serotype and virulence gene carrying status of E. coli isolates. The most virulent serotypic endemic strain QH1 / L was screened out. The LD50 of the chicken was 3.16 脳 106CFU / mL1.07 脳 107CFU / mL1.07 脳 107CFU / mL / mL, 5 脳 108CFU / mL-1 respectively. The bacteriostasis test of single Chinese herbal medicine in vitro was carried out by using plate Agar punching method and modified microdilution plate method. The Chinese herbal medicines Flos Lonicerae, Coptis chinensis, Fraxinus chinensis, Fructus Schisandrae and Schisandra chinensis, which are sensitive to the local strain QH1 / OH7, QH2O89, QH4O1, have been screened from 26 Chinese herbal medicines. The inhibitory circle diameter of Flos Lonicerae, Coptis chinensis, Fructus Schisandra and Fructus Schisandrae is between 20.32mm and 22.7mm, and the MIC is 15.653.31.25 mg/mL. Five compound Chinese herbal medicines were combined by using the theory of Chinese veterinary medicine dialectical theory of pathogenic Escherichia coli disease in chicken and the pharmacological properties of each kind of Chinese herbal medicine. The in vitro bacteriostasis and artificial infection of animal were tested. Compound 鈪，

本文編號(hào)：1539588