本文選題：犢牛腹瀉　切入點：大腸桿菌　出處：《內(nèi)蒙古農(nóng)業(yè)大學》2015年碩士論文

【摘要】：長期使用抗生素會引發(fā)許多如畜產(chǎn)品內(nèi)藥物殘留、耐藥性等現(xiàn)象。蒙藥是一種純天然的藥物,在課題組前期研究中發(fā)現(xiàn),蒙藥巴布-7抑菌作用強,療效顯著。為了篩選出該復方諸多成分中,有效抑菌成分以及最佳體內(nèi)外抑菌組方,并揭示其作用機理,為此進行了本研究。試驗結(jié)果如下：1.在調(diào)查呼和浩特市周邊致病性大腸桿菌的血清學分布時,從呼和浩特市周邊某牛場采集了117份糞樣,進行分離培養(yǎng)后得到97株疑似大腸桿菌,進行形態(tài)學觀察與生化鑒定,確定其中65株為大腸桿菌。進行“O”血清型鑒定,分離出0145、O152、0167三種血清型,分別占65株大腸桿菌的1.96%、1.96%、7.84%。將這65株大腸桿菌進行PCR鑒定后發(fā)現(xiàn)又多了新的血清型026和083,這兩種血清型在血清型試驗中沒有檢測到,該結(jié)果說明了PCR方法的敏感性。將這65株大腸桿菌進行動物致病性試驗,其中有51株為強致病性大腸桿菌,占采樣量的40%。說明強致病性大腸桿菌分布較廣。2.采用牛津杯法,初步篩選出蒙藥巴布-7的6種抑菌有效成分中的5種對致犢牛腹瀉病大腸桿菌均有抑菌作用,測定最低抑菌濃度(MIC)與分級抑菌濃度(FIC)指數(shù),確定5種藥物互相都無拮抗作用,適宜配伍。5種藥物對大腸桿菌的MIC分別是茜草素為5mg/mL,大黃酸為20mg/mL,大黃素為20mg/mL,槲皮素為7.5mg/mL,沒食子酸為7.5mg/mL3.首先得出對大腸桿菌具有最佳抑菌作用的藥物,針對大腸桿菌5種藥物均用藥。之后確定各藥物之間的劑量配比關(guān)系。得出對大腸桿菌抑菌效果最好的藥物組合為A2B1C3D2E2.4.采用微量稀釋法測定抑菌組方的MIC,得出蒙藥抑菌組方對大腸桿菌的最低抑菌濃度為0.675mg/mL,通過測定小鼠體內(nèi)保護率得出該抑菌組方保護率為50%,說明蒙藥抑菌成分組方體內(nèi)抑菌保護率與陽性對照組環(huán)丙沙星66.7%接近。5.為了給優(yōu)化后的抑菌組方的抑菌作用提供理論依據(jù),試驗通過鄰硝基苯β-D-半乳吡喃糖苷(ONPG)法測定其對E. coli細胞膜滲透性的影響,并測定其對E. coli電導率、細胞壁的影響,進一步研究了抑菌組方對大腸桿菌細胞膜的影響。結(jié)果表明,大腸桿菌經(jīng)過抑菌組方作用后,細胞膜的通透性、溶液的電導率與堿性磷酸酶(AKP)的含量均增加。證明藥物可能是通過破壞細菌細胞膜導致其滲透性增加,從而發(fā)揮其抑菌作用。6.為了測定蒙藥抑菌成分復方的抗菌譜,通過體外抑菌直徑的測定與最低抑菌濃度的測定,得出該抑菌組方對大腸桿菌、金黃色葡萄球菌等細菌均有抑制作用。
[Abstract]:Long-term use of antibiotics can lead to many phenomena such as drug residues in animal products, drug resistance, etc. Mongolian medicine is a pure natural drug, in the previous research group found that the Mongolian drug Babu-7 strong bacteriostatic effect, In order to screen out the effective antimicrobial components and the best in vitro and in vivo antimicrobial components of the compound, and to reveal the mechanism of its action, The results are as follows: 1.When investigating the serological distribution of pathogenic Escherichia coli around Hohhot, 117 feces were collected from a cattle farm around Hohhot. After isolation and culture, 97 strains of suspected Escherichia coli were obtained, and 65 of them were identified as E. coli by morphological observation and biochemical identification. The new serotypes 026 and 083 were found after the PCR identification of the 65 strains of Escherichia coli. These two serotypes were not detected in the serotype test. The results showed the sensitivity of the PCR method. Among the 65 strains of Escherichia coli, 51 were strong pathogenic Escherichia coli, accounting for 40% of the sample size. 2. 2. Oxford Cup method was used to test the pathogenicity of Escherichia coli. Five of the six antimicrobial active components of Mongolian drug Babu-7 had bacteriostatic effect on Escherichia coli caused by diarrhea disease in calves. The index of minimal inhibitory concentration (MIC) and fractionated inhibitory concentration (FICI) were determined, and the five drugs had no antagonistic effect on each other. The suitable MIC for E. coli was 5 mg / mL for alizarin, 20 mg / mL for Rhein, 20 mg / mL for emodin, 7.5 mg / mL for quercetin and 7.5 mg / mL for Gallic acid, respectively. The dose-ratio relationship between each drug was determined. The best drug combination for Escherichia coli was A _ 2B _ 1C _ 3D _ 2E _ 2.4.The microdilution method was used to determine the antimicrobial composition and the Mongolian medicine was obtained. The minimum inhibitory concentration of bacteriostasis prescription against Escherichia coli was 0.675 mg / mL. The protective rate of the antimicrobial component was 50 by measuring the protective rate in vivo of mice, indicating that the bacteriostatic protection rate in Mongolian medicine was 66.7% compared with ciprofloxacin in positive control group. In order to provide a theoretical basis for the bacteriostatic effect of the optimized antimicrobial prescription, The effects of o-nitrobenzene 尾 -Dgalactopyranoside on the permeability of E. coli cell membrane, the conductivity of E. coli and the cell wall were measured by the method of o-nitrobenzene 尾 -D- galactopyranoside. The effect of antimicrobial prescription on the membrane of Escherichia coli was further studied. Both the electrical conductivity of the solution and the content of alkaline phosphatase (AKP) increased. It was proved that the drug may cause the increase of its permeability by destroying the bacterial cell membrane, thus exerting its bacteriostatic effect .6. in order to determine the antibacterial spectrum of the compound of the bacteriostasis component of Mongolian medicine, Through the determination of the antibacterial diameter in vitro and the determination of the minimum inhibitory concentration, it was concluded that the antimicrobial compound had inhibitory effect on Escherichia coli, Staphylococcus aureus and other bacteria.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S852.61;S853.7

【參考文獻】

相關(guān)期刊論文 前10條

1 劉程惠;胡文忠;何煜波;姜愛麗;徐國凱;;中華民族醫(yī)藥——蒙藥的發(fā)展歷史與研究進展[J];安徽農(nóng)業(yè)科學;2012年22期

2 蔡世X,張瑛;犢牛腹瀉的原因與防治[J];黃牛雜志;2004年02期

3 胡先春,金利峰,李彩紅,李蓉;動物急性感染性腹瀉的病原、發(fā)病機制和治療[J];黃牛雜志;2005年04期

4 葉賀佳,葉萬樹,黃東璋;細菌耐藥性的產(chǎn)生機理及其控制對策[J];動物醫(yī)學進展;2005年10期

5 菅永峰;王瑞;徐昌領(lǐng);喬虹;王林川;;鴨致病性大腸桿菌的分離與鑒定[J];廣東畜牧獸醫(yī)科技;2006年02期

6 孫鵬舉;新生牛的殺手——犢牛腹瀉[J];中國畜牧獸醫(yī);2003年04期

7 荔霞;王勝義;劉永明;齊志明;董書偉;劉世祥;;犢牛腹瀉病因及其藥物防治研究進展[J];中國畜牧獸醫(yī);2010年10期

8 易檢祥;;提高犢牛頑固性腹瀉病治愈率的關(guān)鍵技術(shù)[J];中國牛業(yè)科學;2014年06期

9 秦曉蓉;張銘金;高緒娜;林義;馬利;何思宜;;槲皮素抗菌活性的研究[J];化學與生物工程;2009年04期

10 黃啟相;;黃芩湯加減治療犢牛腹瀉[J];畜牧與飼料科學(畜牧版);2006年01期

相關(guān)會議論文 前1條

1 石玉江;吳桂梅;;蒙藥基本性能[A];全國民族醫(yī)藥�？茖２W術(shù)研討會論文選編[C];2001年

相關(guān)碩士學位論文 前4條

1 方光遠;犢牛腹瀉大腸桿菌LEE和HPI毒力島相關(guān)基因的檢測及特性分析[D];南京農(nóng)業(yè)大學;2007年

2 劉延麟;沒食子單體化合物分離鑒定及其藥理活性研究[D];吉林大學;2009年

3 金婷婷;免疫捕獲—實時熒光LAMP法快速檢測大腸埃希氏O157：H7的研究[D];河北農(nóng)業(yè)大學;2013年

4 姚媛媛;致犢牛腹瀉病腸道大腸桿菌的分離鑒定及蒙藥復方對其抗菌效果研究[D];內(nèi)蒙古農(nóng)業(yè)大學;2014年

，

本文編號：1685831