本文選題：生長抑素 + 基因疫苗��； 參考：《華中農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：前期研究證明,重組菌C500(p VGS/2SS-asd)基因疫苗免疫可以促進(jìn)小鼠生長,并且對(duì)小動(dòng)物無不良影響。為了進(jìn)一步了解該疫苗對(duì)大動(dòng)物的免疫效果及其機(jī)制和安全性,本研究選擇78頭80日齡的杜長大育肥豬,隨機(jī)分成6組,用同一劑量(5×1011CFU/頭)三種免疫方式(口腔+鼻腔、口腔、鼻腔)、同一免疫方式(口腔+鼻腔)三種劑量(高5×1011 CFU/頭、中5×1010 CFU/頭、低5×109 CFU/頭)進(jìn)行臨床試驗(yàn),試驗(yàn)期90 d,探討最適免疫方法和免疫劑量。采用美國奧飼本(OSBORNE)公司生產(chǎn)的投料器和種豬生產(chǎn)性能WINFIRE測(cè)定系統(tǒng)每天自動(dòng)記錄實(shí)驗(yàn)豬的飼料消耗量、測(cè)定豬體重,飼料報(bào)酬以評(píng)定基因疫苗免疫效果;從免疫后豬的日常行為及體溫是否正常、應(yīng)用目的基因擴(kuò)增等方法從基因整合、環(huán)境擴(kuò)散等方面評(píng)價(jià)疫苗的安全性;應(yīng)用ELISA方法測(cè)定抗體和激素水平,以探討其作用機(jī)制。結(jié)果如下:1.促生長效果及飼料報(bào)酬采用美國奧飼本公司生產(chǎn)的種豬生產(chǎn)性能WINFIRE測(cè)定系統(tǒng)每天全自動(dòng)記錄豬采食量和體重?cái)?shù)據(jù),免疫前期育肥豬生長速度較快,后期增重減緩;探究免疫方式對(duì)育肥豬的促生長效果,結(jié)果表明,各時(shí)期試驗(yàn)組日增重均高于對(duì)照組,免疫后4-7 w,噴鼻組日增重顯著高于對(duì)照組(P0.05);探究免疫劑量對(duì)育肥豬的促生長效果,結(jié)果表明,與對(duì)照組相比,初免疫后7 w,中劑量、低劑量組日增重均提高19.94%,差異極顯著(P0.01),整個(gè)試驗(yàn)階段,各免疫組的平均日增重均高于對(duì)照組,而低劑量組的日增重比對(duì)照組提高9.41%,差異顯著(P0.05)。免疫方式組、免疫劑量組的日采食量相比對(duì)照組均有所增加,但無明顯差別;不同免疫方式組間,鼻腔組的料肉比低于對(duì)照組(P0.05),不同免疫劑量組間,低劑量組的料肉比低于對(duì)照組(P0.05)。2.安全性研究本研究從免疫后豬的日常行為、基因整合情況、環(huán)境污染三個(gè)方面評(píng)估重組菌C500(p VGS/2SS-asd)的安全性,結(jié)果顯示,所有試驗(yàn)豬免疫后采食、飲水、糞便情況、精神狀態(tài)等日常行為和體溫均正常,初步證明該疫苗的安全性;免疫后12 w(即出欄時(shí))采集每頭豬的心、肝、肺、肌肉、脂肪組織并提取DNA,經(jīng)PCR方法檢測(cè),所有組織中均未檢測(cè)到目的基因GS/2SS,結(jié)果提示,該質(zhì)粒沒有整合至動(dòng)物的基因組中;在每次免疫后第1、3、5、7、10、15、20、30 d采集所有試驗(yàn)豬的糞樣、水樣、豬舍前后5 m的土壤樣本,用麥康凱選擇培養(yǎng)基培養(yǎng),挑選可疑菌落進(jìn)行PCR檢測(cè),所有樣本均未擴(kuò)增出目的基因GS/2SS,表明生長抑素基因疫苗沒有造成環(huán)境污染。3.免疫應(yīng)答反應(yīng)探究免疫方式(口腔+鼻腔、口腔、鼻腔)對(duì)育肥豬的免疫效果,研究表明,三種免疫方式均誘導(dǎo)機(jī)體產(chǎn)生免疫應(yīng)答,口腔+鼻腔組和口腔組的抗體水平顯著高于對(duì)照組(P0.05),且口腔+鼻腔組顯著高于口腔組,提示口腔+鼻腔的免疫方式可以誘導(dǎo)更高水平的免疫應(yīng)答;探究免疫劑量(高5×1011 CFU/頭、中5×1010 CFU/頭、低5×109 CFU/頭)對(duì)育肥豬免疫效果的結(jié)果顯示,低劑量組體現(xiàn)了更好的免疫效果,其抗體水平顯著(P0.05)高于對(duì)照組,生長激素水平高于對(duì)照組,但差異不顯著(P0.05)。
[Abstract]:Previous studies have shown that the immunization of recombinant C500 (P VGS/2SS-asd) gene vaccine can promote the growth of mice and have no adverse effects on small animals. In order to further understand the immune effect, mechanism and safety of the vaccine to large animals, this study chose 78 80 days old Duchang big pig, randomly divided into 6 groups, using the same dose (5 x 1011CFU). Three kinds of immune methods (oral cavity + nasal cavity, oral cavity, nasal cavity), three kinds of doses (high 5 * 1011 CFU/ head, 5 x 1010 CFU/ head and low 5 * 109 CFU/ head) for the same immune system (5 x 1010 CFU/ head, 5 * 109 CFU/ head) for clinical trial, and the test period 90 d to explore the optimum immunization and immunization dose. The performance WINFIRE measurement system automatically records the feed consumption of experimental pigs every day, determines the weight of pigs and the feed reward to evaluate the immune effect of the gene vaccine, and evaluates the safety of the vaccine from the aspects of gene integration and environmental spread from the daily behavior and body temperature of the pig, and the application of the target gene amplification to evaluate the safety of the vaccine. The ELISA method is applied. Determine the level of antibody and hormone to explore its mechanism of action. The results are as follows: 1. the effect of growth promoting and feed reward is used to record the data of pig feed intake and weight automatically every day by the WINFIRE system of the production performance of the sows produced by the American Austrian forage company. The growth rate of the pre immunization fattening pig is faster and the later weight gain slows down, and the immunization method is explored. The results showed that the growth promoting effect of the fattening pig was higher than that of the control group. The daily weight gain of the group was 4-7 w after immunization, and the daily weight gain of the nasal spray group was significantly higher than that of the control group (P0.05). The results showed that the effect of the immunization dose on the growth of the fattening pigs was 7 W after the initial immunization, and the daily weight gain of the medium dose and low dose group increased by 19.94%. The average daily gain of each immune group was higher than that of the control group, while the average daily gain of each immune group was higher than the control group, while the daily gain of the low dose group was 9.41% higher than that of the control group (P0.05). The daily intake of the immunization group and the immune dose group increased, but there was no significant difference between the immunization group and the control group, but there was no significant difference between the immunization group and the P0.01 group. Compared with the control group (P0.05), the meat meat ratio in the low dose group was lower than that of the control group (P0.05).2. safety study. This study evaluated the safety of the recombinant bacteria C500 (P VGS/2SS-asd) from three aspects of the daily behavior, gene integration and environmental pollution of the immune pigs. The results showed that all the test pigs were fed, drinking, and feces after immunization. The daily behavior and body temperature are normal, and the safety of the vaccine is preliminarily proved. 12 W after immunization (when out of column) collect the heart, liver, lung, muscle, fat tissue and DNA of each pig. The gene GS/2SS is not detected in all tissues by PCR method. The results suggest that the plasmid is not integrated into the gene of the animal. In the group, the feces samples of all test pigs, water samples, 5 m soil samples before and after the pig house were collected after each immunization, and the culture medium was selected by Kang Kai selection medium before and after the pig house, and the suspicious colonies were selected for PCR detection. All the samples did not amplify the target gene GS/2SS, indicating that the vaccine did not cause environmental pollution.3. immune response. In answer to the response, the immunological effects of immunization (oral cavity, nasal cavity, mouth, nasal cavity) on fattening pigs were investigated. The results showed that the immune responses were induced by the three immune modes. The antibody level in oral + nasal group and oral group was significantly higher than that of the control group (P0.05), and the oral + nasal cavity group was significantly higher than that in the oral group, suggesting that the oral + nasal cavity was immune to the oral cavity. In order to induce a higher level of immune response, the immune effect of immunization dose (5 x 1011 CFU/ head, 5 x 1010 CFU/ head and low 5 x 109 CFU/ head) showed that the low dose group showed better immune effect, its antibody level was significantly higher than that of the control group, and the level of growth hormone was higher than that of the control group, but the difference was not significant (P0.05).
【學(xué)位授予單位】：華中農(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)】：S828

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 滕家波,蘇國富;載體—宿主平衡致死系統(tǒng)在減毒傷寒沙門氏活菌苗中的應(yīng)用[J];國外醫(yī)學(xué)(免疫學(xué)分冊(cè));1999年03期

2 舒鄧群;王開云;曾檢華;魏金剛;吳紅翔;韓曉萍;;生長抑素基因疫苗對(duì)肉用雞生產(chǎn)性能、屠宰性能及肌肉品質(zhì)的影響[J];揚(yáng)州大學(xué)學(xué)報(bào)(農(nóng)業(yè)與生命科學(xué)版);2011年02期

3 舒翠莉,高杰英;鼻粘膜相關(guān)淋巴組織的研究進(jìn)展[J];免疫學(xué)雜志;2000年S1期

4 曹少先,楊利國,茆達(dá)干,張文偉,邢朝芳;生長抑素DNA疫苗pEGS/2SS在大鼠體內(nèi)的表達(dá)與分布[J];南京農(nóng)業(yè)大學(xué)學(xué)報(bào);2005年02期

5 舒鄧群;茆達(dá)干;陳榮達(dá);吳志敏;楊利國;;粒細(xì)胞-巨噬細(xì)胞集落刺激因子與生長抑素融合表達(dá)質(zhì)粒對(duì)斷奶仔豬生長及相關(guān)激素的影響[J];南京農(nóng)業(yè)大學(xué)學(xué)報(bào);2007年04期

6 孫迎基,汪興生,張新元,沈子龍;生長抑素DNA疫苗在COS細(xì)胞中的瞬時(shí)表達(dá)[J];藥物生物技術(shù);2002年02期

相關(guān)博士學(xué)位論文 前1條

1 雷姍;生長抑素對(duì)腸上皮細(xì)胞屏障功能的保護(hù)作用及機(jī)制研究[D];南方醫(yī)科大學(xué);2014年

，

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