【作者】 徐志远；

【导师】 刘阳；

【作者基本信息】 中国农业科学院 ， 食品科学与工程， 2011， 硕士

【摘要】 本研究以鲤鱼(Cyprinus carpio)为实验动物,利用转Cry1Ac/sck基因糙米制作饲料,以亲本糙米饲料作为对照,选取健康鲤鱼240尾,随机分为两组,即转基因糙米组和亲本糙米组,进行56d鲤鱼喂养实验,系统进行了转Cry1Ac/sck基因糙米对鲤鱼营养安全性、免疫安全性、肠道微生态安全性和外源基因表达蛋白在体内的残留情况的研究,以全面评价转Cry1Ac/sck基因糙米作为鲤鱼日粮原料的安全性。实验结果如下:1、糙米营养成分分析转Cry1Ac/sck基因糙米与亲本糙米相比,水分、粗蛋白、粗脂肪、粗灰分、粗纤维、无氮浸出物、氨基酸、钙和磷等的含量均无显著性差异。2、转Cry1Ac/sck基因糙米作为鲤鱼日粮原料的营养安全性评价在0-28d、29-56d及整个0-56d生长过程中,饲喂转Cry1Ac/sck基因糙米饲料的鲤鱼与亲本饲料组相比,其生长性能和肌肉粗蛋白、粗脂肪、粗灰分、氨基酸、钙和磷等组成成分的含量均无显著性差异。鲤鱼对转Cry1Ac/sck基因糙米日粮总能、干物质、粗蛋白、粗脂肪、粗灰分、氨基酸、钙及磷表观消化率指标与亲本糙米日粮相比均无显著差异。转Cry1Ac/sck基因糙米及亲本糙米的营养物质表观消化基本等同,未发现转Cry1Ac/sck基因糙米对鲤鱼消化机能造成任何显著的不良影响。饲喂转基因组糙米日粮组鲤鱼与亲本糙米日粮组相比,肝胰腺、脾脏、肾脏、前肠、中肠和后肠的器官指数指标均无显著差异;肝胰腺、脾脏和肾脏未见明显可见的病理变化,肠段绒毛长度和隐窝深度均无显著差异。结果表明转Cry1Ac/sck基因糙米对鲤鱼器官发育未造成任何显著的不良影响。3、转Cry1Ac/sck基因糙米作为鲤鱼日粮原料的免疫安全性评价在0-28d、29-56d及整个0-56d生长过程中,饲喂转基因饲料对鲤鱼的免疫器官指数、血液生理生化、血清溶菌酶活性、血清激素、血清抗氧化能力指标、免疫因子(TNFα,IL-1β,IFNγ,iNOS2a,IL-10和TLR5)等指标未见显著的不良影响。4、转Cry1Ac/sck基因糙米作为鲤鱼日粮原料对鲤鱼肠道微生物菌群的影响在0-28d、29-56d及整个0-56d生长过程中,饲喂转基因饲料对鲤鱼前肠、中肠、后肠内的大肠杆菌、乳酸杆菌和双歧杆菌菌群数量未见显著不良影响,不影响肠道菌群结构。5、转Cry1Ac/sck基因糙米Cry1Ac蛋白在鲤鱼肠道内的残留检测饲喂转基因糙米日粮与亲本糙米日粮的鲤鱼前肠、中肠和后肠的肠道内容物均未检测到Cry1Ac蛋白的存在,Cry1Ac外源蛋白在鲤鱼体内未见残留。本实验结果表明,在实验期内,鲤鱼生长良好,转Cry1Ac/sck基因糙米未对鲤鱼营养、免疫、肠道微生态产生明显可见的不良影响,在鲤鱼体内未检测到Cry1Ac蛋白残留。可见,实验期内转Cry1Ac/sck基因糙米作为鲤鱼日粮原料与亲本糙米具有实质性等同,未对鲤鱼产生显著的有害影响。更多还原

【Abstract】 A 56 days feeding study was used to evaluate whether a carp diet prepared with brown rice derived from transgenic rice modified with Cry1Ac gene and sck gene had any unanticipated adverse effects on carp (Cyprinus carpio) as compared with diet prepared with brown rice derived from non-transgenic control rice. 240 healthy carps were divided into two treatment groups randomly, and each group was divided into four replicates, furthermore every replicate fed with 30 fish. The control group (86) fed with ordinary 86 brown rice diet, the test group (FM86) was fed transgenic brown rice diet. Effects of transgenic brown rice on nutritional safety, immunotoxicological safety, intestinal microflora were studie and Cry1Ac gene protein was detected in foregut, midgut and hindgut of carp.The test results were as follows:Experiment 1: The nutrient composition of the brown riceThe content of Moisture, Crude protein, Crude fat, Ash, Crude fiber, Nitrogen-free-extract, Amino acid, Calcium and Phosphorus between non-transgenic control rice and transgenic rice had no statistic difference.Experiment 2: Nutritional safety assessment of rice genetically modified with Cry1Ac and sck by feeding studies on carpDuring the period of 0 to 28d, the period of 29 to 56d and the whole 0 to 56d period, no significant statistically differences in the growth performance and Muscle content were observed between carps reared on the diets prepared with transgenic brown rice or control rice.With 0.4% Cr2O3 as the exogenous indicator, the apparent digestibility of total energy, dry material, crude protein, crude fat, crude ash, amino acid, Calcium and Phosphorus between non-transgenic control rice and transgenic rice had no statistic difference. The feeding value of transgenic rice and parental rice were similar in this study, and no unexpected effects were observed in rice genetically modified with Cry1Ac gene and sck gene.During the period of 0 to 28 d, the period of 29 to 56d and the whole 0 to 56d period, the carp in both groups grew well without marked differences in food intake and body weight. the organ index of liver pancreas, spleen, kidney, foregut, midgut and hindgut between non-transgenic control rice and transgenic rice had no statistic difference. Necropsy indicated neither pathological symptoms nor histopathological abnormalities in the liver, spleen, kidney, foregut, midgut and hindgut. Length of intestine villus and crypt depth of intestinalsegment were observed no significantly different. Judging from these results: no detrimental unexpected effects were observed in rice genetically modified with Cry1Ac gene and sck gene.Experiment 3: Immunotoxicological safety assessment of rice genetically modified with Cry1Ac and sck by feeding studies on carpDuring the period of 0 to 28d, the period of 29 to 56d and the whole 0 to 56d period, the carp in both groups grew well without marked differences in food intake, body weight and the growth of immune organs. There were also no significant differences in the physiological indexes of whole blood, serum regular parameters, serum lysozyme activity, serum hormone and antioxidant capacity indexs and immune factors in serum (TNFα, IL-1β, IFNγ, iNOS2a, IL-10 and TLR5) of carp. Necropsy at the end of the experiment indicated neither pathological symptoms nor histopathological abnormalities in the liver, spleen, foregut, midgut and hindgut. Judging from these results, the rice genetically modified with Cry1Ac and sck is considered to have been essentially the same in nutritional and Immunotoxicological characteristics as the control rice.Experiment 4: Intestinal microflora safety assessment of rice genetically modified with Cry1Ac and sck by feeding studies on carpDuring the period of 0 to 28d, the period of 29 to 56d and the whole 0 to 56d period, no significant differences were observed between carps reared on the diets prepared with transgenic brown rice or control rice.The Number of colonies for E. coli, Lactobacillus and Bifidobacterium which were residing in foregut, midgut and hindgut had no significant differences. The carps were not observed statistically significant difference in the impact of Intestinal microbial flora of carp. Experiment 5: Cry1Ac gene protein was detected in foregut, midgut and hindgut of carpExogenous Cry1Ac protein was not detected in foregut, midgut and hindgut. The transgenic protein will not spread to nature by way of excreta.To sum up, transgenic brown rice has substantial equivalence as compared with ordinary brown rice, and has no negative effects to carp.更多还原