摘要
本课题组制备出肌肉中富含ω-3不饱和脂肪酸的.sFat-1转基因猪,对改善人类的饮食健康具有重要的意义。虽然sFat-1转基因猪具有广泛的应用前景,但其生物安全性正日益成为人们关注的焦点。
转基因动物体内微生物安全评价,本部分研究的目的主要是从转基因猪肠道内微生物菌群数量变化的角度去评估sFat-1基因的表达对转基因猪本身健康和生态环境安全的影响。为了监测sFat-1转基因猪不同段肠道和粪便中微生物菌群数量的改变,选取总需氧菌、总厌氧菌、特异有益菌和有害菌进行培养分析。结果表明,与对照组相比,sFat-1转基因猪不同段肠道和粪便中微生物菌群没有发生明显改变。此部分研究表明,外源基因sFat-1基因的表达对转基因猪肠道和粪便中的微生物菌群数量未产生明显影响。
水平漂移风险评估,本部分研究的目的是评价sFat-1转基因猪的外源基因是否会水平漂移至其肠道内微生物菌群中,以及是否会通过哺乳和交配的方式将外源基因sFat-1基因水平漂移至非转基因猪的基因组内。哺乳介导基因水平漂移实验包括两部分：sFat-1转基因奸娠母猪分别哺乳非转基因仔猪和sFat-1转基因仔猪吸允非转基因母猪：交配介导基因水平漂移实验包括sFat-1转基因公猪与非转基因母猪交配和sFat-1转基因母猪与非转基因公猪交配。通过PCR方法检测转基因猪体内微生物以及非转基因猪不同组织中是否含有sFat-1基因片段。结果表明,转基因猪体内微生物菌群基因组DNA和非转基因猪各组织器官基因组DNA中都未检测到外源目的基因sFat-1基因。此部分研究表明,sFat-1转基因猪外源基因未发生水平漂移至其肠道内微生物菌群中,同时也未通过哺乳和交配方式将外源基因水平漂移至非转基因猪体内。
sFat-1转基因猪肉30天喂养小鼠毒理评价研究,实验共分七组包括对照猪肉低剂量组、对照猪肉高剂量组、sFat-1转基因猪肉低剂量转组、sFat-1转基因猪肉高剂量组、三文鱼肉低剂量组、三文鱼肉高剂量组和基础饲料组,对各组小鼠分别开展血生化、血常规、免疫指标、微生物菌群、病理组织切片等研究,实验结果表明,饲喂含低剂量与高剂量sFat-1转基因猪肉的饲料对小鼠血生化、血常规、T细胞亚群分布、免疫球蛋白及肠道内和粪便中微生物菌群未产生明显影响；病理组织切片结果表明,饲喂含低剂量与高剂量sFat-1转基因猪肉的饲料对小鼠肝脏的免疫系统具有一定的改善作用,对其它各组织器官均未产生明显影响。综合结果表明,未发现sFat-1转基因猪肉对小鼠健康具有毒副作用,并初步确定对提高机体的免疫系统具有一定的促进作用。
综合sFat-1转基因猪生物安全评价工作,未发现sFat-1转基因猪及其可食产品具有潜在的生物安全危害。
We previously developed and characterized sFat-1(synthesized fatty acid desaturase-1) transgenic pigs that contain high ω-3unsaturated fatty acid content in their meat which is beneficial to the health of human being. Although sFat-1transgenic pig has a broad perspective in application, its bio-safety is gaining more and more public attention.
Safety assessment of in vivo microbe of transgenic animals, the aim of this part study is to assess the effects of expression of sFat-1in pig on the health of animal themselves and on the safety of environment from the angle of the changes in microbe population of pig intestinal tract. For inspection of any change in populations of microbial flora in various parts of intestinal tract and feces of sFat-1transgenic pigs total aerobe, total anaerobe. main beneficial bacteria and main harmful bacteria were cultured and analyzed. Results indicated that in comparison with control pig microbe population of various species in different parts of intestinal tract and feces of sFat-1transgenic pig remained unchanged. In conclusion, the data of this study suggest that microbe population in intestinal tract and feces of transgenic pig was not influenced by the expression of transferred foreign gene sFat-1gene.
Risk assessment of horizontal gene transfer, the aim of this part study was to assess whether the inserted foreign gene of sFat-1transgenic pigs could transfer into the genome of in vivo microbial or non-transgenic pigs by suckling or mating. The tests for suckling mediated horizontal gene transfer included sFat-1transgenic sows nursing non-transgenic piglets and sFat-1transgenic piglets suckling non-transgenic sows. The tests for mating mediated horizontal gene transfer involved male sFat-1transgenic pigs mating with non-transgenic females and female sFat-1transgenic pigs mating with non-transgenic males. Foreign gene drift was assessed by PCR amplifying foreign fragment in samples of in vivo microbes of transgenic pigs and the various tissues sampled from non-transgenic pigs. The results showed that the foreign target gene sFat-1could not be detected in the microbial genome of transgenic pigs and the genomic DNA of various tissues and organs of these sampled non-transgenic pigs. In conclusion, this study suggest that foreign gene transferring from transgenic animal genome to in vivo microbial genome and horizontal gene transfer of transgenic pigs via suckling or mating unlikely occurs.
Toxicological evaluation of sFat-1transgenic pork in mice for30days, the experiment including the low-dose control pork group, the high dose control pork group, the low dose sFat-1transgenic pork group, the high dose sFut-1transgenic pork group, the low dose salmon group, the high dose salmon group and the basal diet group. Each group of mice was carried out blood chemistry, hacmatology. immune indicators, bacteriological quantification and histopathology research. The results showed that the mice fed diets with low-dose and high dose sFat-1transgenic pork did not produce significant impact on blood chemistry, haematology. the distribution of T lymphocyte subsets, immune globulin and intestinal and fecal microbial flora. The histopathology results showed that the mice fed diets with low-dose and high dose sFat-1transgenic pork may improve the immune system of liver, and have no significant impact on other tissues and organs. The results indicated that sFat-1transgenic pork have no toxic effects on mouse health while improve the mouse immune system.
In conclusion, no potential adverse effects of sFat-1transgenic pigs and its edible products have been detected through the comprehensive bio-safety assessment research.
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