Influence of Genetically modified Soya on the BirthWeight and Survival of Rat Pups.docx

1、Influence of Genetically modified Soya on the BirthWeight and Survival of Rat PupsInfluence of Genetically modified Soya on the Birth-Weight and Survival of Rat Pups 转基因大豆对幼鼠出生重量和生存状况的影响By Irina V.EmakovaAbstract Investigation of the influence of GM soya on the birthrate and survival of the offsprin

2、g后代of Wistar rats2 were performed. A group of female rats were fed GM soya flour before mating 交配and pregnancy. The control group of females were fed traditional soya and the third group of females, the positive control group, received feed without any soya. The weight and the mortality死亡 rate of th

3、e newborn pups were analyzed. The study showed that there was a very high rate of pup mortality (55.6%) in the GM soya group in comparison with the control group and the positive control group (9% and 6.8% respectively. Moreover, death in the first group continued during lactation哺乳, and the weights

4、 of the survivors are lower those from the other two groups. It was revealed in these experiments, that GM soya could have a negative influence on the offspring of Wistar rats.摘要我们进行了转基因大豆对大白鼠后代出生率和生存状况的影响研究。一组母鼠在交配和怀孕前被喂食转基因大豆粉。母鼠的控制组被喂食传统大豆，第三组母鼠，即积极控制组，吃的食物没有任何大豆。我们分析了新生幼鼠的体重和死亡率。研究表明相比控制组合积极控制组的

5、死亡率（分别是9%和6.8%），转基因大豆组幼鼠死亡率（55.6%）很高。另外，转基因大豆组的幼鼠死亡率在哺乳期还在继续，存活幼鼠的体重也比其他两组幼鼠要轻。这些实验表明，转基因大豆对大白鼠后代可能有消极的影响。IntroductionThe term genetically modified organism (GMO3s) refers to plants, microbes and animals with genes transferred from other species in order to produce certain novel characteristics (for

6、 example resistance to pests, or herbicides) and are produced by recombinant DNA technology. Four main sources of the hazards危险 of GMO are discussed by scientists worldwide: 1) those due to the new genes, and gene products introduced; 2) unintended effects inherent内在的 to the technology; 3) interacti

7、ons between foreign genes and host genes; and 4) those arising from the spread of the introduced genes by ordinary cross-pollination异花授粉 as well as by horizontal gene transfer (World Scientists Statement 2000).介绍术语“转基因生物”指的是拥有从其他物种传递来的基因的植物、微生物和动物，转移基因是为了产生某些新的特点（比如抗病虫，或抗除草剂），并且是由重组DNA技术产生的。全世界的科学家讨

8、论了四种转基因生物风险的主要来源：1）那些因为新的基因，以及引入的基因产品导致的风险；2）技术内在的无意间的影响；3）外来基因和宿主基因的相互作用；4）因为普通的异花传粉和水平基因传递导致的引入基因传播产生的风险（世界科学家声明，2000年）GM crops contain material, which is not present in them under natural conditions, and they form a part of our daily diet. To understand what effect they can have on us and on our

9、animals it is vitally important to study the influence of these GM plants in different organisms for several generations. At the present, these studies are lacking from the scientific literature. Also, several detrimental effects of GM crops had been showed on the metabolism of animals. The hazard o

10、f genetically modified organisms (GMO) was shown for animals and the environment in many investigations (Traavik 1995; Ho and Tappeser 1997; Pusztai 1999 and 2001; Kuznetcov et al. 2004 and others).Earlier it was shown that consumption of GM food by animals led to the negative changes in their organ

11、isms. Experiments, conducted by A. Pusztai showed that potatoes modified by the insertion of the gene of the snowdrop lectin4 (an insecticidal proteins), stunted the growth of rats, significantly affected some of their vital organs, including the kidneys, thymus, gastrocnemius muscle and others (199

12、8) and damaged their intestines and their immune system (Ewen and Pusztai 1999). Similar effect of GM potatoes on rats was obtained at Institute of Nutrition in Russia (Ermakova 2005).转基因作物包含一些材料，在自然条件下这些材料不会出现在作物中，转基因作物成为了我们日常食物的一部分。为了理解它们对我们和动物有什么影响，研究这些转基因植物在几代不同生物体中的影响是非常重要的。目前，这些研究还缺乏科学文献。同时，转基

13、因作物对动物新陈代谢的几种不利影响已被揭示。在很多调查中转基因生物对动物和环境的危险也被揭示。（Traavik1995;）。早期有研究表明，动物食用转基因食品会导致有机体负面变化。A.Pustzai做的实验表明，嵌入雪花莲凝集素（一种杀虫剂蛋白质）基因之后的转基因土豆，阻碍了老鼠的生长，极大地影响了它们的一些重要器官，包括肾脏、胸腺、腓肠肌和其它器官（1998）并且损伤了它们的肠子和免疫系统（EwenandPustzai1999）。俄罗斯营养学院也得到了类似的转基因土豆对老鼠影响的结果。 It is put forward in the risk assessment documents th

14、at the GM components of transformed plants are completely destroyed in the digestive tract of humans and animals, together with the other genetic material found in them. However, foreign DNA plasmids5 are steadier against the digestion than it was originally believed. Plasmid DNA and GM DNA were fou

15、nd in microorganisms of the intestine and in saliva (Mercer et al. 1998; Coghlan 2002). Experimental researches in mice showed that ingested foreign DNA can persist in fragmented form in the gastrointestinal tract, penetrate the intestinal wall, and reach the nuclei of leukocytes, spleen and liver c

16、ells (Schubbert et al. 1994). In another research of Schubbert et al. (1998) the plasmid containing the gene for the green fluorescent protein6 (pEGFP-C1) or bacteriophage M13 DNA were fed to pregnant mice. Foreign DNA, orally ingested by pregnant mice, was discovered in blood (leukocytes), spleen,

17、liver, heart, brain, testes and other organs of foetuses and newborn animals. The authors considered that maternally ingested foreign DNA could be potential mutagens for the developing fetus. At the same time Brake and Evenson (2004) analyzing the testis in mice as a sensitive biomonitor of potentia

18、l toxic, didnt find negative effects of transgenic soybean diet on fetal, postnatal, puber talor adult testicular development. 在风险评估文件中提出已经转变的植物中转基因成分在人类和动物的消化道中被完全破坏，还有其它被发现的转基因材料也是如此。但是，外来DNA质粒比最初人们认为地更能抵御消化作用。在肠道和唾液微生物中发现了质粒DNA和转基因DNA(Merceretal1998;)。对老鼠的实验研究表明被吸收的外来DNA可以以片段的形式持续存在于胃肠道中，穿透肠壁，到达白

19、细胞、脾细胞和肝细胞的细胞核。（Schubbertetal.1994）。在Schubbert等人做的另一项研究中（1998），包含绿色荧光蛋白（pEGFP-c1）基因或噬菌体M13DNA的质粒被喂给怀孕的老鼠。外来DNA被怀孕的老鼠口服后，在胚胎和新生幼鼠的血液（白细胞）、脾、肝、心、脑、睾丸和其它器官都有发现。作者们认为被雌性老鼠吸收的外来DNA可能是正在发育的胚胎的潜在诱变因素。同时Brake和Evenson(2004)分析了老鼠中的睾丸，它是敏感的潜在毒性物质的生物监控器，他们并未发现转基因大豆食物对胎儿、产后、青年和成年睾丸器官的发展有消极影响。 There is a lack of

20、investigations on the influence of GM crops on mammals, especially on their reproductive function. Therefore, it was decided that we undertake a study to see the effect of the most commonly used GM crop on the birth rate, mortality and weight gain of rat pups, whose mother were fed diets supplemente

21、d with the Roundup- Ready soya, a kind of GM food.转基因作物对哺乳动物，尤其对他们的繁殖功能的影响还缺乏调查。因此，我们决定进行一项研究，以确认最常用的转基因作物对幼鼠的出生率、死亡率和体重增加的影响，而生这些幼鼠的母鼠被喂食的食物补充了“抗农达”转基因大豆，是一种转基因食物。MethodsAnimals Wistar rats were used in the experiment. The animals were brought up to sexual maturity on laboratory rat feed. When thei

22、r weight reached about 180-200g, the female rats were divided into 3 groups, and housed in groups (3 rat/cage), and kept under normal laboratory conditions.方法在实验中被用到大白鼠。这些大白鼠被喂食实验室老鼠饲料，养育到性成熟阶段。当它们的重量达到180-200g,母鼠被分成三组，按组放在不同的笼子里（每笼三只老鼠），并且处于正常的实验室条件下。 The feeding scheme was as follows. Females in e

23、very cage daily received dry pellets from a special container placed on the top of their cage. Those rats receiving soya flour supplement, were given the soya flour in a small container placed inside their cage (20g x 40 ml water) for three rats and, so 5-7g flour for each rat every day.喂食计划如下：每个笼子里

24、的母鼠每天从位于笼子顶部的特殊容器接受干的颗粒。那些接受大豆粉补充物的老鼠从放在笼子里的小容器中得到大豆粉（20g和40ml水），笼子里有三只老鼠，所以每天每只老鼠5-7克大豆粉。Experiment One group of female rats of 180-200g weight was allocated to the experimental group, and received 5-7 soy a flour/rat/day prepared from Roundup-Ready soya, added to the rat feed for two weeks. Anothe

25、r group females (3) were allocated to the control group, but their diet was supplemented with the same amount of soya flour, prepared from the traditional soya in which only traces (0.08+ 0.04%) of the GM construct was present, most likely resulting from cross-contamination. We also introduced a pos

26、itive control group (in two cages: 3x3), which had not been exposed to soya flour. Therefore females have only got the standard laboratory feed without any supplementation, although it is acknowledged that the energy and protein content of this diet was less than in the other two groups.实验一组重180-200

27、g的母鼠被分配到实验组，每只老鼠每天接受5-7g来自“抗农达”的转基因大豆粉，这些大豆粉加到老鼠食物中，持续2周。另一组母鼠（3只）被分配到控制组，但是它们的食物补充的是同量的大豆粉，从传统的大豆磨制，其中只有微量的（0.08+0.04%）的转基因构成物出现，最有可能是因为交叉感染。我们还引入了一个积极控制组（两个笼子：每个笼子3只老鼠），这一组没有接触大豆粉。所以这些母鼠只有标准的实验室食物，没有任何补充物。尽管我们承认这种食物的能量和蛋白质含量比其它两组少。 After two weeks on the diets all groups of 3 females were mated wi

28、th two healthy males of the same age, who have never been exposed to soya flour supplements. First the one, then the other male was put into the cage for 3 days. In order to avoid infection of females, the sperm count and quality has not been determined. We carried on with feeding the respective die

29、ts to all females during mating and pregnancy. Upon delivery, all females were transferred to individual cages, and the amount of soya supplement was increased by an additional g for every pup born. Lab feed and water was available for all animals during the experimental period. When rat pups opened

30、 their eyes and could feed themselves (from 13-14 days of age), the daily dose of soya supplement was increased till 2-3g for every pup, although all rats had free approach to the soya. All rats ate their soya portions well. Organs of some pups were taken out and weighed. The level of mortality was

31、analyzed by the one-way ANOVA7, using of Newman-Keuls test8 for share distribution. The pups weight and its distribution were checked by Mann-Whitney test9 and Chi-square10 in StatSoft Statistica v6.0 Multilingua (Russia).食用相应的食物两周后，所有各组的3只母鼠与两只同龄的健康公鼠交配，这两只公鼠从没接触过大豆粉补充物。首先是第一只，然后是另外一只被放进笼子三天。为了避免母鼠

32、感染，精子的计数和质量没有被确定。在交配和怀孕期间，我们继续给母鼠喂食各自的食物。分娩之后，所有的母鼠被转移到单独的笼子。对于每只刚出生的幼鼠，大豆补充物的数量额外增加了一克。在实验期间对于所有动物实验室的食物和水都可得到。当幼鼠睁开眼睛并且可以自己进食（从13-14天开始），每只幼鼠每天大豆补充物的量增加到2-3g,尽管所有的老鼠都可以自由靠近大豆。所有老鼠都很好地食用了自己份内的大豆。有些小鼠的器官被取出来并且称重。利死亡率由单方方差分析进行，对于份额分布使用N-K测试。小鼠的重量和分布有曼-惠特尼测试进行检验，还有卡方检验，都由俄罗斯的Statsoft统计V6.0多语软件进行。Resul

33、ts By the end of the experiment, from the 15 females included in the experiment, 11 gave birth and produced a total of 132 rat pups. The 4 rats who became pregnant from 6 females on the positive control diet gave birth to 44 pups (an average of 11 pups/female), while the four females, from the six on GM so