Effects of bar-transgenic rice on the intestinal microflora of the mice (Mus musculus)

Jin Liu, Yi Huang, Yanbo Sun, Hengmei Yan

College of Life Science, Hunan Normal University, Changsha, China.

DOI: 10.4236/ajmb.2012.23022                

Abstract

Microbial molecular ecology approaches were used to the effects of Bar-transgenic rice on Intestinal Micro-flora of the Mice (Mus musculus). Kunming mice (Mus musculus) of 100 SPF-grade (20 g ± 2 g), half of which were male and the other half female, were randomly divided into five groups with four replications per group and five mice per replication to assess the safety of Bar-transgenic rice. Five diets meetinging or exceeding the minimum nutrient requirement were fed for 180 days. After 90 days, parental generation (P) was bred to produce the first filial generation (F₁). Each generation was fed for 180 days. On the 180^th day, five mice from each group were randomly sampled, and their intestinal contents were collected for DNA isolation. The V3 region of the 16S rDNA was amplified by polymerase chain reaction (PCR) and analyzed via denaturing gradient gel electrophoresis (DGGE). The resulting PCR-DGGE band number (bacterial species) was counted, and the banding patterns were analyzed by calculating the Sorenson’s pairwise similarity coefficients (Cs), an index used to measure bacterial species found among all samples. The sequence analysis of bands was performed to identify the intestinal predominant microflora of the mice. The intergroup Cs values of the samples across all groups did not differ (P > 0.05) from each other. The effect of Bar-transgenic rice on the intestinal microflora of the mice was considered insignificant.

Keywords

Bar-Transgenic Rice; Mus musculus;Intestinal; Microflora; Denaturing Gradient Gel Electrophoresis

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Liu, L.F., Cao, J.S., Wu, W.H., et al. (2009) Safety assessment of transgenic food and the present situation of management in China. Acta Agriculturae Jiangxi, 21(4), 155-158.
[2]	Kuipera, H.A., Konig, A., Kletera, G.A., et al. (2004) Concluding remarks. Food and Chemical Toxicology, 42, 1195-1202.
[3]	Anthony, J.C., Jeanne, M.E.J. (1999) Genetic engineering of crops as potential source of genetic hazard in the human diet. Mutation Research, 443, 223-234.
[4]	Wang, Z.H., Wang, Y., Cui, H.R., et al. (2002) Toxicological evaluation of transgenic rice flour with a synthetic cry1Ab gene from Bacillus thuringiensis. Journal of the Science of Food and Agriculture, 82, 738-744.
[5]	Wang, Y., Lai, W.Q., Chen, J.G., et al. (2000) Toxicity of anti-herbicide gene(Bar) transgenic rice. Journal of Hygiene Research, 29(3), 141-142.
[6]	Zhao, Z.H., Yang, L.T., Ai, X.J., et al. (2005) Influence of genetically modified rice containing codA gene on physiological metabolism and genetic horizontal transformation in fed rats. Journal of Agricultural Biotechnology, 13(3), 341-346.
[7]	Momma, K., Hashimoto, W., Yoon, H., et al. (2000) Safety assessment of rice genetically modified with soybean glycinin by feeding studies rats. Bioscience Biotechnology and Biochemistry, 64(9), 1881-1886.
[8]	Cromwell, G.L., Henry, B.J., Scott, A.L., et al. (2005) Glufosinate herbicide-tolerant (Liberty Link) rice vs. conventional rice indiets for growing-finishing swine. Journal of Animal Science, 83, 1068-1074.
[9]	Wang, Z.H., Wang, Y., Cui, H.R., et al. (2002) A Preliminary study on toxicological evaluation of transgenic rice flour with a synthetic cry1Ab gene from bacillus thuringiensis. Scientia Agricultura Sinica, 35(12), 1487-1492.
[10]	Li, Y.H., Piao, J.H., Zhuo, Q., et al. (2004) Subchronic toxicity test of Xa21 transgenic rice. Journal of Hygiene Research, 33(5), 575-578.
[11]	Li, Y.H., Piao, J.H., Zhuo, Q., et al. (2004) Study on the teratogenicity effects of genetically modified rice with Xa21 on rats. Journal of Hygiene Research, 33(6), 710-712.
[12]	[12] Zhuo, Q., Chen, X.P., Piao, J.H., et al. (2004) Study on the teratogenicity effects of genetically modified rice which expressed cowpea trypsin inhibitor on rats. Journal of Hygiene Research, 33(1), 74-77.
[13]	Wang, Z.H., Wang, Y., Shu, Q.Y., et al. (2004) Study on mutagenicity of transgenic rice flour with a synthetic cry1Ab gene from bacillus thuringiensis. Scientia Agricultura Sinica, 37(12), 2043-2046.
[14]	Mackie, R.I., White, B.A., Isaacson, R.E. (1977) Gastrointestinal microbes and host interactions. Gastrointestinal Microbiology. Chapman ＆ Hall, New York.
[15]	Savory, C.J. (1992) Enzyme supplementation degradation and metabolism of three U-14C-labelled cell-wall substrates in the fowl. British Journal of Nutrition, 67(1), 91-102.
[16]	Reid, C.A., and Hillman, K. (1999) The effects of retrogradation and amylose/amylopectin ratio of starches on carbohydrate fermentation and microbial populations in the porcine colon. Animal Science, 68, 503-510.
[17]	Wang, H.F., Zhu, W.Y., Yao, W., et al. (2007) DGGE and 16S rDNA sequencing analysis of bacterial communities in colon content and feces of pigs fed whole crop rice. Anaerobe, 13, 127-133.
[18]	Zhu, W.Y., Yao, W., and Mao, S.Y. (2003) Development of bacterial community in feces of weaning piglets as revealed by denaturing gradient gel electrophoresis. Acta Microbiologica Sinica, 43(4), 503-508.
[19]	Ni, X.Q., Gong, J., Hai, Y., et al. (2008) The Bacterial Community and Diversity in the Layer Gastrointestinal Tract: From Crop to Cecum Analyzed by PCR-DGGE. Chinese Journal of Animal and Veterinary Sciences, 39(7), 955-961.
[20]	Ministry of Agriculture of the People's Republic of China. (2006) Safety assessment of genetically modified plant and derived products 90-day feeding test on rats. Agriculture industry standard of the People's Republic of China NY/T 11022006
[21]	Gong, J.H., Si, W., Forster, R.J., et al. (2007) 16S rRNA genebased analysis of mucosa-associated bacterial community and phylogeny in the chicken gastrointestinal tracts: from crops to ceca. FEMS Microbiology Ecology, 59, 147-157.
[22]	Yu, Z. and Morrison, M. (2004) Improved extraction of PCR 2 quality community DNA from digesta and fecal samples. Bio-Techniques, 36, 808-812.
[23]	Muyzer, G., de Waal, E.C., and Uitterlinden, A.G. (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology, 59(3), 695-700.
[24]	Simpson, J.M., McCracken, V.J., Gaskins, H.R., et al. (2000) Denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA amplicons to monitor changes in fecal bacterial populations of weaning pigs after introduction of Lactobacillus reuteri strain MM53. Applied and Environmental Microbiology, 66(11), 4705-4714.
[25]	Zhu, X.F., Xiong, D.X., Li, X.Y., et al. (1995) Comparison of membrane microflora and intestinal microflora in Several Animals. Chinese Journal of Laboratory Animal Science, 5(1), 30-32.
[26]	Wohlleben, W., Amold, W., Broer, I., et al. (1988) Nucleotide　Sequence of the Phosphinothricin N-acetyltransferase　Gene from the Streptomyces Hygroscopicus Tu 494 and its expression in Nicotiana tobacum. Gene, 70(1), 25-37
[27]	Liu, H.Y., Mi, X.J., and Cui, J.Z. (2007) Characteristics and safety of bar gene, PAT proteins and glufosinate. Chinese Journal of Ecology, 26(6), 938-942.