A 90-DAY SUBCHRONIC TOXICOLOGICAL ASSESSMENT of DEINOCOCCUS GRANDIS FERMENTED SOYMILK IN SPRAGUE-DAWLEY RATS
Objective: Despite the fact that there was no adverse effect observed in previous animal safety studies of Deinococcus grandis (D. grandis) fermented soymilk, including acute oral toxicity assay, 3 different test systems of genotoxicity test and teratogenicity study, whether D. grandis fermented soymilk is safe for long-term use remains unknown. Therefore, the study was conducted further to clarify the edible safety of D. grandis fermented soymilk for long term use.
Methods: Eighty Sprague-Dawley (SD) rats were divided into four groups, each consisting of ten male and ten female rats. Rats were orally administrated with reverse osmosis water (control) or 1,000, 2,000 and 3,000 mg/kg b.w./d freeze dried D. grandis fermented soymilk powder for 90 consecutive days. Clinical observation of the rats was carried out daily. The body weight and feed intake of the rats were recorded weekly. At the end of the study, all rats were sacrificed and the blood and organs were collected for hematology, clinical biochemistry and histopathological examination.
Results: During the study period, no abnormality occurred in clinical signs, body weight, and ophthalmological examination. There were no significant differences in urinalysis, hematology and clinical biochemistry parameters between the treatment and control group. Necropsy and histopathological examination showed no treatment-related change.
Conclusion: According to the results, the no-observed-adverse-effect level (NOAEL) of D. grandis fermented soymilk was greater than 3,000 mg/kg b.w./d in SD rats.
Keywords: Deinococcus grandis (D. grandis), 90-day subchronic toxicity, NOAEL, Safety assessment, GKB-Aid 1995
2. Lee GA, Crawford GW, Liu L, Sasaki Y, Chen X. Archaeological soybean (Glycine max) in East Asia: does size matter. PloS One 2011;6:e26720. Doi:10.1371/journal.pone.0026720. [Article in Press]
3. American Heart Association. Dietary guidelines for healthy American Adults: A statement for physicians and health professionals. American Heart Association; 1996.
4. Wang Y, Jones PJ, Ausman LM, Lichtenstein AH. Soy protein reduces triglyceride levels and triglyceride fatty acid fractional synthesis rate in hypercholesterolemic subjects. Atherosclerosis 2004;173:269-75.
5. Anderson JW, Smith BM, Washnock CS. Cardiovascular and renal benefits of dry bean and soybean intake. Am J Clin Nutr 1999;70:464-74.
6. Song Y, Paik HY, Joung H. Soybean and soy isoflavone intake indicate a positive change in bone mineral density for 2 y in young Korean women. Nutr Res 2008;28:25-30.
7. Clarkson TB, Utian WH, Barnes S, Gold EB, Basaria SS, Aso T, et al. The role of soy isoflavones in menopausal health: report of The North American Menopause Society/Wulf H. Utian Translational Science Symposium in Chicago, IL (October 2010). Menopause 2011;18:732-53.
8. Zhou JR, Gugger ET, Tanaka T, Guo Y, Blackburn GL, Clinton SK. Soybean phytochemicals inhibit the growth of transplantable human prostate carcinoma and tumor angiogenesis in mice. J Nutr 1999;129:1628-35.
9. Liu CJ, Chen CC, Jhang LN, Yeh SX, Lee KM. The immunomodulatory effect of soybean fermented fluid. J Testing Qual Assur 2014;3:175-84.
10. Huang MT, Ferraro T. Phenolic compounds in food and cancer prevention. In ACS symposium series (USA); 1992.
11. Cassady JM, Baird WM, Chang CJ. Natural products as a source of potential cancer chemotherapeutic and chemopreventive agents. J Nat Prod 1990;53:23-41.
12. Bringe NA. U. S. Patent No. 6,171,640. Washington, DC: U. S. Patent and Trademark Office; 2011.
13. Lokuruka MN. Effects of processing on soybean nutrients and potential impact on consumer health: an overview. Afr J Food Agric Nutr Dev 2011;11;5000-17.
14. Hattori T, Ohishi H, Yokota T, Ohoami H, Watanabe K. Antioxidative effect of a crude antioxidant preparation from soybean fermented by Bacillus natto. LWT-Food Sci Technol 1995;28:135-8.
15. Lai MN, Lo YK, Chen JC, Hu CK, Hwang CC. Identification of N2-fixing bacteria and determination of its antioxidative properties. J Chin Agric Chem Soc 1995;33:85-93.
16. Woo PCY, Lau SKP, Teng JLL, Tse H, Yuan KY. Then and now: use of 16S rDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories. Clin Microbiol Infect 2008;14:908-34.
17. Renaud FNR, Bergeron E, Tigaud S, Fuhrmann C, Gravagna B, Freney J. Evaluation of the new Vitek 2 GN card for the identification of gram-negative bacilli frequently encountered in clinical laboratories. Eur J Clin Microbiol Infect Dis 2005;24:671-6.
18. Wu SY, Li IC, Lin YC, Chen CC. Characterization and safety evaluation of Denococcus grandis as feed additive for hens. Regul Toxicol Pharmacol 2016;76:121-7.
19. Liu CJ, Chen CC, Jiang LN, Yeh SX, Lee KM. The immunomodulatory effect of soybean fermented fluid. J Testing Qual Assur 2014;3:175-84.
20. Oyaizu H, Stackebrandt E, Schleifer KH, Ludwig W, Pohla H, Ito H, et al. A radiation-resistant rod-shaped bacterium, Deinobacter grandis gen. nov., sp. nov., with peptidoglycan containing ornithine. Int J Syst Bacteriol 1987;37:62-7.
21. Krasinski SD, Russell RM, Furie BC, Kruger SF, Jacques PF, Furie B. The prevalence of vitamin K deficiency in chronic gastrointestinal disorders. Am J Clin Nutr 1985;41:639-43.
22. Petterino C, Argentino-Storino A. Clinical chemistry and haematology historical data in control Sprague-Dawley rats from pre-clinical toxicity studies. Exp Toxicol Pathol 2006;57:213-9.