British Journal of Nutrition


Anticarcinogenic effect of probiotic fermented milk and chlorophyllin on aflatoxin-B1-induced liver carcinogenesis in rats

M. Kumara1 p1 c1, V. Vermaa2, R. Nagpala3, A. Kumara4, P. V. Beharea5, B. Singha6 and P. K. Aggarwala1

a1 Dairy Microbiology Division, National Dairy Research Institute, Karnal 132 001, Haryana, India

a2 Department of Biochemistry, National University of Singapore, MD 7 #02-03, 8 Medical Drive, Singapore 117597, Singapore

a3 Department of Biotechnology, JMIT Institute of Engineering and Technology, Radaur, Haryana, India

a4 Department of Zoology, MLK Post-Graduate College, Balrampur, UP, India

a5 College of Dairy Technology, Udgir (Latur), Maharashtra, India

a6 Indian Veterinary Research Institute, Regional Station, Palampur 176 061, Himachal Pradesh, India


The present investigation was carried out to evaluate the hepatoprotective effect of probiotic fermented milk (FM) containing Lactobacillus rhamnosus GG and Lactobacillus casei strain Shirota, alone as well as in combination with chlorophyllin (CHL) as an antioxidant agent in male Wistar rats administered aflatoxin-B1 (AFB1). AFB1 was injected intraperitoneally at the rate of 450 μg/kg body weight per animal twice a week for 6 weeks, maintaining an equal time interval between the two consecutive AFB1 administrations. A total of 125 male Wistar rats were randomly allocated to five groups, each group having twenty-five animals. Group I was offered FM containing L. rhamnosus GG and L. casei strain Shirota. Group II was administered AFB1 and served as the control group; group III was administered FM-AFB1, in which besides administering AFB1, FM was also offered. Group IV was offered CHL and AFB1, and group V was offered both FM and CHL along with AFB1. The rats were euthanised at the 15th and 25th week of the experiment and examined for the biochemical and hepatopathological profile. A significant reduction in thiobarbituric acid-reactive substances (TBARS) was observed in the FM–CHL–AFB1 group compared with the AFB1 control group. FM alone or in combination with CHL was found to show a significant (P < 0·05) hepatoprotective effect by lowering the levels of TBARS and by enhancing the activities of antioxidant enzymes such as glutathione peroxidase, superoxide dismutase, catalase and glutathione-S-transferase, indicating that probiotic FM alone or in combination with CHL possesses a potent protective effect against AFB1-induced hepatic damage.

(Received January 20 2011)

(Revised June 06 2011)

(Accepted June 22 2011)

(Online publication August 05 2011)


c1 Corresponding author: M. Kumar, email

p1 Present address: Department of Microbiology and Immunology, National Institute of Nutrition, Hyderabad, India.


Abbreviations: AFB1, aflatoxin-B1; CAT, catalase; CHAPS, 3-((3-chloramidopropyl)dimethylammonio)-1-propane sulfonate; CHL, chlorophyllin; FM, fermented milk; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GST, glutathione-S-transferase; MDA, malonyl dialdehyde; NDRI, National Dairy Research Institute; ROS, reactive oxygen species; SOD, superoxide dismutase; TBARS, thiobarbituric acid-reactive substances