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Margaret W. Toussaint,¹ Alan B. Rosencrance,² Linda M. Brennan,¹ Joseph R. Beaman,¹ Marilyn J. Wolfe,³ Florence J. Hoffmann,² and Henry S. Gardner, Jr.²

¹GEO-CENTERS, INC., Fort Detrick, Maryland, USA
²U.S. Army Center for Environmental Health Research, Fort Detrick, Maryland, USA
³Experimental Pathology Laboratory, Herndon, Virginia, USA

Abstract

Japanese medaka (Oryzias latipes) were continually exposed in a flow-through diluter system for 9 months to measured chloroform concentrations of 0.017, 0.151, or 1.463 mg/L. Parameters evaluated were hepatocarcinogenicity, hepatocellular proliferation, hematology, and intrahepatic chloroform concentration. Histopathology was evaluated at 6 and 9 months. Chloroform was not hepatocarcinogenic to the medaka at the concentrations tested. Chronic toxicity was evidenced at these time points by statistically significant ( = 0.05) levels of gallbladder lesions and bile duct abnormalities in medaka treated with 1.463 mg/L chloroform. We assessed hepatocellular proliferation by exposing test fish to 5-bromo-2´-deoxyuridine in the aquarium water for 72 hr after 4 and 20 days of chloroform exposure ; we then quantified area-labeling indices of the livers using computer-assisted image analysis. We observed no treatment-related increases in cellular proliferation. We analyzed cells in circulating blood in medaka after 6 months of chloroform exposure. Hematocrit, leukocrit, cell viability, and cell counts of treated fish were not significantly different from those of control fish. Using gas chromatography (GC) , we evaluated intrahepatic concentrations of chloroform in fish after 9 months of exposure. Livers from the 0.151 and 1.463 mg/L chloroform-treated fish had detectable amounts of chloroform, but these levels were always lower than the aquaria concentrations of chloroform. Thus, it appeared that chloroform did not bioaccumulate in the liver. Unidentified presumptive metabolite peaks were found in the GC tracings of these fish livers. Key words: aquatic toxicology, 5-bromo-2´-deoxyuridine, carcinogenicity, cell proliferation, chloroform, fish, hematology, histopathology, medaka, Oryzias latipes. Environ Health Perspect 109:35-40 (2001) . [Online 1 December 2000]

http://ehpnet1.niehs.nih.gov/docs/2001/109p35-40toussaint/ abstract.html

Address correspondence to M.W. Toussaint, GEO-CENTERS, INC., @ USACEHR, 568 Doughten Drive, Fort Detrick, MD 21702-5010. Telephone: (301) 619-7209. Fax: (301) 619-2569. E-mail: Margaret.Toussaint@amedd.army.mil

We thank R. Miller, R. Bishoff, M. Hennessey, and W. Dennis for technical assistance.

This project was sponsored by the NIEHS under Interagency Agreement Y1-ES-8051-02 (formerly Y1-ES-7096-02) , the U.S. Army Medical Research and Materiel Command, and the U.S. Army Corps of Engineers.

The views, opinions, and/or findings contained in this report are those of the authors and should not be construed as official Department of the Army position, policy, or decision, unless so designated by other official documentation. Citations of commercial organizations or trade names in this report do not constitute an official Department of the Army endorsement or approval of the products or services of these organizations.

Received 2 June 2000 ; accepted 7 September 2000.