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. 2001 Jan;109(1):35–40. doi: 10.1289/ehp.0110935

Chronic toxicity of chloroform to Japanese medaka fish.

M W Toussaint 1, A B Rosencrance 1, L M Brennan 1, J R Beaman 1, M J Wolfe 1, F J Hoffmann 1, H S Gardner Jr 1
PMCID: PMC1242048  PMID: 11171522

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 ([alpha] = 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.

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Selected References

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  1. Christensen J. M., Rasmussen K., Køppen B. Automatic headspace gas chromatographic method for the simultaneous determination of trichloroethylene and metabolites in blood and urine. J Chromatogr. 1988 Jun 17;442:317–323. doi: 10.1016/s0021-9673(00)94479-0. [DOI] [PubMed] [Google Scholar]
  2. Connolly K. M., Bogdanffy M. S. Evaluation of proliferating cell nuclear antigen (PCNA) as an endogenous marker of cell proliferation in rat liver: a dual-stain comparison with 5-bromo-2'-deoxyuridine. J Histochem Cytochem. 1993 Jan;41(1):1–6. doi: 10.1177/41.1.7678022. [DOI] [PubMed] [Google Scholar]
  3. Cunningham M. L., Matthews H. B. Relationship of hepatocarcinogenicity and hepatocellular proliferation induced by mutagenic noncarcinogens vs carcinogens. II. 1- vs 2-nitropropane. Toxicol Appl Pharmacol. 1991 Sep 15;110(3):505–513. doi: 10.1016/0041-008x(91)90050-o. [DOI] [PubMed] [Google Scholar]
  4. Eldridge S. R., Tilbury L. F., Goldsworthy T. L., Butterworth B. E. Measurement of chemically induced cell proliferation in rodent liver and kidney: a comparison of 5-bromo-2'-deoxyuridine and [3H]thymidine administered by injection or osmotic pump. Carcinogenesis. 1990 Dec;11(12):2245–2251. doi: 10.1093/carcin/11.12.2245. [DOI] [PubMed] [Google Scholar]
  5. Gardner H. S., Jr, Brennan L. M., Toussaint M. W., Rosencrance A. B., Boncavage-Hennessey E. M., Wolfe M. J. Environmental complex mixture toxicity assessment. Environ Health Perspect. 1998 Dec;106 (Suppl 6):1299–1305. doi: 10.1289/ehp.98106s61299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gemma S., Faccioli S., Chieco P., Sbraccia M., Testai E., Vittozzi L. In vivo CHCl3 bioactivation, toxicokinetics, toxicity, and induced compensatory cell proliferation in B6C3F1 male mice. Toxicol Appl Pharmacol. 1996 Dec;141(2):394–402. doi: 10.1006/taap.1996.0305. [DOI] [PubMed] [Google Scholar]
  7. Golden R. J., Holm S. E., Robinson D. E., Julkunen P. H., Reese E. A. Chloroform mode of action: implications for cancer risk assessment. Regul Toxicol Pharmacol. 1997 Oct;26(2):142–155. doi: 10.1006/rtph.1997.1161. [DOI] [PubMed] [Google Scholar]
  8. Hatanaka J., Doke N., Harada T., Aikawa T., Enomoto M. Usefulness and rapidity of screening for the toxicity and carcinogenicity of chemicals in medaka, Oryzias latipes. Jpn J Exp Med. 1982 Oct;52(5):243–253. [PubMed] [Google Scholar]
  9. Ishikawa T., Shimamine T., Takayama S. Histologic and electron microscopy observations on diethylnitrosamine-induced hepatomas in small aquarium fish (Oryzias latipes). J Natl Cancer Inst. 1975 Oct;55(4):909–916. doi: 10.1093/jnci/55.4.909. [DOI] [PubMed] [Google Scholar]
  10. Jamison K. C., Larson J. L., Butterworth B. E., Harden R., Skinner B. L., Wolf D. C. A non-bile duct origin for intestinal crypt-like ducts with periductular fibrosis induced in livers of F344 rats by chloroform inhalation. Carcinogenesis. 1996 Apr;17(4):675–682. doi: 10.1093/carcin/17.4.675. [DOI] [PubMed] [Google Scholar]
  11. Kato M., Popp J. A., Conolly R. B., Cattley R. C. Relationship between hepatocyte necrosis, proliferation, and initiation induced by diethylnitrosamine in the male F344 rat. Fundam Appl Toxicol. 1993 Feb;20(2):155–162. doi: 10.1006/faat.1993.1021. [DOI] [PubMed] [Google Scholar]
  12. Koza R. A., Moore M. J., Stegeman J. J. Elevated ornithine decarboxylase activity, polyamines and cell proliferation in neoplastic and vacuolated liver cells of winter flounder (Pleuronectes americanus) Carcinogenesis. 1993 Mar;14(3):399–405. doi: 10.1093/carcin/14.3.399. [DOI] [PubMed] [Google Scholar]
  13. Køppen B., Dalgaard L., Christensen J. M. Determination of trichloroethylene metabolites in rat liver homogenate using headspace gas chromatography. J Chromatogr. 1988 Jun 17;442:325–332. doi: 10.1016/s0021-9673(00)94480-7. [DOI] [PubMed] [Google Scholar]
  14. Larson J. L., Templin M. V., Wolf D. C., Jamison K. C., Leininger J. R., Méry S., Morgan K. T., Wong B. A., Conolly R. B., Butterworth B. E. A 90-day chloroform inhalation study in female and male B6C3F1 mice: implications for cancer risk assessment. Fundam Appl Toxicol. 1996 Mar;30(1):118–137. doi: 10.1006/faat.1996.0049. [DOI] [PubMed] [Google Scholar]
  15. Pitot H. C., Goldsworthy T. L., Moran S., Kennan W., Glauert H. P., Maronpot R. R., Campbell H. A. A method to quantitate the relative initiating and promoting potencies of hepatocarcinogenic agents in their dose-response relationships to altered hepatic foci. Carcinogenesis. 1987 Oct;8(10):1491–1499. doi: 10.1093/carcin/8.10.1491. [DOI] [PubMed] [Google Scholar]
  16. Powers D. A. Fish as model systems. Science. 1989 Oct 20;246(4928):352–358. doi: 10.1126/science.2678474. [DOI] [PubMed] [Google Scholar]
  17. Stone R. A molecular approach to cancer risk. Science. 1995 Apr 21;268(5209):356–357. doi: 10.1126/science.7716533. [DOI] [PubMed] [Google Scholar]
  18. Templin M. V., Jamison K. C., Sprankle C. S., Wolf D. C., Wong B. A., Butterworth B. E. Chloroform-induced cytotoxicity and regenerative cell proliferation in the kidneys and liver of BDF1 mice. Cancer Lett. 1996 Nov 29;108(2):225–231. doi: 10.1016/s0304-3835(96)04427-8. [DOI] [PubMed] [Google Scholar]
  19. Templin M. V., Jamison K. C., Wolf D. C., Morgan K. T., Butterworth B. E. Comparison of chloroform-induced toxicity in the kidneys, liver, and nasal passages of male Osborne-Mendel and F-344 rats. Cancer Lett. 1996 Jun 24;104(1):71–78. doi: 10.1016/0304-3835(96)04234-6. [DOI] [PubMed] [Google Scholar]
  20. Toussaint M. W., Wolfe M. J., Burton D. T., Hoffmann F. J., Shedd T. R., Gardner H. S., Jr Histopathology of Japanese medaka (Oryzias latipes) chronically exposed to a complex environmental mixture. Toxicol Pathol. 1999 Nov-Dec;27(6):652–663. doi: 10.1177/019262339902700607. [DOI] [PubMed] [Google Scholar]
  21. Walker W. W., Manning C. S., Overstreet R. M., Hawkins W. E. Development of aquarium fish models for environmental carcinogenesis: an intermittent-flow exposure system for volatile, hydrophobic chemicals. J Appl Toxicol. 1985 Aug;5(4):255–260. doi: 10.1002/jat.2550050407. [DOI] [PubMed] [Google Scholar]
  22. Weghorst C. M., Henneman J. R., Ward J. M. Dose response of hepatic and renal DNA synthetic rates to continuous exposure of bromodeoxyuridine (BrdU) via slow-release pellets or osmotic minipumps in male B6C3F1 mice. J Histochem Cytochem. 1991 Feb;39(2):177–184. doi: 10.1177/39.2.1987261. [DOI] [PubMed] [Google Scholar]
  23. van der Hoeven J. C., Bruggeman I. M., Alink G. M., Koeman J. H. The killifish Nothobranchius rachowi, a new animal in genetic toxicology. Mutat Res. 1982 Feb;97(1):35–42. doi: 10.1016/0165-1161(82)90017-6. [DOI] [PubMed] [Google Scholar]

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