Assessing contamination levels of amprolium and tylosin using the embryo toxicity assay and three biomarkers

Authors

  • Jun Hu
  • Yong Liang School of Medicine, Jianghan University, Hubei Province, P.R China 430056
  • Minjie Chen School of Medicine, Jianghan University, Hubei Province, P.R China 430056
  • Xiaorong Wang School of the Environment, Nanjing University, Nanjing 210093, P.R. China

Keywords:

veterinary pharmaceutical, zebrafish embryo, oxidative stress, heat shock protein, risk assessment

Abstract

This study was conducted to examine the adverse biological effects of two veterinary pharmaceuticals (Amprolium and tylosin) on aquatic organisms. The zebrafish embryo toxicity assays were performed in combination with three biomarkers, including superoxide dismutase (SOD), lipid peroxidation (LPO) and heat shock protein 70(Hsp70). The LPO extent was expressed by malondialdehyde (MDA) in this paper. Logit Loglinear analysis was used to study the effect of exposure concentration and time on survival rate and percent hatched in embryo toxicity tests. The data on SOD, MDA and Hsp70 were analyzed by one-way ANOVA followed by Dunnett's test. The results showed that both pharmaceuticals had no effect on Hsp70 levels in zebrafish. Amprolium and tylosin had significant effect on the survival rate, the percent hatched, SOD and MDA in the range of the experimental concentrations. Furthermore, there was a sharp increase, followed by a decrease in the survival rate, as the exposure concentrations of amprolium or tylosin increased; while SOD and MDA indicated a sharp decrease followed by an increase in survival rates. These results indicated that both pharmaceuticals can be useful to the embryos at low concentrations, while causing acute toxicity at high concentrations. However, two pharmaceuticals cause an acute toxicity effect at concentrations several orders of magnitude higher than detected in the aquatic environments. Both do not result in acute toxicity to zebrafish, but it is recommended that the proper strategy and approach be found to perform chronic low-dose exposure studies.

References

Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K., eds. 1999. Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, New York: John Wiley & Sons.

Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1–2): 248–254.

Brain, R. A., Bestari, K., Sanderson, H., Hanson, M. L., Wilson, C. J., Johnson, D. J., Sibley, P. K. and Solomon, K. R. 2005. Aquatic microcosm assessment of the effects of tylosin on Lemna gibba Myriophyllum spicatum. Environmental Pollution, 133(3): 389–401.

Cleuvers, M. 2003. Aquatic ecotoxicity of pharmaceuticals including the assessment of combination effects. Toxicology Letters, 142(3): 185–194.

Dussault, E. B., Balakrishnan, V. K., Sverko, E., Solomon, K. R. and Sibley, P. K. 2008. Toxicity of human pharmaceuticals and personal care products to benthic invertebrates. Environ Toxicol Chem., 27(2): 425–432.

Enick, O. V. and Moore, M. M. 2007. Assessing the assessments: Pharmaceuticals in the environment. Environmental Impact Assessment Review, 27(8): 707–729.

Escher, B. I., Bramaz, N., Eggen, R. I. L. and Richter, M. 2005. In Vitro Assessment of Modes of Toxic Action of Pharmaceuticals in Aquatic Life. Environ. Sci. Technol., 39(9): 3090–3100.

Hallare, A. V., Kohler, H. R. and Triebskorn, R. 2004. Developmental toxicity and stress protein responses in zebrafish embryos after exposure to diclofenac and its solvent, DMSO. Chemosphere, 56(7): 659–666.

Hallare, A. V., Kosmehl, T., Schulze, T., Hollert, H., Kohler, H. R. and Triebskorn, R. 2005. Assessing contamination levels of Laguna Lake sediments (Philippines) using a contact assay with zebrafish (Danio rerio) embryos. Science of The Total Environment, 347(1-3): 254–271.

Hallare, A. V., Pagulayan, R., Lacdan, N., Köhler, H. R. and Triebskorn, R. 2005. Assessing water quality in a tropical lake using biomarkers in zebrafish embryos: developmental toxicity and stress protein responses. Environmental Monitoring and Assessment, 104(1): 171–187.

Hollert, H., Keiter, S., König, N., Rudolf, M., Ulrich, M. and Braunbeck, T. 2003. A new sediment contact assay to assess particle-bound pollutants using zebrafish (Danio rerio) embryos. Journal of Soils and Sediments, 3(3): 197–207.

Ingerslev, F., Torang, L., Loke, M.-L., Halling-Sorensen, B. and Nyholm, N. 2001. Primary biodegradation of veterinary antibiotics in aerobic and anaerobic surface water simulation systems. Chemosphere, 44(4): 865–872.

Kakkar, P. and Jaffery, F. N. 2005. Biological markers for metal toxicity. Environmental Toxicology and Pharmacology, 19(2): 335–349.

Kammann, U., Biselli, S., Hühnerfuss, H., Reineke, N., Theobald, N., Vobach, M. and Wosniok, W. 2004. Genotoxic and teratogenic potential of marine sediment extracts investigated with comet assay and zebrafish test. Environmental Pollution, 132(2): 279–287.

Kemper, N. 2008. Veterinary antibiotics in the aquatic and terrestrial environment. Ecological Indicators, 8(1): 1–13.

Kolpin, D. W., Furlong, E. T., Meyer, M. T., Thurman, E. M., Zaugg, S. D., Barber, L. B. and Buxton, H. T. 2002. Pharmaceuticals, Hormones, and Other Organic Wastewater Contaminants in U. S. Streams, 1999–2000: A National Reconnaissance. Environ. Sci. Technol., 36(6): 1202–1211.

Kummerer, K., Al-Ahmad, A. and Mersch-Sundermann, V. 2000. Biodegradability of some antibiotics, elimination of the genotoxicity and affection of wastewater bacteria in a simple test. Chemosphere, 40(7): 701–710.

Laville, N., Ait-Aissa, S., Gomez, E., Casellas, C. and Porcher, J. M. 2004. Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes. Toxicology, 196(1-2): 41–55.

Lissemore, L., Hao, C., Yang, P., Sibley, P. K., Mabury, S. and Solomon, K. R. 2006. An exposure assessment for selected pharmaceuticals within a watershed in Southern Ontario. Chemosphere, 64(5): 717–729.

Liu, F., Ying, G.-G., Tao, R., Zhao, J.-L., Yang, J.-F. and Zhao, L.-F. 2009. Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities. Environmental Pollution, 157(5): 1636–1642.

Luckenbach, T., Kilian, M., Triebskorn, R. and Oberemm, A. 2001. Fish early life stage tests as a tool to assess embryotoxic potentials in small streams. Journal of Aquatic Ecosystem Stress and Recovery (Formerly Journal of Aquatic Ecosystem Health), 8(3): 355–370.

McArdell, C. S., Molnar, E., Suter, M. J. F. and Giger, W. 2003. Occurrence and Fate of Macrolide Antibiotics in Wastewater Treatment Plants and in the Glatt Valley Watershed, Switzerland. Environ. Sci. Technol., 37(24): 5479–5486.

Nikolaou, A., Meric, S. and Fatta, D. 2007. Occurrence patterns of pharmaceuticals in water and wastewater environments. Analytical and Bioanalytical Chemistry, 387(4): 1225–1234.

Pyza, E., Mak, P., Kramarz, P. and Laskowski, R. 1997. Heat Shock Proteins (HSP70) as Biomarkers in Ecotoxicological Studies. Ecotoxicology and Environmental Safety, 38(3): 244–251.

Richards, S. M., Wilson, C. J., Johnson, D. J., Castle, D. M., Lam, M., Mabury, S. A., Sibley, P. K. and Solomon, K. R. 2004. Effects of pharmaceutical mixtures in aquatic microcosms. Environ Toxicol Chem., 23(4): 1035–1042.

Sanderson, H., Brain, R. A., Johnson, D. J., Wilson, C. J. and Solomon, K. R. 2004. Toxicity classification and evaluation of four pharmaceuticals classes: antibiotics, antineoplastics, cardiovascular, and sex hormones. Toxicology, 203(1-3): 27–40.

Sarmah, A. K., Meyer, M. T. and Boxall, A. B. A. 2006. A global perspective on the use, sales, exposure pathways, occurrence, fate and effects of veterinary antibiotics (VAs) in the environment. Chemosphere, 65(5): 725–759.

Seiler, J. P. 2002. Pharmacodynamic activity of drugs and ecotoxicology–can the two be connected?. Toxicology Letters, 131(1-2): 105–115.

Tong, L., Li, P., Wang, Y. and Zhu, K. 2009. Analysis of veterinary antibiotic residues in swine wastewater and environmental water samples using optimized SPE-LC/MS/MS. Chemosphere, 74(8): 1090–1097.

Vaclavik, E., Halling-Sorensen, B. and Ingerslev, F. 2004. Evaluation of manometric respiration tests to assess the effects of veterinary antibiotics in soil. Chemosphere, 56(7): 667–676.

van der Oost, R., Beyer, J. and Vermeulen, N. P. E. 2003. Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environmental Toxicology and Pharmacology, 13(2): 57–149.

Wollenberger, L., Halling-Sorensen, B. and Kusk, K. O. 2000. Acute and chronic toxicity of veterinary antibiotics to Daphnia magna. Chemosphere, 40(7): 723–730.

Zurita, J. L., Jos, A., del Peso, A., Salguero, M., Lopez-Artiguez, M. and Repetto, G. 2007. Ecotoxicological effects of the antioxidant additive propyl gallate in five aquatic systems. Water Research, 41(12): 2599–2611.

Downloads

Published

2010-08-31

Issue

Vol. 13 No. 3 (2010)

Section

Research article

License

Manuscripts must be original. They must not be published or be under consideration for publication elsewhere, in whole or in part. It is required that the lead author of accepted papers complete and sign the MSU Press AEHM Author Publishing Agreement and provide it to the publisher upon acceptance.