Evaluation of Chlortetracycline Impact on Earthworms (Eisenia fetida) by Quantitative Real-Time PCR

Abstract:

Chlortetracycline (CTC), antimicrobial chemical, is widely used to treat disease and protect the health of animals. As CTC is poorly absorbed during digestion, the majority of the bioactive compound or metabolites are excreted in faeces and urine. Toxicogenomics approach was used to examine the toxicological effects of CTC on earthworms (E. fetida). Quantification of gene expression levels encoding antioxidative system and biotransformation enzymes were analyzed in E. fetida. The results showed that GST transcript level was significantly inhibited at 300 mg CTC kg-1, with an approximately a 65% decrease compared to the control. Ferritin and serine were marked by high level transcripts at the lowest concentration and followed by decreasing levels with the increased concentraion. Hsp70 transcript appeared to be disrupted at the lowest concentration. Thus, environmental assessment using these genes expression levels in earthworms might be sensitive and helpful in the early diagnosis of potentially polluted soil.

• Periodical: Applied Mechanics and Materials (Volume 108)
• Pages: 230-237
• DOI: https://doi.org/10.4028/www.scientific.net/AMM.108.230
• Online since: October 2011
• Authors: Da Song Lin, Kang Xin He, Qi Xing Zhou, Ye Li, Xiu Jie Xie, Yao Liu
• Keywords: Biomarkers, Chlortetracycline, Earthworm, Gene Expression, Quantitative Real Time PCR

References

[1] G.K. Elmund, S.M. Morrison, D.W. Grant, M.P. Nevins, Role of excreted chlortetracycline in modifying the decomposition process in feedlot waste, Bull. Environ. Contam. Toxicol. 1971, 6, 129-132.

DOI: 10.1007/bf01540093

[2] G. Hamscher, A. Abu-Quare, S. Sczesny, H. Hoper, H. Nau. Determination of tetracyclines and tylosin in soil and water samples from agricultural areas in Lower Saxony, in: L.A. van Ginkel, A. Ruiter (Eds. ), Proceedings of the Euroside IV Conference, National Institute of Public Health and the Environment (RIVM), Bilthoven, Netherlands, 2000, pp.522-526.

[3] X.J. Xie, Q.X. Zhou, Z.C. He, Y.Y. Bao, Physiological and potential genetic toxicity of chlortetracycline as an emerging pollutant in wheat (Tritium aestivum L. ), Environ. Toxicol. Chem. 2010, 29, 922-928.

DOI: 10.1002/etc.79

[4] US Department of Agriculture Economic Research Service (USDAERS). Agricultural chemicals and production technology: nutrient management. 2006, 16, http: /www. ers. usda. gov/briefing/AgChemicals/nutrientmangement.

[5] S.R. Sturzenbaum, P. Kille, A.J. Morgan,. Heavy metal-induced molecular responses in the earthworm, Lumbricus rubellus: genetic fingerprinting by directed differential display, Appl. Soil Ecol. 1998a, 9, 495-500.

DOI: 10.1016/s0929-1393(98)00111-5

[6] F. Bernard, F. Brulle, F. Douay, S. Lemiere, S. Demuynck, F. Vandenbulcke, Metallic trace element body burdens and gene expression analysis of biomarker candidates in Eisenia fetida, using an exposure/depuration experimental scheme with field soils, Ecotoxicol. Environ. Saf. 2010, 73, 1034-1045.

DOI: 10.1016/j.ecoenv.2010.01.010

[7] H.J. Ricketts, A.J. Morgan, D.J. Spurgeon, P. Kille, Measurement of annetocin gene expression: a new reproductive biomarker in earthworm ecotoxicology, Ecotoxicol. Environ. Saf. 2004, 57, 4-10.

DOI: 10.1016/j.ecoenv.2003.08.008

[8] Y.G. Xue, X.Y. Gu, X.R. Wang, C. Sun, X.H. Xu, J. Sun, B.G. Zhang, The hydroxyl radical generation and oxidative stress for the earthworm Eisenia fetida exposed to tetrabromobisphenol A, Ecotoxicology. 2009, 18, 693-699.

DOI: 10.1007/s10646-009-0333-2

[9] F. Corpet, Multiple sequence alignment with hierarchical clustering, Nucleic Acids Res. 1998, 16, 10881-10890.

DOI: 10.1093/nar/16.22.10881

[10] J.D. Thompson, D.G. Higgins, T.J. Gibson, CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment throught sequence weighting, positions-specific gap penalties and weight matrix choice, Nucleic Acids Res. 1994, 22, 4673-4680.

DOI: 10.1093/nar/22.22.4673

[11] T.M. Rose, E.R. Schultz, J.G. Henikoff, S. Pietrokovski, C.M. McCallum, S. Henikov, "Consensus-degenerate hybrid oligonucleotide primers for amplification of distantly related sequences. Nucleic Acids Res. 1998, 26, 1628-1635.

DOI: 10.1093/nar/26.7.1628

[12] F. Brulle, A.J. Morgan, C. Cocquerelle, F. Vandenbulcke, Transcriptomic underpinning of toxicant-mediated physiological function alterations in three terrestrial invertebrate taxa: A review, Environ. Pollu. 2010, 158, 2793-3808.

DOI: 10.1016/j.envpol.2010.06.019

[13] K.J. Livak, T.D. Schmittgen, Analysis of relative gene expression data using realtime quantitative PCR and the 2(-Delta Delta C(T) method, Methods. 2001, 25, 402-408.

DOI: 10.1006/meth.2001.1262

[14] B. Nota, M. Bosse, B. Ylstra, N.M. Straalen, D. Roelofs, Transcriptomics reveals extensive inducible biotransformation in the soil-dwelling invertebrate Folsomia candida exposed to phenanthrene, BMC Genomics 2009, 10, 236-249.

DOI: 10.1186/1471-2164-10-236

[15] D.S. Lin, Q.X. Zhou, X.J. Xie, Y. Liu, Potential biochemical and genetic toxicity of triclosan as an emerging pollutant on earthworms (Eisenia fetida), Chemosphere, 2010, 81, 1328-1333.

DOI: 10.1016/j.chemosphere.2010.08.027

[16] P.J. Brown, S.M. Long, D.J. Spurgeon, C. Svendsen, P.K. Hankard, Toxicological and biochemical responses of the earthworm Lumbricus rubellus to pyrene, a non-carcinogenic polycyclic aromatic hydrocarbon, Chemosphere 2004, 57, 1675-1681.

DOI: 10.1016/j.chemosphere.2004.05.041

[17] A. Bigot, P. Vasseur, F. Rodius, SOD and CAT cDNA cloning, and expression pattern of detoxification genes in the reshwater bivalve Unio tumidus transplanted into the moselle river, Ecotoxicology, 2010, 19, 369-376.

DOI: 10.1007/s10646-009-0419-x

[18] P. Gong, N. Basu, A.M. Scheuhammer, E.J. Perkins, Neurochemical and electrophysiological diagnosis of reversible neurotoxicity in earthworms exposed to sublethal concentrations of CL-20, Environ Sci Pollut Res. 2010, 17, 181-186.

DOI: 10.1007/s11356-009-0117-5

[19] A.M. Lescure, D. Proudhon, H. Peser, M. Ragland, E.C. Theil, J.F. Briat, Ferritin gene transcription is regulated by iron in soybean cell cultures, Proc. Natl. Acad. Sci. 1991, 88, 8222-8226.

DOI: 10.1073/pnas.88.18.8222

[20] S. Lewis, R.D. Handy, B. Cordi, Z. Billinghurst, M.H. Depledge, Stress proteins (HSP's): methods of detection and their use as an environmental biomarker, Ecotoxicology 1999, 8, 351-368.

DOI: 10.1023/a:1008982421299

[21] D. Nadeau, S. Corneau, I. Plante, G. Morrow, R.M. Tanguay, Evaluation for Hsp70 as a biomarker of effect of pollutants on the earthworm Lumbricus terrestris, Cell Stress Chaperones 2001, 6, 153-163.

DOI: 10.1379/1466-1268(2001)006<0153:efhaab>2.0.co;2

[22] Y.R. Liu, Y.M. Zheng, L.M. Zhang, Y.X. Luan, J.Z. He, Effects of mercury on reproduction, avoidance, and heat shock protein gene expression of the soil springtail folsomia candida, Environ. Toxicol. Chem. 2009, 29, 654-659.

DOI: 10.1002/etc.59