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A rapid method of species identification of wild chironomids (Diptera: Chironomidae) via electrophoresis of hemoglobin proteins in sodium dodecyl sulfate polyacrylamide gel (SDS–PAGE)

Published online by Cambridge University Press:  13 June 2014

J.T. Oh ,
J.H. Epler  and
C.S. Bentivegna
J.T. Oh
Affiliation:
Department of Biological Sciences, Seton Hall University, South Orange, New Jersey 07079, USA
J.H. Epler
Affiliation:
461 Tiger Hammock Road, Crawfordville, Florida 32327, USA
C.S. Bentivegna*
Affiliation:
Department of Biological Sciences, Seton Hall University, South Orange, New Jersey 07079, USA
*
*Author for correspondence Fax: +973 275-2905 E-mail: [email protected]
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Abstract

Studying aquatic benthic macroinvertebrates (BMIs) in the field requires accurate taxonomic identification, which can be difficult and time consuming. Conventionally, head capsule morphology has been used to identify wild larvae of Chironomidae. However, due to the number of species and possible damage and/or deformity of their head capsules, another supporting approach for identification is needed. Here, we provide hemoglobin (Hb) protein in hemolymph of chironomids as a new biomarker that may help resolve some of the ambiguities and difficulties encountered during taxonomic identification. Chironomids collected from two locations in Maine and New Jersey, USA were identified to the genus level and in some cases to the species-level using head capsule and body morphologies. The head capsule for a particular individual was then associated with a corresponding Hb protein profile generated from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). Distinct Hb profiles were observed from one group (Thienemannimyia) and four genera (Chironomus, Cricotopus, Dicrotendipes, and Glyptotendipes) of chironomids. Several species were polymorphic, having more than one Hb profile and/or having bands of the same size as those of other species. However, major bands and the combination of bands could distinguish individuals at the genus and sometimes species-level. Overall, this study showed that Hb profiles can be used in combination with head capsule morphology to identify wild chironomids.

Keywords

Chironomidaehead capsule morphologyhemoglobin proteinSDS–PAGE
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 104 , Issue 5 , October 2014 , pp. 639 - 651
Copyright
Copyright © Cambridge University Press 2014 

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References

Armitage, P., Cranston, P.S. & Pinder, L.C.V. (1995) The Chironomidae. Biology and Ecology of Nonbiting Midges. London, Chapman & Hall, pp. 572.Google Scholar
Bentivegna, C.S., Alfano, J-E., Bugel, S.M. & Czechowicz, K. (2004) Influence of sediment characteristics on heavy metal toxicity in an urban marsh. Urban Habitats 2(1), 91–111. Available online at: URL http://www.urbanhabitats.org/v02n01/index.html. (accessed on 1 February 2013).Google Scholar
Bentivegna, C.S., Oh, J.T., Doan, K. & DiPietro, C. (2009) Hemoglobin as a biomarker for heavy metals using aquatic midge fly larvae, Chironomidae. The Bulletin, MDI Biological Laboratory 48, 116–119.Google Scholar
Bergtrom, G., Laufer, H. & Rogers, R. (1976) Fat body: a site of hemoglobin synthesis in Chironomus thummi (Diptera). The Journal of Cell Biology 69, 264–274.CrossRefGoogle ScholarPubMed
Carew, M.E., Pettigrove, V. & Hoffmann, A.A. (2003) Identifying chironomids (Diptera: Chironomidae) for biological monitoring with PCR-RFLP. Bulletin of Entomological Research 93, 483–490.CrossRefGoogle ScholarPubMed
Cartier, V., Claret, C., Garnier, R. & Franquet, E. (2011) How salinity affects life cycle of a brackish water species, Chironomus salinarius KIEFFER (Diptera: Chironomidae). Journal of Experimental Marine Biology and Ecology 405, 93–98.CrossRefGoogle Scholar
Chetelat, J., Amyot, M., Cloutier, L. & Poulain, A. (2008) Metamorphosis in chironomids, more than mercury supply, controls methylmercury transfer to fish in high Arctic Lakes. Environmental Science and Technology 42(24), 9110–9115.CrossRefGoogle ScholarPubMed
Cranston, P.S. (2000) Electronic identification guide to the Australian Chironomidae. Available online at: URL http://entomology.ucdavis.edu/chiropage/The_Electronic_Guide_to_the_Chironomidae_of_Australia/. (accessed on 24 January 2013).Google Scholar
Cranston, P.S., Hardy, N.B. & Morse, G.E. (2012) A dated molecular phylogeny for the Chironomidae (Diptera). Systematic Entomology 37, 172–188.CrossRefGoogle Scholar
Das, R. & Handique, R. (1996) Hemoglobin in Chironomus ramosus (Insecta, Diptera): an electrophoretic study of polymorphism, developmental sequence and interspecific relationship. Hydrobiologia 318, 43–50.CrossRefGoogle Scholar
Ebrahimnezhad, M. & Fakhri, F. (2005) Taxonomic study of Chironomidae (Diptera) larvae of Zayandehrood River, Iran, and effects of selected ecological factors on their abundance and distribution. Iranian Journal of Science and Technology, Transaction A 29, 89–105.Google Scholar
Epler, J.H. (2001) Identification Manual for the Larval Chironomidae (Diptera) of North and South Carolina. A guide to the taxonomy of the midges of the southeastern United States, including Florida. Special Publication SJ2001-SP13. Raleigh (NC) and Palatka (FL): North Carolina Department of Environment and Natural Resources, and St. Johns River Water Management District. 526 pp.Google Scholar
Guryev, V., Makarevitch, I., Blinov, A. & Martin, J. (2001) Phylogeny of the genus Chironomus (Diptera) inferred from DNA sequences of mitochondrial cytochrome b and cytochrome oxidase I. Molecular Phylogenetics and Evolution 19, 9–21.CrossRefGoogle ScholarPubMed
Ha, M-H. & Choi, J. (2008) Effects of environmental contaminants on hemoglobin of larvae of aquatic midge, Chironomus riparius (Diptera: Chironomidae): a potential biomarker for ecotoxicity monitoring. Chemosphere 71, 1928–1936.CrossRefGoogle ScholarPubMed
Hemes, B.D. (1998) Gel Electrophoresis of Proteins: A Practical Approach. Oxford, Oxford University Press.CrossRefGoogle Scholar
Jensen, O.N. (2004) Modification-specific proteomics: characterization of post-translational modifications by mass spectrometry. Current Opinion in Chemical Biology 8, 33–41.CrossRefGoogle ScholarPubMed
Lee, S-M., Lee, S-B., Park, C-H. & Choi, J. (2006) Expression of heat shock protein and hemoglobin genes in Chironomus tentans (Diptera, Chironomidae) larvae exposed to various environmental pollutants: a potential biomarker of freshwater monitoring. Chemosphere 65, 1074–1081.CrossRefGoogle ScholarPubMed
Madden, C.P., Suter, P.J., Nicholson, B.C. & Austin, A.D. (1992) Deformities in chironomid larvae as indicators of pollution (Pesticide) stress. Netherlands Journal of Aquatic Ecology 26(2–4), 551–557.CrossRefGoogle Scholar
Mann, M. & Jensen, O.N. (2003) Proteomic analysis of post-translational modifications. Nature Biotechnology 21, 255–261.CrossRefGoogle ScholarPubMed
Martinez, E.A., Moore, B.C., Schaumloffel, J. & Dasgupta, N. (2003) Morphological abnormalities in Chironomus tentans exposed to cadmium – and copper-spiked sediments. Ecotoxicology and Environmental Safety 55, 204–212.CrossRefGoogle ScholarPubMed
Nair, P.M.G., Park, S.Y. & Choi, J. (2011) Expression of catalase and glutathione S-transferase genes in Chironomus riparius on exposure to cadmium and nonylphenol. Comparative Biochemistry and Physiology, Part C 154, 399–408.Google ScholarPubMed
Osmulski, P.A. & Leyko, W. (1986) Structure, function and physiological role of Chironomus haemoglobin. Comparative Biochemistry and Physiology 85B, 701–722.Google Scholar
Pfenninger, M., Nowak, C., Kley, C., Steinke, D. & Streit, B. (2007) Utility of DNA taxonomy and barcoding for the inference of larval community structure in morphologically cryptic Chironomus (Diptera) species. Molecular Ecology 16, 1957–1968.CrossRefGoogle ScholarPubMed
Pfrender, M.E., Hawkins, C.P., Bagley, M., Courtney, G.W., Creutzburg, B.R., Epler, J.H., Fend, S., Ferrington, L.C., Hartzell, P.L., Jackson, S., Larsen, D.P., Levesque, C.A., Morse, J.C., Petersen, M.J., Ruiter, D., Schindel, D., Whiting, M. (2010) Assessing macroinvertebrate biodiversity in freshwater ecosystems: advances and challenges in DNA-BASED approaches. The Quarterly Review of Biology 85, 319–340.CrossRefGoogle ScholarPubMed
Ribbing, W. & Ruterjans, H. (1980) Isomeric incorporation of the haem into monomeric haemoglobins of Chironomus thummi thummi: 2. The Bohr effect of the component III explained on a molecular basis and functional differences between the two isomeric structures. European Journal of Biochemistry 108, 89–102.CrossRefGoogle ScholarPubMed
Schin, K., Poluhowich, J.J., Gamo, T. & Laufer, H. (1974) Degradation of haemoglobin in Chironomus during metamorphosis. Journal of Insect Physiology 20, 561–571.CrossRefGoogle Scholar
Schmidt, E.R., Keyl, H-G. & Hankeln, T. (1988) In situ localization of two haemoglobin gene clusters in the chromosomes of 13 species of Chironomus. Chromosoma 96, 353–359.CrossRefGoogle Scholar
Swansburg, E.O., Fairchild, W.L., Fryer, B.J. & Ciborowski, J.H. (2002) Mouthpart deformities and community composition of Chironomidae (Diptera) larvae downstream of metal mines in New Brunswick, Canada. Environmental Toxicology and Chemistry 21(12), 2675–2684.CrossRefGoogle ScholarPubMed
Tichy, H. (1975) Nature genetic basis and evolution of the haemoglobin polymorphism in Chironomus. Journal of Molecular Evolution 6, 39–50.CrossRefGoogle ScholarPubMed
U.S. Environmental Protection Agency. (1996) Ecological Effects Test Guidelines: OPPTS 850.1790 Chironomid Sediment Toxicity Test. EPA/712/C-96/313.Google Scholar
U.S. Environmental Protection Agency. (2010 a) Watershed Assessment, Tracking & Environmental Results: Maine Water Quality Assessment Report. Available online at: URL http://ofmpub.epa.gov/waters10/attains_state.control?p_state=ME. (accessed on 1 February 2013).Google Scholar
U.S. Environmental Protection Agency. (2010 b) Watershed Assessment, Tracking & Environmental Results: New Jersey Water Quality Assessment Report. Available online at: URL http://ofmpub.epa.gov/waters10/attains_state.control?p_state=NJ. (accessed on 1 February 2013).Google Scholar
Vafopoulou-Mandalos, X., Laufer, H. (1982) The ontogeny of multiple hemoglobins in Chironomus thummi (Diptera): the effects of a compound with juvenile hormone activity. Developmental Biology 92, 135–143.CrossRefGoogle ScholarPubMed
Vafopoulou-Mandalos, X., Laufer, H. (1984) Regulation of hemoglobin synthesis by ecdysterone and juvenile hormone during development of Chironomus thummi (Diptera). Differentiation 27, 94–105.CrossRefGoogle ScholarPubMed
Warwick, W.F. (1985) Morphological abnormalities in Chironomidae (Diptera) larvae as measures of ioxic stress in freshwater ecosystems: indexing antennal deformities in Chironomus Meigen. Canadian Journal of Fisheries and Aquatic Sciences 42, 1881–1914.CrossRefGoogle Scholar
Watts, M.M., Pascoe, D. (2000) A comparative study of Chironomus riparius Meigen and Chironomus tentans Fabricius (Diptera:Chironomidae) in aquatic toxicity tests. Archives of Environmental Contamination and Toxicology 39, 299–306.Google ScholarPubMed
Wollmer, A., Buse, G., Sick, H. & Gersonde, K. (1972) A myoglobin-type haemoglobin of Chironomus thummi thummi. European Journal of Biochemistry 24, 547–552.CrossRefGoogle ScholarPubMed