Methemoglobin in Chironomus Larvae as Potential Biomarker of Nitrate Contamination in Water

  • Judith A. Khazenzi School of Environmental Studies Moi University, P.O Box 3900-30100, Eldoret, Kenya.
  • Johnstone Wakhisi School of Medicine Moi University, P.O Box 3900-30100, Eldoret, Kenya.
  • Odipo Osano School of Environmental Studies Moi University, P.O Box 3900-30100, Eldoret, Kenya.
  • Phillip Raburu School of Natural Resource Management Moi University, P.O Box 3900-30100, Eldoret, Kenya.
Keywords: Chironomus, Biomarker, Methemoglobin, Nitrate


Nitrate contamination of groundwater and surface water has been found to be high in various areas. Nitrates are known to have health impacts if consumed in different concentrations. The study set out to investigate the potential of using methemoglobin within chironomid larva e as a biomarker for nitrates in water. The ubiquitous and hemoglobin – containing chironomid larvae from Lake Victoria basin were identified using morphological characteristics and Chironomuswas found to bethe most common genera. The larvae can also withstand polluted waters. The Chironomus was therefore chosen for the study and was exposed to different concentrations of nitrate in water in the laboratory. The Chironomus larvae were analyzed for methemoglobin using a spectrophotometer and the levels compar ed to the exposure nitrate concentration of the test solution. Acute toxicity test was carried out by exposing the larvae to different concentrations of nitrate and determining the LC 50. Results showed a positive correlation between nitrate concentration and hemoglobin absorbance in the tested cases. The LC 50 after 48hrs was found to be 34.2 (30-39; 95% confidence limit) mgL-1 NO3-N for third instar larvae and for first instar larvae after 96hrs was 41.3 (35.9-50.0; 95% confidence limit) mgL- 1 NO3-N. From the results it can be seen that chironomid larvae have the potential to be used to indicate differences in nitrate concentration in water containing nitrate concentrations of upto 40 mgL-1 NO3-N. The study can help in the development of a bioassessment tool for nitrates in water. However, further work needs to be carried out on effect of age of the larvae on methemoglobin formation.


Aime, S., Dastrú, W., Fasano, M., Arnelli, A., Castagnola, M., & Giardina, B., (1992). Quantitative Determination of Methemoglobin by Measuring the Solvent-Water Proton-Nuclear Magnetic Resonance Relaxation Rate. Clin. Chem. 38, 2401-2404.

Almasri, M. N. (2007). Nitrate contamination of groundwater: A conceptual management framework. Environmental Impact Assessment Review, 27, 220–242.

Al-Qudaha, K. M., Rousan, L. M., & Ereifej, K. I. (2009). Nitrate/nitrite p oisoning in dairy cattle associated with consumption of forages irrigated with municipally treated wastewater. Toxicol. Environ. Chem . 91, 163–170.

APHA. (1992). Standard Methods For the Examination of Water and Wastewater. Washington DC: American Public Health Association.

Avilez, I. M., Altran, A. E., Aguiar, L. H., & Moraes, G. (2004). Hematological responses of the Neotropical teleost matrinxã (Brycon cephalus) to environmental nitrite. Comparative Biochemistry and Physiology, Part C, 139 , 135–139.

Carew, M. E., Pettigrove, V., & Hoffmann, A. A. (2003). Identifying chironomids (Diptera: Chironomidae) for biological monitoring with PCR-RFLP. Bull. Entomol. Res. 93, 483-490.

Eggermont, H., Verschuren, D., & Dumont, H. (2005). Taxonomic diversity and biogeo graphy of Chironomidae (Insecta: Diptera) in lakes of tropical West Africa using subfossil remains extracted from surface sediments. J. Biogeogr. 32, 1063–1083.

Gruener, N., & T oeplitz, R. (1975). T he Effect Of Changes In Nitrate Concentration In Drinking Water On Methemoglobin Levels

In Infants. International Journal of Environmental Studies, 71 , 61-163.

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.

Hankeln, T., Jaenicke, V., Kiger, L., Dewilde, S., Ungerechts, G., & Schmidt, M., et al. (2002). Characterization of Drosophila Hemoglobin: Evidence for Hemoglobin -Mediated Respiration in Insects. J. Biol. Chem . 277, 29012-29017.

Hudson, L. A., & Ciborowski, J. J. (1996). Teratogenic and Genotoxic Responses of Larval Chironomus salinarius group (Diptera :
Chironomidae) to Contaminated Sediment . Environmental Toxicology and Chemistry, 15:(8), 1375–1381.

Hyne, R. V., & Maher, W. A. (2003). Invertebrate biomarkers: links to toxicosis that predict population decline. Ecotoxicol. Environ. Safety. 54, 366–374.

Khazenzi, J. A., Osano, O., Wakhisi, J., & Raburu, P. (2011). Potential Use of Chironomid Larvae in Methemoglobinemia Risk Assessment: A Case of Groundwater Users in Langas, Eldoret. Journal of technology socio-economic development 1(1), 63-69.

Leyko, W., & Osmulski, P. A. (1985). Seasonal variability of hemoglobin content and compon ent composition of Chironomus thummi thummi larvae. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 80(3), 613-616.

Marziali, L., Armanini, D. G., Cazzola, M., Erba, S., & T oppi, E. (2010). Responses of Chironomid Larvae (insecta, diptera) to Ecological Quality in Mediterranean River Mesohabitats (South Italy). River Res. Applic. 26,, 1036-1051.

McGavin, G. C. (1992). Insects of the Northern Hemisphere. Limpsfied: Dragons World Limited.

Panis, L. I., Goddeeris, B., & Verheyen, R. (1995). T he hemoglobin concentration of Chironomus cf.Plumosus l. (Diptera:
Chironomidae) larvae from two lentic habitats. Aquatic Ecology, 29, 1-4.

Park, K., & Kwak, I. -S. (2010). Molecular effects of endocrine-disrupting chemicals on the Chironomus riparius estrogen-related receptor gene. Chemosphere 79, 934–941.

Péry, A. R., Mons, R., & Garric, J. (2005). Modelling of the life cycle of Chironomus species using an energy -based model.
Chemosphere 59, 247–253.

Republic of Kenya. (2006). Environmental Management and Co-ordination (Water Quality) Regulations,. Kenya Gazette supplement No 68. Legal Notice No. 120 .

Rouse, J. D., Bishop, C. A., & Struger, J. (1999). Nitrogen Pollution: An Assessment of Its Threat to Amphibian Survival. Environ. Health Perspect. 107, 799-803.

Sadeq, M. M., Attarassi, B., Cherkaoui, I., ElAouad, R., & Idrissi, L. (2008). Drinking water nitrate and prevalence of methemoglobinemia among infants and children aged 1 –7 years in Moroccan areas. International Journal of Hygiene and Environmental Health, 211, 546–554.

Weber, R. E., & Vinogradov, S. N. (2001). Nonvertebrate Hemoglobins: Functions and Molecular Adaptations. Physiol. Rev. 81, 569-628.

Wylie, B. R., & Lovric, A. (1988). Hemoglobinometry: Evaluation Of A New Method With A Stable Primary Standard. Pathology,
20:, 152-155.

Zijlstra, W. G., & Buursma, A. (1997). Spectrophotometry of Hemoglobin: Absorption Spectra of Bovine Oxyhemoglobin, Deoxyhemoglobin, Carboxyhemoglobin, and Methemoglobin. Comparative Biochemical and Physiology, 118B:( 4), 743–749.
How to Cite
Khazenzi, J. A., Wakhisi, J., Osano, O., & Raburu, P. (2018, August 27). Methemoglobin in Chironomus Larvae as Potential Biomarker of Nitrate Contamination in Water. African Journal of Education,Science and Technology, 1(2), pp 84-90. Retrieved from