Acute Toxicity of Nitrate and Nitrite to Sensitive Freshwater Insects, Mollusks, and a Crustacean



Both point- and nonpoint-sources of pollution have contributed to increased inorganic nitrogen concentrations in freshwater ecosystems. Although numerous studies have investigated the toxic effects of ammonia on freshwater species, relatively little work has been performed to characterize the acute toxicity of the other two common inorganic nitrogen species: nitrate and nitrite. In particular, to our knowledge, no published data exist on the toxicity of nitrate and nitrite to North American freshwater bivalves (Mollusca) or stoneflies (Insecta, Plecoptera). We conducted acute (96-h) nitrate and nitrite toxicity tests with two stonefly species (Allocapnia vivipara and Amphinemura delosa), an amphipod (Hyalella azteca), two freshwater unionid mussels (Lampsilis siliquoidea and Megalonaias nervosa), a fingernail clam (Sphaerium simile), and a pond snail (Lymnaea stagnalis). Overall, we did not observe a particularly wide degree of variation in sensitivity to nitrate, with median lethal concentrations ranging from 357 to 937 mg NO3-N/l; furthermore, no particular taxonomic group appeared to be more sensitive to nitrate than any other. In our nitrite tests, the two stoneflies tested were by far the most sensitive, and the three mollusks tested were the least sensitive. In contrast to what was observed in the nitrate tests, variation among species in sensitivity to nitrite spanned two orders of magnitude. Examination of the updated nitrite database, including previously published data, clearly showed that insects tended to be more sensitive than crustaceans, which were in turn more sensitive than mollusks. Although the toxic mechanism of nitrite is generally thought to be the conversion of oxygen-carrying pigments into forms that cannot carry oxygen, our observed trend in sensitivity of broad taxonomic groups, along with information on respiratory pigments in those groups, suggests that some other yet unknown mechanism may be even more important.


  1. Alonso A, Camargo JA (2003) Short-term toxicity of ammonia, nitrite, and nitrate to the aquatic snail Potamopyrgus antipodarum (Hydrobiidae, Mollusca). Bull Environ Contam Toxicol 70:1006–1012CrossRefGoogle Scholar
  2. Alonso A, Camargo JA (2006) Toxicity of nitrite to three species of freshwater invertebrates. Environ Toxicol 21:90–94CrossRefGoogle Scholar
  3. Alonso A, Camargo JA (2008) Ameliorating effect of chloride on nitrite toxicity to freshwater invertebrates with different physiology: a comparative study between amphipods and planarians. Arch Environ Contam Toxicol 54:259–265CrossRefGoogle Scholar
  4. American Public Health Association (2005) Standard methods for the examination of water and wastewater, 21st ed. APHA-AWWA-WPCF, Washington, DCGoogle Scholar
  5. American Society for Testing and Materials (2002) Standard guide for conducting acute toxicity testing on test materials with fishes, macroinvertebrates, and amphibians. Designation: E729-96. ATSM, West Conshocken, PAGoogle Scholar
  6. American Society for Testing, Materials (2006) Standard guide for conducting laboratory toxicity tests with freshwater mussels. Designation: E2455-06. ATSM, West Conshocken, PAGoogle Scholar
  7. Beitinger TL, Huey DW (1981) Acute toxicity of nitrite to crayfish Procambarus simulans in varied environmental conditions. Environ Pollut 26:305–311CrossRefGoogle Scholar
  8. Borgmann U (1996) Systematic analysis of aqueous ion requirements of Hyalella azteca: a standard artificial medium including the essential bromide ion. Arch Environ Contam Toxicol 30:356–363CrossRefGoogle Scholar
  9. Camargo JA, Alonso A (2006) Ecological and toxicological effects of inorganic nitrogen pollution in aquatic ecosystems: a global assessment. Environ Int 32:831–849CrossRefGoogle Scholar
  10. Camargo JA, Ward JV (1992) Short-term toxicity of sodium nitrate (NaNO3) to non-target freshwater invertebrates. Chemosphere 24:23–28CrossRefGoogle Scholar
  11. Camargo JA, Alonso A, Salamanca A (2005) Nitrate toxicity to aquatic animals: a review with new data for freshwater invertebrates. Chemosphere 58:1255–1267CrossRefGoogle Scholar
  12. Chandler JH Jr, Marking LL (1979) Toxicity of fishery chemicals to the Asiatic clam Corbicula manilensis. Prog Fish Cult 41:148–151CrossRefGoogle Scholar
  13. Corrao NM, Darby PC, Pomory CM (2006) Nitrate impacts on the Florida apple snail, Pomacea paludosa. Hydrobiologia 568:135–143CrossRefGoogle Scholar
  14. Douda K (2010) Effects of nitrate nitrogen pollution on Central European unionid bivalves revealed by distributional data and acute toxicity testing. Aquat Conserv Mar Freshw Ecosyst 20:189–197CrossRefGoogle Scholar
  15. Ewell WS, Gorsuch JW, Kringle RO, Robillard KA, Spiegel RC (1986) Simultaneous evaluation of the acute effects of chemicals on seven aquatic species. Environ Toxicol Chem 5:831–840CrossRefGoogle Scholar
  16. Galloway JN, Cowling EB (2002) Reactive nitrogen and the world: 200 years of change. Ambio 31:64–71Google Scholar
  17. Gutzmer MP, Tomasso JR (1985) Nitrite toxicity to the crayfish Procambarus clarkii. Bull Environ Contam Toxicol 34:369–376CrossRefGoogle Scholar
  18. Hall RL, Pearson JS, Wood EJ (1975) The haemocyanin of Lymnaea stagnalis L. (Gastropoda: Pulmonata). Comp Biochem Physiol 52B:211–218Google Scholar
  19. Hamilton MA, Russo RC, Thurston RV (1977) Trimmed Spearman–Karber method for estimating lethal concentrations in toxicity bioassays. Environ Sci Technol 1:714–719CrossRefGoogle Scholar
  20. Harris RR, Coley S (1991) The effects of nitrite on chloride regulation in the crayfish Pacifastacus leniusculus Dana (Crustacea: Decapoda). J Comp Physiol B 161:199–206CrossRefGoogle Scholar
  21. Hymel TM (1985) Water quality dynamics in commercial crayfish ponds and toxicity of selected water quality variables to Procambarus clarkii. Master’s thesis. School of Forestry, Wildlife and Fisheries, Louisiana State University, Baton Rouge, LAGoogle Scholar
  22. Jayasankar P, Muthu MS (1983) Toxicity of nitrite to the larvae of Penaeus indicus. Indian J Fish 30:231–240Google Scholar
  23. Jensen FB (1990) Sublethal physiological changes in freshwater crayfish, Astacus astacus, exposed to nitrite: Haemolymph and muscle tissue electrolyte status, and haemolymph acid–base balance and gas transport. Aquat Toxicol 18:51–60CrossRefGoogle Scholar
  24. Jensen FB (1996) Uptake, elimination and effects of nitrite and nitrate in freshwater crayfish (Astacus astacus). Aquat Toxicol 34:95–104CrossRefGoogle Scholar
  25. Kelso BHL, Glass DM, Smith RV (1999) Toxicity of nitrite to freshwater invertebrates. In: Wilson WS, Ball AS, Hinton RH (eds) Managing risks of nitrates to humans and the environment. Royal Society of Chemistry, Cambridge, UK, pp 175–188Google Scholar
  26. Kemble NE, Dwyer FJ, Ingersoll CG, Dawson TD, Norberg-King TJ (1999) Tolerance of freshwater test organisms to formulated sediments for use as control materials in whole-sediment toxicity tests. Environ Toxicol Chem 18:222–230CrossRefGoogle Scholar
  27. Kozák P, Máchová J, Policar T (2005) The effect of chloride content in water on the toxicity of sodium nitrite for spiny-cheek crayfish (Orconectes limosus Raf.). Bull Fr Pêche Piscic 376–377:705–714CrossRefGoogle Scholar
  28. Liu H, Avault W Jr, Medley P (1995) Toxicity of ammonia and nitrite to juvenile redclaw crayfish, Cherax quadricarinatus (von Martens). Freshw Crayfish 10:249–255Google Scholar
  29. McMahon RF, Bogan AE (2001) Mollusca: bivalvia. In: Thorp JH, Covich AP (eds) Ecology and classification of North American freshwater invertebrates, 2nd edn. Academic, San Diego, CA, pp 331–429CrossRefGoogle Scholar
  30. Meade ME, Watts SA (1995) Toxicity of ammonia, nitrite, and nitrate to juvenile Australian crayfish, Cherax quadricarinatus. J Shellfish Res 14:341–346Google Scholar
  31. Monson P (2010) Aquatic life water quality standards technical support document for nitrate. Minnesota Pollution Control Agency.
  32. Newton TJ, Bartsch MR (2007) Lethal and sublethal effects of ammonia to juvenile lampsilis mussels (Unionidae) in sediment and water-only exposures. Environ Toxicol Chem 26:2057–2065CrossRefGoogle Scholar
  33. Newton TJ, Allran JW, O’Donnell JA, Bartsch MR, Richardson WB (2003) Effects of ammonia on juvenile unionid mussels (Lampsilis cardium) in laboratory sediment toxicity tests. Environ Toxicol Chem 22:2554–2560CrossRefGoogle Scholar
  34. Pandey RB, Adams GL, Warren LW (2011) Survival and precopulatory behavior of Hyalella azteca (Amphipoda) exposed to nitrate in the presence of atrazine. Environ Toxicol Chem 30:1170–1177CrossRefGoogle Scholar
  35. Resh VH, Buchwalter DB, Lamberti GA, Eriksen CH (2008) Aquatic insect respiration. In: Merritt RW, Cummins KW, Berg MB (eds) An introduction to the aquatic insects of North America, 4th edn. Kendall Hunt, Dubuque, IAGoogle Scholar
  36. Rouse DB, Kastner RJ, Reddy KS (1995) Toxicity of ammonia and nitrite to hatchling redclaw crayfish, Cherax quadricarinatus. Freshw Crayfish 10:298–303Google Scholar
  37. Scott G, Crunkilton RL (2000) Acute and chronic toxicity of nitrate to fathead minnows (Pimephales promelas), Ceriodaphnia dubia, and Daphnia magna. Environ Toxicol Chem 19:2918–2922Google Scholar
  38. Smith DG (2001) Pennak’s freshwater invertebrates of the United States, 4th edn. Wiley, New York, NY, pp 327–400Google Scholar
  39. Soucek DJ (2007) Comparison of hardness- and chloride-regulated acute effects of sodium sulfate on two freshwater crustaceans. Environ Toxicol Chem 26:773–779CrossRefGoogle Scholar
  40. Soucek DJ, Dickinson A, Koch B (2011) Acute and chronic toxicity of boron to various freshwater organisms. Environ Toxicol Chem 30:1906–1914CrossRefGoogle Scholar
  41. Stanley EH, Maxted JT (2008) Changes in the dissolved nitrogen pool across land cover gradients in Wisconsin streams. Ecol Appl 18:1579–1590CrossRefGoogle Scholar
  42. Sugano H, Hoshi T (1971) Purification and properties of blood hemoglobin from fresh-water cladocera, Moina macrocopa and Daphnia magna. Biochim Biophys Acta 229:349–358Google Scholar
  43. Thorp JH, Covich AP (2001) Introduction to the subphylum Crustacea. In: Thorp JH, Covich AP (eds) Ecology and classification of North American freshwater invertebrates, 2nd edn. Academic, San Diego, CA, pp 777–809Google Scholar
  44. Thurston RV, Russo RC, Smith CE (1978) Acute toxicity of ammonia and nitrite to cutthroat trout fry. Trans Am Fish Soc 107:361–368CrossRefGoogle Scholar
  45. Tucker CS, Schwedler TE (1983) Acclimation of channel catfish (Ictalurus punctatus) to nitrite. Bull Environ Contam Toxicol 30:516–521CrossRefGoogle Scholar
  46. United States Environmental Protection Agency (1971) Method 354.1: determination of nitrite by spectrophotometry. USEPA, Washington, DCGoogle Scholar
  47. United States Environmental Protection Agency (1978) Method 353.1: determination of nitrate-nitrite by colorimetry. USEPA, Washington, DCGoogle Scholar
  48. United States Environmental Protection Agency (2000) Methods for measuring the toxicity and bioaccumulation of sediment-associated contaminants with freshwater invertebrates, 2nd ed. EPA/600/R-99/064. USEPA, Washington, DCGoogle Scholar
  49. United States Environmental Protection Agency (2002) Short-term methods for estimating the chronic toxicity of effluents and receiving waters to freshwater organisms, 4th ed. EPA-821-R-02-013. USEPA, Washington, DCGoogle Scholar
  50. United States Environmental Protection Agency (2009) Draft 2009 update aquatic life. Ambient water quality criteria for ammonia—Freshwater. EPA-822-D-09-001. USEPA, Washington, DCGoogle Scholar
  51. Vitousek PM, Aber JD, Howarth RW, Likens GE, Matson PA, Schindler DW et al (1997) Human alteration of the global nitrogen cycle: sources and consequences. Ecol Appl 7:737–750Google Scholar
  52. Wang N, Ingersoll CG, Hardesty DK, Ivey CD, Kunz JL, May TW et al (2007a) Contaminant sensitivity of freshwater mussels: acute toxicity of copper, ammonia, and chlorine to glochidia and juveniles of freshwater mussels (Unionidae). Environ Toxicol Chem 26:2036–2047CrossRefGoogle Scholar
  53. Wang N, Ingersoll CG, Greer IE, Hardesty DK, Ivey CD, Kunz JL et al (2007b) Chronic toxicity of copper and ammonia to juvenile freshwater mussels (Unionidae). Environ Toxicol Chem 26:2048–2056CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Illinois Natural History SurveyUniversity of Illinois at Urbana-ChampaignChampaignUSA

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