Abstract
New explosive formulations are being developed to be less sensitive to impact and inadvertent explosion, increasing safety for the warfighter. Since testing and training make environmental releases imminent, the toxicity of 3-nitro-1,2,4-triazol-5-one (NTO), a component of Insensitive Munitions eXplosive (IMX) formulations, was assessed in a one-generation study to the northern leopard frog (Lithobates ( = Rana) pipiens). Because NTO in water creates acidic conditions, acute studies were conducted with non-pH-adjusted NTO, while a long-term (70-d) study was conducted with neutralized NTO. In the acute study, 48-h and 7-d LC50s were ~250 mg NTO/L. In the long-term study, tadpoles were dead by day 2 in 11,350 mg/L NTO, and by day 63 in 8382 mg/L. The 70-d LC50 was 3670 mg (neutralized) NTO/L. The number of organisms reaching complete metamorphosis was reduced by NTO; the lowest IC25 was 1999 mg NTO/L for the Number Completing Metamorphosis. The NOECs for Time to Front Limb Eruption or Time to Metamorphosis were the same at 1346 mg/L. Histopathology did not significantly distinguish between NTO-exposed and unexposed animals, although possible effects on the density of spermatogonia in NTO-exposed males was suggested. The test data indicate that acute toxicity to ambient NTO can be attributed primarily to its acidic nature; relatively low chronic toxicity of neutralized NTO is due to delays in metamorphosis. The consequence from this latter observation may be ecologically significant as delays of even a few days could increase mortality through predation and/or loss of the aquatic medium in temporary water bodies.
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References
Analytical Software (2008) Statistix 9. Analytical Software, Tallahassee, FL
Barrie A (2013) New generation of explosives for the army. Military Daily News, 1 April 2013. http://www.military.com/daily-news/2013/04/01/new-generation-of-explosives-for-the-army.html. Accessed 9 June 2017
Chai L, Wang H, Zhao H, Deng H (2016) Chronic effects of triclosan on embryonic development of Chinese toad, Bufo gargarizans. Ecotoxicology 25:1600–1608
Crouse LCB, Lent EM, Leach GJ (2015) Oral toxicity of 3-Nitro-1,2,4-triazol-5-one in rats. Int J Toxicol 34(1):55–66
DeBoer JA, Webber CM, Dixon TA, Pope KL (2016) The influence of severe reservoir drawdown on springtime zooplankton and larval fish assemblages in Red Willow Reservoir, Nebraska. J Freshw Ecol 31:131–146
Freda J, Dunson WA (1984) Sodium balance of amphibian larvae exposed to low environmental pH. Physiol Zool 57:435–443
Gosner KL (1960) A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 16:183–190
Gray N (2008) Insensitive munitions – new explosives on the horizon. Army Acquisition Logistics Tech Magazine Jan-Mar: 34-35. http://asc.army.mil/docs/pubs/alt/archives/2008/Jan-Mar-2008.pdf. Accessed 9 June 2017
Haley MV, Kuperman RG, Checkai RT (2009) Aquatic toxicity of 3-nitro-1,2,4-triazol-5-one. Edgewood Chemical Biological Center, U.S. Army Research Development and Engineering Command, Report ECBC-TR-726, Aberdeen Proving Ground, MD
Hayes TB, Falso P, Gallipeau S, Stice M (2010) The cause of global amphibian declines: a developmental endocrinologist’s perspective. J Exp Biol 213(6):921–933
Horne MT, Dunson WA (1995) Effects of low pH, metals, and water hardness on larval amphibians. Arch Environ Contam Toxicol 29:500–505
James SM, Little EE, Semlitsch RD (2005) Metamorphosis of two amphibian species after chronic cadmium exposure in outdoor aquatic mesocosms. Environ Toxicol Chem 24:1994–2001
Johnson MS, Salice CJ (2009) Toxicity of energetic compounds to wildlife species. In: Sunahara GI, Lotufo G, Kuperman RG, Hawari J (eds) Ecotoxicology of explosives. CRC Press, Boca Raton, FL, p 157–175
Kerby JL, Richards-Hrdlicka KL, Storfer A, Skelly DK (2010) An examination of amphibian sensitivity to environmental contaminants: are amphibians poor canaries? Ecology Lett 13:60–67
Lotufo GR, Biedenbach JM, Sims JG, Chappell P, Stanley JK, Gust KA (2015) Bioaccumulation kinetics of the conventional energetics TNT and RDX relative to insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles. Environ Toxicol Chem 34(4):880–886
Nipper M, Carr RS, Lotufo GR (2009) Aquatic toxicology of explosives. In: Sunahara GI, Lotufo G, Kuperman RG, Hawari J (eds) Ecotoxicology of explosives. CRC Press, Boca Raton, FL, p 77–115
Norberg-King T (1993) A linear interpolation method for sublethal toxicity: the Inhibition Concentration (ICp) approach. Version 2.0. National Effluent Toxicity Assessment Center, United States Environmental Protection Agency, Duluth, MN
Ortiz-Santaliestra ME, Sparling DW (2007) Alteration of larval development and metamorphosis by nitrate and perchlorate in southern leopard frogs (Rana sphenocephala). Arch Environ Contam Toxicol 53:639–646
Paden NE, Smith EE, Maul JD, Kendall RJ (2011) Effects of chronic 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, and 2,6-dinitrotoluene exposure on developing bullfrog (Rana catesbeiana) tadpoles. Ecotoxicol Environ Safety 74:924–928
Reddy G, Song J, Kirby P, Lent EM, Crouse LCB, Johnson MS (2011) Genotoxicity assessment of an energetic propellant compound, 3-nitro-1,2,4-triazol-5-one (NTO). Mutat Res 719:35–40
Rosenshield ML, Jofré MB, Karasov WH (1999) Effects of polychlorinated biphenyl 126 on green frog (Rana clamitans) and leopard frog (Rana pipiens) hatching success, development, and metamorphosis. Environ Toxicol Chem 18:2478–2486
Rumrill CT, Scott DE, Lance SL (2016) Effects of metal and predator stressors in larval southern toads (Anaxyrus terrestris). Ecotoxicology 25:1278–1286
Russell AL, Seiter JM, Coleman JG, Winstead B, Bednar AJ (2014) Analysis of munitions constituents in IMX formulations by HPLC and HPLC-MAS. Talanta 128:524–530
Semlitsch RD (1987) Relationship of pond drying to the reproductive success of the salamander Ambystoma talpoideum. Copeia 1987(1):61–69
Stanley JK, Lutufo GR, Biedenback JM, Chappell P, Gust KA (2015) Toxicity of conventional energetics TNT and RDX relative to new insensitive munitions constituents DNAN and NTO in Rana pipiens tadpoles. Environ Toxicol Chem 34(4):873–879
Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues ASL, Fischman DL, Waller RW (2004) Status and trends of amphibian declines and extinctions worldwide. Science 306:1783–1786
Theodrakis CW, Rinchard J, Carr JA, Park JW, McDaniel L, Liu FJ, Wages M (2006) Thyroid endocrine disruption in stonerollers and cricket frogs from perchlorate-contaminated streams in east-central Texas. Ecotoxicology 15:31–50
Tidepool Scientific (2014) Comprehensive Environmental Toxicity Information System (CETIS); User Guide (version 1.8.7). Tidepool Scientific, LLC, McKinleyville, CA
USEPA (2002a) Short-term methods for estimating the chronic toxicity of effluents and receiving waters to freshwater organisms. 4th edn, United States Environmental Protection Agency, Washington D.C., USA
USEPA (2002b) Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms. 5th edn, United States Environmental Protection Agency, Washington D.C., USA
Werner EE (1986) Amphibian metamorphosis: Growth rate, predation risk, and the optimal size at transformation. Amer Nat 128(3):319–341
Zohary T, Ostrovsky I (2011) Ecological impacts of excessive water level fluctuations in stratified freshwater lakes. Inland Waters 1:47–59
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This research was supported by the Strategic Environmental Research and Development Program (SERDP), Contract ER-2223, Development of Environmental Health Criteria for Insensitive Munitions.
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Pillard, D.A., Eck, W.S., Johnson, M.S. et al. Effects of 3-Nitro-1,2,4-triazol-5-one on Survival, Growth and Metamorphosis in the Northern Leopard Frog, Lithobates pipiens . Ecotoxicology 26, 1170–1180 (2017). https://doi.org/10.1007/s10646-017-1842-z
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DOI: https://doi.org/10.1007/s10646-017-1842-z