ASTM E 2455:2022

1.1 This standard guide describes methods for conducting laboratory toxicity tests with early life stages of freshwater mussels including glochidia and juvenile mussels in water-only and effluent exposures ( Annex A1 ). Future revisions to this standard may describe methods for conducting toxicity tests with endpoints of reproduction, behaviors, and biomarkers. 1.2 Freshwater mussels (order Unionida) are one of the most imperiled groups of animals in the world, and environmental contamination has been linked as a contributing factor to the decline of mussel populations (Lydeard et al. 2004 ( 1 ) ; Strayer et al. 2004 ( 2 ) ; Haag 2012 ( 3 ) ; Lopes-Lima et al. 2017 ( 4 ) ). 2 Three critical life stages (glochidia, juvenile mussels, and adults) have been used in toxicity assessments and the toxicity studies are separated according to the medium of exposure (water, sediment, and host fish (Ingersoll et al. 2007 ( 5 ) ). Recent studies on early life stages of mussels have demonstrated that the mussels are among the most sensitive freshwater species to a variety of contaminants, including ammonia, some metals (for example, aluminum, copper, nickel, and zinc), and major ions (for example, chloride, nitrate, potassium, and sulfate) (Bringolf et al. 2007 ( 6 ) ; Newton et al. 2007 ( 7 ) ; Wang et al. 2007ab, 2010, 2011ab, 2016, 2017ab, 2018abc, 2020ab ( 8- 20 ) ; Cope et al. 2008 ( 21 ) ; Gillis et al. 2008, 2010, 2011, 2021 ( 22- 25 ) ; Miao et al. 2010 ( 26 ) ; Salerno et al. 2020 ( 27 ) ). These studies indicate that environmental guideline values for individual chemicals established for the protection of aquatic organisms may not be adequately protective of sensitive stages of freshwater mussels. For example, when freshwater mussel toxicity data were included in an update to the United States Environmental Protection Agency (USEPA) ambient water quality criteria (WQC) for ammonia, the acute criterion decreased by about a 1.4 fold and the chronic criterion decreased by 2.4 fold (USEPA 2013) ( 28 ) . 1.3 Summary of Life History of Freshwater Mussels: 1.3.1 Freshwater mussels are bivalve mollusks belonging to the taxonomic Order Unionida (section 10.1 ). Like most bivalves, mussels are totally aquatic, relatively sedentary, filter-feeding animals, and spend most of their lives partially or completely burrowed in the substrate of streams, rivers, or lakes. Freshwater mussels have an unusual and complex life cycle that includes a larval stage, the glochidium, that is briefly parasitic on fish ( Fig. 1 ). 1.6 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information only. 1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements are given in Section 7 . 1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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