Standard Guide for Acute Toxicity Test with the Rotifer Brachionus

SIGNIFICANCE AND USE
5.1 An important goal of aquatic toxicology is to determine the effects of toxic compounds on species that play a central role in aquatic communities. Rotifers have a major impact on several important ecological processes in freshwater and coastal marine environments. As filter-feeders on phytoplankton and bacteria, rotifers exert substantial grazing pressure that at times exceeds that of the larger crustacean zooplankton  (1, 2).4 Rotifer grazing on phytoplankton is highly selective (2-4) and can influence phytoplankton composition, the coexistence of competitors, and overall water quality (5). The contribution of rotifers to the secondary production of many aquatic communities is substantial (6-9). In fresh water, rotifers often account for the major fraction of zooplankton biomass at certain times of the year (10, 11) . Rotifers and other zooplankton are a significant food source for many larval fish, planktivorous adult fish (12, 13), and several invertebrate predators (14-16). The high metabolic rates of rotifers contribute to their role in nutrient cycling, which might make rotifers more important than crustaceans in certain communities (17, 18).  
5.2 In addition to their important ecological role in aquatic communities, rotifers are attractive organisms for toxicological studies because an extensive database exists on the basic biology of this group. Techniques have been published for the culture of many rotifer species (3, 19). The rotifer life cycle is well defined (20, 21), and the factors regulating it are reasonably well understood (22-25). Several aspects of rotifer behavior have been examined closely (26-29). The biogeography of many rotifer species has been characterized (30, 31), and the systematics of the group are well described (32, 33).  
5.3 Toxicity tests with rotifers of the genus Brachionus are more easily performed than with many other aquatic animals because of their rapid reproduction, short generation times, sensitivity (34), and the c...
SCOPE
1.1 This guide describes procedures for obtaining laboratory data concerning the acute toxicity of chemicals and aqueous effluents released into fresh, estuarine or marine waters. Acute toxicity is measured by exposing  Brachionus  newly hatched from cysts to a series of toxicant concentrations under controlled conditions. This guide describes a test for using  B. calyciflorus, a freshwater rotifer, and the Appendix describes modifications of this test for estuarine and marine waters using  B. plicatilis. These procedures lead to an estimation of acute toxicity, including the concentration expected to kill 50 % of the test rotifers (LC50) in 24 h. Procedures not specifically stated in this guide should be conducted in accordance with Guide E729 and Guide E1192.  
1.2 Modifications of these procedures might be justified by special needs or circumstances. Although using appropriate procedures is more important than following prescribed procedures, the results of tests conducted using modified procedures might not be comparable to rotifer acute tests that follow the protocol described here. Comparison of the results using modified procedures might provide useful information concerning new concepts and procedures for conducting acute toxicity tests on chemicals and aqueous effluents.  
1.3 This guide is organized as follows:    
Section    
Scope  
1    
Referenced Documents  
2    
Terminology  
3    
Summary of Guide  
4    
Significance and Use  
5    
Apparatus  
6    
Dilution Water  
7    
Hazards  
8    
Test Material  
9    
Test Organisms  
10    
Test Procedure  
11    
Calculation of Results  
12    
Acceptability of the Test  
13    
Report  
14    
Keywords  
15  
1.4 These procedures are applicable to most chemicals, either individually or in formulations, commercial products, or mixtures. This guide can also be used to investigate the effects on rotifer su...

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ASTM E1440-23 - Standard Guide for Acute Toxicity Test with the Rotifer Brachionus
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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.
Designation: E1440 − 23
Standard Guide for
1
Acute Toxicity Test with the Rotifer Brachionus
This standard is issued under the fixed designation E1440; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
Calculation of Results 12
Acceptability of the Test 13
1.1 This guide describes procedures for obtaining laboratory
Report 14
data concerning the acute toxicity of chemicals and aqueous Keywords 15
effluents released into fresh, estuarine or marine waters. Acute
1.4 These procedures are applicable to most chemicals,
toxicity is measured by exposing Brachionus newly hatched
either individually or in formulations, commercial products, or
from cysts to a series of toxicant concentrations under con-
mixtures. This guide can also be used to investigate the effects
trolled conditions. This guide describes a test for using B.
on rotifer survival of pH, hardness, and salinity and on
calyciflorus, a freshwater rotifer, and the Appendix describes
materials such as aqueous effluents, leachates, oils, particulate
modifications of this test for estuarine and marine waters using
matter, sediments, and surface waters. This guide might not be
B. plicatilis. These procedures lead to an estimation of acute
appropriate for materials with high oxygen demand, with high
toxicity, including the concentration expected to kill 50 % of
volatility, subject to rapid biological or chemical transforma-
the test rotifers (LC50) in 24 h. Procedures not specifically
tion or those readily sorb to test chambers.
stated in this guide should be conducted in accordance with
1.5 This standard does not purport to address all of the
Guide E729 and Guide E1192.
safety concerns, if any, associated with its use. It is the
1.2 Modifications of these procedures might be justified by
responsibility of the user of this standard to establish appro-
special needs or circumstances. Although using appropriate
priate safety, health, and environmental practices and deter-
procedures is more important than following prescribed
mine the applicability of regulatory limitations prior to use.
procedures, the results of tests conducted using modified
For specific hazards statements, see Section 8.
procedures might not be comparable to rotifer acute tests that
1.6 This international standard was developed in accor-
follow the protocol described here. Comparison of the results
dance with internationally recognized principles on standard-
using modified procedures might provide useful information
ization established in the Decision on Principles for the
concerning new concepts and procedures for conducting acute
Development of International Standards, Guides and Recom-
toxicity tests on chemicals and aqueous effluents.
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
1.3 This guide is organized as follows:
Section
2. Referenced Documents
2
Scope 1
2.1 ASTM Standards:
Referenced Documents 2
D1129 Terminology Relating to Water
Terminology 3
Summary of Guide 4
D4447 Guide for Disposal of Laboratory Chemicals and
Significance and Use 5
Samples
Apparatus 6
E729 Guide for Conducting Acute Toxicity Tests on Test
Dilution Water 7
Hazards 8
Materials with Fishes, Macroinvertebrates, and Amphib-
Test Material 9
ians
Test Organisms 10
E943 Terminology Relating to Biological Effects and Envi-
Test Procedure 11
ronmental Fate
E1192 Guide for Conducting Acute Toxicity Tests on Aque-
ous Ambient Samples and Effluents with Fishes,
Macroinvertebrates, and Amphibians
1
This guide is under the jurisdiction of ASTM Committee E50 on Environmental
Assessment, Risk Management and Corrective Action and is the direct responsibil-
ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.
2
Current edition approved Sept. 1, 2023. Published October 2023. Originally For referenced ASTM standards, visit the ASTM website, www.astm.org, or
approved in 1991. Last previous edition approved in 2012 as E1440 – 91 (2012) contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
which was withdrawn January 2021 and reinstated in September 2023. DOI: Standards volume information, refer to the standard’s Document Summary page on
10.1520/E1440-23. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

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E1733 Guide for Use of Li
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