National Toxicology Program

National Toxicology Program
http://ntp.niehs.nih.gov/go/iccvam-recs

Summaries of ICCVAM Test Method Recommendations

This page has summaries of all ICCVAM recommendations that have been formally transmitted to ICCVAM member agencies. Links lead to pages with more details about the test method evaluations and ICCVAM recommendations.

Select links to jump to test method area of interest:


Acute Systemic Toxicity

Revised Up-and-Down Procedure for Acute Oral Toxicity Testing

ICCVAM concluded that use of the revised up-and-down procedure (UDP) would reduce and refine animal use. Therefore, ICCVAM recommended that the revised UDP should: (1) replace the Organisation for Economic Cooperation and Development (OECD) UDP test guideline from 1998 and (2) be used instead of the conventional LD50 test to determine the acute oral toxicity hazard of chemicals.

In Vitro Cytotoxicity Test Methods for Estimating Starting Doses for Acute Oral Systemic Toxicity Tests

ICCVAM recommended that the in vitro basal cytotoxicity test methods tested in the joint ICCVAM/ECVAM validation study should be considered before using animals for acute oral systemic toxicity testing, and that the methods should be used where determined appropriate. Data from the test methods should be used in a weight-of-evidence approach for determining starting doses for in vivo studies. Using these in vitro methods where appropriate is expected to reduce the number of animals required for each toxicity test.


Endocrine Disruptors

Review of the Validation Status of In Vitro Endocrine Disruptor Screening Assays

ICCVAM conducted a comprehensive review of four types of in vitro test methods for detecting substances with potential endocrine disrupting activity, and concluded that there were no adequately validated in vitro endocrine disruptor screening assays. ICCVAM developed recommendations for minimum procedural standards and test methods for future validation of in vitro estrogen receptor and androgen receptor binding and transactivation assays.

Use of the BG1Luc Estrogen Receptor Tranactivation Test Method to Identify Substances With the Potential to Interact With the Estrogen Receptor

ICCVAM evaluated the BG1Luc estrogen receptor (ER) transactivation (TA) agonist and antagonist assays and recommended how they could be used to identify substances that induce or inhibit human ER activity in vitro. The U.S. Environmental Protection Agency (EPA) responded that they regard the BG1Luc ER TA test method as an alternative to the current ER TA test method currently used in their Endocrine Disruptor Screening Program.


Eye Corrosion and Irritation

In Vitro Test Methods for Detecting Ocular Corrosives and Severe Irritants

ICCVAM recommended that the bovine corneal opacity and permeability and isolated chicken eye test methods can be used in a tiered-testing strategy to identify substances with the potential to cause severe, permanent eye injury, with specific limitations for certain chemical classes and/or physical properties. Substances that test positive in these assays can be classified as ocular corrosives or severe irritants without further testing in animals. ICCVAM also recommended that these in vitro test methods should be considered before using animals for ocular testing and used when determined appropriate.

In Vitro Testing Strategy for Ocular Hazard Classification and Labeling of Antimicrobial Cleaning Products

ICCVAM conducted an evaluation of a proposed testing strategy that used three in vitro test methods to determine the EPA hazard category and labeling requirements for antimicrobial cleaning products. ICCVAM recommended further studies to characterize the usefulness and limitations of the proposed strategy.

Evaluation of the Low Volume Eye Test

ICCVAM recommended that the low volume eye test, an alternative approach to the rabbit eye test, should not be used for future regulatory testing due to performance issues.

In Vitro Test Methods Proposed for Identifying Eye Injury Hazard Potential of Chemicals and Products

ICCVAM recommended that the cytosensor microphysiometer (CM) test method can be used as a screening test to identify some types of water-soluble substances that may cause permanent or severe eye injuries. Substances that are appropriate for testing using the CM test method and yield positive results can be classified as having the potential to cause severe or permanent eye injuries without additional testing using animals. ICCVAM also recommended that the CM test method can be used, for a limited range of substances (water-soluble surfactants and surfactant-containing formulations such as cosmetics and personal care products), to determine that chemicals and products do not present sufficient potential to cause eye injuries to require eye hazard labeling.

Routine Use of Topical Anesthetics, Systemic Analgesics, and Humane Endpoints to Avoid or Minimize Pain and Distress in Ocular Safety Testing

ICCVAM recommends that alternative in vitro test methods should be considered and used where appropriate for eye safety testing. When it is determined necessary to use the rabbit eye test for regulatory safety assessments, ICCVAM recommends that pain management procedures should always be used. The report on the ICCVAM evaluation of the use of anesthetics, analgesics, and humane endpoints in ocular safety testing includes a test method protocol that describes how to use topical anesthetics and systemic analgesics prior to and after test article administration in order to avoid or minimize animal pain and distress. The report also identifies specific clinical signs and lesions that can be used as humane endpoints to allow the investigator to end a study early in order to avoid or minimize animal pain and distress.

Identifying Chemical Eye Hazards With Fewer Animals

ICCVAM recommended that using a classification criterion of one or more positive animals in a three-animal test to identify chemicals and products that are eye hazards will maintain hazard classification equivalent to that provided by current testing procedures (16 CFR 1500.42), while using up to 50% to 83% fewer animals. ICCVAM recommended consideration of the use of this classification criterion together with eye safety testing procedures that use a maximum of three animals per test substance.


Pyrogenicity

In Vitro Test Methods Proposed for Assessing Pyrogenicity of Pharmaceuticals and Other Products

ICCVAM evaluated the validation status of five in vitro test methods proposed for assessing the potential pyrogenicity (i.e., ability to induce fever) of pharmaceuticals and other products. ICCVAM recommended that, although none of these test methods can be considered a complete replacement for the rabbit pyrogen test for the detection of Gram-negative endotoxin, they can be considered for use to detect Gram-negative endotoxin in human parenteral drugs on a case-by-case basis, subject to validation for each specific product to demonstrate equivalence to the rabbit pyrogen test, in accordance with applicable U.S. Federal regulations. When used in this manner, these methods should be able to reduce the number of animals used for pyrogenicity testing.


Skin Corrosion and Irritation

Corrositex Assay for Identification of Substances With the Potential to Cause Severe, Permanent Skin Injury

ICCVAM recommended that for certain testing circumstances, such as testing performed to comply with the U.S. Department of Transportation regulations, Corrositex is useful as a stand-alone assay for evaluating the corrosivity or noncorrosivity of acids, bases, and acid derivatives. Used in this manner, Corrositex can replace the use of animals for corrosivity testing of qualified chemicals in some chemical classes.

EPISKIN™, EpiDerm™(EPI-200) and the Rat Skin Transcutaneous Electrical Resistance Assay for Identification of Substances With the Potential to Cause Severe, Permanent Skin Injury

ICCVAM recommended that these assays may be used for assessing the dermal corrosion potential of chemicals in a weight-of-evidence approach in an integrated testing scheme. In this approach, positive in vitro corrosivity responses do not generally require further testing and can be used for classification and labeling.

Performance Standards for In Vitro Test Methods for Skin Corrosion

ICCVAM established performance standards for in vitro test methods for skin corrosion based on four in vitro test methods evaluated by ICCVAM for the identification of substances with the potential to cause skin corrosion. These performance standards can be used to evaluate the reliability and accuracy of other test methods that are based on similar scientific principles and that measure or predict the same biological or toxic effect.


Skin Sensitization

Murine Local Lymph Node Assay for Assessing Allergic Contact Dermatitis Hazard Potential

ICCVAM recommended that the murine local lymph node assay (LLNA) is a valid alternative to currently accepted guinea pig test methods, and that the LLNA reduces the number of animals required for testing and eliminates animal pain and distress. Compared to guinea pig tests, the LLNA can also be completed in a shorter timeframe and provides dose-response information.

Reduced Murine Local Lymph Node Assay

ICCVAM concluded that the performance of the reduced LLNA (rLLNA) is sufficient to distinguish between skin sensitizers and nonsensitizers in cases that do not require dose-response information or if the test substance is expected to have no allergic contact dermatitis hazard potential. Therefore, ICCVAM recommended that the rLLNA test method be used routinely to determine the allergic contact dermatitis potential of chemicals and products before conducting the multidose LLNA.

Performance Standards for the Murine Local Lymph Node Assay

ICCVAM developed recommended performance standards for the LLNA that could be used to more rapidly and efficiently determine the validity of nonradioactive and other modified versions of the LLNA.

Evaluation of the Applicability Domain for the Murine Local Lymph Node Assay

ICCVAM recommended that the LLNA may be used to test any chemical or product for the potential to cause allergic contact dermatitis unless the chemical or product to be tested has properties that may interfere with the ability of the LLNA to detect sensitizing substances.

Nonradiolabeled Murine Local Lymph Node Assay Methods for Assessing Allergic Contact Dermatitis Hazard Potential

ICCVAM recommended that two modified versions of the LLNA that do not use radioactive isotopes can be used to identify substances as potential skin sensitizers or nonsensitizers, with certain limitations. The availability of LLNA methods that do not use radioactivity is expected to allow more institutions to take advantage of the animal welfare benefits of the LLNA and provides environmental benefits as well.

Use of the Murine Local Lymph Node Assay for Potency Categorization of Chemicals Causing Allergic Contact Dermatitis in Humans

ICCVAM recommended that the LLNA can be used to categorize substances as strong sensitizers (Globally Harmonized System of Classification and Labelling of Chemicals Subcategory 1A). However, substances that are not identified as strong sensitizers using the LLNA require additional information to categorize them as other than strong sensitizers (Globally Harmonized System of Classification and Labelling of Chemicals Subcategory 1B).

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