National Toxicology Program

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

Project Milestones

 

This page summarizes the status of ongoing and completed ICCVAM alternative test method evaluation projects, as well as projects to which NICEATM, ICCVAM, and ICCVAM agency scientists are contributing.

Alternative test methods:

  • Replace animals with non-animal systems or one animal species with a less highly developed one (for example, replacing a mouse with a fish)
  • Reduce the number of animals used to the minimum number required to obtain scientifically valid data
  • Refine procedures to lessen or eliminate animal pain and distress

 

Acute Oral Systemic Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Up-and-Down Procedure (oral) 2001 2001, revised 2008 (OECD TG 425)
Fixed Dose Procedure (oral) 2001 2002 (OECD TG 420)
Acute Toxic Class Method (oral) 2001 2002 (OECD TG 423)
In Vitro Cytotoxicity Test Methods: 3T3 Cells 2008 2010 (OECD GD 129)
In Vitro Cytotoxicity Test Methods: NHK Cells 2008 2010 (OECD GD 129)

Acute Inhalation Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Acute Toxic Class Method (inhalation) (Not applicable - adopted via OECD Test Guideline) 2009 2009 (OECD TG 436)
Fixed Concentration Procedure (inhalation) In progress (OECD) (Not applicable - to be adopted via OECD Test Guideline)   Anticipated 2016 (OECD)

Biologics and Vaccines

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Use of Humane Endpoints in Animal Testing of Veterinary Products 2002 1986 (European Union)
Use of Humane Endpoints in Animal Testing of Rabies Vaccines 2004 2002 (European Pharmacopoeia)

Dermal Absorption

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In Vitro Dermal Absorption (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 428)

Dermal Corrosivity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Corrositex® Assay 2000 2006 (OECD TG 435)
EpiDermTM Assay 2004 2004 (OECD TG 431)
EPISKINTM Assay 2004 2004 (OECD TG 431)
Rat Transcutaneal Electrical Resistance Assay 2004 2004 (OECD TG 430)
SkinEthic RHE Assay 2004 2004 (OECD TG 431)
LabCyte EPI-MODEL24 Assay In progress      

Dermal Irritation

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
EpiDermTM Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
EPISKINTM Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
SkinEthic RHE Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
LabCyte EPI-MODEL24 Assay In progress      

Dermal Phototoxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
3T3 NRU Phototoxicity Test (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 432)
3T3 NRU Phototoxicity Test: Application to UV Filter Chemicals (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 432)

Developmental Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Frog Embryo Teratogenesis Assay: Xenopus (FETAX) ICCVAM evaluation in 2000 found that FETAX was not sufficiently reliable for regulatory use and provided recommendations for improving accuracy and reliability

Endocrine Disruptor

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In vitro Androgen Receptor (AR) Binding ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro AR Transcriptional Activation (TA) ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro Estrogen Receptor (ER) Binding ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro ER TA ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
BG1Luc ER TA Assay Evaluation 2012 (OECD TG 457)
MCF-7 Cell Proliferation Assay Evaluation Results of NICEATM validation study indicated that the MCF-7 cell proliferation assay protocols require additional development to improve interlaboratory reproducibility

Genetic Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In Vitro Micronucleus Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 487)
In Vivo Comet Assay (Not applicable - adopted via OECD Test Guideline) 2014 2014 (OECD TG 489)
In Vitro Comet Assay In progress (JaCVAM)        
In Vitro Syrian Hamster Embryo Cell Transformation Assays (Not applicable - to be adopted via OECD Guidance Document)   Anticipated 2015 (OECD)
BHAS Cell Transformation Assay (Not applicable - to be adopted via OECD Test Guideline)   Anticipated 2015 (OECD)

Immunotoxicity: Allergic Contact Dermatitis

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Murine Local Lymph Node Assay (LLNA) 1999 2002 (OECD TG 429)
Updated LLNA Protocol (20% animal reduction) 2010 2010 (Updated OECD TG 429)
Reduced LLNA Test Method 2010 2010 (Updated OECD TG 429)
Development of LLNA Performance Standards 2010 2010 (Updated OECD TG 429)
Nonradioactive LLNA Method: LLNA:DA 2011 2010 (OECD TG 442A)
Nonradioactive LLNA Method: LLNA:BrdU-ELISA 2011 2010 (OECD TG 442B)
Nonradioactive LLNA Method: LLNA:BrdU-Flow Cytometry Interlaboratory validation study required        
Use of the LLNA for Testing Pesticide Formulations, Metals, Substances in Aqueous Solutions, and Other Products 2011 2010 (Updated OECD TG 429)
Use of the LLNA for Skin Sensitization Potency Categorization 2009 (UN GHS)
Direct Peptide Reactivity Assay In progress (OECD)      
Human Cell Line Activation Test In progress (OECD)      
Myeloid U937 Skin Sensitization Test EURL ECVAM review found that additional development was required to improve reproducibility.

Metabolism and Toxicokinetics

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Provision of a Standard for Human Hepatic Metabolism and Toxicity by Assessing as an Indicator Biotransformation Enzyme Induction Using HepaRG® Cells and Cryopreserved Human Hepatocytes In progress (OECD) (Not applicable - to be adopted via OECD Test Guideline)   Anticipated 2015 (OECD)

Ocular Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Bovine Corneal Opacity and Permeability (BCOP) - Ocular Corrosivity/Severe Irritation 2008 2009 (OECD TG 437)
Isolated Chicken Eye (ICE) - Ocular Corrosivity/Severe Irritation 2008 2009 (OECD TG 438)
Hen's Egg Test/Choriallantoic Membrane (HET-CAM) - Ocular Corrosivity/Severe Irritation ICCVAM review in 2007 found that this test method was not sufficiently accurate for regulatory use and recommended additional studies
Isolated Rabbit Eye (IRE) - Ocular Corrosivity/Severe Irritation ICCVAM review in 2007 found that this test method was not sufficiently accurate for regulatory use and recommended additional studies
Integrated Non-animal Testing Strategy for Eye Irritation Potential of Antimicrobial Cleaning Products ICCVAM review in 2009 concluded that data on the testing strategy were not sufficient to demonstrate its utility for the proposed application and recommended additional studies
BCOP - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
HET-CAM - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
ICE - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
IRE - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
Cytosensor Microphysiometer Test Method 2011 In progress (OECD)
Routine use of topical anesthestics, systemic analgesics, and humane endpoints in in vivo testing 2011 2012 (Updated OECD TG 405)
In vivo low volume eye test ICCVAM reviewed the LVET in 2009 and does not recommend its use for prospective ocular safety testing
Fluorescein leakage test method - ocular corrosivity/severe irritation (Not applicable - adopted via OECD Test Guideline) 2012 2012 (OECD TG 460)
Neutral red release test method ICCVAM review in 2009 found that this test method was not adequately validated for regulatory use
Red blood cell haemolysis test method ICCVAM review in 2009 found that this test method was not adequately validated for regulatory use

Pyrogenicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
The Human Whole Blood/IL-1 In Vitro Pyrogen Test 2009 2010 (European Pharmacopoeia)
The Human Whole Blood/ IL-1 In Vitro Pyrogen Test Using Cryopreserved Human Whole Blood 2009 2010 (European Pharmacopoeia)
The Human Whole Blood/IL-6 In Vitro Pyrogen Test (WB/IL-6) 2009 2010 (European Pharmacopoeia)
In Vitro Pyrogen Test Using Human Peripheral Blood Mononuclear Cells (PBMC/IL-6) 2009 2010 (European Pharmacopoeia)
An Alternative In Vitro Pyrogen Test Using the Human Monocytoid Cell Line MONO MAC 6 (MM6/IL-6) 2009 2010 (European Pharmacopoeia)

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Project Details and Links

Oral Up-And-Down Procedure

Acute oral systemic toxicity testing identifies substances that are poisonous when ingested so that they may be appropriately labeled and packaged. The up-and-down procedure reduces animal use for this purpose by up to 70%.

Milestone Date Document Link or
More Information
Nomination by the U.S. Environmental Protection Agency (EPA) August 1999 -
Peer Review Panel Meeting July 2000 View webpage
Peer Review Panel Follow-up Teleconference August 2001 View webpage
Publication of OECD Test Guideline 425 October 2001 -
Test Method Evaluation Report Published November 2001 View webpage
Recommendations Made to Federal Agencies March 2003 View webpage
Federal Agency Responses Received December 2003 View webpage
Publication of Updated OECD Test Guideline 425 October 2008 PDF

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Fixed Dose Procedure (oral)

Acute oral systemic toxicity testing identifies substances that are poisonous when ingested so that they may be appropriately labeled and packaged. The fixed dose procedure reduces animal use for this purpose by up to 70%.

Milestone Date Document Link or
More Information
Publication of OECD Test Guideline 420 2001 PDF

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Acute Toxic Class Method (oral) 

Acute oral systemic toxicity testing identifies substances that are poisonous when ingested so that they may be appropriately labeled and packaged. The acute toxic class method reduces animal use for this purpose by up to 70%.

Milestone Date Document Link or
More Information
Publication of OECD Test Guideline 423 2001 PDF

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In Vitro Cytotoxicity Test Methods 

Acute oral systemic toxicity testing identifies substances that are poisonous when ingested so that they may be appropriately labeled and packaged. In vitro cytotoxicity test methods replace animal use for initial dose setting for these tests.

Milestone Date Document Link or
More Information
EPA Nomination August 1999 -
Workshop Held October 2000 View webpage
Guidance Document Published August 2001 PDF
Workshop Report Published August 2001 PDF
Recommendations Made to Federal Agencies March 2003 View webpage
Federal Agency Responses Received December 2003 View webpage
Peer Review Panel Meeting May 2006 View webpage
Peer Review Panel Report Published June 2006 PDF
Background Review Document Published November 2006 View webpage
Test Method Evaluation Report Published November 2006 View webpage
Recommendations Made to Federal Agencies March 2008 View webpage
Federal Agency Responses Received September 2008 View webpage
OECD Guidance Document 129 Published July 2010 PDF

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Acute Toxic Class Method (inhalation)

Acute inhalation systemic toxicity testing identifies substances that are poisonous when inhaled so that they may be appropriately labeled and packaged. The acute toxic class method reduces animal use for this purpose.

Milestone Date Document Link or
More Information
Publication of OECD Test Guideline 436 September 2009 PDF

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Humane Endpoints in Animal Testing of Veterinary Products and Rabies Vaccines 

Animal testing of biological products and rabies vaccines is performed to ensure that each lot of these products is safe and effective. A USDA directive refines animal use for this purpose.

Milestone Date Document Link or
More Information
USDA Directive Published April 2004 PDF

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In Vitro Method for Measuring Skin Absorption 

Dermal toxicity testing identifies substances that may be poisonous when absorbed through the skin so that they may be appropriately labeled and packaged. Use of this test method may reduce animal use for this purpose.

Milestone Date Document Link or
More Information
OECD Test Guideline 428 Issued April 2004 PDF

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Corrositex® for Identification of Dermal Corrosives 

Dermal corrosivity testing identifies substances that cause chemical burns to the skin so that they may be appropriately labeled and packaged. Corrositex® reduces and refines animal use for this purpose.

Milestone Date Document Link or
More Information
Corrositex® Submission May 1998 -
Peer Review Panel Meeting January 1999 View webpage
Peer Review Panel Report Published June 1999 PDF
Recommendations Made to Federal Agencies June 1999 -
Federal Agency Responses Received October 1999 View webpage
Publication of OECD Test Guideline 435 July 2006 PDF

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In Vitro Test Methods for Identification of Dermal Corrosives  

Dermal corrosivity testing identifies substances that cause chemical burns to the skin so that they may be appropriately labeled and packaged. Use of EpiSkin™, EPIDERM™ and the rat skin transcutaneous electrical resistance assay tests reduces and refines animal use for this purpose.

Milestone Date Document Link or
More Information
Background Review Document Published August 2001 PDF
Test Method Evaluation Report Published June 2002 View webpage
Recommended Performance Standards Published May 2004 PDF
Publication of OECD Test Guidelines 430 and 431 July 2006 -
Updates submitted to OECD Test Guidelines 430 and 431 July 2009 -
Publication of Updated OECD Test Guidelines 430 and 431 July 2013 TG 430 - PDF
TG 431 - PDF

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In Vitro Test Methods for Identification of Dermal Irritants 

Dermal irritant testing identifies substances that irritate the skin so that they may be appropriately labeled and packaged. Use of EpiSkin™, EPIDERM™ and the SkinEthic RHE assay reduces animal use for this purpose.

Milestone Date Document Link or
More Information
Adoption of OECD Test Guideline 439 July 2010 -
Update of OECD Test Guideline 439 July 2013 PDF

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In Vitro 3T3 NRU Phototoxicity Test 

Dermal phototoxicity testing identifies substances that may increase skin sensitivity to light so that they may be appropriately labeled and packaged. Use of the in vitro 3T3 NRU phototoxicity test will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
OECD Test Guideline 432 Issued April 2004 PDF

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Frog Embryo Teratogenesis Assay: Xenopus (FETAX) 

Developmental toxicity testing identifies substances that may cause birth defects so that they may be appropriately labeled. Use of FETAX could reduce or replace animal use for this purpose.

Milestone Date Document Link or
More Information
EPA Nomination May 1998 -
Background Review Document Published March 2000 View webpage
Expert Panel Meeting May 2000  PDF

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In Vitro ER and AR Binding and TA Assays 

Endocrine disruptor testing identifies substances that may interfere with normal human and animal development. Use of in vitro tests could reduce animal use for this purpose in the EPA Endocrine Disruptor Screening Program (EDSP).

Milestone Date Document Link or
More Information
EPA Nomination April 2000 -
Expert Panel Meeting May 2002 View webpage
Expert Panel Report September 2002 PDF
Background Review Documents Published October 2002 View webpage
Test Method Evaluation Report Published May 2003  View webpage
Addendum to Test Method Evaluation Report Published September 2006  PDF

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BG1Luc ER TA Test Method 

Endocrine disruptor testing identifies substances that may interfere with normal human and animal development. Use of the BG1Luc ER TA test method could reduce animal use for this purpose in the EPA EDSP.

Milestone Date Document Link or
More Information
BG1Luc ER TA Test Method Nomination January 2004 PDF
Draft Pre-Screen Evaluation Released August 2004 PDF
Validation Study Initiated November 2007 View webpage
Peer Review Panel Meeting March 2011 -
ICCVAM Recommendations on the Use of the BG1Luc ER TA Test Method Transmitted to U.S. Federal Agencies February 2012 View recommendations
ICCVAM Recommendations on the Use of the BG1Luc ER TA Test Method Accepted by U.S. Federal Agencies August 2012 View recommendations and agency responses
Test Guideline 457 Adopted by OECD October 2012 PDF

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CertiChem Inc. MCF-7 Cell Proliferation Assay 

Endocrine disruptor testing identifies substances that may interfere with normal human and animal development. Use of the MCF-7 Cell Proliferation Assay could reduce animal use for this purpose in the EPA EDSP.

Milestone Date Document Link or
More Information
MCF-7 Cell Proliferation Assay Nomination June 2004 PDF
Pre-Screen Evaluation Released October 2006 PDF
Validation Study Initiated June 2010 View webpage
Validation Study Completed 2011 -
Draft Validation Study Report June 2012 PDF

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In Vitro Mammalian Cell Micronucleus Test

Genetic toxicity testing identifies substances that may cause DNA damage and increase risk of cancer or birth defects. Use of the in vitro mammalian cell micronucleus test could reduce animal use for this purpose.

Milestone Date Document Link or
More Information
Test Guideline 487 Adopted by OECD July 2010 PDF

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Murine Local Lymph Node Assay (LLNA) 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. Use of the LLNA reduces and refines animal use for this purpose.

Milestone Date Document Link or
More Information
Nomination by Sponsors January 1998 -
Peer Review Panel Meeting September 1998 View webpage
Peer Review Panel Report February 1999 PDF
Recommendations Made to Federal Agencies February 1999 View webpage
Adoption of OECD Test Guideline 429 April 2002 PDF
Update to OECD Test Guideline 429 Adopted July 2010 PDF

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Updated Protocol for the LLNA 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. The ICCVAM-recommended updated protocol will reduce the number of animals used by 20% when the LLNA is used for this purpose.

Milestone Date Document Link or
More Information
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
Recommendations on ICCVAM-recommended Performance Standards Transmitted to U.S. Federal Agencies; Revised Protocol is Appendix A of Performance Standards September 2009 View webpage
Recommmendations to U.S. Federal Agencies Accepted March 2010 View webpage
Update to OECD Test Guideline 429 Adopted July 2010 PDF

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Performance Standards for the LLNA 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. Accepted performance standards will enable the development of new versions of the LLNA for this purpose.

Milestone Date Document Link or
More Information
Draft Performance Standards Published September 2007 -
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
Recommendations on ICCVAM-recommended Performance Standards Transmitted to U.S. Federal Agencies September 2009 View webpage
Recommmendations to U.S. Federal Agencies Accepted March 2010 View webpage
Update to OECD Test Guideline 429 Adopted July 2010 PDF

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Reduced Murine Local Lymph Node Assay (rLLNA) 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. The reduced LLNA will reduce animal use for this purpose.

Milestone Date Document Link or
More Information
CPSC Nomination January 2007 PDF
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
ICCVAM Recommendations on rLLNA Transmitted to U.S. Federal Agencies September 2009 View webpage
Recommmendations to U.S. Federal Agencies Accepted March 2010 View webpage
Update to OECD Test Guideline 429 Adopted (containing rLLNA procedure) July 2010 PDF

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Nonradioactive Versions of the LLNA 

  • LLNA:DA
  • LLNA:BrdU-ELISA
  • LLNA:BrdU-Flow Cytometry

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. Nonradioactive version of the LLNA will reduce and refine animal use for this purpose by enabling more widespread use of the LLNA.

Milestone Date Document Link or
More Information
CPSC Nomination January 2007 PDF
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
Revised Background Review Documents and ICCVAM Recommendations Published March 2009 -
Second Peer Review Panel Meeting April 2009 -
Second Peer Review Panel Report Published June 2009 PDF
ICCVAM Recommendations on Nonradioactive LLNA: DA and LLNA: BrdU-ELISA Methods Transmitted to U.S. Federal Agencies June 2010 -
New OECD Test Guidelines for the LLNA: DA and LLNA: BrdU-ELISA Methods Adopted July 2010 PDF: OECD TG 442A (DA)
PDF: OECD TG 442B (ELISA)
Federal Agency Responses Received February 2011 View webpage

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Evaluation of the LLNA for Testing Pesticide Formulations, Metals, Substances in Aqueous Solutions, and Other Products 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. Increasing the number of substance types that can be tested using the LLNA will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
CPSC Nomination January 2007 PDF
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
Revised Background Review Documents and ICCVAM Recommendations Published March 2009 -
Second Peer Review Panel Meeting April 2009 -
Second Peer Review Panel Report Published June 2009 PDF
ICCVAM Recommendations on the LLNA Applicability Domain Transmitted to U.S. Federal Agencies June 2010 -
Update to OECD Test Guideline 429 Adopted July 2010 PDF
Federal Agency Responses Received February 2011 View webpage

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Evaluation of the LLNA for Skin Sensitization Potency Categorization 

Skin sensitization testing identifies substances that may act as sensitizers and cause the development of allergic contact dermatitis. Use of the LLNA to identify "strong sensitizers" will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
CPSC Nomination January 2007 PDF
Peer Review Panel Meeting March 2008 -
Peer Review Panel Report Published May 2008 PDF
ICCVAM Recommendations on the Use of the LLNA for Skin Sensitization Potency Categorization Transmitted to U.S. Federal Agencies June 2011 View webpage
Federal Agency Responses Received February 2012 View webpage

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In Vitro Ocular Test Methods for Identification of Eye Corrosives and Severe Irritants 

  • Bovine corneal opacity and permeability (BCOP)
  • Hen's eggs test - chorioallantic membrane (HET-CAM)
  • Isolated chicken eye (ICE)
  • Isolated rabbit eye (IRE)

Ocular toxicity testing identifies substances that may cause permanent or temporary blindness. Use of these in vitro test methods will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
EPA Nomination October 2003 -
Expert Panel Meeting January 2005 View webpage
Expert Panel Report March 2005 PDF
Expert Panel Teleconference September 2005 View webpage
Expert Panel Report Addendum November 2005 PDF
Test Method Evaluation Report Published October 2007  View webpage
Recommendations Made to Federal Agencies October 2007  View webpage
Federal Agency Responses Received May 2008  View webpage
Adoption of OECD Test Guidelines 437 (BCOP) and 438 (ICE) September 2009  View webpage
Adoption of Guidance Document Supporting OECD Test Guidelines 437 and 438: Collection of Tissues for Histopathology Evalution and Data on Nonsevere Irritants April 2011 PDF

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Routine Use of Topical Anesthestics, Systemic Analgesics and Humane Endpoints in In Vivo Testing 

Ocular toxicity testing identifies substances that may cause irritation to the eye. Routine use of topical anesthestics, systemic analgesics and humane endpoints will refine animal use for this purpose.

Milestone Date Document Link or
More Information
Peer Review Panel Meeting May 2009 -
Peer Review Panel Report Published July 2009 Read report
Recommendations Made to U.S. Federal Agencies September 2010 View webpage
Federal Agency Responses Received March 2011 View webpage
Updated Test Guideline 405 Adopted by OECD October 2012 PDF

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In Vitro Ocular Test Methods for Identification of Eye Irritants and Substances Not Required Eye Irritation Hazard Labeling 

  • Bovine corneal opacity and permeability (BCOP)
  • Hen's egg test - chorioallantoic membrane (HET-CAM)
  • Cytosensor microphysiometer (CM)
  • Isolated chicken eye (ICE)
  • Isolated rabbit eye (IRE)

Ocular toxicity testing identifies substances that may cause irritation to the eye. Use of these in vitro test methods will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
EPA Nomination October 2003 -
Peer Review Panel Meeting May 2009 -
Peer Review Panel Report Published July 2009 Read report
Recommendations Made to U.S. Federal Agencies September 2010 View webpage
Federal Agency Responses Received March 2011 View webpage

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In Vitro Ocular Test Methods for Identification of Eye Irritants and Substances Not Required Eye Irritation Hazard Labeling 

  • Fluorescein leakage
  • Neutral red release
  • Red blood cell haemolysis

Ocular toxicity testing identifies substances that may cause irritation to the eye. Use of these in vitro test methods will reduce and refine animal use for this purpose.

Milestone Date Document Link or
More Information
ICCVAM Working Group Comment on Background Review Documents submitted to the European Committee for the Validation of Alternative Methods July 2008 -
Test Guideline 460 for Fluorescein Leakage Test Method Adopted by OECD October 2012 PDF

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Non-animal Assessment Approach for Evaluating Eye Irritation Potential of Antimicrobial Cleaning Products 

Ocular toxicity testing identifies substances that may cause irritation to the eye. Use of this in vitro approach could replace animal use for this purpose to meet EPA labeling requirements.

Milestone Date Document Link or
More Information
Submission by IIVS January 2008 PDF
Peer Review Panel Meeting May 2009 -
Peer Review Panel Report Published July 2009 Read report
Recommendations Made to U.S. Federal Agencies September 2010 View webpage
Federal Agency Responses Received March 2011 View webpage

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In Vivo Low Volume Eye Test 

This method is an alternative to the traditional in vivo rabbit eye test that is also used as a reference method for in vitro methods under evaluation.

Milestone Date Document Link or
More Information
Peer Review Panel Meeting May 2009 -
Peer Review Panel Report Published July 2009 Read report
Recommendations Made to U.S. Federal Agencies September 2010 View webpage
Federal Agency Responses Received March 2011 View webpage

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In Vitro Pyrogen Test Methods 

  • The Human Whole Blood/IL-1 In Vitro Pyrogen Test
  • The Human Whole Blood/ IL-1 In Vitro Pyrogen Test Using Cryopreserved Human Whole Blood
  • The Human Whole Blood/IL-6 In Vitro Pyrogen Test (WB/IL-6)
  • In Vitro Pyrogen Test Using Human Peripheral Blood Mononuclear Cells (PBMC/IL-6)
  • An Alternative In Vitro Pyrogen Test Using the Human Monocytoid Cell Line MONO MAC 6 (MM6/IL-6)

Pyrogenicity testing is performed on products to be administered by injection to ensure that each lot of these products is free of substances that could induce a dangerous fever reaction. Use of these in vitro tests could replace animal use for this purpose.

Milestone Date Document Link or
More Information
Submission by ECVAM June 2005 PDF
Peer Review Panel Meeting February 2007 -
Peer Review Panel Report Published April 2007 PDF
Test Method Evaluation Report Published October 2008 View webpage
Recommendations to Federal Agencies November 2008 View webpage
Acceptance of Test Methods by European Pharmacopeia March 2009 -
Acceptance by Federal Agencies May 2009 View webpage
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