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 agency scientists are contributing.

Alternative test methods:

  • 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
  • Replace animals with non-animal systems or one animal species with a less highly developed one (for example, replacing a mouse with a fish).

View status of test method evaluation projects grouped by project area

Acute Oral Systemic Toxicity Dermal Irritation Genetic Toxicity
Acute Inhalation Toxicity Ocular Toxicity Developmental Toxicity
Acute Dermal Systemic Toxicity Immunotoxicity Endocrine Disruptor
Dermal Corrosivity Dermal Phototoxicity Pyrogenicity
In Vitro Metabolism and Toxicokinetics Dermal Absorption Biologics and Vaccines

Select project name in summary table to jump down the page to details and links

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 2002 (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 (OECD TG 436)
Fixed Dose Procedure (inhalation) Anticipated 2013 (OECD) (Not applicable - to be adopted via OECD Test Guideline) Anticipated 2014 (OECD)
Acute Dermal Systemic Toxicity
Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Up-and-Down Procedure (dermal) In progress Anticipated 2013 Anticipated 2013 Anticipated 2014 Anticipated 2014 (OECD)
In Vitro 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 Anticipated 2013 (OECD) (Not applicable - to be adopted via OECD Test Guideline) Anticipated 2014 (OECD)
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 (OECD TG 439)
EPISKINTM Assay (Not applicable - adopted via OECD Test Guideline) 2010 (OECD TG 439)
SkinEthic RHE Assay (Not applicable - adopted via OECD Test Guideline) 2010 (OECD TG 439)
LabCyte EPI-MODEL24 Assay In progress      
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 Anticipated 2013 (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 (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
Immunotoxicity
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)
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 (OECD TG 432)
3T3 NRU Phototoxicity Test: Application to UV Filter Chemicals (Not applicable - adopted via OECD Test Guideline) 2004 (OECD TG 432)
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 (OECD TG 428)
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 (OECD TG 487)
In Vivo Comet Assay In progress (JaCVAM)      
In Vitro Comet Assay In progress (JaCVAM)        
Cell Transformation Assays (Not applicable - to be adopted via OECD Test Guideline) Anticipated 2013 (OECD)
BHAS Cell Transformation Assay In progress (JaCVAM)      
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 Andogen 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
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)
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)
Alternative Methods for Botulinum Toxin Potency Testing ICCVAM workshop in 2006 assessed the state of the science and concluded that additional development and/or validation of these methods were needed prior to evaluation by ICCVAM
Alternative Methods for Vaccine Potency and Safety Testing ICCVAM workshop in September 2010 assessed the state of the science and identified priorities for research, development, and validation efforts: report detailing conclusions and recommendations published in 2011
Alternative Methods for Rabies Vaccine Potency Testing ICCVAM workshop in October 2011 reviewed the available methods and approaches to reduce, refine, and replace animals used in rabies vaccine potency testing and developed an implementation strategy to achieve global acceptance and use of these alternatives; report detailing conclusions and recommendations published in 2012
Alternative Methods for Leptospira Vaccine Potency Testing An ICCVAM workshop in September 2012 reviewed the available methods and approaches to reduce, refine, and replace animals used in Leptospira vaccine potency testing and developed an implementation strategy to achieve global acceptance and use of these alternatives: report detailing conclusions and recommendations published in 2013
Alternatives to the HIST Test for Acellular Pertussis Vaccines A workshop in November 2012 reviewed available alternatives to the murine histamine sensitization (HIST) test, reviewed results of an international validation study, and considered an implementation strategy to achieve global acceptance and use of these alternatives
Report detailing conclusions and recommendations to be published in 2013

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Acute Oral Systemic Toxicity

Up-And-Down Procedure (oral)

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
Publication of OECD Test Guideline 425 1998 PDF
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
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

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 1998 PDF

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 1998 PDF

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Evaluation of 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

Validation Study of 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. Neutral Red Uptake Test Methods replace animal use for initial dose setting for these tests.

Milestone Date Document Link or
More Information
Peer Review Panel Meeting May 2006 HTML
Peer Review Panel Report Published June 2006 PDF
Background Review Document Published November 2006 View webpage
Test Method Evaluation Report Published November 2006 HTML
Recommendations Made to Federal Agencies March 2008 HTML
Federal Agency Responses Received September 2008 HTML
OECD Guidance Document 129 Published July 2010 PDF


Developing and Advancing In Vitro Alternatives to Acute Chemical Systemic Toxicity Testing

Acute oral systemic toxicity testing identifies substances that are poisonous when ingested so that they may be appropriately labeled and packaged. An ICCVAM-sponsored workshop explored ways to reduce, refine, or replace animal use for this purpose.

Milestone Date Document Link or
More Information
Workshop Held February 2008 HTML
Workshop Report Published May 2009 HTML

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Acute Inhalation Toxicity

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

Dermal Corrosivity

Evaluation of Corrositex® for the Identification of Substances Potentially Corrosive to Human Skin

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 HTML
Peer Review Panel Report Published June 1999 PDF
Recommendations Made to Federal Agencies June 1999 -
Federal Agency Responses Received October 1999 HTML
Publication of OECD Test Guideline 435 July 2006 PDF


Evaluation of EpiSkinTM, EPIDERMTM and the Rat Skin Transcutaneous Electrical Resistance Assay (TER) for the Identification of Substances Potentially Corrosive to Human Skin

Dermal corrosivity testing identifies substances that cause chemical burns to the skin so that they may be appropriately labeled and packaged. Use of these in vitro 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 HTML
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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Dermal Irritation

Evaluation of EpiSkinTM, EPIDERMTM and the SkinEthic RHE Assay for the Identification of Substances Potentially Irritating to Human Skin

Dermal irritation testing identifies substances that cause irritation to the skin so that they may be appropriately labeled and packaged. Use of these in vitro tests 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

Ocular Toxicity

In Vitro Test Methods for Detecting Ocular Corrosives and Severe Irritants

  • Bovine Corneal Opacity and Permeability Test Method
  • Isolated Chicken Eye Test Method
  • Isolated Rabbit Eye Test Method
  • Hen's Egg Test - Chorioallantoic Membrane Test Method

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 HTML
Expert Panel Report March 2005 PDF
Expert Panel Teleconference September 2005 HTML
Expert Panel Report Addendum November 2005 PDF
Test Method Evaluation Report Published October 2007 HTML
Recommendations Made to Federal Agencies October 2007 HTML
Federal Agency Responses Received May 2008 HTML
Adoption of OECD Test Guidelines 437 (BCOP) and 438 (ICE) September 2009 HTML
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


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 HTML
Federal Agency Responses Received March 2011 HTML
Updated Test Guideline 405 Adopted by OECD October 2012 PDF


In Vitro Test Methods for Detecting Nonsevere Irritants and Substances Not Requiring Ocular Hazard Labeling

  • Bovine Corneal Opacity and Permeability Test Method
  • Isolated Chicken Eye Test Method
  • Cytosensor Microphysiometer Test Method
  • Isolated Rabbit Eye Test Method
  • Hen's Egg Test - Chorioallantoic Membrane Test Method

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 HTML
Federal Agency Responses Received March 2011 HTML

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In Vitro Test Methods for Detecting Nonsevere Ocular Irritants and Substances Not Requiring Ocular Hazard Labeling

  • Fluorescein Leakage Test Method
  • Neutral Red Release Test Method
  • Red Blood Cell Haemolysis Test Method

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


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 HTML
Federal Agency Responses Received March 2011 HTML


The In Vivo Low Volume Eye Test

The low volume eye test 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 HTML
Federal Agency Responses Received March 2011 HTML

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Immunotoxicity

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 HTML
Peer Review Panel Report February 1999 PDF
Recommendations Made to Federal Agencies February 1999 HTML
Adoption of OECD Test Guideline 429 April 2002 PDF
Update to OECD Test Guideline 429 Adopted July 2010 PDF


Updated Protocol for the Murine Local Lymph Node Assay (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 HTML
Recommmendations to U.S. Federal Agencies Accepted March 2010 HTML
Update to OECD Test Guideline 429 Adopted July 2010 PDF


Development of Performance Standards for the Murine Local Lymph Node Assay (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
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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 HTML
Recommmendations to U.S. Federal Agencies Accepted March 2010 HTML
Update to OECD Test Guideline 429 Adopted July 2010 PDF


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 HTML
Recommmendations to U.S. Federal Agencies Accepted March 2010 HTML
Update to OECD Test Guideline 429 Adopted (containing rLLNA procedure) July 2010 PDF

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Nonradioactive Murine Local Lymph Node Assay (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 refine and reduce 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 HTML


Use of the Murine Local Lymph Node Assay (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. Use of the LLNA will refine and 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
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 HTML


Use of the Murine Local Lymph Node Assay (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 will refine and 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 the Use of the LLNA for Skin Sensitization Potency Categorization Transmitted to U.S. Federal Agencies June 2011 View recommendations
Federal Agency Responses Received February 2012 View agency responses


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Phototoxicity

Use of the 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 refine and reduce animal use for this purpose.

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


Dermal Absorption

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


Genetic Toxicity

Draft Test Guideline for the 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
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Test Guideline 487 Adopted by OECD July 2010 PDF

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Developmental Toxicity

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 HTML
Expert Panel Meeting May 2000 PDF

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Endocrine Disruptor

Evaluation of 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 HTML
Expert Panel Report September 2002 PDF
Background Review Documents Published October 2002 HTML
Test Method Evaluation Report Published May 2003 HTML
Addendum to Test Method Evaluation Report Published September 2006 PDF

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BG1Luc ER TA Test Method Validation Study

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 HTML
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


CertiChem Inc. MCF-7 Cell Proliferation Assay Evaluation

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 HTML
Validation Study Completed 2011 -
Draft Validation Study Report June 2012 PDF

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Pyrogenicity

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 HTML
Recommendations to Federal Agencies November 2008 HTML
Acceptance of Test Methods by European Pharmacopeia March 2009 -
Acceptance by Federal Agencies May 2009 HTML


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Biologics

Use of 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
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USDA Directive Published April 2004 PDF


Alternative Methods for Botulinum Toxin Potency Testing

Animal testing of botulinum toxin is performed to ensure that each lot of this product is safe and effective. An ICCVAM-sponsored workshop explored ways to reduce, refine, or replace animal use for this purpose.

Milestone Date Document Link or
More Information
Nomination by the Humane Society of the United States October 2005 PDF
Workshop Held November 2006 HTML
Workshop Report Published February 2008 PDF

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Alternative Methods for Vaccine Potency and Safety Testing

Animal testing of vaccines is performed to ensure that each lot of these products is safe and effective. An ICCVAM-sponsored workshop explored ways to reduce, refine, or replace animal use for this purpose.

Milestone Date Document Link or
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Workshop Held September 2010 HTML
Workshop Proceedings Published December 2011 Workshop Report Exiting the ICCVAM website


Alternative Methods for Rabies Vaccine Potency Testing

Animal testing of rabies vaccines is performed to ensure that each lot of these products is effective for its intended use. An ICCVAM-sponsored workshop will explore ways to reduce, refine, or replace animal use for this purpose.

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Workshop Held October 2011 View webpage
Workshop Report Published September 2012 Workshop Report Exiting the ICCVAM website


Alternative Methods for Leptospira Vaccine Potency Testing

Animal testing of Leptospira vaccines is performed to ensure that each lot of these products is effective for its intended use. An ICCVAM-sponsored workshop will explore ways to reduce, refine, or replace animal use for this purpose.

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Workshop Held September 2012 View webpage


Alternatives to the HIST Test for Acellular Pertussis Vaccines

Animal testing of acellular pertussis vaccines is performed to ensure that each lot of these products is safe for use. An ICCVAM-sponsored workshop will explore ways to reduce, refine, or replace animal use for this purpose.

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Workshop Held November 2012 View webpage
NTP is located at the National Institute of Environmental Health Sciences, part of the National Institutes of Health.