266 • Alternatives to Animal Use in Research, Testing, and Education 
Table 12-2.— Alternative Tests Under Development 
at the Food and Drug Administration 
• Genetic probes for toxigenic strains of Campylobacter 
jejuni 
• Genetic probes for invasive Escherichia coli 
• In vitro invasiveness test based on siderophore avidity 
for iron 
• Enzymatic and chemical in vitro evaluation of infant 
formula protein quality 
• Development of an assay for genetic transposition in 
bacteria 
• Cultures of rat embryos to detect agents that cause 
developmental toxicity and to determine the 
mechanism by which effects are produced 
• Porcine kidney explant cultures for screening 
potentially nephrotoxic agents 
• In vitro micromolecular biosynthesis as an index of 
potential tissue damage by chemical agents 
• In vitro determination of effects of chemical agents of 
T- and B-lymphocyte function 
• Improved procedures for use of unscheduled DNA 
synthesis for genotoxic effects 
• In vitro use of renal cortex tissue to determine 
biochemical correlates for evaluating toxicity of 
natural toxicants 
• In vitro assays to assess biological vaccine potency 
and safety (diphtheria antitoxin, rabies, polio vaccines) 
• In vitro assays to assess drug potency (gonadotropin, 
lactogenic hormone, corticotropin, oxytocin, insulin) 
• In vitro methods to determine percutaneous absorption 
of hydrophobic compounds 
• In vitro immunoassay methods (RIA, ELISA) for 
assessment of immunotoxic effects of drugs and 
environmental pollutants 
• Liquid and thin-layer chromatographic methods for 
ciguatera and paralytic shellfish toxins 
• In vitro immunoassay methods (RIA, ELISA) for 
assessment of seafood toxins 
SOURCE: A.P. Borsetti, Staff Scientist, Office of Science Coordination, Food and 
Drug Administration, U.S. Department of Health and Human Services, 
Rockville, MD, personal communication, 1985. 
testing protocols. A desire for improved tests and 
responsiveness to public concern over animal use 
also drive the search for alternatives. As public 
relations tools, nonanimal methods have proved 
valuable in reassuring the public that these cor- 
porations share their concern about animal use 
and are exploring other systems, while being care- 
ful not to jeopardize public health and safety. 
In 1980, Revlon Research Center, Inc., awarded 
a 3-year, $750,000 grant to Rockefeller University 
to establish the Rockefeller Laboratory for In Vitro 
Toxicology Assay. Revlon’s investment was the first 
serious, publicly taken step by industry in the 
search for alternatives. The Revlon award has been 
extended into a fifth year and totals more than 
$1.25 million (5). The laboratory employs four 
scientific staff, working on projects including alter - 
Table 12-3. —Alternative Tests in Use 
at the Food and Drug Administration 
• Genetic probes for heat-labile and heat-stable 
enterotoxin of Escherichia coli 
• Genetic probes for invasive strains of Yersinia 
enterocolitica 
• Genetic probes for classical 01 cholera toxin 
• Genetic probes for pathogenic organisms (01 and 
Non-01 Vibrio cholerae, Vibrio parahemolyticus, Vibrio 
vulnificus) 
• In vitro tests for percutaneous absorption of cosmetic 
ingredients 
• In vitro cell transformation assay 
• Unscheduled DNA synthesis in primary rat hepatocytes 
• Salmonella microsome assay for gene mutations 
• Limulus amebocyte lysate test for pyrogenicity of 
drugs and biologies 
• Sister-chromatid exchange for assessing mutagenic 
potential 
• Use of primary myocytes and endothelial cells from 
neonatal rat heart ventricles for identification of 
potential cardiotoxic agents 
• High-performance liquid chromatography as a screen 
for the vitamin D assay (used for products other than 
infant formula) 
• Instrumental analysis assay for potency of three 
anticancer drugs for batch release (Dactinomycin, 
Doxorubicin hydrochloride, and Plicamycin) 
• Instrumental analysis assays to determine the potency 
of biological vaccines 
• Genetic probes for invasive Shigella 
• In vitro assays for tumor-producing potential (HL 60 
differentiation, V-79 metabolic cooperativity, and 
Epstein-Barr virus activation 
• Assays for detection of mycotoxins (mass 
spectrometry, instrumental methods, brine shrimp 
assay) 
SOURCE: A.P. Borsetti, Staff Scientist, Office of Science Coordination, Food and 
Drug Administration, U.S. Department of Health and Human Services, 
Rockville, MD, personal communication, 1985. 
natives to the Draize eye irritancy test and other 
animal cell culture applications . Prior to the estab- 
lishment of this facility, there were no laboratories 
committed to alternatives research. 
The Johns Hopkins Center for Alternatives to 
Animal Testing has committed $2.1 million to the 
search for alternatives since 1981, funding 30 
grants for research (19). The Center has both an 
information program (consisting of a regularly 
published newsletter, symposia, and a book series) 
and a research program (focused on in vitro acute 
and chronic toxicity testing and acute irritancy of 
the skin and eye). The center’s enabling sponsor, 
the Cosmetic, Toiletry, and Fragrance Association 
(CTFA), is joined by other corporate donors, in- 
cluding the Bristol Myers Company, as well as by 
consumer and industrial groups and private indi- 
viduals. CAAT solicits projects from scientists by 
