Biosensor, ELISA, and frog embryo teratogenesis assay: Xenopus (FETAX) analysis of water associated with frog malformations in Minnesota

Between 1995 and 1997 over 62% of the counties in Minnesota reported the presence of malformed frogs. While most sites have recently shown a decline in malformed frog populations, one site in northeastern Minnesota with no prior history of containing malformed frogs was recently discovered to contain > 67% malformed Rana pipiens (northern leopard frogs). As part of an effort to study the presence of hormonally active agents in fresh water sources, water samples were collected from lakes in Minnesota containing malformed frogs and analyzed for the presence of hormonally active compounds using a novel evanescent field fluorometric biosensor and the frog embryo teratogenesis assay: Xenopus (FETAX) bioassay. The waveguide based biosensor developed by ThreeFold Sensors (TFS biosensor, Ann Arbor, MI) detects the presence of estrogenic compounds capable of interacting with free human ER-a and by inhibiting binding to an immobilized estrogen. The FETAX bioassay is a developmental assay, which measures teratogenicity, mortality, and inhibition of growth during the first 96 hours of organogenesis and thereby provides a universal screen for endocrine disruptors. TFS biosensor and FETAX screening of the water samples suggest a relationship between estrogenic activity, mineral supplementation, and the occurrence of malformed frogs.

[1]  K. Gallagher,et al.  Effects of pond water, sediment, and sediment extracts from minnesota and vermont, USA, on early development and metamorphosis of xenopus , 1999, Environmental toxicology and chemistry.

[2]  C. Lyttle,et al.  A Transcriptional Coactivator, Steroid Receptor Coactivator-3, Selectively Augments Steroid Receptor Transcriptional Activity* , 1998, The Journal of Biological Chemistry.

[3]  J. Bantle,et al.  Fetax interlaboratory validation study: Phase II testing , 1994 .

[4]  C. Carey,et al.  Amphibian declines: an immunological perspective. , 1999, Developmental and comparative immunology.

[5]  M. Shelby,et al.  Induction of mortality and malformation in Xenopus laevis embryos by water sources associated with field frog deformities. , 1998, Environmental health perspectives.

[6]  S. Sessions,et al.  Explanation for naturally occurring supernumerary limbs in amphibians. , 1990, The Journal of experimental zoology.

[7]  J. Helgen,et al.  PROGRESS TOWARD IDENTIFYING CAUSES OF MALDEVELOPMENT INDUCED IN XENOPUS BY POND WATER AND SEDIMENT EXTRACTS FROM MINNESOTA, USA , 1999, Environmental toxicology and chemistry.

[8]  Kevin B. Lunde,et al.  The effect of trematode infection on amphibian limb development and survivorship. , 1999, Science.

[9]  J. Bantle,et al.  FETAX Interlaboratory Validation Study: Phase III–Part 1 Testing , 1996, Journal of applied toxicology : JAT.

[10]  T. Schultz,et al.  Toxic and teratogenic effects of selected aromatic amines on embryos of the amphibianXenopus laevis , 1981, Archives of environmental contamination and toxicology.

[11]  J. Bantle,et al.  Initial interlaboratory validation study of FETAX: Phase I testing , 1994, Journal of applied toxicology : JAT.

[12]  D Neumeier,et al.  Biotinylated steroid derivatives as ligands for biospecific interaction analysis with monoclonal antibodies using immunosensor devices. , 2000, Analytical biochemistry.

[13]  C. Meteyer,et al.  Hind limb malformations in free-living northern leopard frogs (Rana pipiens) from Maine, Minnesota, and Vermont suggest multiple etiologies. , 2000, Teratology.

[14]  J. Bantle,et al.  Phase III interlaboratory study of FETAX part 3. FETAX validation using 12 compounds with and without an exogenous metabolic activation system , 1999, Journal of applied toxicology : JAT.

[15]  S. Sessions,et al.  Morphological clues from multilegged frogs: are retinoids to blame? , 1999, Science.

[16]  C. Lyttle,et al.  Estrogen Receptor Ligands Modulate Its Interaction with DNA* , 1997, The Journal of Biological Chemistry.

[17]  J. Bonin,et al.  HINDLIMB DEFORMITIES (ECTROMELIA, ECTRODACTYLY) IN FREE-LIVING ANURANS FROM AGRICULTURAL HABITATS , 1997, Journal of wildlife diseases.

[18]  G. Ankley,et al.  Effects of ultraviolet light and methoprene on survival and development of Rana pipiens , 1998 .

[19]  J. Kaiser Deformed Frogs Leap Into Spotlight at Health Workshop , 1997, Science.

[20]  G. Ankley,et al.  Effects of water quality on development of Xenopus laevis: A frog embryo teratogenesis assay—Xenopus assessment of surface water associated with malformations in native anurans , 2000 .

[21]  J. Bantle,et al.  Analysis of the mechanism of isoniazid-induced developmental toxicity with frog embryo teratogenesis assay: Xenopus (FETAX). , 1990, Teratogenesis, carcinogenesis, and mutagenesis.

[22]  J. Faber,et al.  Normal table of Xenopus laevis (Daudin). A systematical and chronological survey of the development from the fertilized egg till the end of metamorphosis. , 1956 .

[23]  C. Scott Findlay,et al.  Quantitative evidence for global amphibian population declines , 2000, Nature.

[24]  P. Daszak,et al.  Perspectives Perspectives Perspectives Perspectives Perspectives Amphibian Population Declines Emerging Infectious Diseases and Amphibian Population Declines , 2022 .

[25]  D. Chivers,et al.  Ambient UV-B radiation causes deformities in amphibian embryos. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Maciej Adamczyk,et al.  Evaluation of chemiluminescent estradiol conjugates by using a surface plasmon resonance detector , 2000, Steroids.

[27]  R. Mann,et al.  Application of the FETAX protocol to assess the developmental toxicity of nonylphenol ethoxylate to Xenopus laevis and two Australian frogs. , 2000, Aquatic toxicology.

[28]  Ulrich,et al.  Development of Kinetic Ligand-binding Assays Using a Fiber Optic Sensor. , 1999, Clinical chemistry.

[29]  R. Finch,et al.  Phase III interlaboratory study of FETAX, Part 2: interlaboratory validation of an exogenous metabolic activation system for frog embryo teratogenesis assay--Xenopus (FETAX). , 1998, Drug and chemical toxicology.

[30]  K. Dumpert Embryotoxic effects of environmental chemicals: tests with the South African clawed toad (Xenopus laevis). , 1987, Ecotoxicology and environmental safety.

[31]  M. Seifert,et al.  In vitro analysis of xenoestrogens by enzyme linked receptor assays (ELRA). , 1998, Advances in experimental medicine and biology.

[32]  W. J. Davies,et al.  Frog embryo teratogenesis assay. Xenopus (FETAX). , 1995, Methods in molecular biology.