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Environ Toxicol Chem
1999 Oct 01;1810:2305-2315. doi: 10.1002/etc.5620181027.
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Effects of pond water, sediment, and sediment extracts from minnesota and vermont, USA, on early development and metamorphosis of xenopus.
Fort DJ
,
Propst TL
,
Stover EL
,
Helgen JC
,
Levey RB
,
Gallagher K
,
Burkhart JG
.
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In recent studies, a high incidence of amphibian mortality and malformation has been reported in the field, suggesting that toxic and/or bioactive agents are present in the environment of the affected amphibians. This study provides evidence for this hypothesis, because it applies to several affected ponds in Minnesota and Vermont, USA. Three developmental bioassays were carried out on samples from three reference and three test sites in Minnesota and one reference and three test sites, in Vermont. The bioassays utilized Xenopus as a model system, measuring altered developmental patterns during the first 4 d of development (frog embryo teratogenesis assay-Xenopus [FETAX]), hind-limb development over a 30-d period, and tail length resorption over a 14-d period. Strong correlations were observed among the results for all three in vitro bioassays, as well as between adverse developmental effects in vitro and in the field.
Fig. 1. Frog embryo teratogenesis assay—Xenopus(FETAX) tests ofpond samples from Minnesota sites. Results of FETAX assays of pondwater, sediment, and aqueous sediment extract samples from referenceand test sites are shown. Reference sites are indicated. Standard erroris shown by error bars. (A) Percent mortality results. (B) Percentmalformation results. An asterisk indicates that percent malformationwas not determined for these samples because of the absence of sur-viving larvae (i.e., 100% mortality was observed).
Fig. 2. Frog embryo teratogenesis assay—Xenopus(FETAX) tests ofpond samples from Vermont sites. Results of FETAX assays of pondwater, sediment, and aqueous sediment extract samples from referenceand test sites are shown. Reference sites are indicated. (A) Percentmortality results. (B) Percent malformation results. An asterisk in-dicates that percent malformation was not determined for these sam-ples because of the absence of surviving larvae (i.e., 100% mortalitywas observed).
Effects from Minnesota and Vermont ponds onXenopus Environ. Toxicol. Chem.18, 1999 2309Fig. 3. Morphology of 4-dXenopusembryos reared in affected and reference site pond water from Minnesota and Vermont. (A) DB site waterand reference. (B) DR site water and reference. (C) NE site water and reference. (D) PN site water and reference.
Effects from Minnesota and Vermont ponds onXenopus Environ. Toxicol. Chem.18, 1999 2311Fig. 4. Example of hind-limb development inXenopus30-d larvae. (A) Normal 30-d limb development. (B) Development at 30 d in water fromthe DB site in Minnesota.
Fig. 5. Effects of Minnesota pond water samples on the rate of tailresorption. Tail length was measured at stages 63 to 66 during in-cubation in the presence of pond water from the indicated site. Ad-ditions to water were as indicated and at the following concentrations:1T45100mg/L thyroxine;1I50.1% (w/v) iodine. For the control,incubation was in the presence of frog embryo teratogenesis assay—Xenopus(FETAX) solution. Standard error is shown by error bars.
Fig. 6. Effects of Vermont pond water samples on the rate of tailresorption. Tail length was measured at stages 63 to 66 during in-cubation in the presence of pond water from the indicated site. Forthe control, incubation was in the presence of frog embryo terato-genesis assay—Xenopus(FETAX) solution. Standard error is shownby error bars. (A) Pond water with no additions. (B) Additions towater were as indicated and at the following concentrations:1T45100mg/L thyroxine;1I50.1% (w/v) iodine;1M5100mg/L me-thimazole.
