The North American Reporting Center for Amphibian Malformations
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Flow cytometry (FC) is a laboratory method used to detect genetic damage induced by environmental contaminants and other stressors in animals, including amphibians. We tested FC methods on three species of ranid frogs collected from farm ponds and natural wetlands in southeastern Minnesota. We compared FC metrics for Rana clamitans between ponds with direct exposure to agricultural contaminants and reference (unexposed) ponds. Concentrations of atrazine in water from our farm ponds ranged from 0.04 to 0.55 ppb. We found that R. clamitans from exposed ponds had DNA content similar to frogs from unexposed ponds. Pond-averaged C-values (a measure of DNA content) ranged from 6.53 to 7.08 for R. pipiens (n . 13), 6.55 to 6.60 for R. clamitans (n . 40) and 6.74 for R. palustris (n . 5). Among all species, the mean sample CVs ranged from 1.91 (R. palustris) to 6.31 (R. pipiens). Deformities were observed in only 2 of 796 individuals among all species and occurred in both reference and exposed ponds. Although we did not detect evidence of DNA damage associated with agriculture in our study, we demonstrated the potential of FC for screening amphibian populations for genetic damage. Metrics from a variety of amphibian species and locations as well as laboratory studies are needed to further assess the value of FC for monitoring amphibian genetic integrity in contaminated sites.
Abstract has not been submitted
Abstract has not been submitted
The national distribution of information on the discovery of malformations in Minnesota frogs in 1995 stimulated collection and examination of newly metamorphosed frogs during 1996. By late summer and early fall of 1996, malformed frogs and toads were reported on U.S. Fish and Wildlife Service (USFWS) lands in Vermont (Northeast, Region 5) and Minnesota (Midwest, Region 3). In response to these reports, biologists in USFWS Regions 3 and 5 conducted a survey, during the summer of 1997 to determine the distribution and type of malformations in frogs and toads on selected federal lands. Region 3 personnel surveyed 38 field stations at National Wildlife Refuges (NWR's) and Wetland Management Districts. Malformed frogs and toads were collected at 23 (61%) of the Region 3 sites. External malformations were detected in 110 of 6632 individuals representing seven of 13 frog species and one of three toad species examined for an overall of 1.7% affected (percentages for affected species ranged from 0.4-5.2%). In Region 5, 17 NWR's and one National Park were surveyed. Malformed frogs were collected at 10 (56%) of the Region 5 sites. External malformations were detected in 58 of 2267 individuals representing six of 11 frog species and one of two toad species examined for an overall total of 2.6% affected (percentages for affected species ranged from 1.8-15.6%). The majority of malformations observed in frogs and toads collected in Regions 3 and 5 were partially or completely missing hind limbs and digits (50%)or malformed hind limbs and digits (14%). A few individuals had an extra limb or toe, missing or malformed front limb, missing eye, or malformation of the mandible. Despite small sample sizes at some sites, malformations were confirmed to be present in eight species of frogs and two species of toads on Federal lands in USFWS Regions 3 and 5. Further study is needed to determine the extent and distribution of amphibian malformations in these Regions. Data from this study were provided to the national database on distribution of malformed amphibians.
This paper summarizes terminology and general concepts involved in animal development for the purpose of providing background for the study and understanding of frog malformations. The results of our radiographic investigation of rear limb malformations in Rana pipiens provide evidence that frog malformations are the product of early developmental errors. Although bacteria, parasites and viruses were identified in these metamorphosed frogs, the relevant window to look for the teratogenic effect of these agents is in the early tadpole stage during limb development. As a result, our microbiological findings must be regarded as inconclusive relative to determining their contribution to malformations because we conducted our examinations on metamorphosed frogs not tadpoles. Future studies need to look at teratogenic agents (chemical, microbial, physical or mechanical) that are present in the embryo, tadpole, and their environments at the stages of development that are relevant for the malformation type. The impact of these teratogenic agents then needs to be assessed in appropriate animal models using studies that are designed to mimic field conditions. The results of these laboratory tests should then be analyzed in such a way that will allow comparison with the findings in the wild-caught tadpoles and frogs.
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Research has recently documented the prevalence of a wide variety of pharmaceuticals and other emerging contaminants (ECs) in streams across the United States. Wastewater treatment plants (WWTPs) have been found to be an important source and collection point of ECs to streams as many ECs are incompletely removed during treatment. To investigate the complex in-stream processes (e.g., dilution, sorption, degradation, dispersion, etc.) that can affect ECs following their input from a WWTP and determining if such input is having an effect on the aquatic ecosystem requires the integration of multi-disciplinary efforts at a carefully selected field site. Preliminary work has identified an 8-km reach of Fourmile Creek in central Iowa as an ideal research site to investigate such important research questions pertaining to ECs. Unique aspects of Fourmile Creek included: (1) it single source effluent-dominated stream, (2) background data document the input of a wide variety of ECs from WWTP discharge, (3) small basin size, (4) relatively simple flow system, (5) background data suggest that undefined processes are taking place decreasing the level of select ECs during stream transport, (6) the WWTP uses a treatment technology (activated sludge) typical of many towns in Iowa and the United States (7) a hydrogeologic setting of a low-gradient, small stream (average discharge less than 1.41 m³/s) in glacial drift is typical of many areas in Iowa and across the Midwest, and (8) the existence of a low-head clam approximately 2 km upstream of the WWTP outfall allowing more accurate "above WWTP" and "below WWTP" comparisons in aquatic ecosystems. Furthermore, the WWTP is scheduled to close by 2011 providing a unique opportunity to determine how stream hydrology, water chemistry and aquatic biota react to the removal of the primary source of flow and ECs in this system. This will allow a novel "before" and "after" assessment not previously available in EC research. Research to date at the site has included installation of a streamflow gauging station, dye-tracing tests (to determine water travel times), Lagrangian water-quality sampling at two flow/water temperature regimes, and sampling for ECs in bed sediment. Selected fish have been collected for analysis and identification. In addition, basic fish community and fish health assessment for different seasons and spawning conditions are being analyzed. The research "framework" is unique at Fourmile Creek for investigating the important question of how ECs are transported through the environment and if the presence of such compounds is having a deleterious effect on aquatic ecosystems.