Annotated Bibliography

This is the first report of data from the Canadian Wildlife Service field surveys that began in 1992. In this paper the authors report finding frogs with missing limb parts. Since that time they have found the other malformations that have been observed elsewhere, including extra limbs and limb parts. Their data indicate a correlation with agricultural activity, and have established the baseline frequency of malformations in pristine environments to be much less than 1%. The typical frequency of malformations in affected sites is 10% or greater.

There are three general hypotheses currently proposed to account for the observed amphibian malformations:

(1) Trauma/mechanical:

This paper presents the theoretical basis for the formation of supernumerary limb structures as a consequence of tissue rearrangements that stimulate growth and pattern formation. This paper is where the parasite hypothesis was first presented.  To date is the only publication presenting data in support of this idea. Beads implanted as mimics of parasite cysts induced some limbs with extra toes, providing indirect evidence that parasites could cause one type of malformation (extra distal skeletal elements).
 

(2) UV-Irradiation:

This paper demonstrates that UV-B can induce malformations at early stages of development. The relationship between these malformations, and those observed in metamorphic frogs is unclear. Unpublished observations from the EPA lab in Duluth, MN, indicate that UV can induce truncated limbs in metamorphic frogs; however, these truncations differ significantly from many of those on animals from the field. The relevance of UV as a causal agent in metamorphic frogs in the environment has yet to be determined.

(3) Chemical Agents:

There are thousands of peer-reviewed publications demonstrating the ability of chemical agents to induce limb malformations. Research in this area is included in the field of toxicology (Developmental Toxicology), and has given rise to the scientific discipline of teratology. It is noteworthy that there is one category of chemical agents, the retinoids, that are known to induce all of the observed limb malformations in deformed frogs.

There are hundreds of publications on the effects of retinoids on limb development and regeneration. This paper is a comprehensive review of that literature through the early 90's. Both of these papers report the same, dramatic and disturbing finding that retinoic acid induces the formation of extra hindlimbs in mouse embryos. Retinoic acid is fed to the pregnant mother, the mice are born with this malformation that is identical to the extra hindlimb malformation that is observed in some of the frogs from the wild. The molecular mechanisms of retinoid activity has been well studied, and there are hundreds of publications. The information is well reviewed in a series of articles published in this issue of Cell. One suspect pesticide that has been mentioned repeatedly in the media is methoprene, which is a juvenile hormone mimic. Although there is no evidence that its use is related to the metamorphic frog malformations, a break down product (methoprene acid) will bind to and activate a retinoid receptor. In this paper, the authors report that derivatives of methoprene can induce early stage developmental abnormalities in Xenopus (FETAX assay). It is unclear at this point how, or if, these results pertain to the types of frog malformations observed in frogs from the wild.

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