Ecological Archives E093-107-A2

Sarah A. Orlofske, Robert C. Jadin, Daniel L. Preston, and Pieter T. J. Johnson. 2012. Parasite transmission in complex communities: Predators and alternative hosts alter pathogenic infections in amphibians. Ecology 93:1247–1253.

Appendix B. Methods for collection and maintenance of amphibians, fish, and aquatic invertebrates used in laboratory experiments.

We collected alternative hosts and predators in June-August 2009 and 2010 in central California during field surveys, with the exception of backswimmers, which we collected from cattle tank mesocosms in Boulder, Colorado in June 2010. We shipped California specimens to the laboratory at University of Colorado Boulder where we conducted the experiments. We maintained invertebrates in 1-L containers with dechlorinated tap water (AmQuel plus and NovAqua plus, Kordon®) and fed algae (clam shrimp) or Daphnia (damselflies, dragonflies, and backswimmers). We fed newt larvae chopped California blackworms (Lumbriculus variegatus) and mosquitofish commercial fish food (TetraMin, Tetra®). The number of individuals of each species examined laboratory trials (N) and the range of body size of the specimens are presented in Table A1.

In both 2009 and 2010, we obtained chorus frog eggs from field sites in California. We maintained tadpoles in 40-L plastic containers with airstones and fed a 1:1 mixture of commercial fish food (TetraMin and Spirulina) with 50 percent water changes occurring every 2–3 days until tadpoles reached approximately stage 33 (Gosner 1960).

Literature Cited

Gosner, K. L. 1960. A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 16:183–190.

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