SURVEY OF ARCHIVED AMPHIBIANS FOR CHYTRID FUNGI

Chytrid fungi are killing amphibians in the wild (Berger et al. 1998). If the microorganisms is a primary parasite, we hypothesise that it has entered populations at particular points in time possibly just prior to the decline of the population. The alternative hypothesis is that the fungus was present in the population previously and other factors have allowed it to manifest as a pathogenic agent. By identifying when the pathogen first occurs in a population and the realationship to deaths of frogs, we hope to be able to:

1. Generate data to support either hypothesis
2. Determine the areas and frog populations where the fungus currently occurs
3. Determine the prevalence of the amphibian chytrid in "normal" frogs.

We are interested in examining any wild amphibians that have been collected and preserved. The chytrids are prevalent on ventral surfaces of the frog and on the feet, especially the webbing. They can be detected in specimens fixed in 10% formalin or 70% alcohol. If you submit fixed skin samples, we prepare histological slides stained with haematoxylin and eosin, and examine the superficial epidermis for fungi.

Protocol for collecting skin samples

The following protocol was developed by Lee Berger and David Earl Green.

Cut a piece of skin as in the diagram. For anurans or salamanders, also send a whole hind foot, or just a toe cut off at the base (ie. toe III or V for anurans, toe IV or V for salamanders). Alternatively, remove skin from the ventral surface of the foot. Place skin samples in labelled container with formalin or alcohol. Record the information requested below (if known) for every sample submitted.

Record sheet for specimens

Name and address of sender: .................................................................................................
...............................................................................................................................................

Phones: .......................... ....................... Fax: ............................ Email: ..............................
Animal ID no: ................ Species: ...................................... Sex: .........
Date collected: ................. Location collected: .................................................................
Type of environment: ..............................................................................................................
Weight................ Snout-Vent length............
Method of euthanasia .......................................................................
Healthy or sick?......................
If sick, describe any signs of disease: ..................................................................................
...................................................................................................................................................
...................................................................................................................................................

ADDRESS TO SEND SAMPLES:

Rick Speare School of Public Health and Tropical Medicine James Cook University Townsville 4811 AUSTRALIA Phone: Fax: Email: 07-47225777 07-47225788 richard.speare@jcu.edu.au

Dr David Earl Green National Institutes of Health Bldg 28A, Room 115, MSC 5230 28 Liberty Drive Bethesda MD 20892-5230 USA Phone: Fax: Email: 1-301-496-4465 1-301-402-1068 greend@exchange.nih.gov

Updated 19 January, 1999
Rick Speare