Int. J. Environ. Res. Public Health 2010, 7 3227 be one of the sources of the microfungal propagules recovered from the mains and reservoir water during the identification and enumeration study mentioned earlier. 2. Materials and Methods 2.1. The Reservoir Samples were collected from the interior stairway and accessible structures of a roofed water service reservoir (R3) in a city in Central Queensland, Australia. Reservoir R3, which was the site for this study, is eminently suitable for that purpose. Its walls and floor are cast concrete and it is roofed with galvanised sheeting on galvanised beams supported by concrete columns. It has a maximum capacity of ca. 10.2 megalitres and is filled solely by an underground trunk main fed directly from another reservoir, designated R2, located ca. 2 km away. Reservoir R2 is automatically re-chlorinated to ca. 0.60 ppm. During the 18 month period over which the identification and enumeration study was conducted the average recovery of yeasts from reservoir R2 was <1 cell per litre and filamentous microfungi was only 3 cells per litre. Consequently the most likely explanation for the existence of the microfungal population in reservoir R3 was through introduction of propagules at the reservoir itself. Reservoir R3 was not re-chlorinated at the time of this study, and free chlorine readings during the identification and enumeration study period averaged less than 0.10 ppm. 2.2. Surface Growth of Microfungi Twenty aged faecal pellets of the Australian green tree frog L. caerulea were collected from the internal stairway and accessible infrastructure of reservoir R3. These were placed in individual sterile 30 mL polycarbonate screw-top tubes (Sarstedt) which were capped immediately after each sample was collected. At the laboratory, the surfaces of the pellets were examined for the presence of microfungal colonies under a Wild M3Z stereomicroscope. Fragments of sporulating colonies were then picked with a dissecting needle from the surfaces of five randomly selected pellets. These fragments were placed in drops of fuchsin acid/lactic acid stain (0.1 g acid fuchsin in 100 mL lactic acid) on glass microscope slides and examined under a Leica DMLB light microscope. Sporulating microfungal colonies were identified by their reproductive structures and spores to genus level by reference to Carmichael et al. [3], Ellis [4,5], Gilman [6], and Kendrick and Carmichael [7]. 2.3. Microfungal Analysis of Frog Excrement Ten freshly deposited L. caerulea faecal pellets were collected for further microfungal analysis. Each pellet was weighed then carefully broken apart and a needle-point sample was taken from the interior of the pellet, placed in a drop of acid fuchsin/lactic acid stain on a glass microscope slide, and examined under a Leica DMLB light microscope for visual evidence of the presence of microfungal spores. Each pellet was then placed in a sterile 30 mL polycarbonate screw-top tube (Sarstedt), 20 mL of sterile reverse osmosis water (SROW) was added, and a faecal suspension was produced by mixing the contents thoroughly for 1 minute on a CSV90 Auto vortex mixer at maximum setting No. 8. Each