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Ninety-three isolates of Metarhizium anisopliae, mostly derived from a survey of termite material, were screened for activity against Nasutitermes exitiosus and Coptotermes frenchi or C. acinaciformis using a grooming assay technique. Twenty-six of the most promising isolates were further evaluated by bioassay against N. exitiosus and C. acinaciformis. All isolates were pathogenic with Coptotermes spp. being more susceptible than N. exitiosus. A group of nine isolates, chosen for their level of pathogenicity for one or other genus of termites and to represent a genetically diverse group, was finally compared in a minicolony test using termite colonies in 1 liter jars. The isolate, code-named FI-610 (derived from nest-mound material of C. lacteus in SE New South Wales), was one of the most effective isolates against termites from both of the two colonies tested. This isolate also grew relatively well on agar plates at 36°C. FI-610 was thus selected for field trials and was found to be effective in killing colonies of C. acinaciformis when 10 g (=3 × 10 11 conidia) or more of conidial powder was blown into the center of the large mound colonies.

A total of 479 samples of nests or feeding sites of 38 species of Australian termite were plated onto a medium selective for isolation of Metarhizium spp. Sixty-seven samples were positive for Metarhizium spp. and a total of 97 isolates of M. anisopliae were obtained. Very few isolates were obtained directly from infected termites. Most isolates were obtained from nest-mound material from eastern Australia. Termite-associated material from the two common mound-building species of termite, Nasutitermes exitiosus and Coptotermes lacteus, provided 75 of the isolates. Similar material from 26 species of termites revealed no Metarhizium. A detailed study of two sites found that some of the Metarhizium isolates found in nest-mound material, including also some M. flavoviride and M. album, were of morphological types also present in adjacent soil. The DNA of isolates from mounds and the adjacent soil were compared using RAPDs and sequence analysis of the ITS region of the nuclear rDNA and the same types were found from both sources. The possible role of the fungus in termite ecology is discussed and it is thought most likely that Metarhizium is only opportunistically a pathogen of termites. Thus, isolates obtained from termite nest material are probably there because of the incorporation of soil into termite nests.

The effect of relative humidities (r.h.) from 90 to 100% on germination of a termite-active isolate of Metarhizium anisopliae (isolate FI25 and FI610) was studied using a liquid germinating medium to which the appropriate amount of glycerol had been added. Germination was increasingly delayed at water activities equivalent to 99, 98, and 96% r.h. and completely inhibited at 94, 92, and 90%. Twenty-one isolates were then screened for germination at 96 and 100% r.h. All isolates showed delayed germination at 96% r.h. but most isolates eventually gave a high final percentage germination at this humidity. Two isolates, FI527 and FI638, were markedly slower to germinate at both humidities. The susceptibility of two species of termites, Nasutitermes exitiosus and Coptotermes acinaciformis, to FI610 was tested at r.h. down to 86%-the lowest humidity at which the insects would survive. No consistent effect of humidity on pathogenicity was detected. Mortality occurred over the range range of humidities tested; sporulation from the disease-killed termites, however, occurred only at r.h. above 93%. It is concluded that the microclimate around living termites is usually sufficiently humid to ensure infection under most field conditions and that humidity is unlikely to limit the efficacy of the fungus in controlling termites.