Collection and Isolation
A spider infected by the fungus Gibellula pulchra was collected in May 2015, from a bottomland hardwood forest near Lafayette, Louisiana (30°15′28.7″N, 91°59′17.6″W). The cadaver was collected along with the leaf to which it was attached and transported to the lab in a clean plastic container (Figure 1e).
The fungus was isolated on Sabouraud Dextrose Agar plus 2% yeast extract (SDAY+) by holding the specimen over the open plate and gently tapping the leaf to which the infected spider was attached, causing fungal spores to be released onto the surface of the medium. Cultures were maintained at room temperature and ambient light and checked daily for fungal growth. Subcultures were made from the germinating spores and pure cultures were established and used for isolation of genomic DNA. The ITS region was sequenced (primers; ITS1F and ITS4) to further confirm the identity of the fungus.
The host was difficult to identify due to the extent of fungal growth, though this fungus has been recovered from several common species in the area including Colonus spp ., Anasaitis canosa , andPlatycryptus sp .. Due to their commonality, individuals of the genus Colonus (previously Thiodina sp.) were chosen as hosts for an ad hoc experiment intending to confirm the cause of mortality as being due to the fungus G. pulchra .