Factors affecting germination of ascospores in Aspergillus fumigatus
Background and objective: Aspergillus fumigatus is the most common airborne fungal pathogen of humans. It is an opportunistic human pathogen in immunocompromised hosts. Following the recent discovery of the sexual reproductive cycle ascospores of this fungus were available and their germination was studied in the present study. This study aimed to study factors influencing germination of ascospores of A. fumigatus.
Methods: A total of 12 environmental A. fumigatus were chosen in both mating type, examined for sexual reproduction by crossing complementary mating type, following removing of cleistothecia, preparation of ascospore suspensions in 0.05% Tween 20, then heat treated to kill any remaining conidia.
Results: Ascospore germination of greater than 75% occurred at 28 °C on a wide variety of mycological media, 0.5% (ACM) was chosen for use in subsequent studies. Ascospores did not require a heat treatment prior to germination; however, heat treatment of ascospores at 75 °C for 1 hour previous to incubation enhanced germination to 89% and also served to inactivate contaminating conidia. Ascospores also exhibited a psychrotolerance following exposure at -80 °C for 96 hours. Inoculum concentration did not have a significant effect on germination rates between the range of 0.125×106 to 4×106 ascospores ml-1 (P >0.05).
Conclusion: This study demonstrated that ascospore germination of greater than 75% occurred at 28 °C on a wide variety of mycological media. Ascospores of A. fumigatus are thermotolerant also exhibited a psychrotolerance whilst at the same time retaining viable sexual ascospores, to assess percentage germination a drop of Lactophenol Cotton Blue was added.
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