An immature merocyst of Hepatocystis malayensis and gametocytes of H. brayi were studied with the electron microscope. The merocyst consisted of a highly complex cytoplasmic reticulum ramifying through an amorphous matrix: the entire complex was enclosed by a simple unit membrane. The host cell was apparently destroyed completely during growth of the cyst. Immature gametocytes were highly amoeboid and showed extensive vacuolisation or attenuation of the cytoplasm. The nucleus contained one or two prominent nucleoli. Mature gametocytes had compact cytoplasm and contained pyriform osmiophilic bodies which were believed to function in the release of the parasites from the host cells. Macrogametocytes were distinguished from microgametocytes by cytoplasmic differences in numbers of ribosomes, and cristate mitochondria and in the extent of development of the smooth endoplasmic reticulum. The compact nuclei of the macrogametocytes had inconspicuous DNA but prominent nucleoli whereas those of the microgametocytes were irregular and showed a central aggregate of DNA. In microgametogenesis karyokinesis of the parent nucleus was delayed until axoneme formation was complete. Then the nuclear buds were extruded into emerging microgametes. At fertilisation the plasmalemmas of the two gametes fused and the single axoneme and nucleus of the microgamete moved into the cytoplasm of the macrogamete.
The development of H. brayi was followed mainly in C. variipenis up to the stage of mature oocysts. Unlike H. kochi, the oocysts of H. brayi develop at the same site as those of Plasmodium between the epithelium and the basal membrane of the stomach.
The microsporidian genus Nosema is characterized by development in direct control with host cell cytoplasm, diplokaryotic nuclei throughout development and disporous sporogony. The genus Vairimorpha exhibits the same features plus an octoporous sporogony producing uninucleate spores in a sporophorous vesicle. A microsporidium from diamondback moth, Plutella xylostella, falls between Nosema and Vairimorpha in that it initiates but fails to complete the octosporous sequence in this host. The name Vairimorpha imperfecta n.sp. is proposed. Merogony is mainly by formation of buds from multinucleate meronts, the buds remaining attached in chains. Diplokaryotic spores measure 4.3 x 2.0 microns (fresh) and have 15.5 coils of the polar tube in 1 rank. The octosporous sporogony is aborted owing to irregular formation of nuclear spindles, incomplete cytoplasmic fission and bizarre deposition of electron-dense episporontal secretions. Phylogenetic analyses of the sequences of the small subunit rRNA genes of V. imperfecta and of several Nosema and Vairimorpha spp. place V. imperfecta in a clade with Nosema spp. from Lepidoptera rather than in the clade containing the more typical species of Vairimorpha. It is suggested that the ancestors of the Vairimorpha/Nosema complex of species exhibited both disporous and octosporous sporogonies, as does the type species of Vairimorpha, Vairimorpha necatrix. It would follow that true Nosema spp. have lost the ability to express an octosporous sequence and that V. imperfecta is in the process of losing it. It is proposed that the genera Nosema and Vairimorpha be placed in the same family Nosematidae Labbé 1899, rather than in separate families and orders as at present.