Parathelohania iranica sp. nov. (Microsporidia: Amblyosporidae) infecting malaria mosquito Anopheles superpictus (Diptera: Culicidae): Ultrastructure and molecular characterization.

Microsporidia are common pathogens of insects and sometimes are considered as a candidate in the biological control of mosquitoes. Recently a microsporidium infection was discovered in Anopheles superpictus (Diptera: Culicidae) larvae, in Iran. The responsible agent belonged to the genus Parathelohania (Microsporidia: Amblyosporidae). This study has been carried out to identify its identity at the species level. Fresh infected larvae were collected from the type locality, Kiar district, in Chahar Mahal and Bakhtiari province, at the central western of Iran. Superficial and the internal ultrastructure of the recovered spores were explored by scanning and transmission electron microscopy, respectively. Molecular techniques were also employed to amplify parts of its ssu rDNA. The obtained data were compared with the available information of congener species and other closely related microsporidia to elucidate evolutionary relationship. A small apical depression and two posterolateral ridges extending backward from a pear shaped anterior body mass were notable under scanning electron microscopy. Transmission electron microscopy revealed 2 broad and 3-4 narrow coils in the either side of spores, respectively. The sequence of a 1062 nucleotide fragment of ssu rDNA was determined by means of PCR technique. This study indicates that the microsporidium infecting An. superpictus differs from other previously described species in the genus Parathelohania. It means that the microsporidium infecting An. superpictus is a new species and hereby it is called Parathelohania iranica. Further work is necessary to clarify its life cycle and probable value in the biological control of mosquitoes.

Factors affecting horizontal transmission of the microsporidium Amblyospora albifasciati to its intermediate copepod host Mesocyclops annulatus.

Factors that directly impact horizontal transmission of the microsporidium Amblyospora albifasciati to its intermediate copepod host, Mesocyclops annulatus were examined in laboratory bioassays. Results were evaluated in relation to life history strategies that facilitate persistence of the parasite in natural populations of its definitive mosquito host, Ochlerotatus albifasciatus. A moderately high quantity of meiospores from mosquito larvae was required to infect adult female copepods; the IC50 was estimated at 3.6 x 10(4) meiospores/ml. Meiospore infectivity following storage at 25 degrees C was detected up to 30 days, while meiospores stored at 4 degrees C remained infectious to copepods for 17 months with virtually no decline in infectivity. Uninfected female M. annulatus are long-lived; no appreciable mortality was observed in field-collected individuals for 26 days, with a few individuals surviving up to 70 days. The pathological impact of A. albifasciati infection on M. annulatus resulted in a 30% reduction in survivorship after 7 days followed by gradual progressive mortality with no infected individuals surviving more than 40 days. This moderate level of pathogenicity allows for a steady continual release of spores into the environment where they may be ingested by mosquito larvae. Infected female copepods survived in sediment under conditions of desiccation up to 30 days, thus demonstrating their capacity to function as a link for maintaining A. albifasciati between mosquito generations following periods of desiccation. The susceptibility of late stage copepodid M. annulatus to meiospores of A. albifasciati and subsequent transstadial transmission of infection to adult females was established.

Epizootiological studies of Amblyospora camposi (Microsporidia: Amblyosporidae) in Culex renatoi (Diptera: Culicidae) and Paracyclops fimbriatus fimbriatus (Copepoda: Cyclopidae) in a bromeliad habitat.

The epizootiology of Amblyospora camposi was studied in a natural population of Culex renatoi, a bromeliad-inhabiting mosquito, and its intermediate host, Paracyclops fimbriatus fimbriatus, over a 2-year period. Twenty Eryngium cabrerae plants were sampled monthly from January 2003 to January 2005 and the prevalence of A. camposi in P.f. fimbriatus and Cx. renatoi populations was determined. The monthly prevalence rates of meiospore infections in Cx. renatoi larvae never exceeded 5.5% and was detected in 50% of the monthly samples. Meiospores were available in plants over the course of the study at a mean concentration of 2 x 10(4) meiospores/ml. Within each plant the parasite was maintained by horizontal transmission. P.f. fimbriatus with vegetative stages and mature spores were found regularly in bromeliads suggesting efficient meiospore infectivity to field copepod populations. The mean concentration of spores from copepods found in plants was 8 x 10(2) spores/ml. Infections in copepods were detected in 54% of the monthly samples with a prevalence rate ranging from 0.55 to 17.4% and an overall average of 5.1%. Vegetative stages in fourth instar mosquito larvae (probably derived from the horizontal pathway via spores formed in copepods) were detected in 12.5% of the monthly samples with an overall prevalence rate of 1.1%. Infections in female and male adults were detected in 20.8% of the monthly samples with an overall average of 4.1% and 6.8%, respectively.