Huntiella decorticans sp. nov. (Ceratocystidaceae) associated with dying Nothofagus in Patagonia.

During a survey of ophiostomatoid fungi in native forests of southern Argentina, several isolates of Huntiella species were obtained from Nothofagus trees. Sequences of multiple gene regions were used to identify these fungi, and their pathogenicity was tested on N. pumilio and N. dombeyi. Phylogenetic analyses revealed a novel taxon described here as H. decorticans sp. nov. Inoculations on N. dombeyi and N. pumilio in the forest showed that H. decorticans is able to produce localized lesions on healthy Nothofagus trees.

Age-related pyrethroid resistance is not a function of P450 gene expression in the major African malaria vector, Anopheles funestus (Diptera: Culicidae).

Anopheles funestus is a major vector of malaria in most of the African region. Resistance to pyrethroid and carbamate insecticides has been recorded in populations of this species in South Africa and Mozambique. The P450 gene, CYP6P9, has been shown to be highly transcribed in a permethrin (pyrethroid)-resistant laboratory strain, FUMOZ-R, originating from southern Mozambique. We examined the relationship between pyrethroid resistance and gene transcription levels of two closely related genes, CYP6P9 and CYP6P13, in FUMOZ-R. Levels of resistance to 0.75% permethrin were determined based on standard WHO insecticide susceptibility assays using females and males of different ages, ranging from 3 to 30 days old. The transcription levels of the two genes were quantified using qPCR for each age cohort. In the WHO insecticide susceptibility assays, survival of both males and females significantly decreased as age increased. Quantitative analysis of the two genes CYP6P9 and CYP6P13 showed the highest levels of expression at 10 days of age. There was no correlation between expression of these two genes and pyrethroid survival by age. We conclude that the resistance of this mosquito strain to permethrin is not directly related to age-mediated differences in CYP6P9 and CYP6P13 expression.

Sequence characterization of cytochrome P450 CYP6P9 in pyrethroid resistant and susceptible Anopheles funestus (Diptera: Culicidae).

Anopheles funestus, one of the main African malaria vectors, caused a major malaria outbreak in South Africa during 1999/2000, even though South Africa had an effective vector control program in place. The outbreak was due to pyrethroid resistant An. funestus invading KwaZulu/Natal. Increased activity of cytochrome P450 (monooxygenase) was responsible for the pyrethroid resistance in this species. A monooxygenase gene, CYP6P9, was highly overexpressed in the pyrethroid-resistant strain compared with a susceptible strain. Characterization of this gene as well as the redox partners involved in the catalytic cycle of P450s was investigated. The full length of the CYP6P9 sequence was isolated, sequenced and compared between the pyrethroid-resistant and -susceptible strains. Sequence identity between the two strains was 99.3%; the sequence differences were mainly outside of the conserved regions. The functional significance is still unknown, but it is feasible that these variations are associated with differences in insecticide metabolism. A second CYP6 gene (CYP6P13) was also isolated; it shared close similarities with CYP6P9. The putative redox partners, cytochrome b(5) (cyt b(5)) and NADPH-cytochrome P450 reductase (CPR), were isolated from An. funestus (resistant strain) and showed high levels of sequence identity to other insect cyt b(5) and CPRs. Isolation of the coding sequences CYP6P9 and its cognate redox partners enables expression of functional recombinant protein for biochemical and structural analysis.

Cytogenetic evidence for a species complex within Anopheles pseudopunctipennis theobald (Diptera: Culicidae).

Anopheles pseudopunctipennis was collected from Acapulco, Mexico and Sallee River, Grenada, West Indies and used in cross-mating experiments. Larvae from the cross, Mexico female X Grenada male, died in the third instar. However, adult progeny were obtained from the reciprocal cross Grenada female x Mexico male. These hybrid males had testes with apparently normal appearance but some without viable sperm. Polytene chromosomes obtained from hybrid females exhibited extensive asynapsis of the X chromosomes. Previously undescribed fixed inversion differences between the two populations were noted on the X chromosome. It is concluded that the two populations belong to different species. The Grenada population is designated An. pseudopunctipennis species C, since it is the third taxon recognized in this species complex.