Historically, members of the Anopheles gambiae complex from Ethiopia have been identified chromosomally as either A. arabiensis or A. quadriannulatus. Recent collections from the Jimma area in Ethiopia, southwest of Addis Ababa, revealed 29 specimens of A. quadriannulatus based on the standard polymerase chain reaction (PCR) identification method. 'Wild' females were induced to lay eggs and the progeny reared as individual families. Resulting adults were cross-mated to a laboratory colony strain of A. quadriannulatus originating from the Kruger National Park, South Africa. Hybrid progeny were obtained only from the colony female x Ethiopian male cross. This cross produced a female/male sex ratio of 0.48. Male offspring were sterile and ovarian polytene chromosomes from hybrid females showed typical asynapsis as expected in interspecific crosses within the A. gambiae complex. The X chromosomes, although apparently having homosequential banding patterns, were usually totally asynapsed. All autosomes were homosequential. The lack of inversion heterozygotes, in both the wild and hybrid samples, may simply be a reflection of the small sample size. Until such time as the Ethiopian species can be formally described and assigned a scientific name, it is provisionally designated Anopheles quadriannulatus species B because of its close similarity to this species.

Background: Malaria remains a major public health problem in Ethiopia. Pyrethroid-treated mosquito nets are one of the major tools available for the prevention and control of malaria transmission. PermaNet ® is a long-lasting insecticide-treated net (LLIN) recommended by WHO for malaria control.

The status of insecticide susceptibility of Anopheles arabiensis populations was monitored in northern and southern Ethiopia using one-to three-day-old mosquitoes reared from larval collections. Anopheles gambiae complex mosquitoes, identified by PCR as An. arabiensis, were exposed to diagnostic concentrations of DDT, malathion, fenitrothion, bendiocarb, propoxur and deltamethrin according to the standard WHO procedure. A heterogeneous and focal distribution of resistance phenotypes was observed in surveyed districts of the country. In Tach Armacho in northern Ethiopia, only resistance to DDT was detected. In the other two northern sites, An. arabiensis populations were susceptible to fenitrothion, resistant to malathion, propoxur and DDT and showed low levels of survival when exposed to bendiocarb requiring further investigations. In three areas of southern Ethiopia, An. arabiensis was susceptible to bendiocarb, propoxur and malathion, with low levels of survival on fenitrothion needing further confirmation. These samples were resistant to DDT and deltamethrin. Analyses for the knockdown resistant (kdr) mutations showed only the L1014F mutation was present with frequencies ranging from 68 to 100 %. The need for routine monitoring and surveillance as part of an insecticide resistance management programme is highlighted.

Anopheles quadriannulatus Theobald historically has been reported from southern Africa, Zanzibar islands, and Ethiopia. However, based on evidences of genetic incompatibility between crosses of South African and Ethiopian populations, the population from Ethiopia was recently reported as a distinct species designated as An. quadriannulatus sp. B. An. quadriannulatus sp. A, denoted the southern African population. To distinguish the two populations, the IGS (intergenic spacer) region of rDNA was sequenced to design a primer specific for An. quadriannulatus sp. B. A cocktail polymerase chain reaction (PCR) involving Anopheles gambiae Giles universal (UN) primer, the new primer and other primers specific for members of the An. gambiae complex produced the expected diagnostic products for the respective species. Using extracted DNA and crushed body parts as sources of template DNA, this assay was reliably used to identify samples of An. quadriannulatus sp. B.

Abstract. Sibling species A and B of Anopheles quadriannulatus (Theobald) are recognized as allopatric members of the Anopheles gambiae Giles complex of Afrotropical mosquitoes (Diptera: Culicidae). Species A represents An. quadriannulatus sensu stricto, widespread in southern Africa, whereas An. quadriannulatus species B occurs in Ethiopia. Because of difficulty of identification, distribution of An. quadriannulatus sensu lato remains poorly known. Cytotaxonomy and the standard DNA polymerase chain reaction (PCR) assay do not distinguish between species A and B of An. quadriannulatus. By optimizing the standard PCR assay (Scott et al., 1993) for identification of members of the An. gambiae complex, we identified two discriminant fragments of 153 bp and 900 bp from DNA of An. quadriannulatus species B, whereas only the 153 bp fragment was amplified for species A from South Africa. This modified PCR assay can therefore be used to distinguish between species A and B of An. quadriannulatus s.l. as well as other members of the An. gambiae complex.