The effects of pyrethroid resistance on transcription of metabolic enzymes in a major African Malaria vector, Anopheles funestus

Anopheles funestus is a major vector of malaria in the southern African region. Insecticide resistance to pyrethroid and carbamate insecticides has been recorded in populations of this species in South Africa and Mozambique. This study aimed to determine the relationship between pyrethroid resistance and gene expression of two closely related genes, CYP6P9 and CYP6P13, by age and sex in a resistant strain An. funestus from southern Africa, FUMOZ-R. The insecticide susceptibility assays showed that percentage survival in both FUMOZ-R sexes significantly decreased as age increased. The mRNA expressions of CYP6P9 and CYP6P13 were higher in FUMOZ-R relative to the insecticide susceptible strain (FANG). The expression of permethrin resistance varies with age in An. funestus FUMOZ-R. The results indicate that other genes may also be involved in insecticide resistance. In addition to this, the expression profile of other metabolic genes involved in insecticide resistance was also investigated. A microarray based approach was used to identify genes differentially expressed in FUMOZ-R and FANG. As the full set of detoxification genes in An. funestus are unknown, this study investigated the utility of the An. gambiae detox chip to screen for differentially expressed detoxification genes in An. funestus. After optimization of the hybridisation conditions, over 90% of the probes showed a positive signal. Only three genes were significantly (P<0.001) differentially expressed in the females, CYP6P9, COI and CYP6M7. The same genes were also significantly differentially expressed in the adult males, together with an additional 21 transcripts. The third part of this study investigated the gene expression in the first instar, fourth instar and 3-day old adults in FUMOZ-R using the An. gambiae detox chip. The variation in metabolic enzyme gene transcription at the different developmental stages in An. funestus are not known. The identification of differentially transcribed genes at the different life stages provides some insight into the role and function of these genes. A large number of cytochrome P450s (monooxygenases), esterases, glutathione S-transferases (GSTs) and other additional genes were differentially expressed in all life stages. This study provided vital information regarding genes potentially involved in pyrethroid resistant and is the first to provide metabolic or detoxifying transcription gene information in An. funestus

Detoxification enzymes associated with insecticide resistance in laboratory strains of Anopheles arabiensis of different geographic origin

Background The use of insecticides to control malaria vectors is essential to reduce the prevalence of malaria and as a result, the development of insecticide resistance in vector populations is of major concern. Anopheles arabiensis is one of the main African malaria vectors and insecticide resistance in this species has been reported in a number of countries. The aim of this study was to investigate the detoxification enzymes that are involved in An. arabiensis resistance to DDT and pyrethroids. Methods The detoxification enzyme profiles were compared between two DDT selected, insecticide resistant strains of An. arabiensis, one from South Africa and one from Sudan, using the An. gambiae detoxification chip, a boutique microarray based on the major classes of enzymes associated with metabolism and detoxification of insecticides. Synergist assays were performed in order to clarify the roles of over-transcribed detoxification genes in the observed resistance phenotypes. In addition, the presence of kdr mutations in the colonies under investigation was determined. Results The microarray data identifies several genes over-transcribed in the insecticide selected South African strain, while in the Sudanese population, only one gene, CYP9L1, was found to be over-transcribed. The outcome of the synergist experiments indicate that the over-transcription of detoxification enzymes is linked to deltamethrin resistance, while DDT and permethrin resistance are mainly associated with the presence of the L1014F kdr mutation. Conclusions These data emphasise the complexity associated with resistance phenotypes and suggest that specific insecticide resistance mechanisms cannot be extrapolated to different vector populations of the same species

DDT and pyrethroid resistance in Anopheles arabiensis from South Africa

BACKGROUND: Pyrethroid resistance has been well documented in Anopheles arabiensis, one of the major African malaria vectors, and the predominant malaria vector in South Africa. METHODS: In this study, the genetic basis of pyrethroid resistance in a selected laboratory strain of An. arabiensis from South Africa was investigated using a custom-made microarray, known as the An. gambiae detoxification chip. RESULTS: A large number of P450 genes were over-transcribed, as well as a suite of redox genes and glutathione S-transferases. The five genes that showed the highest level of gene transcription when compared with an insecticide susceptible strain were: CYP6AG2, CYPZ1, TPX2, CYPZ2 and CYP6P1. CONCLUSIONS: Permethrin resistance in South African An. arabiensis is associated with increased transcription of multiple genes, and a large proportion of these genes were also previously recorded as over-transcribed in another An. arabiensis strain selected for resistance to DDT with cross-resistance to deltamethrin. The deltamethrin resistance developed de novo in the DDT-selected strain and is most likely due to increased transcription of those genes associated with DDT resistance. However, of particular interest was the fact that the strain selected for resistance to pyrethroids did not develop de novo resistance to DDT. These differences are compared and discussed.National Research Foundation Scarce Skills Scholarships and DAAD (LN), National Research Foundation and the National Health Laboratory Service-Research Trust to LLK, partial funding from the Department of Science and Technology, National Research Foundation Research Chair Initiative grant to Maureen Coetzee.http://www.parasitesandvectors.com/content/6/1/229am201

DDT and pyrethroid resistance in Anopheles arabiensis from South Africa

BACKGROUND: Pyrethroid resistance has been well documented in Anopheles arabiensis, one of the major African malaria vectors, and the predominant malaria vector in South Africa. METHODS: In this study, the genetic basis of pyrethroid resistance in a selected laboratory strain of An. arabiensis from South Africa was investigated using a custom-made microarray, known as the An. gambiae detoxification chip. RESULTS: A large number of P450 genes were over-transcribed, as well as a suite of redox genes and glutathione S-transferases. The five genes that showed the highest level of gene transcription when compared with an insecticide susceptible strain were: CYP6AG2, CYPZ1, TPX2, CYPZ2 and CYP6P1. CONCLUSIONS: Permethrin resistance in South African An. arabiensis is associated with increased transcription of multiple genes, and a large proportion of these genes were also previously recorded as over-transcribed in another An. arabiensis strain selected for resistance to DDT with cross-resistance to deltamethrin. The deltamethrin resistance developed de novo in the DDT-selected strain and is most likely due to increased transcription of those genes associated with DDT resistance. However, of particular interest was the fact that the strain selected for resistance to pyrethroids did not develop de novo resistance to DDT. These differences are compared and discussed.National Research Foundation Scarce Skills Scholarships and DAAD (LN), National Research Foundation and the National Health Laboratory Service-Research Trust to LLK, partial funding from the Department of Science and Technology, National Research Foundation Research Chair Initiative grant to Maureen Coetzee.http://www.parasitesandvectors.com/content/6/1/229am201

An African perspective on the genetic diversity of Toxoplasma gondii: A systematic review

The study of Toxoplasma gondii genotypes is beneficial for detecting strains linked to increased disease severity and uncovering the processes involved in the transmission and distribution of this zoonotic parasite. A systematic review of literature was conducted to investigate the present status of T. gondii genetic diversity in African countries and among host species on the continent. Data from the results in the included studies were sorted, reviewed and descriptively analysed using tables, graphs and maps. Results indicate that there is a relative amount of genetic diversity with a clear difference in the population structure between geographical regions and the propensity for unique and regional genotypes to be predominant in tropical rainforest biomes, near the equator. From a clinical perspective, connections between specific T. gondii genotypes and disease manifestations were found. Theories are outlined on the dissemination of African T. gondii genotypes to other continents. The overrepresentation of samples from one geographical area and dissimilar genotyping methodologies creates challenges when concluding on the genetic diversity of T. gondii in Africa. The need for uniform genotyping methods with a continent-wide sampling of an extensive host range involving humans, domestic animals and wildlife is emphasized

Lethal and Pre-Lethal Effects of a Fungal Biopesticide Contribute to Substantial and Rapid Control of Malaria Vectors

Rapidly emerging insecticide resistance is creating an urgent need for new active ingredients to control the adult mosquitoes that vector malaria. Biopesticides based on the spores of entomopathogenic fungi have shown considerable promise by causing very substantial mortality within 7–14 days of exposure. This mortality will generate excellent malaria control if there is a high likelihood that mosquitoes contact fungi early in their adult lives. However, where contact rates are lower, as might result from poor pesticide coverage, some mosquitoes will contact fungi one or more feeding cycles after they acquire malaria, and so risk transmitting malaria before the fungus kills them. Critics have argued that ‘slow acting’ fungal biopesticides are, therefore, incapable of delivering malaria control in real-world contexts. Here, utilizing standard WHO laboratory protocols, we demonstrate effective action of a biopesticide much faster than previously reported. Specifically, we show that transient exposure to clay tiles sprayed with a candidate biopesticide comprising spores of a natural isolate of Beauveria bassiana, could reduce malaria transmission potential to zero within a feeding cycle. The effect resulted from a combination of high mortality and rapid fungal-induced reduction in feeding and flight capacity. Additionally, multiple insecticide-resistant lines from three key African malaria vector species were completely susceptible to fungus. Thus, fungal biopesticides can block transmission on a par with chemical insecticides, and can achieve this where chemical insecticides have little impact. These results support broadening the current vector control paradigm beyond fast-acting chemical toxins

Optimisation of adult Anopheles funestus blood-feeding on an artificial membrane feeding system

Malaria is one of the most severe vector-borne diseases caused by Plasmodium parasites and transmitted by Anopheles mosquitoes. Laboratory-reared anophelines are essential to advance research needed to reduce or eliminate malaria. The success of laboratory rearing as well as studies on parasite-mosquito transmission, is advanced by using an artificial membrane feeding systems. These require the optimisation of mosquito feeding to ensure that an optimal number of mosquitoes feed, thereby enabling successful reproduction or research sample sizes. In this study, various parameters such as the type of artificial membrane, density of adults in the feeding cup, age of the mosquito, duration of starvation, method of starvation, the volume of blood meal, duration of feeding, feeding in the light or dark and the effect of lactic acid were evaluated to determine their impact on the feeding rate of a main African malaria vector, Anopheles funestus. By optimising the artificial membrane feeding parameters, an increase in the feeding rate of the An. funestus mosquitoes was observed. The results obtained from these parameters increased the feeding rate of An. funestus above 50%. However, feeding rates were not significantly increased by the type of membrane, mosquito density, the volume of blood meal, duration of feeding and the addition of lactic acid to the cattle intestine membrane. Therefore, this study provides information on suitable conditions for adult mosquito feeding that allows for successful laboratory rearing and colony maintenance. Furthermore, it provides additional information for research studies that are dependent on blood-feeding, such as transmission blocking studies, endectocide studies etc

An African perspective on the genetic diversity of Toxoplasma gondii: A systematic review

A systematic review on available literature on the genotypes of Toxoplasma gondii that have been found in African countrie