Population Genomics of Two Closely Related Anhydrobiotic Midges Reveals Differences in Adaptation to Extreme Desiccation.

The sleeping chironomid Polypedilum vanderplanki is capable of anhydrobiosis, a striking example of adaptation to extreme desiccation. Tolerance to complete desiccation in this species is associated with emergence of multiple paralogs of protective genes. One of the gene families highly expressed under anhydrobiosis and involved in this process is protein-L-isoaspartate (D-aspartate) O-methyltransferases (PIMTs). Recently, another closely related midge was discovered, Polypedilum pembai, which is able not only to tolerate desiccation but also to survive multiple desiccation-rehydration cycles. To investigate the evolution of anhydrobiosis in these species, we sequenced and assembled the genome of P. pembai and compared it with P. vanderplanki and also performed a population genomics analysis of several populations of P. vanderplanki and one population of P. pembai. We observe positive selection and radical changes in the genetic architecture of the PIMT locus between the two species, including its amplification in the P. pembai lineage. In particular, PIMT-4, the most highly expressed of these PIMTs, is present in six copies in the P. pembai; these copies differ in expression profiles, suggesting possible sub- or neofunctionalization. The nucleotide diversity of the genomic region carrying these new genes is decreased in P. pembai, but not in the orthologous region carrying the ancestral gene in P. vanderplanki, providing evidence for a selective sweep associated with postduplication adaptation in the former. Overall, our results suggest an extensive relatively recent and likely ongoing adaptation of the mechanisms of anhydrobiosis.

Serological and molecular evidence of chikungunya virus infection among febrile outpatients seeking healthcare in Northern Malawi.

Introduction: Despite global evidence of chikungunya fever (CHIKF) in humans that is caused by chikungunya virus (CHIKV), little is known about the occurrence of CHIKF in Malawi. This study was conducted to determine the seroprevalence of CHIKF and to molecularly confirm the presence of CHIKV ribonucleic acid (RNA) among febrile outpatients seeking health care at Mzuzu Central Hospital in the Northern Region of Malawi. Methods: Enzyme-immunosorbent assay (ELISA) was used to detect the presence or absence of specific antibodies against CHIKV. Reversetranscription polymerase chain reaction (RT-PCR) was conducted on randomly selected anti-CHIKV IgM-positive samples to detect CHIKV RNA. Results: Out of 119 CHIKF suspected samples analyzed, 73 tested positive for anti-CHIKV IgM antibodies, with an overall seroprevalence of 61.3%. Most of the CHIKV infected individuals presented with joint pain, abdominal pain, vomiting and nose bleeding with seroprevalence of 45.2%, 41.1%, 16.4% and 12.3%, respectively. All the randomly selected samples that were positive for CHIKV anti-IgM by ELISAhad detectable CHIKV RNA by RT-PCR. Conclusion: The presence of anti-CHIKV IgM antibodies suggests the presence of recent CHIKV infection. We therefore recommend for the inclusion of CHIKF as the differential diagnosis in febrile ill patients in Mzuzu city, Malawi.

Effects of Indoor Environmental Factors and House Structures on Vaporization of Active Ingredient from Spatial Repellent Devices in Rural Houses in Malawi.

The use of a metofluthrin-impregnated spatial repellent device (MSRD) is a new and effective method for preventing mosquito blood feeding. Indoor environmental factors such as room temperature and ventilation rate are thought to be important for MSRD activity. Measurements of room temperature and vaporization of metofluthrin from MSRD in typical rural metal-roof and thatched-roof houses in southeastern Malawi were conducted. The relationship between house structure and the number of collected Anopheline mosquitoes with and without MSRD treatment was also investigated. The difference between daytime and nighttime room temperature was significantly higher in metal-roof houses than in thatched-roof houses. The vaporization of metofluthrin from the MSRD was not accelerated by the high room temperature, but by the high indoor air flow by ventilation. The number of mosquito collections was significantly higher in thatched-roof houses than in metal-roof houses. MSRD-treated thatched-roof houses have a higher probability of mosquito infestation, but the vaporization of metofluthrin is also higher because of indoor air flow, resulting in a reduction in mosquito numbers. Metal-roof houses with closed eaves reduce the probability of mosquito invasion, and a longer predicted effectiveness occurs with MSRD because of the controlled release of metofluthrin through lower indoor air flow.

DNA barcoding of mosquitoes collected through a nationwide survey in 2011 and 2012 in Malawi, Southeast Africa.

We conducted a nationwide survey of mosquito distribution in Malawi from November 2011 to April 2012, and from July to September 2012. Using dried specimens of mosquito adults collected during the survey, we analyzed their cytochrome c oxidase subunit I (COI) gene sequences, prepared specimens, and registered the genetic information (658 bp) of 144 individuals belonging to 51 species of 10 genera in GenBank. Using the obtained genetic information, we analyzed the degree of intraspecific variation and investigated the various species from morphological and genetic perspectives. Moreover, we conducted phylogenetic analysis of the medically important species distributed from Africa to Asia and explored their geographical differentiation. Results showed that individuals morphologically classified as Culex univittatus complex included a individual of Cx. perexiguus which, to date, have not been reported in southern Africa. Furthermore, Mansonia uniformis, distributed in Africa and Asia, was revealed to belong to genetically distinct populations, with observed morphological differences of the samples suggesting that they are separate species. The results of genetic analysis further suggested that Cx. ethiopicus is not a synonym of Cx. bitaeniorhynchus, but that it is an independent species; although, in this study, the only definite morphological difference observed was in the shape of the wing scales. Further morphological and genetic investigation of individuals of these species, including larvae, is highly recommended.

Effect of Metofluthrin-Impregnated Spatial Repellent Devices Combined with New Long-Lasting Insecticidal Nets (Olyset® Plus) on Pyrethroid-Resistant Malaria Vectors and Malaria Prevalence: Field Trial in South-Eastern Malawi.

Long-lasting insecticidal nets (LLINs) experience some operational problems that reduce their effectiveness, such as limited spaces for hanging, biting of mosquitoes outdoors, a shift of key biting time from midnight to dawn or dusk, and development of pyrethroid resistance in mosquitoes. The concept of spatial repellency may be a countermeasure to overcome the above issues. The effect of the combined use of metofluthrin-impregnated spatial repellent devices (MSRDs) and LLINs (Olyset® Plus) on malaria prevalence and vector mosquitoes were examined in malaria endemic villages in south-eastern Malawi. The intervention reduced the infection rate in children as well as the number of pyrethroid-resistant vector mosquitoes. To achieve effective malaria control, continued intervention using MSRDs with 2 strips per 10 m2 at 3-month intervals to reduce the density of malaria mosquitoes is recommended.

Karyotypical characteristics of two allopatric African populations of anhydrobiotic Polypedilum Kieffer, 1912 (Diptera, Chironomidae) originating from Nigeria and Malawi.

The African chironomid Polypedilumvanderplanki Hinton, 1951 is the only chironomid able to withstand almost complete desiccation in an ametabolic state known as anhydrobiosis. The karyotypes of two allopatric populations of this anhydrobiotic chironomid, one from Nigeria and another from Malawi, were described according to the polytene giant chromosomes. The karyotype from the Nigerian population was presented as the reference chromosome map for Polypedilumvanderplanki. Both populations, Nigerian and Malawian, showed the same number of chromosomes (2n=8), but important differences were found in the band sequences of polytene chromosomes, and in the number and the arrangement of active regions between the two populations. Such important differences raise the possibility that the Malawian population could constitute a distinct new species of anhydrobiotic chironomid.

Beyond buzzing: mosquito watching stimulates malaria bednet use-a household-based cluster-randomized controlled assessor blind educational trial.

Malaria remains a severe health problem in Sub-Saharan Africa, with approximately one million deaths and 365 million cases each year. In terms of malaria control, insecticide-treated bednets are an effective tool, and many organizations have distributed free or highly subsidized bednets in malaria endemic areas. Nevertheless, some recipients do not use bednets because of social, environmental or cultural factors. Making vulnerable populations aware of the presence of mosquitoes may improve bednet use among people owning but not using a bednet. We hypothesized that showing freshly collected mosquitoes from the vicinity could improve bednet use in households owning but not using bednets. To test this hypothesis, we applied a household-based cluster-randomized controlled assessor blind educational trial. Indirect observation of mosquitoes, via educational leaflets, produced no change in bednet use, while showing freshly captured mosquitoes led to a 13-fold increase in bednet use. Our results suggest that direct observation of freshly captured mosquitoes can encourage bednet use and may potentially improve effective bednet coverage for malaria control and elimination.

Influence on sensitivity to insecticides: a case study of a settled area and a game park in Liwonde.

The close proximity of Liwonde National Park to Liwonde town creates a unique situation of a large human population adjacent to a natural undisturbed animal reserve. The closeness of the two ecosystems has an impact on biology of mosquitoes of the area, such as susceptibility to insecticides. Susceptibility to insecticide was determined using knockdown bioassays. The mosquito, Anopheles gambiae, was exposed to 0.05% deltamethrin and 0.75% permethrin giving LT 50 and LT 90. The LT50 values for A. gambiae from the town was 17.23 minutes and those from the park, 14.7 minutes (p < 0.05). The calculated LT 90 values were 32.8 and 28.3 minutes respectively. These results suggest that human settlements using insecticides in mosquito control reduce susceptibility of mosquitoes to regularly used insecticides such as deltamethrin and permethrin in this study.

Comparison of deltamethrin as indoor residual spray or on insecticide treated nets for mosquito control in Lake Chilwa.

We conducted a study on the control of mosquitos on Chisi Island in Lake Chilwa from August to November, 2006. The aim was to compare the cost and efficacy of deltamethrin, a pyrethroid based insecticide, when used in insecticide treated nets (ITN) and when used in indoor residual spray (IRS). Thirty village huts were enrolled in the study. Fifteen were systematically selected in a stratified manner and sprayed with deltamethrin following manufacturers' standard application procedures of 0.02g/m2. The remaining fifteen were provided with ITNs. In both groups deltamethrin KO tabs were used. Pyrethroid knockdown (PKD) spray was used for indoor rest captures in the houses monthly for three months. Houses treated with IRS had significantly reduced number of mosquitoes resting indoors than houses provided with nets (p<0.05). Based on the prevailing market price of MK550 each, the cost of five nets was calculated as MK2750.00 per house hold of five compared to five 20 g knock-out tablets costing MK300.00 using IRS. The cost of IRS is 10 times cheaper than ITN. These results suggest that it is cheaper and more effective to use deltamethrin in IRS than in ITN.