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Background: To date there has been no evidence of mosquito-borne virus transmission of public health concern in the UK, despite the occurrence of more than 30 species of mosquito, including putative vectors of arboviruses. The saltmarsh mosquito Ochlerotatus detritus [syn. Aedes (Ochlerotatus) detritus] is locally common in parts of the UK where it can be a voracious feeder on people., Methods: Here, we assess the competence of O. detritus for three major arboviruses: dengue virus (DENV), chikungunya virus (CHIKV) and West Nile virus (WNV) using adult mosquitoes reared from wild, field-obtained immatures., Results: We demonstrate laboratory competence for WNV at 21 °C, with viral RNA detected in the mosquito's saliva 17 days after oral inoculation. By contrast, there was no evidence of laboratory competence of O. detritus for either DENV or CHIKV., Conclusions: To our knowledge, this is the first study to demonstrate competence of a UK mosquito for WNV and confirms that O. detritus may present a potential risk for arbovirus transmission in the UK and that further investigation of its vector role in the wild is required.

Background: The Republic of Yemen has the highest incidence of malaria in the Arabian Peninsula, yet little is known of its vectors or transmission dynamics., Methods: A 24-month study of the vectors and related epidemiological aspects of malaria transmission was conducted in two villages in the Taiz region in 2004-2005., Results: Cross-sectional blood film surveys recorded an overall malaria infection rate of 15.3 % (250/1638), with highest rates exceeding 30 % in one village in May and December 2005. With one exception, Plasmodium malariae, all infections were P. falciparum. Seven Anopheles species were identified among 3407 anophelines collected indoors using light traps (LT) and pyrethrum knockdown catches (PKD): Anopheles arabiensis (86.9 %), An. sergentii (9 %), An. azaniae, An. dthali, An. pretoriensis, An. coustani and An. algeriensis. Sequences for the standard barcode region of the mitochondrial COI gene confirmed the presence of two morphological forms of An. azaniae, the typical form and a previously unrecognized form not immediately identifiable as An. azaniae. ELISA detected Plasmodium sporozoites in 0.9 % of 2921 An. arabiensis (23 P. falciparum, two P. vivax) confirming this species as the primary malaria vector in Yemen. Plasmodium falciparum sporozoites were detected in An. sergentii (2/295) and a single female of An. algeriensis, incriminating both species as malaria vectors for the first time in Yemen. A vector in both wet and dry seasons, An. arabiensis was predominantly anthropophilic (human blood index = 0.86) with an entomological inoculation rate of 1.58 infective bites/person/year. Anopheles sergentii fed on cattle (67.3 %) and humans (48.3; 20.7 % mixed both species), but only 14.7 % were found in PKDs, indicating predominantly exophilic behaviour. A GIS analysis of geographic and socio-economic parameters revealed that An. arabiensis were significantly higher (P < 0.001) in houses with televisions, most likely due to the popular evening habit of viewing television collectively in houses with open doors and windows., Conclusions: The predominantly indoor human biting vectors recorded in this study could be targeted effectively with LLINs, indoor residual spraying and/or insecticide-treated window/door curtains reinforced by education to instil a perception that effective and affordable malaria prevention is achievable.

Dengue prevention efforts rely on control of virus vectors. We investigated use of insecticide-treated screens permanently affixed to windows and doors in Mexico and found that the screens significantly reduced infestations of Aedes aegypti mosquitoes in treated houses. Our findings demonstrate the value of this method for dengue virus vector control.

Despite decades of research, there is still no agreement on which indices of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) presence and abundance better quantify entomological risk for dengue. This study reports the results of a multi-scale, cross-sectional entomological survey carried out in 1160 households in the city of Merida, Mexico to establish: (a) the correlation between levels of Ae. aegypti presence and abundance detected with aspirators and ovitraps; (b) which immature and egg indices correlate with the presence and abundance of Ae. aegypti females, and (c) the correlations amongst traditional Aedes indices and their modifications for pupae at the household level and within medium-sized geographic areas used for vector surveillance. Our analyses show that ovitrap positivity was significantly associated with indoor adult Ae. aegypti presence [odds ratio (OR) = 1.50; P = 0.03], that the presence of pupae is associated with adult presence at the household level (OR = 2.27; P = 0.001), that classic Aedes indices are informative only when they account for pupae, and that window screens provide a significant level of protection against peridomestic Ae. aegypti (OR = 0.59; P = 0.02). Results reinforce the potential of using both positive collections in outdoor ovitraps and the presence of pupae as sensitive indicators of indoor adult female presence., (© 2014 The Royal Entomological Society.)

Background: In order to sustain the gains achieved by current malaria control strategies, robust surveillance systems that monitor dynamics of vectors and their roles in malaria transmission over time are essential. This longitudinal study demonstrates the trends in malaria vector dynamics and their relative contribution to malaria transmission in hyperendemic transmission settings in Tanzania., Methods: The study was conducted in two villages within the Kilombero Valley, in rural Tanzania for five consecutive years (2008-2012). Seventy-two houses were selected per village and each house was sampled for mosquitoes monthly using a CDC light trap. Collected mosquitoes were assessed for species identity and sporozoite infection status using PCR and ELISA, respectively. Anopheles funestus and Anopheles arabiensis susceptibility to insecticides was assessed using WHO guidelines., Results: A total of 100,810 malaria vectors were collected, of which 76% were Anopheles gambiae s. l. and 24% were An. funestus. Of all An. funestus samples that amplified with PCR (n = 2,737), 97% were An. funestus s.s., 2% were Anopheles rivorulum and 1% Anopheles leesoni. Whereas for An. gambiae s.l. (n = 8,117), 93% were An. arabiensis and 7% were Anopheles gambiae s.s. The proportion of An. gambiae s.s. identified by PCR (2,924) declined from 0.2% in the year 2008 to undetectable levels in 2012. Malaria transmission intensity significantly decreased from an EIR of 78.14 infectious bites/person/year in 2008 to 35 ib/p/yr in 2011 but rebounded to 226 ib/p/yr in 2012 coinciding with an increased role of An. funestus in malaria transmission. Insecticide susceptibility tests indicated high levels of resistance in An. funestus against deltamethrin (87%), permethrin (65%), lambda cyhalothrin (74%), bendiocarb (65%), and DDT (66%). Similarly, An. arabiensis showed insecticide resistance to deltamethrin (64%), permethrin (77%) and lambda cyhalothrin (42%) in 2014., Conclusion: The results indicate the continuing role of An. arabiensis and the increasing importance of An. funestus in malaria transmission, and pyrethroid resistance development in both species. Complementary vector control and surveillance tools are needed that target the ecology, behaviour and insecticide resistance management of these vector species, in order to preserve the efficacy of LLINs.

Background: Despite doubts about methods used and the association between vector density and dengue transmission, routine sampling of mosquito vector populations is common in dengue-endemic countries worldwide. This study examined the evidence from published studies for the existence of any quantitative relationship between vector indices and dengue cases., Methodology/principal Findings: From a total of 1205 papers identified in database searches following Cochrane and PRISMA Group guidelines, 18 were included for review. Eligibility criteria included 3-month study duration and dengue case confirmation by WHO case definition and/or serology. A range of designs were seen, particularly in spatial sampling and analyses, and all but 3 were classed as weak study designs. Eleven of eighteen studies generated Stegomyia indices from combined larval and pupal data. Adult vector data were reported in only three studies. Of thirteen studies that investigated associations between vector indices and dengue cases, 4 reported positive correlations, 4 found no correlation and 5 reported ambiguous or inconclusive associations. Six out of 7 studies that measured Breteau Indices reported dengue transmission at levels below the currently accepted threshold of 5., Conclusions/significance: There was little evidence of quantifiable associations between vector indices and dengue transmission that could reliably be used for outbreak prediction. This review highlighted the need for standardized sampling protocols that adequately consider dengue spatial heterogeneity. Recommendations for more appropriately designed studies include: standardized study design to elucidate the relationship between vector abundance and dengue transmission; adult mosquito sampling should be routine; single values of Breteau or other indices are not reliable universal dengue transmission thresholds; better knowledge of vector ecology is required.

Background: Malaria vector control strategies that target adult female mosquitoes are challenged by the emergence of insecticide resistance and behavioural resilience. Conventional larviciding is restricted by high operational costs and inadequate knowledge of mosquito-breeding habitats in rural settings that might be overcome by the juvenile hormone analogue, Pyriproxyfen (PPF). This study assessed the potential for Anopheles arabiensis to pick up and transfer lethal doses of PPF from contamination sites to their breeding habitats (i.e. autodissemination of PPF)., Methods: A semi-field system (SFS) with four identical separate chambers was used to evaluate PPF-treated clay pots for delivering PPF to resting adult female mosquitoes for subsequent autodissemination to artificial breeding habitats within the chambers. In each chamber, a tethered cow provided blood meals to laboratory-reared, unfed female An. arabiensis released in the SFS. In PPF-treated chambers, clay pot linings were dusted with 0.2 - 0.3 g AI PPF per pot. Pupae were removed from the artificial habitats daily, and emergence rates calculated. Impact of PPF on emergence was determined by comparing treatment with an appropriate control group., Results: Mean (95% CI) adult emergence rates were (0.21 ± 0.299) and (0.95 ± 0.39) from PPF-treated and controls respectively (p < 0.0001). Laboratory bioassay of water samples from artificial habitats in these experiments resulted in significantly lower emergence rates in treated chambers (0.16 ± 0.23) compared to controls 0.97 ± 0.05) (p < 0.0001). In experiments where no mosquitoes introduced, there were no significant differences between control and treatment, indicating that transfer of PPF to breeding sites only occurred when mosquitoes were present; i.e. that autodissemination had occurred. Treatment of a single clay pot reduced adult emergence in six habitats to (0.34 ± 0.13) compared to (0.98 ± 0.02) in the controls (p < 0.0001), showing a high level of habitats coverage amplification of the autodissemination event., Conclusion: The study provides proof of principle for the autodissemination of PPF to breeding habitats by malaria vectors. These findings highlight the potential for this technique for outdoor control of malaria vectors and call for the testing of this technique in field trials.

The efficacy of insecticide-treated window curtains (ITCs) for dengue vector control was evaluated in Thailand in a cluster-randomized controlled trial. A total of 2,037 houses in 26 clusters was randomized to receive the intervention or act as control (no treatment). Entomological surveys measured Aedes infestations (Breteau index, house index, container index, and pupae per person index) and oviposition indices (mean numbers of eggs laid in oviposition traps) immediately before and after intervention, and at 3-month intervals over 12 months. There were no consistent statistically significant differences in entomological indices between intervention and control clusters, although oviposition indices were lower (P < 0.01) in ITC clusters during the wet season. It is possible that the open housing structures in the study reduced the likelihood of mosquitoes making contact with ITCs. ITCs deployed in a region where this house design is common may be unsuitable for dengue vector control.

As vector control of dengue typically targets individual households to eliminate breeding sites or control adult mosquitoes during epidemics, public spaces are frequently neglected. To investigate the importance of such places as sources of dengue vectors, a study of Aedes aegypti productivity in a cemetery in northern Venezuela, using standard entomological indices for assessing dengue vector infestations, was carried out in the wet season in May 2003. Containers were found on 72.8% of graves; 44% of the containers held water and in 46.9% of these we found A. aegypti larvae and/or pupae. The average density of Aedes-infested containers was 39 per hectare. There were no significant differences in infestation rates between container types (vases, planters or others) or materials (cement, plastic, glass, etc.). Containers with 1-5 l of water held the greatest proportion of the pupae found in the cemetery (46.7%); containers with <100 ml of water contained no pupae. The mean number of pupae produced in the cemetery was estimated at 4185 pupae/ha per 48 h and the daily output of potential vector mosquitoes from the entire cemetery was calculated at approximately 3000 females per day. These mosquitoes presumably left the cemetery to feed in the nearby communities, thus thwarting the environmental management and health education programmes that had reduced household dengue vector infestations. The study shows the importance of public places as sources of dengue vectors in urban areas, and the need to include such areas in vector control programmes.

Background: A single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. The screening is commonly performed using a multiplex Polymerase Chain Reaction (PCR) which, since it is reliant on a single nucleotide polymorphism, can be unreliable. Here we present a reliable and potentially high throughput method for screening An. gambiae for the kdr mutation., Methods: A Hot Ligation Oligonucleotide Assay (HOLA) was developed to detect both the East and West African kdr alleles in the homozygous and heterozygous states, and was optimized for use in low-tech developing world laboratories. Results from the HOLA were compared to results from the multiplex PCR for field and laboratory mosquito specimens to provide verification of the robustness and sensitivity of the technique., Results and Discussion: The HOLA assay, developed for detection of the kdr mutation, gives a bright blue colouration for a positive result whilst negative reactions remain colourless. The results are apparent within a few minutes of adding the final substrate and can be scored by eye. Heterozygotes are scored when a sample gives a positive reaction to the susceptible probe and the kdr probe. The technique uses only basic laboratory equipment and skills and can be carried out by anyone familiar with the Enzyme-linked immunosorbent assay (ELISA) technique. A comparison to the multiplex PCR method showed that the HOLA assay was more reliable, and scoring of the plates was less ambiguous., Conclusion: The method is capable of detecting both the East and West African kdr alleles in the homozygous and heterozygous states from fresh or dried material using several DNA extraction methods. It is more reliable than the traditional PCR method and may be more sensitive for the detection of heterozygotes. It is inexpensive, simple and relatively safe making it suitable for use in resource-poor countries.

An investigation of lymphatic filariasis vectors in Malawi is reported. Anopheles funestus, A. arabiensis, and A. gambiae sensu stricto had high rates of filarial infection (2.2-3.1%) and carried infective larvae. Anopheles funestus was the predominant species collected (77.6%) and was the primary vector during the study period of April to May 2002.

Onchocerciasis is endemic on the island of Bioko, Equatorial Guinea, where it is transmitted by the 'Bioko form' of the Simulium damnosum complex, a cytospecies unique to the island. To determine the distribution of vector breeding, three dry season and two wet season expeditions were made in 1989, 1996 and 1997, and 226 of the island's 247 rivers (91.5%) were visited. Of these 226 rivers, 130 (58%) were flowing during the dry season, forty-five (20%) supported aquatic stages of Simuliidae of any species and twenty-five (11%) contained larvae or pupae of the S. damnosum complex. The twenty-one rivers not prospected were in the mountainous south of the island, where an additional seventeen rivers were reached but not satisfactorily prospected. Of these thirty-eight rivers, twenty-nine were considered highly likely to support vector breeding, bringing the total number of rivers which could harbour the vector during the dry season to fifty-four (21.9% of the island's total). Breeding was believed to be limited to river stretches below 1000 m altitude, and during the dry season the total length of those stretches which could support breeding on Bioko was estimated to be 1020 km. A combination of factors, including low river discharges during the dry season, the relatively low water temperature on Bioko, the suitability of limited stretches of most rivers as vector breeding sites and the close proximity of many rivers within a small geographical area, render the vector vulnerable to eradication by aerial treatment of rivers with insecticide. The isolation of the Bioko form of the S. damnosum complex suggests that reinvasion following treatment would be unlikely, and eradication of the vector might be achieved by a dry season larviciding programme in one or two years.

The transmission of bovine Onchocerca spp. in Britain was investigated by studying the Ceratopogonidae and Simuliidae which attacked bait cattle between April and October at two sites in North Wales, where either Onchocerca lienalis and O. gutturosa (Cynwyd), or O. lienalis alone (Pentrefoelas) occurred. Based on the relative abundance of each species, their seasonal variation in abundance and preferential feeding sites on cattle, the presence of Onchocerca sp. infective larvae and the development of patent infections in bait cattle, vectors were identified as Simulium ornatum s.1. at Pentrefoelas and S. reptans at Cynwyd (0.5% and 1.5% of parous flies infective, respectively). There was no significant difference between the lengths of the infective larvae from vectors at either site (Pentrefoelas: mean = 543.5 microns, SD = 29.45; Cynwyd: mean = 550.86 microns, SD = 35.26; p > 0.1), which were consistent with descriptions of O. lienalis. The identity of a vector for O. gutturosa was not determined and the role of certain candidates is discussed. In this respect, the absence of Culicoides nubeculosus was notable. Transmission of Onchocerca sp. was limited to a short period of about 8 weeks, by a combination of vector abundance and parasite uptake from dermal skin layers, and a mean daily temperature requirement of 15 degrees C or more for complete parasite development in the vector. The effect of this temperature restriction in relation to the geographical distribution of the two species is discussed.

Previous studies of bovine Onchocerca spp. in cattle in Sierra Leone indicated that only O. gutturosa was transmitted in the forest zone at high intensity. To determine its vector(s) and the extent to which Onchocerca-like infections in Simulium damnosum were likely to be of bovine origin, three lines of investigation were pursued. Firstly, a study was made of the biting flies attacking an ox bait animal over a 14-month period at Njala University Campus, near Bo. Secondly, attempts were made to infect the dominant local forest cytospecies of S. damnosum s.l. with O. gutturosa by feeding them on an infected ox under a bed-net. Thirdly, S. damnosum s.l. were infected by intra-thoracic injection of O. gutturosa microfilariae (mff). In 113 collections made at dawn and dusk at weekly intervals from the ox bait, 624 simuliids and 7740 Culicoides spp. were collected. Almost all the simuliids were S. damnosum s.l. which, on the basis of iso-enzyme examination and knowledge of local breeding sites, were identified as S. soubrense 'B'. Although this cytospecies fed readily on the ox at ventral sites where O. gutturosa mff occurred and the bed-net experiments showed that 16.1% of engorged S. soubrense 'B' ingested an average of 3.3 O. gutturosa mff each, no development occurred. The refractoriness of S. damnosum s.l. to O. gutturosa was confirmed by the intra-thoracic injection experiments. The predominant Culicoides spp. attacking the ox bait were C. krameri, C. trifasciellus and C. fulvithorax, with smaller numbers of C. schultzei. In 5803 dissected Culicoides spp., natural infections of Onchocerca-like larvae, presumed to be O. gutturosa, were found in 0.3% of C. fulvithorax, 0.1% of C. trifasciellus and 0.06% of C. krameri. It was concluded that, in the forest zone of Sierra Leone, S. damnosum s.l. is not a vector of O. gutturosa and all Onchocerca-like larvae in S. damnosum are likely to be O. volvulus, while the natural vectors of O. gutturosa are the Culicoides species C. fulvithorax, C. trifasciellus and C. krameri.

A collapsible animal-type silhouette trap was designed to catch zoophilic female blackflies (Diptera: Simuliidae) which feed on the head or ventral surface of cattle. The trap was assessed for its ability to sample attacking blackflies by comparison with simultaneous manual catches from a bait cow, in an enzootic bovine onchocerciasis area in North Wales. In thirteen 1-hour collections, the trap provided a representative sample of the blackfly population in terms of the relative abundance of species. There was a strong correlation between the catch sizes from both methods (r = +0.73), and the body site feeding preferences for all species were similar with both trap and cow. There was no significant difference between the Onchocerca spp. infection rates of flies caught by either method.

As part of a study to assess to what extent Onchocerca-like infections in S. damnosum s.l. in the forest zone of Sierra Leone were likely to be of bovine origin, Onchocerca infections were studied by post mortem examination of cattle killed at Bo abattoir and by examination of skin biopsies from cattle born and reared at Njala, in the forest zone, and Musaia, in the savanna zone. Post mortem studies of 45 cattle which had originated from northern Sierra Leone and Guinea revealed a prevalence of infection with microfilariae (mff) of O. gutturosa of 87%, O. ochengi 18% and O. armillata 7%. Examination of indigenous cattle revealed transmission of O. gutturosa in both forest (Njala) and savanna (Musaia) areas, and age prevalence analysis at Njala indicated a high intensity of transmission. Autochthonous transmission of O. armillata occurred at Njala at low intensity. Onchocerca gutturosa from Sierra Leone resembled U.K. isolates of O. gutturosa in terms of glucose phosphate isomerase electrophoretic mobility and acid phosphatase activity of in situ-stained mff. Hide mapping of O. gutturosa revealed skin concentrations of mff dorsally at the withers area and also ventrally at the dewlap, brisket and upper forelegs. It was concluded that only O. gutturosa was transmitted at high intensity to cattle in the forest zone of Sierra Leone. Hence, subsequent studies were directed towards determining the vector of O. gutturosa.

Laboratory reared nulliparous female flies of six temperate species of Simuliidae were examined for their susceptibility to infection with Onchocerca ochengi by intrathoracic injection of cryopreserved skin microfilariae obtained from cattle in Mali. Three species (S. equinum, S. ornatum and S. erythrocephalum) supported development to the infective stage, one species (S. variegatum) allowed partial development and the remaining two species (S. reptans and S. aureum) were insusceptible to infection. The most suitable surrogate vectors were S. equinum and S. ornatum which had survival rates of 44% and 49%, proportions of microfilariae developing to third stage larvae of 6.4% and 3%, and infection rates with infective larvae of 13.5% and 14% respectively. O. volvulus infective larvae, produced by intrathoracic microfilarial injection in S. ornatum, were 586-760 microns (mean 687 microns) long and were significantly shorter (p less than 0.02) than the O. ochengi infective larvae (645-880 microns, mean 756 microns). No constant differences in the posterior or anterior morphology, or in the acid phosphatase staining patterns between O. ochengi and O. volvulus, were seen. These results raise the possibility that the presence of O. ochengi in a population of infective larvae from vector flies in endemic onchocerciasis zones might be identifiable on the basis of their length alone.