bims-mosdis 2020-04-19 papers

bims-mosdis

Biomed News

on Mosquito distribution and disease

Issue of 2020–04–19
23 papers selected by
Richard Halfpenny, Staffordshire University

Baseline Susceptibility Status of Florida Populations of Aedes aegypti (Diptera: Culicidae) and Aedes albopictus.
The invasive Asian bush mosquito Aedes japonicus found in the Netherlands can experimentally transmit Zika virus and Usutu virus.
Multiple Wolbachia strains provide comparative levels of protection against dengue virus infection in Aedes aegypti.
The 'lifecycle' of human beings: a call to explore vector-borne diseases from an ecosystem perspective.
Vector competence of Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus mosquitoes for Mayaro virus.
Stable establishment of wMel Wolbachia in Aedes aegypti populations in Yogyakarta, Indonesia.
Importation of dengue, Zika and chikungunya infections in Europe: the current situation in Greece.
A standardised method of marking male mosquitoes with fluorescent dust.
Metagenomic analysis of Aedes aegypti and Culex quinquefasciatus mosquitoes from Grenada, West Indies.
Evolution of kdr haplotypes in worldwide populations of Aedes aegypti: Independent origins of the F1534C kdr mutation.
Longevity of the insecticidal effect of three pyrethroid formulations applied to outdoor vegetation on a laboratory-adapted colony of the Southeast Asian malaria vector Anopheles dirus.
Identification and RNAi Profile of a Novel Iflavirus Infecting Senegalese Aedes vexans arabiensis Mosquitoes.
Spatial modelling of the infestation indices of Aedes aegypti: an innovative strategy for vector control actions in developing countries.
The potential distribution and dynamics of important vectors Culex pipiens pallens and Culex pipiens quinquefasciatus in China under climate change scenarios: An ecological niche modelling approach.
Association between the proportion of Plasmodium falciparum and Plasmodium vivax infections detected by passive surveillance and the magnitude of the asymptomatic reservoir in the community: a pooled analysis of paired health facility and community data.
A 70% Reduction in Mosquito Populations Does Not Require Removal of 70% of Mosquitoes.
Impacts of fluorescent powders on survival of different age cohorts, blood-feeding success, and tethered flight speed of Aedes aegypti (Diptera: Culicidae) females.
Low transmission of Wuchereria bancrofti in cross-border districts of Côte d'Ivoire: A great step towards lymphatic filariasis elimination in West Africa.
Seasonal Changes of Host Use by Culiseta melanura (Diptera: Culicidae) in Central Florida.
Molecular and morphological evidence for sibling species within Anopheles (Anopheles) lindesayi Giles (Diptera: Culicidae) in Bhutan.
Persistence of aerially applied mosquito-pesticide, Naled, in fresh and marine waters.
Not Just from Blood: Mosquito Nutrient Acquisition from Nectar Sources.
High levels of imported asymptomatic malaria but limited local transmission in KwaZulu-Natal, a South African malaria-endemic province nearing malaria elimination.

J Med Entomol. 2020 Apr 17. pii: tjaa068. [Epub ahead of print]

Baseline Susceptibility Status of Florida Populations of Aedes aegypti (Diptera: Culicidae) and Aedes albopictus.

Casey Parker, Daviela Ramirez, Carol Thomas, C Roxanne Connelly.

  Resistance to insecticides used to control mosquito vectors threatens the ability of mosquito-control organizations to protect public health. Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) are invasive species widely distributed throughout Florida and have been implicated in recent epidemics of Zika, dengue, and chikungunya viruses. Knowledge of the susceptibility status of these mosquito species to pyrethroid and organophosphate active ingredients (AIs) is needed to inform product selection and treatment decisions. The susceptibility of 37 Ae. aegypti and 42 Ae. albopictus populations from Florida was assessed in response to six pyrethroid and three organophosphate AIs using the CDC bottle bioassay method. Of all bioassays completed with a pyrethroid AI, 95% for Ae. aegypti and 30% for Ae. albopictus resulted in a resistant outcome. For organophosphate AIs, ~31% of assays conducted for both species were classified as resistant. The highest frequency of susceptibility for both species was observed in response to the organophosphate AI, naled. Lambda-cyhalothrin was the only pyrethroid to result in a susceptible status for Ae. aegypti and also had the highest frequency of susceptibility for Ae. albopictus. Resistance was detected to every AI tested for both Ae. aegypti and Ae. albopictus, but there was a pronounced trend of pyrethroid resistance in Florida populations of Ae. aegypti. The results of this work provide evidence for the need to decrease reliance on pyrethroids and to implement different methods of control of Ae. aegypti in Florida.

Keywords:   Aedes aegypti ; Aedes albopictus ; insecticide resistance; organophosphate; pyrethroid

DOI:  https://doi.org/10.1093/jme/tjaa068
PLoS Negl Trop Dis. 2020 Apr;14(4): e0008217

The invasive Asian bush mosquito Aedes japonicus found in the Netherlands can experimentally transmit Zika virus and Usutu virus.

Sandra R Abbo, Tessa M Visser, Haidong Wang, Giel P Göertz, Jelke J Fros, Marleen H C Abma-Henkens, Corinne Geertsema, Chantal B F Vogels, Marion P G Koopmans, Chantal B E M Reusken, Sonja Hall-Mendelin, Roy A Hall, Monique M van Oers, Constantianus J M Koenraadt, Gorben P Pijlman.

BACKGROUND: The Asian bush mosquito Aedes japonicus is invading Europe and was first discovered in Lelystad, the Netherlands in 2013, where it has established a permanent population. In this study, we investigated the vector competence of Ae. japonicus from the Netherlands for the emerging Zika virus (ZIKV) and zoonotic Usutu virus (USUV). ZIKV causes severe congenital microcephaly and Guillain-Barré syndrome in humans. USUV is closely related to West Nile virus, has recently spread throughout Europe and is causing mass mortality of birds. USUV infection in humans can result in clinical manifestations ranging from mild disease to severe neurological impairments.
METHODOLOGY/PRINCIPAL FINDINGS: In our study, field-collected Ae. japonicus females received an infectious blood meal with ZIKV or USUV by droplet feeding. After 14 days at 28°C, 3% of the ZIKV-blood fed mosquitoes and 13% of the USUV-blood fed mosquitoes showed virus-positive saliva, indicating that Ae. japonicus can transmit both viruses. To investigate the effect of the mosquito midgut barrier on virus transmission, female mosquitoes were intrathoracically injected with ZIKV or USUV. Of the injected mosquitoes, 96% (ZIKV) and 88% (USUV) showed virus-positive saliva after 14 days at 28°C. This indicates that ZIKV and USUV can efficiently replicate in Ae. japonicus but that a strong midgut barrier is normally restricting virus dissemination. Small RNA deep sequencing of orally infected mosquitoes confirmed active replication of ZIKV and USUV, as demonstrated by potent small interfering RNA responses against both viruses. Additionally, de novo small RNA assembly revealed the presence of a novel narnavirus in Ae. japonicus.
CONCLUSIONS/SIGNIFICANCE: Given that Ae. japonicus can experimentally transmit arthropod-borne viruses (arboviruses) like ZIKV and USUV and is currently expanding its territories, we should consider this mosquito as a potential vector for arboviral diseases in Europe.

DOI: https://doi.org/10.1371/journal.pntd.0008217
PLoS Pathog. 2020 Apr 13. 16(4): e1008433

Multiple Wolbachia strains provide comparative levels of protection against dengue virus infection in Aedes aegypti.

Heather A Flores, Jyotika Taneja de Bruyne, Tanya B O'Donnell, Vu Tuyet Nhu, Nguyen Thi Giang, Huynh Thi Xuan Trang, Huynh Thi Thuy Van, Vo Thi Long, Le Thi Dui, Huynh Le Anh Huy, Huynh Thi Le Duyen, Nguyen Thi Van Thuy, Nguyen Thanh Phong, Nguyen Van Vinh Chau, Duong Thi Hue Kien, Tran Thuy Vi, Bridget Wills, Scott L O'Neill, Cameron P Simmons, Lauren B Carrington.

The insect bacterium Wolbachia pipientis is being introgressed into Aedes aegypti populations as an intervention against the transmission of medically important arboviruses. Here we compare Ae. aegypti mosquitoes infected with wMelCS or wAlbB to the widely used wMel Wolbachia strain on an Australian nuclear genetic background for their susceptibility to infection by dengue virus (DENV) genotypes spanning all four serotypes. All Wolbachia-infected mosquitoes were more resistant to intrathoracic DENV challenge than their wildtype counterparts. Blocking of DENV replication was greatest by wMelCS. Conversely, wAlbB-infected mosquitoes were more susceptible to whole body infection than wMel and wMelCS. We extended these findings via mosquito oral feeding experiments, using viremic blood from 36 acute, hospitalised dengue cases in Vietnam, additionally including wMel and wildtype mosquitoes on a Vietnamese nuclear genetic background. As above, wAlbB was less effective at blocking DENV replication in the abdomen compared to wMel and wMelCS. The transmission potential of all Wolbachia-infected mosquito lines (measured by the presence/absence of infectious DENV in mosquito saliva) after 14 days, was significantly reduced compared to their wildtype counterparts, and lowest for wMelCS and wAlbB. These data support the use of wAlbB and wMelCS strains for introgression field trials and the biocontrol of DENV transmission. Furthermore, despite observing significant differences in transmission potential between wildtype mosquitoes from Australia and Vietnam, no difference was observed between wMel-infected mosquitoes from each background suggesting that Wolbachia may override any underlying variation in DENV transmission potential.

DOI: https://doi.org/10.1371/journal.ppat.1008433
Infect Dis Poverty. 2020 Apr 16. 9(1): 37

The 'lifecycle' of human beings: a call to explore vector-borne diseases from an ecosystem perspective.

Olav T Muurlink, Andrew W Taylor-Robinson.

   BACKGROUND: Dengue virus, an Aedes mosquito-borne flavivirus, is associated with close to 400 million reported infections per annum worldwide. Reduction of dengue virus transmission depends entirely on limiting Aedes breeding or preventing adult female contact with humans. Currently, the World Health Organization promotes the strategic approach of integrated vector management in order to optimise resources for mosquito control.
MAIN TEXT: Neglected tropical disease researchers focus on geographical zones where the incidence of clinical cases, and prevalence of vectors, are high. In combatting those infectious diseases such as dengue that affect mainly low-income populations in developing regions, a mosquito-centric approach is frequently adopted. This prioritises environmental factors that facilitate or impede the lifecycle progression of the vector. Climatic variables (such as rainfall and wind speed) that impact the vector's lifecycle either causally or by happenstance also affect the human host's 'lifecycle', but in very different ways. The socioeconomic impacts of the same variables that influence vector control impact host vulnerability but at different points in the human lifecycle to those of the vector. Here, we argue that the vulnerability of the vector and that of the host interact in complex and unpredictable ways that are characteristic of (complex and intransigent) 'wicked problems'. Moreover, they are treated by public health programs in ways that may ignore this complexity. This opinion draws on recent evidence showing that the best climate predictors of the scale of dengue outbreaks in Bangladesh cannot be explained through a simple vector-to-host causal model.
CONCLUSIONS: In mapping causal pathways for vector-borne diseases this article makes a case to elevate the lifecycle of the human host to a level closer in equivalence to that of the vector. Here, we suggest value may be gained from transferring Rittel and Webber's concept of a wicked (social) problem to dengue, malaria and other mosquito-transmitted public health concerns. This would take a 'problem definition' rather than a 'solution-finding' approach, particularly when considering problems in which climate impacts simultaneously on human and vector vulnerability.

Keywords:  Climate change; Dengue; Design thinking; Ecosystem; Mosquito; Social innovation; Vector-borne disease; Wicked problem

DOI:  https://doi.org/10.1186/s40249-020-00653-y
PLoS Negl Trop Dis. 2020 Apr 14. 14(4): e0007518

Vector competence of Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus mosquitoes for Mayaro virus.

Thiago Nunes Pereira, Fabiano Duarte Carvalho, Silvana Faria De Mendonça, Marcele Neves Rocha, Luciano Andrade Moreira.

Newly emerging or re-emerging arthropod-borne viruses (arboviruses) are important causes of human morbidity and mortality worldwide. Arboviruses such as Dengue (DENV), Zika (ZIKV), Chikungunya (CHIKV), and West Nile virus (WNV) have undergone extensive geographic expansion in the tropical and sub-tropical regions of the world. In the Americas the main vectors of DENV, ZIKV, and CHIKV are mosquito species adapted to urban environments, namely Aedes aegypti and Aedes albopictus, whereas the main vector of WNV is Culex quinquefasciatus. Given the widespread distribution in the Americas and high permissiveness to arbovirus infection, these mosquito species may play a key role in the epidemiology of other arboviruses normally associated with sylvatic vectors. Here, we test this hypothesis by determining the vector competence of Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus to Mayaro (MAYV) virus, a sylvatic arbovirus transmitted mainly by Haemagogus janthinomys that has been causing an increasing number of outbreaks in South America, namely in Brazil. Using field mosquitoes from Brazil, female mosquitoes were experimentally infected, and their competence for infection and transmission rates of MAYV was evaluated. We found consistent infection rate for MAYV in Ae. aegypti (57.5%) and Ae. albopictus (61.6%), whereas very low rates were obtained for Cx. quinquefasciatus (2.5%). Concordantly, we observed high potential transmission ability in Ae. aegypti and Ae. albopictus (69.5% and 71.1% respectively), in contrast to Cx. quinquefasciatus, which could not transmit the MAYV. Notably, we found that very low quantities of virus present in the saliva (undetectable by RT-qPCR) were sufficiently virulent to guarantee transmission. Although Ae. aegypti and Ae. albopictus mosquitoes are not the main vectors for MAYV, our studies suggest that these mosquitoes could play a significant role in the transmission of this arbovirus, since both species showed significant vector competence for MAYV (Genotype D), under laboratory conditions.

DOI: https://doi.org/10.1371/journal.pntd.0007518
PLoS Negl Trop Dis. 2020 Apr 17. 14(4): e0008157

Stable establishment of wMel Wolbachia in Aedes aegypti populations in Yogyakarta, Indonesia.

Warsito Tantowijoyo, Bekti Andari, Eggi Arguni, Nida Budiwati, Indah Nurhayati, Iva Fitriana, Inggrid Ernesia, Edwin W Daniwijaya, Endah Supriyati, Dedik H Yusdiana, Munasdi Victorius, Dwi S Wardana, Hilmi Ardiansyah, Riris Andono Ahmad, Peter A Ryan, Cameron P Simmons, Ary A Hoffmann, Edwige Rancès, Andrew P Turley, Petrina Johnson, Adi Utarini, Scott L O'Neill.

The successful establishment of the wMel strain of Wolbachia for the control of arbovirus transmission by Aedes aegypti has been proposed and is being implemented in a number of countries. Here we describe the successful establishment of the wMel strain of Wolbachia in four sites in Yogyakarta, Indonesia. We demonstrate that Wolbachia can be successfully introgressed after transient releases of wMel-infected eggs or adult mosquitoes. We demonstrate that the approach is acceptable to communities and that Wolbachia maintains itself in the mosquito population once deployed. Finally, our data show that spreading rates of Wolbachia in the Indonesian setting are slow which may reflect more limited dispersal of Aedes aegypti than seen in other sites such as Cairns, Australia.

DOI: https://doi.org/10.1371/journal.pntd.0008157
New Microbes New Infect. 2020 May;35 100663

Importation of dengue, Zika and chikungunya infections in Europe: the current situation in Greece.

M Emmanouil, M Evangelidou, A Papa, A Mentis.

  Although several arboviruses-such as dengue (DENV), Zika (ZIKV) and chikungunya (CHIKV) viruses-are not endemic in Europe, they have the potential to emerge following importation of the virus, taking advantage of the favourable climate and ecosystem. DENV, ZIKV and CHIKV are transmitted by Aedes species mosquitoes and are amongst the most common travel-associated arboviruses. Furthermore, they are linked to sporadic, local outbreaks, especially in the southern parts of Europe. In this review we present in brief the DENV, ZIKV and CHIKV cases imported to Greece during the last 6 years (2013-2018), and we describe the recent laboratory data obtained from the Hellenic Pasteur Institute and the National Reference Centre for Arboviruses. We report 21 imported cases of DENV, ZIKV and CHIKV infections in travellers arriving in Greece. The probable origins were south-eastern Asian (71%) and north-central American (29%) countries. Furthermore, we stress the importance of early and accurate diagnosis in spite of a plethora of diagnostic challenges that clinicians and virologists have to face. Altogether, with the authorities' awareness and the preventive measures to be applied, local transmission events can be successfully avoided, especially in summer when the temperature is favourable for mosquito-borne infections.

Keywords:  Aedes; Greece; Zika; arbovirus; chikungunya; dengue; diagnosis; imported; outbreak; prevention

DOI:  https://doi.org/10.1016/j.nmni.2020.100663
Parasit Vectors. 2020 Apr 15. 13(1): 192

A standardised method of marking male mosquitoes with fluorescent dust.

Nicole J Culbert, Maria Kaiser, Nelius Venter, Marc J B Vreysen, Jeremie R L Gilles, Jérémy Bouyer.

   BACKGROUND: Prior to a major release campaign of sterile insects, including the sterile insect technique, male mosquitoes must be marked and released (small scale) to determine key parameters including wild population abundance, dispersal and survival. Marking insects has been routinely carried out for over 100 years; however, there is no gold standard regarding the marking of specific disease-transmitting mosquitoes including Anopheles arabiensis, Aedes aegypti and Aedes albopictus. The research presented offers a novel dusting technique and optimal dust colour and quantities, suitable for small-scale releases, such as mark-release-recapture studies.
METHODS: We sought to establish a suitable dust colour and quantity for batches of 100 male An. arabiensis, that was visible both by eye and under UV light, long-lasting and did not negatively impact longevity. A set of lower dust weights were selected to conduct longevity experiments with both Ae. aegypti and Ae. albopictus to underpin the optimal dust weight. A further study assessed the potential of marked male An. arabiensis to transfer their mark to undusted males and females.
RESULTS: The longevity of male An. arabiensis marked with various dust colours was not significantly reduced when compared to unmarked controls. Furthermore, the chosen dust quantity (5 mg) did not negatively impact longevity (P = 0.717) and provided a long-lasting mark. Dust transfer was found to occur from marked An. arabiensis males to unmarked males and females when left in close proximity. However, this was only noticeable when examining individuals under a stereomicroscope and thus deemed negligible. Overall, male Ae. aegypti and Ae. albopictus displayed a greater sensitivity to dusting. Only the lowest dust weight (0.5 mg) did not significantly reduce longevity (P = 0.888) in Ae. aegypti, whilst the lowest two dust weights (0.5 and 0.75 mg) had no significant impact on longevity (P = 0.951 and 0.166, respectively) in Ae. albopictus.
CONCLUSION: We have devised a fast, inexpensive and simple marking method and provided recommended dust quantities for several major species of disease-causing mosquitoes. The novel technique provides an evenly distributed, long-lasting mark which is non-detrimental. Our results will be useful for future MRR studies, prior to a major release campaign.

Keywords:  Diptera; Dust; Mark-release-recapture; Sterile insect technique

DOI:  https://doi.org/10.1186/s13071-020-04066-6
PLoS One. 2020 ;15(4): e0231047

Metagenomic analysis of Aedes aegypti and Culex quinquefasciatus mosquitoes from Grenada, West Indies.

Maria E Ramos-Nino, Daniel M Fitzpatrick, Korin M Eckstrom, Scott Tighe, Lindsey M Hattaway, Andy N Hsueh, Diana M Stone, Julie A Dragon, Sonia Cheetham.

The mosquitoes Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae) and Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) are two major vectors of arthropod-borne pathogens in Grenada, West Indies. As conventional vector control methods present many challenges, alternatives are urgently needed. Manipulation of mosquito microbiota is emerging as a field for the development of vector control strategies. Critical to this vector control approach is knowledge of the microbiota of these mosquitoes and finding candidate microorganisms that are common to the vectors with properties that could be used in microbiota modification studies. Results showed that bacteria genera including Asaia, Escherichia, Pantoea, Pseudomonas, and Serratia are common to both major arboviral vectors in Grenada and have previously been shown to be good candidates for transgenetic studies. Also, for the first time, the presence of Grenada mosquito rhabdovirus 1 is reported in C. quinquefasciatus.

DOI: https://doi.org/10.1371/journal.pone.0231047
PLoS Negl Trop Dis. 2020 Apr 16. 14(4): e0008219

Evolution of kdr haplotypes in worldwide populations of Aedes aegypti: Independent origins of the F1534C kdr mutation.

Luciano Veiga Cosme, Andrea Gloria-Soria, Adalgisa Caccone, Jeffrey Robert Powell, Ademir Jesus Martins.

Aedes aegypti is the primary vector of dengue, chikungunya, Zika, and urban yellow fever. Insecticides are often the most effective tools to rapidly decrease the density of vector populations, especially during arbovirus disease outbreaks. However, the intense use of insecticides, particularly pyrethroids, has selected for resistant mosquito populations worldwide. Mutations in the voltage gated sodium channel (NaV) are among the principal mechanisms of resistance to pyrethroids and DDT, also known as "knockdown resistance," kdr. Here we report studies on the origin and dispersion of kdr haplotypes in samples of Ae. aegypti from its worldwide distribution. We amplified the IIS6 and IIIS6 NaV segments from pools of Ae. aegypti populations from 15 countries, in South and North America, Africa, Asia, Pacific, and Australia. The amplicons were barcoded and sequenced using NGS Ion Torrent. Output data were filtered and analyzed using the bioinformatic pipeline Seekdeep to determine frequencies of the IIS6 and IIIS6 haplotypes per population. Phylogenetic relationships among the haplotypes were used to infer whether the kdr mutations have a single or multiple origin. We found 26 and 18 haplotypes, respectively for the IIS6 and IIIS6 segments, among which were the known kdr mutations 989P, 1011M, 1016I and 1016G (IIS6), 1520I, and 1534C (IIIS6). The highest diversity of haplotypes was found in African samples. Kdr mutations 1011M and 1016I were found only in American and African populations, 989P + 1016G and 1520I + 1534C in Asia, while 1534C was present in samples from all continents, except Australia. Based primarily on the intron sequence, IIS6 haplotypes were subdivided into two well-defined clades (A and B). Subsequent phasing of the IIS6 + IIIS6 haplotypes indicates two distinct origins for the 1534C kdr mutation. These results provide evidence of kdr mutations arising de novo at specific locations within the Ae. aegypti geographic distribution. In addition, our results suggest that the 1534C kdr mutation had at least two independent origins. We can thus conclude that insecticide selection pressure with DDT and more recently with pyrethroids is selecting for independent convergent mutations in NaV.

DOI: https://doi.org/10.1371/journal.pntd.0008219
PLoS One. 2020 ;15(4): e0231251

Longevity of the insecticidal effect of three pyrethroid formulations applied to outdoor vegetation on a laboratory-adapted colony of the Southeast Asian malaria vector Anopheles dirus.

Victor Chaumeau, Praphan Wisisakun, Sunisa Sawasdichai, Prasan Kankew, Gay Nay Htoo, Somsak Saithanmettajit, Sarang Aryalamloed, Naw Yu Lee, Gilles Delmas, François Nosten.

Outdoor residual spraying is proposed for the control of exophilic mosquitoes. However, the residual effect of insecticide mists applied to outdoor resting habitats of mosquitoes is not well characterized. The objective of this study was to assess the longevity of the residual insecticidal effect of three pyrethroid formulations applied to outdoor vegetation against the Southeast Asian malaria vector Anopheles dirus. Lambda-cyhalothrin capsule suspension, deltamethrin emulsifiable concentrate and bifenthrin wettable powder were sprayed on dense bamboo bushes on the Thailand-Myanmar border during the dry season 2018. The duration and magnitude of the residual insecticidal effect were assessed weekly with a standard cone assay, using freshly collected insecticide-treated bamboo leaves and a laboratory-adapted colony of Anopheles dirus sensu stricto susceptible to pyrethroids. The experiment was repeated during the rainy season to assess the persistence of the lambda-cyhalothrin formulation after natural rains and artificial washings. During the dry season (cumulative rainfall = 28 mm in 111 days), mortality and knockdown (KD) rates were >80% for 60 days with bifenthrin and 90 days with lambda-cyhalothrin and deltamethrin. The 50% knockdown time (TKD50) was <15 min with lambda-cyhalothrin and deltamethrin, and <30 min with bifenthrin. During the rainy season (cumulative rainfall = 465 mm in 51 days), mortality and KD rates were >80% for 42 days and TKD50 was <15 min with lambda-cyhalothrin. Additional artificial washing of the testing material with 10L of tap water before performing the cone tests had no significant effect on the residual insecticidal effect of this formulation. Long-lasting residual insecticidal effect can be obtained when spraying pyrethroid insecticides on the outdoor resting habitats of malaria vectors.

DOI: https://doi.org/10.1371/journal.pone.0231251
Viruses. 2020 Apr 14. pii: E440. [Epub ahead of print]12(4):

Identification and RNAi Profile of a Novel Iflavirus Infecting Senegalese Aedes vexans arabiensis Mosquitoes.

Rhys Parry, Fanny Naccache, El Hadji Ndiaye, Gamou Fall, Ilaria Castelli, Renke Lühken, Jolyon Medlock, Benjamin Cull, Jenny C Hesson, Fabrizio Montarsi, Anna-Bella Failloux, Alain Kohl, Esther Schnettler, Mawlouth Diallo, Sassan Asgari, Isabelle Dietrich, Stefanie C Becker.

  The inland floodwater mosquito Aedes vexans (Meigen, 1830) is a competent vector of numerous arthropod-borne viruses such as Rift Valley fever virus (Phenuiviridae) and Zika virus (Flaviviridae). Aedes vexans spp. have widespread Afrotropical distribution and are common European cosmopolitan mosquitoes. We examined the virome of Ae. vexans arabiensis samples from Barkédji village, Senegal, with small RNA sequencing, bioinformatic analysis, and RT-PCR screening. We identified a novel 9494 nt iflavirus (Picornaviridae) designated here as Aedes vexans iflavirus (AvIFV). Annotation of the AvIFV genome reveals a 2782 amino acid polyprotein with iflavirus protein domain architecture and typical iflavirus 5' internal ribosomal entry site and 3' poly-A tail. Aedes vexans iflavirus is most closely related to a partial virus sequence from Venturia canescens (a parasitoid wasp) with 56.77% pairwise amino acid identity. Analysis of AvIFV-derived small RNAs suggests that AvIFV is targeted by the exogenous RNA interference pathway but not the PIWI-interacting RNA response, as ~60% of AvIFV reads corresponded to 21 nt Dicer-2 virus-derived small RNAs and the 24-29 nt AvIFV read population did not exhibit a "ping-pong" signature. The RT-PCR screens of archival and current (circa 2011-2020) Ae. vexans arabiensis laboratory samples and wild-caught mosquitoes from Barkédji suggest that AvIFV is ubiquitous in these mosquitoes. Further, we screened wild-caught European Ae. vexans samples from Germany, the United Kingdom, Italy, and Sweden, all of which tested negative for AvIFV RNA. This report provides insight into the diversity of commensal Aedes viruses and the host RNAi response towards iflaviruses.

Keywords:  Aedes vexans; RNAi; iflavirus; virus discovery

DOI:  https://doi.org/10.3390/v12040440
Parasit Vectors. 2020 Apr 16. 13(1): 197

Spatial modelling of the infestation indices of Aedes aegypti: an innovative strategy for vector control actions in developing countries.

Ana Carolina Policarpo Cavalcante, Ricardo Alves de Olinda, Alexandrino Gomes, John Traxler, Matt Smith, Silvana Santos.

   BACKGROUND: Larval indices such as the house index (HI), Breteau index (BI) and container index (CI) are widely used to interpret arbovirus vector density in surveillance programmes. However, the use of such data as an alarm signal is rarely considered consciously when planning programmes. The present study aims to investigate the spatial distribution pattern of the infestation of Aedes aegypti, considering the data available in the Ae. aegypti Infestation Index Rapid Survey (LIRAa) for the city of Campina Grande, Paraíba State in Brazil.
METHODS: The global and local Moran's indices were used in spatial analysis to measure the effects of spatial dependencies between neighbourhoods, using secondary data related to HI and BI gathered from surveillance service.
RESULTS: Our analysis shows that there is a predominance of high rates of mosquito infestation, placing Campina Grande at a near-constant risk of arbovirus outbreaks and epidemics. A highly significant Moran's index value (P < 0.001) was observed, indicating a positive spatial dependency between the neighbourhoods in Campina Grande. Using the Moran mapping and LISA mapping, the autocorrelation patterns of Ae. aegypti infestation rates among neighbourhoods have revealed hotpots that should be considered a priority to preventive actions of the entomological surveillance services. Predominance of high infestation rates and clearer relationships of these between neighbourhoods were observed between the months of May and July, the period with the highest rainfall in the city.
CONCLUSIONS: This analysis is an innovative strategy capable of providing detailed information on infestation locations to the relevant public health authorities, which will enable a more efficient allocation of resources, particularly for arbovirus prevention.

Keywords:  Aedes; Arbovirus infections; Spatial analysis

DOI:  https://doi.org/10.1186/s13071-020-04070-w
Pest Manag Sci. 2020 Apr 12.

The potential distribution and dynamics of important vectors Culex pipiens pallens and Culex pipiens quinquefasciatus in China under climate change scenarios: An ecological niche modelling approach.

Boyang Liu, Xiang Gao, Keren Zheng, Jun Ma, Zhihui Jiao, Jianhua Xiao, Hongbin Wang.

BACKGROUND: Intense studies have been carried out on the effects of climate change on vector-borne diseases and vectors. Culex pipiens pallens and Culex pipiens quinquefasciatus are two medically concerned mosquito species in temperate and tropical areas, which serve as important disease-transmitting pests of a variety of diseases. The ongoing geographically expansion of these mosquitoes has brought an increasing threat to public health.
RESULTS: Based on mosquito occurrence records and high-resolution environmental layers, an ecological niche model was established to model their current and future potential distribution in China. Our model showed that the current suitable area for Cx. p. pallens is distributed in the central, eastern and northern parts of China, while Cx. p. quinquefasciatus is distributed in vast areas in southern China. Under future climate change scenarios, both species were predicted to expand their range to varying degrees and RCP 8.5 provides the largest expansion. Northward core shifts will occur in ranges of both species. Environmental variables which have significant impact on the distribution of mosquitoes were also revealed by our model.
CONCLUSION: Severe habitat expansion of vectors is likely to occur in the future 21st century. Our models mapped the high-risk areas and risk factors which needs to be paid attention. The results of our study can be referenced in further ecological surveys and will guide the development of strategies for the prevention and control of vector-borne diseases. This article is protected by copyright. All rights reserved.

DOI: https://doi.org/10.1002/ps.5861
Lancet Infect Dis. 2020 Apr 08. pii: S1473-3099(20)30059-1. [Epub ahead of print]

Association between the proportion of Plasmodium falciparum and Plasmodium vivax infections detected by passive surveillance and the magnitude of the asymptomatic reservoir in the community: a pooled analysis of paired health facility and community data.

Gillian Stresman, Nuno Sepúlveda, Kimberly Fornace, Lynn Grignard, Julia Mwesigwa, Jane Achan, John Miller, Daniel J Bridges, Thomas P Eisele, Jacklin Mosha, Pauline Joy Lorenzo, Maria Lourdes Macalinao, Fe Esperanza Espino, Fitsum Tadesse, Jennifer C Stevenson, Antonio M Quispe, André Siqueira, Marcus Lacerda, Shunmay Yeung, Siv Sovannaroth, Emilie Pothin, Joanna Gallay, Karen E Hamre, Alyssa Young, Jean Frantz Lemoine, Michelle A Chang, Koukeo Phommasone, Mayfong Mayxay, Jordi Landier, Daniel M Parker, Lorenz Von Seidlein, Francois Nosten, Gilles Delmas, Arjen Dondorp, Ewan Cameron, Katherine Battle, Teun Bousema, Peter Gething, Umberto D'Alessandro, Chris Drakeley.

BACKGROUND: Passively collected malaria case data are the foundation for public health decision making. However, because of population-level immunity, infections might not always be sufficiently symptomatic to prompt individuals to seek care. Understanding the proportion of all Plasmodium spp infections expected to be detected by the health system becomes particularly paramount in elimination settings. The aim of this study was to determine the association between the proportion of infections detected and transmission intensity for Plasmodium falciparum and Plasmodium vivax in several global endemic settings.
METHODS: The proportion of infections detected in routine malaria data, P(Detect), was derived from paired household cross-sectional survey and routinely collected malaria data within health facilities. P(Detect) was estimated using a Bayesian model in 431 clusters spanning the Americas, Africa, and Asia. The association between P(Detect) and malaria prevalence was assessed using log-linear regression models. Changes in P(Detect) over time were evaluated using data from 13 timepoints over 2 years from The Gambia.
FINDINGS: The median estimated P(Detect) across all clusters was 12·5% (IQR 5·3-25·0) for P falciparum and 10·1% (5·0-18·3) for P vivax and decreased as the estimated log-PCR community prevalence increased (adjusted odds ratio [OR] for P falciparum 0·63, 95% CI 0·57-0·69; adjusted OR for P vivax 0·52, 0·47-0·57). Factors associated with increasing P(Detect) included smaller catchment population size, high transmission season, improved care-seeking behaviour by infected individuals, and recent increases (within the previous year) in transmission intensity.
INTERPRETATION: The proportion of all infections detected within health systems increases once transmission intensity is sufficiently low. The likely explanation for P falciparum is that reduced exposure to infection leads to lower levels of protective immunity in the population, increasing the likelihood that infected individuals will become symptomatic and seek care. These factors might also be true for P vivax but a better understanding of the transmission biology is needed to attribute likely reasons for the observed trend. In low transmission and pre-elimination settings, enhancing access to care and improvements in care-seeking behaviour of infected individuals will lead to an increased proportion of infections detected in the community and might contribute to accelerating the interruption of transmission.
FUNDING: Wellcome Trust.

DOI: https://doi.org/10.1016/S1473-3099(20)30059-1
J Med Entomol. 2020 Apr 17. pii: tjaa066. [Epub ahead of print]

A 70% Reduction in Mosquito Populations Does Not Require Removal of 70% of Mosquitoes.

J Lega, H E Brown, R Barrera.

  We added a vector control component to our existing abundance model to simulate intensive vector control in Puerto Rico. Removing 20-30% of gravid females in the model matches observed 60-80% reductions. The model's capacity to reproduce vector control increases its utility for planning and evaluation strategies.

Keywords:  gravid; modeling; vector control

DOI:  https://doi.org/10.1093/jme/tjaa066
Acta Trop. 2020 Apr 10. pii: S0001-706X(20)30072-3. [Epub ahead of print] 105491

Impacts of fluorescent powders on survival of different age cohorts, blood-feeding success, and tethered flight speed of Aedes aegypti (Diptera: Culicidae) females.

Diana Rojas-Araya, Barry W Alto, Nathan D Burkett-Cadena, Derek At Cummings.

  Fluorescent powders are one of the most common external markers used to study mosquito behavior and ecology. For their reliable and practical use, it is important to evaluate their effect on biological parameters such as survival, blood-feeding, and mobility. We evaluated the effect of five different fluorescent powders (Day-Glo ® ECO Series) on the survival of different age cohorts, blood-feeding success, and tethered flight speed on Aedes aegypti (Linnaeus) adult females. For survival analysis, three cohorts (2-5, 6-9 and 10-13 days old) were marked and mortality recorded until all died. To examine the effect of fluorescent powders on female response to blood-feeding, the proportions of unfed, partially fed, and fully engorged females, after being exposed to host blood under two different time sets (20 and 40 min.), were compared. Their impact on female tethered flight speed was evaluated recording their flight for 30 min. with a flight mill. Survival distributions between treatments were not significantly different within each cohort. Blood-feeding was not significantly different among marked or unmarked females at both times of blood exposure, with the exception of Signal Green-ECO 18 and Ultra Violet- ECO 20 (at 20 and 40 min.), in which a higher proportion of partially fed females was observed compared to control females. In relation to flight performance, no statistically significant difference in mean tethered flight speed (m/s), among marked and unmarked mosquito groups, was observed. Our results indicate that the tested powders and application method have few significant impacts on Ae. aegypti survival, blood-feeding success and flight performance, and are thus suitable for investigations of mosquito biology in the environment.

Keywords:  Culicidae; Flight mill; Fluorescent powder; Marking; Mosquitoes; Vector

DOI:  https://doi.org/10.1016/j.actatropica.2020.105491
PLoS One. 2020 ;15(4): e0231541

Low transmission of Wuchereria bancrofti in cross-border districts of Côte d'Ivoire: A great step towards lymphatic filariasis elimination in West Africa.

Firmain N Yokoly, Julien B Z Zahouli, Aboulaye Méite, Millicent Opoku, Bernard L Kouassi, Dziedzom K de Souza, Moses Bockarie, Benjamin G Koudou.

BACKGROUND: Lymphatic filariasis (LF) is widely endemic in Côte d'Ivoire, and elimination as public health problem (EPHP) is based on annual mass drug administration (MDA) using ivermectin and albendazole. To guide EPHP efforts, we evaluated Wuchereria bancrofti infection indices among humans, and mosquito vectors after four rounds of MDA in four cross-border health districts of Côte d'Ivoire.
METHODOLOGY: We monitored people and mosquitoes for W. bancrofti infections in the cross-border health districts of Aboisso, Bloléquin, Odienné and Ouangolodougou, Côte d'Ivoire. W. bancrofti circulating filarial antigen (CFA) was identified using filariasis test strips, and antigen-positive individuals were screened for microfilaremia. Moreover, filarial mosquito vectors were sampled using window exit traps and pyrethrum sprays, and identified morphologically at species level. Anopheles gambiae s.l. and Culex quinquefasciatus females were analyzed for W. bancrofti infection using polymerase chain reaction (PCR) technique.
PRINCIPAL FINDINGS: Overall, we found a substantial decline in W. bancrofti infection indices after four rounds of MDA compared to pre-MDA baseline data. CFA prevalence fell from 3.38-5.50% during pre-MDA to 0.00-1.53% after MDA interventions. No subjects had detectable levels of CFA in Ouangolodougou. Moreover, post-MDA CFA prevalence was very low, and below the 1% elimination threshold in Aboisso (0.19%) and Odienné (0.49%). Conversely, CFA prevalence remained above 1% in Bloléquin (1.53%). W. bancrofti microfilariae (Mf) were not found in Aboisso, Bloléquin, and Ouangolodougou, except for Odienné with low prevalence (0.16%; n = 613) and microfilaremia of 32.0 Mf/mL. No An. gambiae s.l. and Cx. quinquefasciatus pools were infected with W. bancrofti in Bloléquin and Ouangolodougou, while they exhibited low infection rates in Aboisso (1% and 0.07%), and Odienné (0.08% and 0.08%), respectively.
CONCLUSIONS: In cross-border areas of Côte d'Ivoire, LF infection indices in humans and mosquito vectors substantially declined after four rounds of MDA. CFA prevalence fell under the World Health Organization (WHO)-established threshold (1%) in Aboisso, Ouangolodougou and Odienné. Moreover, W. bancrofti prevalence in mosquitoes was lower than WHO-established threshold (2%) in all areas. This might suggest the interruption of W. bancrofti transmission, and possible MDA cessation. However, a formal transmission assessment survey (TAS) and molecular xenomonitoring in mosquito vectors should be implemented before eventual MDA cessation. However, MDA should pursue in Bloléquin where W. bancrofti infection prevalence remained above 1%. Our results provided important ramifications for LF control efforts towards EPHP in Côte d'Ivoire.

DOI: https://doi.org/10.1371/journal.pone.0231541
J Med Entomol. 2020 Apr 13. pii: tjaa067. [Epub ahead of print]

Seasonal Changes of Host Use by Culiseta melanura (Diptera: Culicidae) in Central Florida.

Richard G West, Derrick K Mathias, Jonathan F Day, Carolina Acevedo, Thomas R Unnasch, Nathan D Burkett-Cadena.

  The mosquito Culiseta melanura (Coquillett) is the primary enzootic vector of eastern equine encephalitis virus (EEEV), a zoonotic Alphavirus endemic to eastern North America. In its northern range, Cs. melanura is considered a strict avian biter, transmitting EEEV among susceptible birds in a cycle of enzootic amplification. In its southern range, however, Cs. melanura is more general in host use, feeding heavily upon birds but also reptiles and mammals. The goal of this study was to better understand how host use of Cs. melanura changes throughout the year in Florida, where year-round EEEV transmission is observed. Mosquitoes were sampled in 2018 from nine sites across three central Florida counties. In total, 213 Cs. melanura bloodmeals were identified by PCR consisting of 39 species of birds, reptiles, and mammals. Avian bloodmeals were prominent throughout the year (range = 30-85%), and songbirds were a large portion of identified bloodmeals (37.1%). Reptiles surpassed birds only in spring (April-June), and brown anole (Anolis sagrei Duméril and Bibron, 1837 [Reptilia: Dactyloidae]) was the most commonly detected single host species (22.1% overall). Mammalian bloodmeals were mainly observed in summer, with humans being the most fed on mammal (12.7% overall). This study reveals that in southern foci of EEEV transmission, Cs. melanura host use varies throughout the year with reptiles providing the majority of blood meals in spring (51.3%), and birds are fed on more than other host groups during all other seasons (50.6-70.1%). In addition, feeding on mammals increases during summer months, which may implicate Cs. melanura in epizootic transmission in Florida.

Keywords:   Anolis ; Culiseta ; arbovirus; host use; seasonal

DOI:  https://doi.org/10.1093/jme/tjaa067
Acta Trop. 2020 Apr 10. pii: S0001-706X(20)30234-5. [Epub ahead of print] 105455

Molecular and morphological evidence for sibling species within Anopheles (Anopheles) lindesayi Giles (Diptera: Culicidae) in Bhutan.

Rinzin Namgay, Dechen Pemo, Tenzin Wangdi, Thanari Phanitchakun, Ralph E Harbach, Pradya Somboon.

  This paper reports the results of a comparative molecular and morphological study of An. lindesayi collected from various districts of Bhutan and An. l. cameronensis from Thailand, compared with GenBank accessions and publications for An. l. japonicus from Japan, South Korea and China, An. l. pleccau from Taiwan, and An. lindesayi from India. Phylogenetic analyses based on ribosomal (ITS2) and mitochondrial (COI) DNA sequences using the Maximum Likelihood method revealed five genetically distinct clades (A, B, C, D and E) in Bhutan. Specimens in Clade A correspond to the original description of An. lindesayi, particularly in wing markings, the pattern of basal pale scales on the hindfemur and the single seta 4-C of larvae, and their COI sequences were closely related to one Indian sequence. Larvae of Clades B, C, D and E are similar in having seta 4-C branched rather than single. The adults of Clades C, D and E (B not available) are distinguishable from those of Clade A and other subspecies. Specimens of Clade C are unique in having a long pale spot on wing vein R and the subcosta, scattered pale scales on several veins and a dark spot at the tip of vein R2. The adults of Clades D and E are similar in having a dark spot at the tip of vein R2 and no scattered pale scales on all other veins. We provisionally recognize mosquitoes of Clades A, B, C, D and E as species A, B, C, D and E, respectively, of the Lindesayi Complex. Species A is An. lindesayi sensu stricto and the others are unnamed species. Concomitantly, the previous concept of the "Lindesayi Complex", which included An. lindesayi, An. menglangensis, An. nilgiricus and An. wellingtonianus, is now recognized as the Lindesayi Subgroup of the Lindesayi Group (Anopheles Series, subgenus Anopheles) with the five sibling species of An. lindesayi comprising a more apposite Lindesayi Complex within the subgroup.

Keywords:  Anopheles lindesayi; Bhutan; COI; ITS2; mosquito; taxonomy

DOI:  https://doi.org/10.1016/j.actatropica.2020.105455
Sci Total Environ. 2020 Apr 02. pii: S0048-9697(20)31904-5. [Epub ahead of print]725 138391

Persistence of aerially applied mosquito-pesticide, Naled, in fresh and marine waters.

Athena S Jones, David Cohen, Francisco Alberdi, Amanda Sanabria, Nakiya Clausell, Matthew Roca, Abelline K Fionah, Naresh Kumar, Helena M Solo-Gabriele, Elsayed M Zahran.

  Naled, an organophosphate pesticide, received considerable attention during 2016 as it was applied aerially to control the first mosquito-borne Zika virus outbreak in the continental United States. Stakeholders living in affected areas raised concerns about its environmental impacts. One factor influencing environmental impacts is the persistence of the chemical applied. The objective of this study was to evaluate the persistence of naled - and its degradation bi-product, dichlorvos - in natural waters. Initial naled concentrations were measured at ground level after full-scale aerial spray activities. Laboratory experiments were designed to evaluate factors (fresh versus marine water chemistry, temperature, and sunlight) that may promote the degradation of naled and dichlorvos in the environment. Results show that natural fresh and marine water chemistry promoted naled degradation as experiments with de-ionized water resulted in half-lives greater than 6 days. The half-life in natural waters without light ranged from 5 to 20 h with lower half lives at higher temperatures. Under light exposure, degradation was accelerated and yielded more dichlorvos. Detectable levels (0.05 μM for naled and 0.10 μM for dichlorvos) were measured in water samples collected from the field during aerial spray events. Results can be used in risk assessments that consider both naled and dichlorvos to better understand ecological impacts and to develop improved public health recommendations.

Keywords:  Aerial spray; Dichlorvos; Naled; Water; Zika

DOI:  https://doi.org/10.1016/j.scitotenv.2020.138391
Trends Parasitol. 2020 May;pii: S1471-4922(20)30040-4. [Epub ahead of print]36(5): 473-484

Not Just from Blood: Mosquito Nutrient Acquisition from Nectar Sources.

Elina Barredo, Matthew DeGennaro.

  Anthropophilic female mosquitoes are well known for their strong attraction to human hosts, but plant nectar is a common energy source in their diets. When sugar sources are scarce, female mosquitoes of some species can compensate by taking larger and more frequent blood meals. Male mosquitoes are exclusively dependent on plant nectar or alternative sugar sources. Plant preference is likely driven by an innate attraction that may be enhanced by experience, as mosquitoes learn to recognize available sugar rewards. Nectar-seeking involves the integration of at least three sensory systems: olfaction, vision and taste. The prevention of vector-borne illnesses, the determination of the mosquitoes' ecological role, and the design of efficient sugar-baited traps will all benefit from understanding the molecular basis of nectar-seeking.

Keywords:  attractive toxic sugar baits (ATSBs); gustation; mosquito; multimodal integration; nectar-seeking; olfaction; sugar-feeding

DOI:  https://doi.org/10.1016/j.pt.2020.02.003
Malar J. 2020 Apr 15. 19(1): 152

High levels of imported asymptomatic malaria but limited local transmission in KwaZulu-Natal, a South African malaria-endemic province nearing malaria elimination.

Jaishree Raman, Laura Gast, Ryleen Balawanth, Sofonias Tessema, Basil Brooke, Rajendra Maharaj, Givemore Munhenga, Power Tshikae, Vishan Lakan, Tshiama Mwamba, Hazel Makowa, Lindi Sangweni, Moses Mkhabela, Nompumelelo Zondo, Ernest Mohulatsi, Zuziwe Nyawo, Sifiso Ngxongo, Sipho Msimang, Nicole Dagata, Bryan Greenhouse, Lyn-Marie Birkholtz, George Shirreff, Rebecca Graffy, Bheki Qwabe, Devanand Moonasar.

   BACKGROUND: KwaZulu-Natal, one of South Africa's three malaria endemic provinces, is nearing malaria elimination, reporting fewer than 100 locally-acquired cases annually since 2010. Despite sustained implementation of essential interventions, including annual indoor residual spraying, prompt case detection using malaria rapid diagnostics tests and treatment with effective artemisinin-based combination therapy, low-level focal transmission persists in the province. This malaria prevalence and entomological survey was therefore undertaken to identify the drivers of this residual transmission.
METHODS: Malaria prevalence as well as malaria knowledge, attitudes and practices among community members and mobile migrant populations within uMkhanyakude district, KwaZulu-Natal were assessed during a community-based malaria prevalence survey. All consenting participants were tested for malaria by both conventional and highly-sensitive falciparum-specific rapid diagnostic tests. Finger-prick filter-paper blood spots were also collected from all participants for downstream parasite genotyping analysis. Entomological investigations were conducted around the surveyed households, with potential breeding sites geolocated and larvae collected for species identification and insecticide susceptibility testing. A random selection of households were assessed for indoor residual spray quality by cone bioassay.
RESULTS: A low malaria prevalence was confirmed in the study area, with only 2% (67/2979) of the participants found to be malaria positive by both conventional and highly-sensitive falciparum-specific rapid diagnostic tests. Malaria prevalence however differed markedly between the border market and community (p < 0001), with the majority of the detected malaria carriers (65/67) identified as asymptomatic Mozambican nationals transiting through the informal border market from Mozambique to economic hubs within South Africa. Genomic analysis of the malaria isolates revealed a high degree of heterozygosity and limited genetic relatedness between the isolates supporting the hypothesis of limited local malaria transmission within the province. New potential vector breeding sites, potential vector populations with reduced insecticide susceptibility and areas with sub-optimal vector intervention coverage were identified during the entomological investigations.
CONCLUSION: If KwaZulu-Natal is to successfully halt local malaria transmission and prevent the re-introduction of malaria, greater efforts need to be placed on detecting and treating malaria carriers at both formal and informal border crossings with transmission blocking anti-malarials, while ensuring optimal coverage of vector control interventions is achieved.

Keywords:  Asymptomatic carriage; Elimination; KAP; KwaZulu-Natal; Malaria; Malaria importation; Rapid diagnostic tests; Residual transmission; South Africa; Vector control

DOI:  https://doi.org/10.1186/s12936-020-03227-3