From eggs to bites: do ovitrap data provide reliable estimates of Aedes albopictus biting females?

Background. Aedes albopictus is an aggressive invasive mosquito species that represents a serious health concern not only in tropical areas, but also in temperate regions due to its role as vector of arboviruses. Estimates of mosquito biting rates are essential to account for vector-human contact in models aimed to predict the risk of arbovirus autochthonous transmission and outbreaks, as well as nuisance thresholds useful for correct planning of mosquito control interventions. Methods targeting daytime and outdoor biting Ae. albopictus females (e.g., Human Landing Collection, HLC) are expensive and difficult to implement in large scale schemes. Instead, egg-collections by ovitraps are the most widely used routine approach for large-scale monitoring of the species. The aim of this work was to assess whether ovitrap data can be exploited to estimate numbers of adult biting Ae. albopictus females and whether the resulting relationship could be used to build risk models helpful for decision-makers in charge of planning of mosquito-control activities in infested areas. Method. Ovitrap collections and HLCs were carried out in hot-spots of Ae. albopictus abundance in Rome (Italy) along a whole reproductive season. The relationship between the two sets of data was assessed by generalized least square analysis, taking into account meteorological parameters. Result. The mean number of mosquito females/person collected by HLC in 150 (i.e., females/HLC) and the mean number of eggs/day were 18.9 ± 0.7 and 39.0 ± 2.0, respectively. The regression models found a significant positive relationship between the two sets of data and estimated an increase of one biting female/person every five additional eggs found in ovitraps. Both observed and fitted values indicated presence of adults in the absence of eggs in ovitraps. Notably, wide confidence intervals of estimates of biting females based on eggs were observed. The patterns of exotic arbovirus outbreak probability obtained by introducing these estimates in risk models were similar to those based on females/HLC (R0 > 1 in 86% and 40% of sampling dates for Chikungunya and Zika, respectively; R0 < 1 along the entire season for Dengue). Moreover, the model predicted that in this case-study scenario an R0 > 1 for Chikungunya is also to be expected when few/no eggs/day are collected by ovitraps. Discussion. This work provides the first evidence of the possibility to predict mean number of adult biting Ae. albopictus females based on mean number of eggs and to compute the threshold of eggs/ovitrap associated to epidemiological risk of arbovirus transmission in the study area. Overall, however, the large confidence intervals in the model predictions represent a caveat regarding the reliability of monitoring schemes based exclusively on ovitrap collections to estimate numbers of biting females and plan control interventions

New adhesive traps to monitor urban mosquitoes with a case study to assess the efficacy of insecticide control strategies in temperate areas

Background: Urban mosquitoes in temperate regions may represent a high nuisance and are associated with the risk of arbovirus transmission. Common practices to reduce this burden, at least in Italian highly infested urban areas, imply calendar-based larvicide treatments of street catch basins – which represent the main non-removable urban breeding site and/or insecticide ground spraying. The planning of these interventions, as well as the evaluation of their effectiveness, rarely benefit of adequate monitoring of the mosquito abundance and dynamics. We propose the use of adhesive traps to monitor Aedes albopictus and Culex pipiens adults and to evaluate the efficacy of insecticide-based control strategies. Methods: We designed two novel types of adhesive traps to collect adult mosquitoes visiting and/or emerging from catch basins. The Mosquito Emerging Trap (MET) was exploited to assess the efficacy of larvicide treatments. The Catch Basin Trap (CBT) was exploited together with the Sticky Trap (ST, commonly used to collect ovipositing/resting females) to monitor adults abundance in the campus of the University of Rome “Sapienza” - where catch basins were treated with Insect Growth Regulators (IGR) bi-monthly and Low-Volume insecticide spraying were carried out before sunset - and in a nearby control area. Results: Results obtained by MET showed that, although all monitored diflubenzuron-treated catch basins were repeatedly visited by Ae. albopictus and Cx. pipiens, adult emergence was inhibited in most basins. Results obtained by ST and CBT showed a significant lower adult abundance in the treated area than in the untreated one after the second adulticide spraying, which was carried out during the major phase of Ae. albopictus population expansion in Rome. Spatial heterogeneities in the effect of the treatments were also revealed. Conclusions: The results support the potential of the three adhesive traps tested in passively monitoring urban mosquito adult abundance and seasonal dynamics and in assessing the efficacy of control measures. ST showed higher specificity for Ae. albopictus and CBT for Cx. pipiens. The results also provide a preliminary indication on the effectiveness of common mosquito control strategies carried out against urban mosquito in European urban areas

Transmission dynamics of the ongoing chikungunya outbreak in Central Italy. From coastal areas to the metropolitan city of Rome, summer 2017

A large chikungunya outbreak is ongoing in Italy, with a main cluster in the Anzio coastal municipality. With preliminary epidemiological data, and a transmission model using mosquito abundance and biting rates, we estimated the basic reproduction number R0 at 2.07 (95% credible interval: 1.47–2.59) and the first case importation between 21 May and 18 June 2017. Outbreak risk was higher in coastal/rural sites than urban ones. Novel transmission foci could occur up to mid-November

Spatial and temporal hot spots of Aedes albopictus abundance inside and outside a South European metropolitan area

Aedes albopictus is a tropical invasive species which in the last decades spread worldwide, also colonizing temperate regions of Europe and US, where it has become a public health concern due to its ability to transmit exotic arboviruses, as well as severe nuisance problems due to its aggressive daytime outdoor biting behaviour. While several studies have been carried out in order to predict the potential limits of the species expansions based on eco-climatic parameters, few studies have so far focused on the specific effects of these variables in shaping its micro-geographic abundance and dynamics. The present study investigated eco-climatic factors affecting Ae. albopictus abundance and dynamics in metropolitan and sub-urban/rural sites in Rome (Italy), which was colonized in 1997 and is nowadays one of the most infested metropolitan areas in Southern Europe. To this aim, longitudinal adult monitoring was carried out along a 70 km-transect across and beyond the most urbanized and densely populated metropolitan area. Two fine scale spatiotemporal datasets (one with reference to a 20m circular buffer around sticky traps used to collect mosquitoes and the second to a 300m circular buffer within each sampling site) were exploited to analyze the effect of climatic and socio-environmental variables on Ae. albopictus abundance and dynamics along the transect. Results showed an association between highly anthropized habitats and high adult abundance both in metropolitan and sub-urban/ rural areas, with “small green islands” corresponding to hot spots of abundance in the metropolitan areas only, and a bimodal seasonal dynamics with a second peak of abundance in autumn, due to heavy rains occurring in the preceding weeks in association with permissive temperatures. The results provide useful indications to prioritize public mosquito control measures in temperate urban areas where nuisance, human-mosquito contact and risk of local arbovirus transmission are likely higher, and highlight potential public health risks also after the summer months typically associated with high mosquito densities

Insertion polymorphisms of SINE200 retrotransposons within speciation islands of Anopheles gambiae molecular forms

<p>Abstract</p> <p>Background</p> <p>SINEs (Short INterspersed Elements) are homoplasy-free and co-dominant genetic markers which are considered to represent useful tools for population genetic studies, and could help clarifying the speciation processes ongoing within the major malaria vector in Africa, <it>Anopheles gambiae </it>s.s. Here, we report the results of the analysis of the insertion polymorphism of a nearly 200 bp-long SINE (<it>SINE200</it>) within genome areas of high differentiation (i.e. "speciation islands") of M and S <it>A. gambiae </it>molecular forms.</p> <p>Methods</p> <p>A <it>SINE</it>-PCR approach was carried out on thirteen <it>SINE200 </it>insertions in M and S females collected along the whole range of distribution of <it>A. gambiae </it>s.s. in sub-Saharan Africa. Ten specimens each for <it>Anopheles arabiensis</it>, <it>Anopheles melas, Anopheles quadriannulatus </it>A and 15 M/S hybrids from laboratory crosses were also analysed.</p> <p>Results</p> <p>Eight loci were successfully amplified and were found to be specific for <it>A. gambiae </it>s.s.: 5 on 2L chromosome and one on X chromosome resulted monomorphic, while two loci positioned respectively on 2R (i.e. <it>S200 </it>2R12D) and X (i.e. <it>S200 </it>X6.1) chromosomes were found to be polymorphic. <it>S200 </it>2R12D was homozygote for the insertion in most S-form samples, while intermediate levels of polymorphism were shown in M-form, resulting in an overall high degree of genetic differentiation between molecular forms (Fst = 0.46 p < 0.001) and within M-form (Fst = 0.46 p < 0.001). The insertion of <it>S200 </it>X6.1 was found to be fixed in all M- and absent in all S-specimens. This led to develop a novel easy-to-use PCR approach to straightforwardly identify <it>A. gambiae </it>molecular forms. This novel approach allows to overcome the constraints associated with markers on the rDNA region commonly used for M and S identification. In fact, it is based on a single copy and irreversible <it>SINE200 </it>insertion and, thus, is not subjected to peculiar evolutionary patterns affecting rDNA markers, e.g. incomplete homogenization of the arrays through concerted evolution and/or mixtures of M and S IGS-sequences among the arrays of single chromatids.</p> <p>Conclusion</p> <p>The approach utilized allowed to develop new easy-to-use co-dominant markers for the analysis of genetic differentiation between M and S-forms and opens new perspectives in the study of the speciation process ongoing within <it>A. gambiae</it>.</p

The sticky resting box, a new tool for studying resting behaviour of Afrotropical malaria vectors

Background: Monitoring densities of adult mosquito populations is a major challenge in efforts to evaluate the epidemiology of mosquito-borne diseases, and their response to vector control interventions. In the case of malaria, collection of outdoor-resting Anophelines is rarely incorporated into surveillance and control, partially due to the lack of standardized collection tools. Such an approach, however, is increasingly important to investigate possible changes in mosquito behaviour in response to the scale up of Insecticide Treated Nets and Indoor Residual Spraying. In this study we evaluated the Sticky Resting Box (SRB) - i.e. a sticky variant of previously investigated mosquito Resting Box, which allows passive collection of mosquitoes entering the box – and compared its performance against traditional methods for indoor and outdoor resting mosquito sampling.&lt;p&gt;&lt;/p&gt; Methods: Daily collections were carried out in two neighbouring villages of Burkina Faso during rainy season 2011 and dry season 2012 by SRB located indoors and outdoors, and by Back-Pack aspiration inside houses (BP) and in ad hoc built outdoor pit-shelters (PIT).&lt;p&gt;&lt;/p&gt; Results: Overall, almost 20,000 Culicidae specimens belonging to 16 species were collected and morphologically identified. Malaria vectors included Anopheles coluzzii (53%), An. arabiensis (12%), An. gambiae s.s. (2.0%) and An. funestus (4.5%). The diversity of species collected in the two villages was similar for SRB and PIT collections outdoors, and significantly higher for SRB than for BP indoors. The population dynamics of An. gambiae s.l. mosquitoes, as obtained by SRB-collections was significantly correlated with those obtained by the traditional methods. The predicted mean estimates of An. gambiae s.l. specimens/sampling-unit/night-of-collections was 6- and 5-times lower for SRB than for BP indoors and PIT outdoors, respectively.&lt;p&gt;&lt;/p&gt; Conclusions: Overall, the daily performance of SRB in terms of number of malaria vectors/trap was lower than that of traditionally used approaches for in- and outdoor collections. However, unlike these methods, SRB could be set up to collect mosquitoes passively over at least a week. This makes SRB a promising tool for passively monitoring anopheline resting populations, with data presented here providing guidance for how to set up SRB-based collections to acquire information comparable to those obtained with other methods.&lt;p&gt;&lt;/p&gt

Enhancement of Aedes albopictus collections by ovitrap and sticky adult trap

Abstract Background: In the last decades, Aedes albopictus has become an increasing public health threat in tropical as well as in more recently invaded temperate areas due to its capacity to transmit several human arboviruses, among which Dengue, Chikungunya and Zika. Enhancing the efficiency of currently used collection approaches, such as ovitraps and sticky traps, is desirable for optimal monitoring of the species abundance, for assessment of the risk of arbovirus transmission and for the optimisation of control activities. Findings: Two sets of 4 × 4 Latin-square experiments were carried out in Tirana (Albania) to test whether modifications in ovitrap shape and size and in oviposition substrate would increase collections of Ae. albopictus eggs and whether hay-infusion would increase adult catches by sticky trap. Generalized Linear Mixed Models with negative binomial error distribution were carried out to analyse the data. Cylindrical ovitraps lined with germination paper yielded significantly higher egg catches than those exploiting either the (commonly used) wooden paddles or floating polystyrene blocks as oviposition substrates. No difference was observed between cylindrical and conical shaped ovitraps. Ovitraps and sticky traps baited with hay infusion yielded significantly higher egg and adult catches than un-baited ones. A significant relationship between ovitrap and sticky trap catches was observed both in the absence and in the presence of attractants, with ovitrap catches increasing more than sticky trap catches at increasing adult female densities. Conclusions: This study provides grounds for optimisation of ovitraps and sticky traps as monitoring tools for Ae. albopictus by (i) supporting use of germination paper as most appropriate oviposition substrate; (ii) suggesting the possible use of stackable conical ovitraps for large scale monitoring; (iii) confirming the use of hay-infusion to increase egg catches in ovitraps, and showing that hay-infusion also significant increases adult catches by sticky traps

Adaptive potential of hybridization among malaria vectors: Introgression at the immune locus TEP1 between Anopheles coluzzii and A. gambiae in 'Far-West' Africa

“Far-West” Africa is known to be a secondary contact zone between the two major malaria vectors Anopheles coluzzii and A. gambiae.We investigated gene-flow and potentially adaptive introgression between these species along a west-to-east transect in Guinea Bissau, the putative core of this hybrid zone. To evaluate the extent and direction of gene flow, we genotyped site 702 in Intron-1 of the para Voltage-Gated SodiumChannel gene, a species-diagnostic nucleotide position throughout most of A. coluzzii and A. gambiae sympatric range. We also analyzed polymorphismin the thioester-binding domain (TED) of the innate immunity-linked thioester-containing protein 1 (TEP1) to investigate whether elevated hybridization might facilitate the exchange of variants linked to adaptive immunity and Plasmodium refractoriness. Our results confirm asymmetric introgression of genetic material from A. coluzzii to A. gambiae and disruption of linkage between the centromeric "genomic islands" of inter-specific divergence. We report that A. gambiae from the Guinean hybrid zone possesses an introgressed TEP1 resistant allelic class, found exclusively in A. coluzzii elsewhere and apparently swept to fixation inWest Africa (i.e. Mali and Burkina Faso). However, no detectable fixation of this allele was found in Guinea Bissau, which may suggest that ecological pressures driving segregation between the two species in larval habitats in this region may be different from those experienced in northern and more arid parts of the species’ range. Finally, our results also suggest a genetic subdivision between coastal and inland A. gambiae Guinean populations and provide clues on the importance of ecological factors in intra-specific differentiation processes

Evaluation of a sticky trap for collecting Aedes (Stegomyia) adults in a dengue-endemic area in Thailand.

Development of new operational techniques for collection and monitoring of adult Stegomyia mosquitoes is considered a pressing need for surveillance and prevention of arboviruses. Here we report the results from a trial carried out in 2 dengue-endemic villages in Thailand to compare the ability to collect Aedes adults of a sticky trap versus a CDC backpack aspirator, which has been used routinely at the study area for entomological/epidemiological surveys.Our comparison was based on a comparable sampling effort required to carry out collections with 2 approaches. Over 19,000 specimens were collected, ∼90% of which were Culex spp. Sticky traps collected significantly more Aedes aegypti and Aedes albopictus females than did backpack aspirators when located outdoors. The percentage of positive sticky-trap catches was double for Ae. aegypti and almost 20 times higher for Ae. albopictus. Operational benefits of the sticky trap are discussed within the context of the results obtained