Integrated taxonomy of black flies (Diptera: Simuliidae) reveals unexpected diversity in the most arid ecosystem of Europe.

The family Simuliidae includes more than 2000 species of black flies worldwide. Their morphological uniformity creates difficulty for species identification, which limits our knowledge of their ecology and vectorial role. We investigated the systematics of black flies in a semi-arid area of the Iberian Peninsula, an ecologically harsh environment for these organisms. Sampling adult black flies in three different habitats (by means of CDC traps) and in avian nest boxes and collecting immature stages in high-salinity rills provided a representative sample of the component species. A combination of approaches, including morphological, chromosomal, and molecular (based on the mitochondrial cytochrome C oxidase subunit I (COI) and internal transcribed spacer 2 (ITS2) genes) revealed five species: four common species (Simulium intermedium, S. petricolum, S. pseudequinum, and S. rubzovianum) and the first European record for S. mellah. Barcoding gap and phylogenetic analyses revealed that ITS2 is a key marker to identify the species, whereas the COI marker does not provide enough resolution to identify some species or infer their phylogenetic relationships. Morphological and chromosomal features are also provided to identify S. mellah unequivocally. Our study highlights the need for integrated studies of black flies in ecologically extreme habitats to increase our knowledge of their distribution, ecology, and potential risks for public health.

Effect of high-volume insecticide spraying on sand fly vectors in household gardens in Spain.

This study investigated the efficacy of high-volume spraying with the adulticide α-cypermethrin alone and in combination with the larvicide diflubenzuron on the density of sand flies in gardens of three detached households in periurban areas in southeast Spain. Treatments were applied four times between June and August 2016, and four nearby sites, two households and two non-urbanized sites, were untreated controls. The number of sand flies collected between May and October 2016 using sticky interception and light attraction traps, was 4446 specimens. Species identified morphologically included Sergentomyia minuta (n = 2101; 48%), Phlebotomus perniciosus (n = 1922; 44%), Phlebotomus papatasi (n = 173; 4%), Phlebotomus sergenti (n = 161; 4%) and Phlebotomus ariasi (n = 36; 1%). Sand flies were detected in both treated and untreated sites. The proportion of positive sticky traps and the median (range) density of sand flies in positive traps were 61% traps and 7 (2-172) sand flies/m2 /day in untreated sites, and 43% traps and 4 (1-56) sand flies/m2 /day in treated sites (p < 0.05). Similarly, for light traps, it was 96% traps and 30 (3-168) flies/trap/day, and 83% traps and 3 (1-12) sand flies/trap/day, respectively (p < 0.05). However, sand fly density followed a comparable seasonal pattern in untreated and treated sites and did not consistently decrease following insecticide applications. These results were confirmed with mixed negative binomial modelling of sand fly density adjusted for time since application, month, environmental setting and site. The limited efficacy of the treatments, added to their cost, the impact of insecticides on non-target organisms and human health, and the risk of development of insecticide resistance, should dissuade similar outdoor applications to control sand fly vector populations in residential areas.

Multiple invasions, Wolbachia and human-aided transport drive the genetic variability of Aedes albopictus in the Iberian Peninsula.

The Asian tiger mosquito, Aedes albopictus, is one of the most invasive species in the world. Native to the tropical forests of Southeast Asia, over the past 30 years it has rapidly spread throughout tropical and temperate regions of the world. Its dramatic expansion has resulted in public health concerns as a consequence of its vector competence for at least 16 viruses. Previous studies showed that Ae. albopictus spread has been facilitated by human-mediated transportation, but much remains unknown about how this has affected its genetic attributes. Here we examined the factors that contributed to shaping the current genetic constitution of Ae. albopictus in the Iberian Peninsula, where the species was first found in 2004, by combining population genetics and Bayesian modelling. We found that both mitochondrial and nuclear DNA markers showed a lack of genetic structure and the presence of worldwide dominant haplotypes, suggesting regular introductions from abroad. Mitochondrial DNA showed little genetic diversity compared to nuclear DNA, likely explained by infection with maternally transmitted bacteria of the genus Wolbachia. Multilevel models revealed that greater mosquito fluxes (estimated from commuting patterns and tiger mosquito population distribution) and spatial proximity between sampling sites were associated with lower nuclear genetic distance, suggesting that rapid short- and medium-distance dispersal is facilitated by humans through vehicular traffic. This study highlights the significant role of human transportation in shaping the genetic attributes of Ae. albopictus and promoting regional gene flow, and underscores the need for a territorially integrated surveillance across scales of this disease-carrying mosquito.

Geographic distribution of the V1016G knockdown resistance mutation in Aedes albopictus: a warning bell for Europe.

BACKGROUND: Colonization of large part of Europe by the Asian tiger mosquito Aedes albopictus is causing autochthonous transmission of chikungunya and dengue exotic arboviruses. While pyrethroids are recommended only to reduce/limit transmission, they are widely implemented to reduce biting nuisance and to control agricultural pests, increasing the risk of insurgence of resistance mechanisms. Worryingly, pyrethroid resistance (with mortality < 70%) was recently reported in Ae. albopictus populations from Italy and Spain and associated with the V1016G point mutation in the voltage-sensitive sodium channel gene conferring knockdown resistance (kdr). Genotyping pyrethroid resistance-associated kdr mutations in field mosquito samples represents a powerful approach to detect early signs of resistance without the need for carrying out phenotypic bioassays which require availability of live mosquitoes, dedicated facilities and appropriate expertise. METHODS: Here we report results on the PCR-genotyping of the V1016G mutation in 2530 Ae. albopictus specimens from 69 sampling sites in 19 European countries. RESULTS: The mutation was identified in 12 sites from nine countries (with allele frequencies ranging from 1 to 8%), mostly distributed in two geographical clusters. The western cluster includes Mediterranean coastal sites from Italy, France and Malta as well as single sites from both Spain and Switzerland. The eastern cluster includes sites on both sides of the Black Sea in Bulgaria, Turkey and Georgia as well as one site from Romania. These results are consistent with genomic data showing high connectivity and close genetic relationship among West European populations and a major barrier to gene flow between West European and Balkan populations. CONCLUSIONS: The results of this first effort to map kdr mutations in Ae. albopictus on a continental scale show a widespread presence of the V1016G allele in Europe, although at lower frequencies than those previously reported from Italy. This represents a wake-up call for mosquito surveillance programs in Europe to include PCR-genotyping of pyrethroid resistance alleles, as well as phenotypic resistance assessments, in their routine activities.

Consolidation of Aedes albopictus Surveillance Program in the Autonomous Community of the Region of Murcia, Spain.

BACKGROUND: Due to the Spanish legal framework, the national program for vector-borne diseases results from the agreement between national and regional governments, and it is the basis for the development of the regional programs, which should include the regional entomological surveillance program. Aedes albopictus was recorded for the first time in the Region of Murcia, in 2011. It gave rise to a new epidemiological scenario due to the presence of a competent vector of several arboviruses, which resulted in autochthonous cases of dengue in 2018. METHODS: 40 out of 45 municipalities participated in the regional entomological surveillance program, and 266 sampling points were established, with two ovitraps at each site as pseudo-replications. The study period was from April 16th to November 26th, with bi-weekly sample collections: 16 regional samplings were carried out. RESULTS: Regional participation was high, and data loss was low (1.26%). Ae. albopictus was detected in 4.9% of samples and 89.4% of points, located in 39 of the 40 municipalities. The intensity of the presence of Ae. albopictus was estimated by a positivity index, that is, the percentage of positive samples over time. The vector phenology was obtained at a regional level, using the number of eggs as estimation of population density and the positivity values of points and municipalities. Every two weeks, real-time results were provided to the municipalities, which could use them as a vector management tool. CONCLUSION: The regional entomological surveillance program for Ae. albopictus in the Region of Murcia was consolidated in 2019, with standardized and comparable methods. Almost all the municipalities of the region have observed the presence of Ae. albopictus, although intensity and spatial and temporal cover vary among them.

Employing Different Traps for Collection of Mosquitoes and Detection of Dengue, Chikungunya and Zika Vector, Aedes albopictus, in Borderline of Iran and Pakistan.

BACKGROUND: Southeastern Iran has been established as an area with the potential to harbor Asian tiger mosquito populations. In 2013, a few numbers of Aedes albopictus were detected in three sampling sites of this region. This field study was aimed to evaluate the efficacy of various traps on monitoring mosquitoes and status of this dengue vector, in five urban and 15 suburban/rural areas. METHODS: For this purpose, four adult mosquito traps (BG-sentinel 2, bednet, Malaise, and resting box trap) were used and their efficacy compared. In addition, large numbers of CDC ovitraps were employed, within 12 months. RESULTS: A total of 4878 adult samples including 22 species covering five genera were collected and identified from traps. It was not revealed any collection of Ae. albopictus. Statistical analysis showed no significant difference in meteorological variables between the two periods, the previous report and the current study. There were significant differences in the total number of mosquitoes collected by various traps in the region across different months. CONCLUSION: The resulting data collected here on the efficiency of the various trap types can be useful for monitoring the densities of mosquito populations, which is an important component of a vector surveillance system. While the presence of Ae. albopictus was determined in this potential risk area, there is no evidence for its establishment and further monitoring needs to be carried out.

Citizen science provides a reliable and scalable tool to track disease-carrying mosquitoes.

Recent outbreaks of Zika, chikungunya and dengue highlight the importance of better understanding the spread of disease-carrying mosquitoes across multiple spatio-temporal scales. Traditional surveillance tools are limited by jurisdictional boundaries and cost constraints. Here we show how a scalable citizen science system can solve this problem by combining citizen scientists' observations with expert validation and correcting for sampling effort. Our system provides accurate early warning information about the Asian tiger mosquito (Aedes albopictus) invasion in Spain, well beyond that available from traditional methods, and vital for public health services. It also provides estimates of tiger mosquito risk comparable to those from traditional methods but more directly related to the human-mosquito encounters that are relevant for epidemiological modelling and scalable enough to cover the entire country. These results illustrate how powerful public participation in science can be and suggest citizen science is positioned to revolutionize mosquito-borne disease surveillance worldwide.

Potential Risk Areas of Aedes albopictus in South-Eastern Iran: A Vector of Dengue Fever, Zika, and Chikungunya.

The possibility of the rapid and global spread of Zika, chikungunya, yellow fever, and dengue fever by Aedes albopictus is well documented and may be facilitated by changes in climate. To avert and manage health risks, climatic and topographic information can be used to model and forecast which areas may be most prone to the establishment of Ae. albopictus. We aimed to weigh and prioritize the predictive value of various meteorological and climatic variables on distributions of Ae. albopictus in south-eastern Iran using the Analytical Hierarchy Process. Out of eight factors used to predict the presence of Ae. albopictus, the highest weighted were land use, followed by temperature, altitude, and precipitation. The inconsistency of this analysis was 0.03 with no missing judgments. The areas predicted to be most at risk of Ae. albopictus-borne diseases were mapped using Geographic Information Systems and remote sensing data. Five-year (2011-2015) meteorological data was collected from 11 meteorological stations and other data was acquired from Landsat and Terra satellite images. Southernmost regions were at greatest risk of Ae. albopictus colonization as well as more urban sites connected by provincial roads. This is the first study in Iran to determine the regional probability of Ae. albopictus establishment. Monitoring and collection of Ae. albopictus from the environment confirmed our projections, though on-going field work is necessary to track the spread of this vector of life-threatening disease.

Implementation and evaluation of the medication management in nursing units of a university hospital by means of a quality improvement cycle.

AIMS: The aim of this study was to improve compliance with protocols for the correct handling and storage of medication, both in medication rooms and on resuscitation trolleys in order to reduce the level of adverse effects on inpatients. BACKGROUND: The most frequent adverse effects in healthcare are due to medication errors. METHODS: Twenty hospital units were studied in 2012. Their medication rooms and resuscitation trolleys share the same medication supply system. Twelve criteria were evaluated and in the first assessment, corrective measures were implemented and criteria were evaluated again. RESULTS: In the first evaluation, the total amount of cases of noncompliance was 153 (65.38%). Seven corrective measures were implemented on the 6 criteria which accumulate 60% of the noncompliance cases. After that, 81 noncompliance cases (34.62%) were obtained. Almost all of evaluated criteria showed an improvement. Nine of the 12 criteria evaluated showed a statistically significant improvement in the second assessment. CONCLUSIONS: A quality improvement cycle is a useful tool to identify safety problems, related to management of medication rooms and resuscitation trolleys of nursing units. It is possible to identify and implement improvement measures without any additional cost.

Review of ten-years presence of Aedes albopictus in Spain 2004-2014: known distribution and public health concerns.

Ten years have gone by since the Asian tiger mosquito Aedes albopictus was recorded for the first time in Spain. In this paper, all relevant published information about this vector in Spain for the period 2004-2014 is reviewed. The known distribution for 2014 is provided, including all historical records (published and unpublished data) and the results from samplings of the last year. The consequences on public health about the presence of the Asian tiger mosquito in Spain are also highlighted. Further, legal aspects and control plans related to the management and diseases transmitted by this invasive vector species are also discussed.

Updated distribution of Aedes albopictus (Diptera: Culicidae) in Spain: new findings in the mainland Spanish Levante, 2013.

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia.

Strepsiptera, phylogenomics and the long branch attraction problem.

Insect phylogeny has recently been the focus of renewed interest as advances in sequencing techniques make it possible to rapidly generate large amounts of genomic or transcriptomic data for a species of interest. However, large numbers of markers are not sufficient to guarantee accurate phylogenetic reconstruction, and the choice of the model of sequence evolution as well as adequate taxonomic sampling are as important for phylogenomic studies as they are for single-gene phylogenies. Recently, the sequence of the genome of a strepsipteran has been published and used to place Strepsiptera as sister group to Coleoptera. However, this conclusion relied on a data set that did not include representatives of Neuropterida or of coleopteran lineages formerly proposed to be related to Strepsiptera. Furthermore, it did not use models that are robust against the long branch attraction artifact. Here we have sequenced the transcriptomes of seven key species to complete a data set comprising 36 species to study the higher level phylogeny of insects, with a particular focus on Neuropteroidea (Coleoptera, Strepsiptera, Neuropterida), especially on coleopteran taxa considered as potential close relatives of Strepsiptera. Using models robust against the long branch attraction artifact we find a highly resolved phylogeny that confirms the position of Strepsiptera as a sister group to Coleoptera, rather than as an internal clade of Coleoptera, and sheds new light onto the phylogeny of Neuropteroidea.

Updated distribution of Aedes albopictus (Diptera: Culicidae) in Spain: new findings in the mainland Spanish Levante, 2013

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia.

Expression of the pupal determinant broad during metamorphic and neotenic development of the strepsipteran Xenos vesparum Rossi.

Derived members of the endoparasitic order Strepsiptera have acquired an extreme form of sexual dimorphism whereby males undergo metamorphosis and exist as free-living adults while females remain larviform, reaching sexual maturity within their hosts. Expression of the transcription factor, broad (br) has been shown to be required for pupal development in insects in which both sexes progress through metamorphosis. A surge of br expression appears in the last larval instar, as the epidermis begins pupal development. Here we ask if br is also up-regulated in the last larval instar of male Xenos vesparum Rossi (Stylopidae), and whether such expression is lost in neotenic larviform females. We clone three isoforms of br from X. vesparum (Xv'br), and show that they share greatest similarity to the Z1, Z3 and Z4 isoforms of other insect species. By monitoring Xv'br expression throughout development, we detect elevated levels of total br expression and the Xv'Z1, Xv'Z3, and Xv'Z4 isoforms in the last larval instar of males, but not females. By focusing on Xv'br expression in individual samples, we show that the levels of Xv'BTB and Xv'Z3 in the last larval instar of males are bimodal, with some males expressing 3X greater levels of Xv'br than fourth instar femlaes. Taken together, these data suggest that neoteny (and endoparasitism) in females of Strepsiptera Stylopidia could be linked to the suppression of pupal determination. Our work identifies a difference in metamorphic gene expression that is associated with neoteny, and thus provides insights into the relationship between metamorphic and neotenic development.

Sperm Cells of a Primitive Strepsipteran.

The unusual life style of Strepsiptera has presented a long-standing puzzle in establishing its affinity to other insects. Although Strepsiptera share few structural similarities with other insect orders, all members of this order share a parasitic life style with members of two distinctive families in the Coleoptera-the order now considered the most closely related to Strepsiptera based on recent genomic evidence. Among the structural features of several strepsipteran families and other insect families that have been surveyed are the organization of testes and ultrastructure of sperm cells. For comparison with existing information on insect sperm structure, this manuscript presents a description of testes and sperm of a representative of the most primitive extant strepsipteran family Mengenillidae, Eoxenos laboulbenei. We compare sperm structure of E. laboulbenei from this family with that of the three other families of Strepsiptera in the other strepsipteran suborder Stylopidia that have been studied as well as with members of the beetle families Meloidae and Rhipiphoridae that share similar life histories with Strepsiptera. Meloids, Rhipiphorids and Strepsipterans all begin larval life as active and viviparous first instar larvae. This study examines global features of these insects' sperm cells along with specific ultrastructural features of their organelles.