Argasid ticks of the Ornithodoros erraticus complex are associated with traditional pig-farming practices on the Iberian Peninsula and are also found elsewhere in North Africa, West Africa, and western Asia. The ticks associated with pig farming on the Iberian Peninsula are the only biological vectors of African swine fever virus (ASFV) known to occur in Europe, and their ecology makes them an extremely effective reservoir of both ASFV and the Borrelia species which cause tick-borne relapsing fever (TBRF) in humans. The recent reappearance of ASFV in the European Union, coupled with evidence that Portuguese tick populations continue to harbor Borrelia despite a lack of confirmed human infections, suggest that these populations merit closer attention. In Portugal, a series of surveys over the last twenty-five years indicates that the number of farm sites with tick infestations has declined and suggest that populations are sensitive to changes in farm management, particularly the use of modern pig housing. Various technologies have been suggested for the control of farm-associated Ornithodoros ticks and related species but, in our opinion, farm management changes are still the most effective strategy for population control. Furthermore, we suggest that this species could probably be eradicated from Iberian pig farms.

Urban wetlands are being created in the UK as part of sustainable urban drainage strategies, to create wetland habitats lost during development, to provide a habitat for protected species, and to increase the public's access to ‘blue-space’ for the improvement of health and well-being. Sewage treatment reedbeds are also being incorporated into newly constructed wetlands to offer an alternative approach to dealing with sewage. This field study aims to provide the first UK evidence of how such newly constructed aquatic habitats are colonized by mosquitoes. A number of new aquatic habitats were surveyed for immature mosquitoes every fortnight over the first two years following wetland construction. The majority of mosquitoes collected were Culex sp. and were significantly associated with the sewage treatment reedbed system, particularly following storm events and sewage inflow. Other more natural aquatic habitats that were subject to cycles of drying and re-wetting contributed the majority of the remaining mosquitoes colonizing. Colonization of permanent habitats was slow, particularly where fluctuations in water levels inhibited emergent vegetation growth. It is recommended that during the planning process for newly constructed wetlands consideration is given on a case-by-case basis to the impact of mosquitoes, either as a cause of nuisance or as potential vectors. Although ornithophagic Culex dominated in this wetland, their potential role as enzootic West Nile virus vectors should not be overlooked.

Bartonellosis is an infectious bacterial disease. The prevalence and genetic characteristics of Bartonella spp. in fleas of wild and domestic animals from Palestinian territories are described. Flea samples (n=289) were collected from 121 cats, 135 dogs, 26 hyraxes and seven rats from northern (n=165), central (n=113), and southern Palestinian territories (n=11). The prevalent flea species were: Ctenocephalides felis (n=119/289; 41.2%), Ctenocephalides canis (n=159/289; 55%), and Xenopsylla sp. (n=7/289; 2.4%). Targeting the Intergenic Transcribed Spacer (ITS) locus, DNA of Bartonella was detected in 22% (64/289) of all fleas. Fifty percent of the C. felis and 57% of the Xenopsylla sp. contained Bartonella DNA. DNA sequencing showed the presence of Bartonella clarridgeiae (50%), Bartonella henselae (27%), and Bartonella koehlerae (3%) in C. felis. Xenopsylla sp. collected from Rattus rattus rats were infected with Bartonella tribocorum, Bartonella elizabethae, and Bartonella rochalimae. Phylogenetic sequence analysis using the 16S ribosomal RNA gene obtained four genetic clusters, B. henselae and B. koehlerae as subcluster 1, B. clarridgeiae as cluster 2, while the rat Bartonella species (B. tribocorum and B. elizabethae) were an outgroup cluster. These findings showed the important role of cat and rat fleas as vectors of zoonotic Bartonella species in Palestinian territories. It is hoped that this publication will raise awareness among physicians, veterinarians, and other health workers of the high prevalence of Bartonella spp. in fleas in Palestinian territories and the potential risk of these pathogens to humans and animals in this region.

A novel multiple membrane blood-feeding system for mosquitoes has been developed for the study and routine maintenance of Aedes aegypti L. and Aedes albopictus Skuse that require a meal of vertebrate blood to produce eggs. This blood-feeding system uses cattle collagen sausage-casing membrane to facilitate feeding. The efficiency of this blood-feeding system was compared to a live mice blood source. We observed that Ae. aegypti that fed on pig whole blood had 89.7% (w/o ATP) and 90.7% (w/ ATP) blood-feeding rates, which were not significantly different from the mice-fed ones (98.0%). Ae. albopictus fed on pig whole blood (w/ ATP) had a success rate of 84.4%, which was significantly different from the mice-fed mosquitoes (51.1%). The feeding rates did not differ between sausage-casing membrane and Parafilm-M^®. The survival rate, fecundity, pupation, and pupal emergence rates of Aedes females fed on pig whole blood were not significantly different from the mice-fed ones. The artificial blood feeder can be applied to replace live animals as blood sources. Considering that this simple, inexpensive, convenient, and efficient feeding device can be built with common laboratory materials for research on Aedes mosquitoes.

Competition bioassays were conducted with the filth fly pupal parasitoids Muscidurax raptor (Girault & Sanders) and M. raptorellus (Kogan & Legner) (Hymenoptera: Pteromalidae) using house fly Musca domestica L. (Diptera: Muscidae) hosts at different host densities. Muscidifurax raptor had a significant impact on M. raptorellus when hosts were limiting in sequential parasitism tests. Fewer than six M. raptorellus adult progeny emerged from groups of 50 fly pupae that were parasitized by M. raptor at the same time or when M. raptor parasitism preceded M. raptorellus by 48 h, respectively, compared with 42–55 M. raptorellus progeny produced when this species was tested alone. Production of M. raptor was significantly lower when parasitism by this species was preceded by M. raptorellus (25) than when M. raptor was tested alone (43). When the two species parasitized hosts at the same time in different proportions at low host:parasitoid densities (5:1), M. raptorellus produced 13 progeny per parent female when it was the sole species present and fewer than two when M. raptor was present. No negative impact of M. raptorellus on M. raptor was observed. Neither species had a substantial effect on the success of the other at higher host:parasitoid densities.

Previous studies have used C and N isotope ratios to investigate the use of different food resources such as plant and animal detritus by container-breeding mosquitoes. This study is the first to report on the potential food resources assimilated by larval mosquitoes in agricultural and reference wetlands. Larval mosquitoes (Diptera: Culcidae) were sampled, along with their potential food resources, from agricultural and reference wetland habitats throughout a seasonal hydroperiod. IsoSource mixing model results indicated that food resources had greater δ¹⁵N isotope values in agricultural wetlands compared with cypress-gum swamps. In February, Aedes vexans (Meigen) and Culex territans Walker larvae fed primarily on lower quality food resources (coarse particulate organic matter and sediment) based on C:N. In contrast, higher quality food resources (fine particulate organic matter) were utilized by Anopheles spp. throughout the study and by Psorophora columbiae (Dyer and Knab) in May. This research contributes to a more comprehensive understanding of the food resources available and assimilated by larval mosquitoes in agricultural wetlands.

This study was designed to compare the behavioral responses of Aedes aegypti to a single essential oil and to a mixture of two or three essential oils using an excito-repellency test chamber. Mixtures were prepared from essential oils extracted from Litsea cubeba (LC), Litsea salicifolia (LS), and Melaleuca leucadendron (ML). In general, the mixture of essential oils produced a much stronger escape response by Ae. aegypti, regardless of the test conditions. No significant difference in escape responses was seen when the mixture of oils was compared with a standard commercial product containing DEET. Greater contact irritancy was seen from mixed oils of LC and LS than with other mixed oils. Mixtures of LC and LS at 0.075% showed the highest synergistic action (65.5% escaped) compared to that with unmixed oil alone at the same concentration (LC=20% and LS=32.2%). In addition, mixtures of LC and LS at 0.075% demonstrated the highest non-contact repellency (62.7%) and showed a greater effect than the use of LC (20%) or LS (20.3%) alone. We conclude that mixtures of two essential oils show potential as active ingredients for mosquito repellents.

Culicoides biting midges play an important role in the epidemiology of many vector-borne infections, including bluetongue virus, an internationally important virus of ruminants. The territory of the Russian Federation includes regions with diverse climatic conditions and a wide range of habitats suitable for Culicoides. This review summarizes available data on Culicoides studied in the Russian Federation covering geographically different regions, as well as findings from adjacent countries. Previous literature on species composition, ranges of dominant species, breeding sites, and host preferences is reviewed and suggestions made for future studies to elucidate vector-virus relationships.

Tick-borne relapsing fever (TBRF) is caused by Borrelia spirochetes transmitted to humans by Argasid soft ticks of the genus Ornithodoros. We investigated the presence of Ornithodoros ticks in rodent burrows in nine sites of the Gharb region of northwestern Morocco where we recently documented a high incidence of TBRF in humans. We assessed the Borrelia infection rate by nested PCR and sequencing. All sites investigated were colonized by ticks of the Ornithodoros marocanus complex and a high proportion of burrows (38.4%) were found to be infested. Borrelia infections were observed in 6.8% of the ticks tested. Two Borrelia species were identified by sequencing: B. hispanica and B. crocidurae. The discovery in northwestern Morocco of Ornithodoros ticks infected by B. crocidurae represents a 350 km range extension of this Sahelo-Saharan spirochete in North Africa. The spread of B. crocidurae may be related to the increasing aridity of northwestern Morocco in relation to climate change.

We investigated the spatial and temporal distribution of blackfly larvae on a mesohabitat scale atfour different breeding sites in the Brazilian savannah “Cerrado.”A total of 2,385 larvae of 12 different species were collected, and three of the sampling sites revealedspecies richness and diversity peaks in April. A principal components analysis was performed to reduce abiotic data into a smaller set of independent variables, such as precipitation and river size, and subsequent linear regressions showed that Simulium subpallidum was positively correlated with precipitation.This species, along with S. nigrimanum, was negatively correlated withriver size, whereasS. perflavum was negatively correlated with precipitation. Beta diversity was negatively correlated withprecipitation, and black fly taxocenoses were markedly different between wet and dry seasons, as shown through nonmetric multidimensional scaling, suggesting that precipitation playsa major role in structuring these communities, probably because of enhanced drift and the dendritic nature of lotic metacommunities.

The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito-repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65–98% escape for contact, 21.4–94.4% escape for non-contact) compared to Ae. aegypti (3.7–72.2% escape (contact), 0–31.7% (non-contact)) and Ae. albopictus (3.5–94.4% escape (contact), 11.2–63.7% (non-contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65–97.8% escape) than non-contact repellents (0-50.8% escape for non-contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.

We investigated the interactions of tadpole shrimp, a mosquito biological control agent, with the juvenile hormone analog methoprene and a monomolecular surface film. In laboratory assays, the tadpole shrimp (TPS) Triops newberryi (Packard) was able to tolerate high concentrations of methoprene without negative impacts on its growth, longevity, and fecundity when exposed to 1 to 10 mg/liter, or 90–900 fold, of the IE₉₀ levels against a laboratory colony of Culex quinquefasciatus Say. The same held true in field trials when the habitats were treated with Altosid^® Liquid Larvicide (Altosid^® LL, 5% methoprene) at 0.3–1.2 liters/ha. or 1–4 fold of the label rates for mosquito control. However, some significant impacts on the TPS occurred when they were exposed to Agnique^® Monomolecular Film (Agnique^® MMF) at the label rates for mosquito control ranging from 1.89–9.45 liters/ha. under laboratory and field conditions. To avoid the negative impact of Agnique MMF on tadpole shrimp, it appears that 1.89 liters/ha. would be the maximum rate when Agnique MMF is used to control mosquitoes in the habitats where the TPS is employed as a biological control agent, or prevailing in the aquatic habitats with potential for suppressing mosquito larval populations.

Mosquitoes of various species mate in swarms comprised of tens of thousands of flying males. In this study, we examined Aedes aegypti swarming behavior and identified associated chemical cues. Novel evidence is provided that Ae. aegypti females aggregate by means of olfactory cues, such as aggregation pheromones. Isolation of Ae. aegypti aggregation pheromones was achieved by aeration of confined mosquitoes and collection of associated volatiles by glass filters. The collected volatiles were identified through gas chromatography mass spectrometry (GCMS). Three aggregation pheromones were collected and identified as 2,6,6-trimethylcyclohex-2-ene-1,4-dione (ketoisophorone) (CAS# 1125-21-9, t_R = 18.75), 2,2,6-trimethylcyclohexane-1,4-dione (the saturated analog of ketoisophorone) (CAS# 20547-99-3, t_R = 20.05), and 1-(4-ethylphenyl) ethanone (CAS# 937-30-4, t_R = 24.22). Our biological studies revealed that the identified compounds stimulated mosquito behavior under laboratory conditions. The mechanism of mosquito swarm formation is discussed in light of our behavioral study findings. A preliminary field trial demonstrated the potential application of the isolated aggregation pheromones in controlling Ae. Aegypti.

Larvae of some species of mosquitoes have been shown to respond to water-borne kairomones from predators by reducing bottom-feeding and replacing it with surface filter-feeding, which uses less movement and is thus less likely to attract a predator. However, if no predator attack takes place, then it would be more efficient to use a risk allocation strategy of habituating their response depending on the predator and the overall risk. The larvae of Culiseta longiareolata Macquart live in temporary rain-filled pools, where they are exposed to a high level of predation. Within one hour, they responded to kairomones from dragonfly or damselfly nymphs, or to the fish Aphanius, by significantly reducing bottom-feeding activity. Continued exposure to the predator kairomones resulted in habituation of their response to damselflies, a slower habituation to fish, but no habituation to dragonflies even after 30 h. In contrast, the larvae of Culex quinquefasciatus Say normally live in highly polluted and thus anaerobic water, where the predation risk will be much lower. They also showed a significant reduction in bottom-feeding after 1 h of exposure to predator kairomones but had completely habituated this response within 6 h of continuous exposure. Some species of mosquito larvae can thus show a very rapid habituation to predator kairomones, while others only habituate slowly depending on the predator and overall predation risk.

A study of species diversity of Anopheles mosquitoes, biting patterns, and seasonal abundance of important mosquito vectors was conducted in two villages of Chang Island, Trat Province, in eastern Thailand, one located along the coast and the other in the low hills of the central interior of the island. Of 5,399 captured female anophelines, 70.25% belong to the subgenus Cellia and remaining specimens to the subgenus Anopheles. Five important putative malaria vectors were molecularly identified, including Anopheles epiroticus, Anopheles dirus, Anopheles sawadwongporni, Anopheles maculatus, and Anopheles minimus. Anopheles epiroticus was the most commonly collected species in the coastal site, whereas An. dirus was found to be most abundant in the forest-hill site. From both locations, a greater number of mosquitoes was collected during the dry season compared to the wet. Anopheles epiroticus showed greater exophagic and zoophilic behavior with the highest blood feeding densities occurring between 18:00 and 19:00. In contrast, An. dirus demonstrated an activity peak between midnight and 01:00. We conclude that An. epiroticus and An. dirus, in coastal and inland areas, respectively, appear to be the most epidemiologically important malaria vectors on Chang Island. As no studies of vector competency specific to Chang Island have been conducted, our conclusions that these two species play a primary role in malaria transmission are based on evidence from other localities in Thailand and mainland Southeast Asia. This information serves as a basis for designing improved vector control programs that target specific species, and if integrated with other interventions could result in the elimination of malaria transmission on the island.

Mosquitoes of both sexes feed on plants to obtain sugar. Nocturnal species probably locate the plants primarily by their volatile semiochemicals that also form the basis for the mosquitoes' innate plant-species preferences. To evaluate these olfactory preferences quantitatively, we used a two-choice wind-tunnel olfactometer to measure the upwind orientation of Anopheles gambiae Giles, an important vector of malaria in equatorial Africa, toward odor plumes produced by nine plant species common where this mosquito occurs. These plants are reported to induce feeding behaviors in An. gambiae and to produce floral or extrafloral nectar. Results presented here demonstrated that the volatiles of S. didymobotrya, P. hysterophorus, S. occidentalis, and L. camara, in descending order of numbers of mosquitoes responding, were all attractive, compared to a control plant species, whereas D. stramonium, R. communis, S. bicapsularis, T. stans, and T. diversifolia were not. As expected, chromatographic analysis of the headspace of attractive plants whose volatiles were captured by stir-bar sorptive extraction revealed a wide range of compounds, primarily terpenoids. Once their bioactivity and attractiveness for An. gambiae, alone and in blends, has been firmly established, some of these semiochemicals may have applications in population sampling and control.

The objective of the present study was to identify mosquito fauna and to evaluate whether different light bulb colors influence the attraction of light traps in the Guapiaçu Ecological Reserve. Samples were obtained monthly during the period of February, 2012 to January, 2013. Centers for Disease Control (CDC) light traps with incandescent light bulbs and LED (ultraviolet, blue, green, and red) bulbs were utilized. In total, 8,170 specimens were captured, including 59 species. The presence of Anopheles nimbus (Theobald 1902) and Orthopodomyia fascipes Coquillet 1906 were recorded for the first time in the state of Rio de Janeiro. The green LED trap attracted the highest number of specimens and presented the highest diversity and mosquito average. The blue and green LED traps attracted the highest number of species. However, the differences between lights were not significant. The most common species were Coquillettidia juxtamansonia (Chagas 1907), Culex declarator Dyar and Knab 1906, and Culex ribeirensis Forattini and Sallum 1985.

The aim of this study was to measure the protective efficacy of Olyset^® Plus, a new long-lasting factory-treated insecticidal net (LLIN) incorporated with 2% permethrin and 1% of the synergist piperonyl butoxide (PBO), against cutaneous leishmaniasis (CL) transmission under field conditions. A village-scale trial, promoting the use of LLIN by the local inhabitants of the study area was conducted as a pilot study in a new hyperendemic focus of CL caused by a Leishmania infantum/L. donovani hybrid parasite transmitted by proven vector species Phlebotomus tobbi in Cukurova Plain, Adana, Turkey, between May, 2013 and May, 2014. The study area comprised eight villages; two of them were selected as an intervention village with Olyset^® Plus net (Kizillar) and a control village without net application (Malihidirli). Six villages with surrounding allopatric barriers were utilized as a buffer zone cluster between intervention and control villages. Monthly entomological surveys were performed in the intervention and control villages and Damyeri, representing the other six villages, to collect adults of Phlebotomus tobbi. Results showed a significant reduction in cutaneous leishmaniasis incidence in the intervention village from 4.78% to 0.37%. The protective efficacy rate of LLIN was 92.2%. In contrast, incidence rates increased in the control village from 3.67% to 4.69%. We also evaluated residual insecticide levels of used nets after six and 12 months of usage. It was determined that the nets had retained full insecticidal strength. These results highlight the value of real-world data on bed net effectiveness and longevity to guide decisions regarding sand fly control strategies. To the best of our knowledge, this is the first field study to evaluate Olyset® Plus efficacy in a hyperendemic cutaneous leishmaniasis area.

Locally acquired dengue cases in the continental U.S. are rare. However, outbreaks of dengue-1 during 2009, 2010, and 2013 in Florida and dengue-1 and -2 in Texas suggest vulnerability to transmission. Travel and commerce between Puerto Rico and the U.S. mainland is common, which may pose a risk for traveler-imported dengue cases. Mosquitoes were collected in Florida and used to evaluate their susceptibility to dengue viruses (DENV) from Puerto Rico. Aedes aegypti and Ae. albopictus were susceptible to virus infection with DENV-1 and -2. No significant differences were observed in rates of midgut infection or dissemination between Ae. aegypti or Ae. albopictus for DENV-1 (6–14%). Aedes aegypti was significantly more susceptible to midgut infection with DENV-2 than Ae. albopictus (Ae. aegypti, ∼28%; Ae. albopictus, ∼9%). The dissemination rate with dengue-2 virus for Ae. aegypti (23%) was greater than Ae. albopictus (0%), suggesting that Ae. albopictus is not likely to be an important transmitter of the DENV-2 isolate from Puerto Rico. These results are discussed in light of Florida's vulnerability to DENV transmission.

Soil pits excavated for home construction are important larval habitats for malaria vectors in certain parts of Africa. Borrow pits in diverse stages of ecological succession in a maize-farming region of Western Ethiopia were surveyed to assess the relationships between stage of succession and the structure and composition of invertebrate and plant communities, with particular attention to Anopheles gambiae s.l. and An. coustani, the primary local malaria vectors. An array of 82 borrow pits was identified in a multi-lobed drainage basin in the community of Woktola. Each pit was evaluated on its physical features and by faunal and floral surveys during August, 2011, at the height of the longer rainy season (kiremt). Anopheles gambiae s.l. and An. coustani were the sole immature anophelines collected, often coexisting with Culex spp. Sedges were the most common plants within these pits, and included Cyperus elegantulus, C. flavescens, C. erectus and C. assimilis. The legume Smithia abyssinica, Nile grass (Acroceras macrum), cutgrass (Leersia hexandra), clover (Trifolium spp.), and the edible herb Centella asiatica, were also common in these habitats. No plant species in particular was strongly and consistently predictive of the presence or absence of mosquito immatures, particularly with regard to An. coustani. The presence of An.gambiae s.l. immatures in borrow pit habitats was negatively correlated with the presence of backswimmers (Notonectidae) (Z = -2.34, P = 0.019). Young (freshly excavated) borrow pits more likely contained immature An. gambiae s.l. (Z =-2.86, P=0.004). Ecological succession was apparent in older pits, and as they aged, they became less likely to serve as habitats for An. gambiae s.l. (Z=0.26, P=0.796), and more likely to support An. coustani (Z=0.728, P=0.007). As borrow pits age they become less suitable for An. gambiae s.l. breeding and more likely to harbor An. coustani. The abundance of notonectids in habitats was a negative indicator for An. gambiae s.l. abundance. Plant species are not reliable indicators for the presence or absence of malaria vectors in borrow pits.

We determined the species diversity, blood-feeding behavior, and host preference of Anopheles mosquitoes in two malaria endemic areas of Tak (Mae Sot District) and Mae Hong Son (Sop Moei District) Provinces, located along the Thai border with Myanmar, during a consecutive two-year period. Anopheline mosquitoes were collected using indoor and outdoor human-landing captures and outdoor cow-baited collections. Mosquitoes were initially identified using morphological characters, followed by the appropriate multiplex AS-PCR assay for the identification of sibling species within Anopheles (Cellia) complexes and groups present. Real-time PCR was performed for parasite-specific detection in mosquitoes (Plasmodium spp. and Wuchereria bancrofti). A total of 7,129 Anopheles females were captured, 3,939 from Mae Sot and 3,190 from Sop Moei, with 58.6% and 37% of all anophelines identified as An. minimus, respectively. All three malaria vector complexes were detected in both areas. One species within the Minimus Complex (An. minimus) was present along with two related species in the Funestus Group, (An. aconitus, An. varuna), two species within the Dirus Complex (An. dirus, An. baimaii), and four species within the Maculatus Group (An. maculatus, An. sawadwongporni, An. pseudowillmori, and An. dravidicus). The trophic behavior of An. minimus, An. dirus, An. baimaii, An. maculatus, and An. sawadwongporni are described herein. The highest An. minimus densities were detected from February through April of both years. One specimen of An. minimus from Mae Sot was found positive for Plasmodium vivax.