Amblyomma americanum (L.) is a three-host tick that spends most of its life off-host and is an important vector of pathogens in the eastern United States. Our objectives were to develop a predictive statistical model that describes the number of active, off-host larvae from 2007 to 2011 and to determine the environmental variables associated with this pattern. Data used in this study came from monitoring conducted in northeast Missouri in which off-host ticks were collected from a permanent plot in a forest and an old field habitat every other week from approximately February to December. Variables examined were day length, degree days, total precipitation prior to sampling, wind speed, saturation deficit, number of adults prior to sampling, and collection site. Of the four regression models tested, the negative binomial model was selected. Fitted candidate models were compared relative to one another using values of eight model selection criteria and model averaging was used to develop a predictive model. The residual plots indicated that the weighted average model performs well in describing the number of larvae. Of the variables considered, the number of larvae was most strongly associated with increasing degree days, the number of active adults prior to sampling, and the forested site.

We investigated the olfactory responses of male mosquitoes to kairomones of vertebrate hosts in a dual-port olfactometer. The behavioral responses of unmated and mated male and female mosquitoes from one to ten days old to human odors were compared to the odors of different human hosts. To evaluate the relationship between the age of male mosquitoes and their responses, we performed experiments with males at different ages. Unmated Ae. aegypti males, one to two days old, did not fly upwind to human odor, whereas between three and ten days old they exhibited increased flight activity. The results showed that unmated and mated females were attracted by human odor, but those mated were more attracted by human odor than when unmated. Mated males were, in general, attracted by human odor, while the unmated males were not attracted but showed increased flight activity in the presence of human odor, suggesting swarming behavior. Further studies should be carried out in order to determine the role of human odors on male Ae. aegypti behavior.

The barcoding of life initiative provides a universal molecular tool to distinguish animal species based on the amplification and sequencing of a fragment of the subunit 1 of the cytochrome oxidase (COI) gene. Obtaining good quality DNA for barcoding purposes is a limiting factor, especially in studies conducted on small-sized samples or those requiring the maintenance of the organism as a voucher. In this study, we compared the number of positive amplifications and the quality of the sequences obtained using DNA extraction methods that also differ in their economic costs and time requirements and we applied them for the genetic characterization of louse flies. Four DNA extraction methods were studied: chloroform/isoamyl alcohol, HotShot procedure, Qiagen DNeasy^® Tissue and Blood Kit and DNA Kit Maxwell^® 16LEV. All the louse flies were morphologically identified as Ornithophila gestroi and a single COI-based haplotype was identified. The number of positive amplifications did not differ significantly among DNA extraction procedures. However, the quality of the sequences was significantly lower for the case of the chloroform/isoamyl alcohol procedure with respect to the rest of methods tested here. These results may be useful for the genetic characterization of louse flies, leaving most of the remaining insect as a voucher.

A phylogeny of anthropophilic and zoophilic anopheline mosquito species was constructed, using the nuclear internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome oxidase subunit I (COI) genes. The ITS2 alignment, typically difficult due to its noncoding nature and large size variations, was aided by using predicted secondary structure, making this phylogenetically useful gene more amenable to investigation. This phylogeny is unique in explicitly including zoophilic, non-vector anopheline species in order to illustrate their relationships to malaria vectors. Two new, cryptic species, Anopheles funestus-like and Anopheles rivulorum-like, were found to be present in Zambia for the first time. Sequences from the D3 region of the 28S rDNA suggest that the Zambian An. funestus-like may be a hybrid or geographical variant of An. funestus-like, previously reported in Malawi. This is the first report of An. rivulorum-like sympatric with An. rivulorum (Leeson), suggesting that these are separate species rather than geographic variants.

Most studies of the sand fly fauna in southeastern Brazil are conducted in the peridomiciliary environment of leishmaniasis endemic regions. Therefore, to increase the knowledge about diversity and richness of sand fly conservation areas, we describe here the sand fly fauna from the National Forest of Goytacazes (NFG), state of Espírito Santo, Brazil, and its surroundings areas. We also used sand fly fauna records from eight conservations units within the state of Espírito Santo to understand the similarity and relationships among them. The sand flies were simultaneously collected from June, 2008 to May, 2009 in two different environments: a preserved environment represented by the NFG and a modified environment represented by a peridomicile. To establish the similarity among the conservation units, we used a method very similar to parsimony analysis of endemism. We collected 2,466 sand fly specimens belonging to 13 species. Pressatia choti and Nyssomyia intermedia were the most abundant sand fly species. Ny. intermedia is a known vector of Leishmania braziliensis and epidemiological surveillance must be conducted in the area. We discuss aspects regarding the diversity of sand flies as well as the risk of transmission of Leishmania parasites in the area. We also provide for the first time a hypothesis of similarity relationships among conservation units within the state of Espírito Santo.

Distribution and abundance of disease vectors are directly related to climatic conditions and environmental changes. Remote sensing data have been used for monitoring environmental conditions influencing spatial patterns of vector-borne diseases. The aim of this study was to analyze the effect of the Normalized Difference Vegetation Index (NDVI) and Land Surface Temperature (LST) obtained from the Moderate Resolution Imaging Spectroradiometer (MODIS), and climatic factors (temperature, humidity, wind velocity, and accumulated rainfall) on the distribution and abundance of Anopheles species in northwestern Argentina using Poisson regression analyses. Samples were collected from December, 2001 to December, 2005 at three localities, Aguas Blancas, El Oculto and San Ramón de la Nueva Orán. We collected 11,206 adult Anopheles species, with the major abundance observed at El Oculto (59.11%), followed by Aguas Blancas (22.10%) and San Ramón de la Nueva Orán (18.79%). Anopheles pseudopunctipennis was the most abundant species at El Oculto, Anopheles argyritarsis predominated in Aguas Blancas, and Anopheles strodei in San Ramón de la Nueva Orán. Samples were collected throughout the sampling period, with the highest peaks during the spring seasons. LST and mean temperature appear to be the most important variables determining the distribution patterns and major abundance of An. pseudopunctipennis and An. argyritarsis within malarious areas.

The catastrophic 2010 earthquake in Port-au-Prince, Haiti, led to the large-scale displacement of over 2.3 million people, resulting in rapid and unplanned urbanization in northern Haiti. This study evaluated the impact of this unplanned urbanization on mosquito ecology and vector-borne diseases by assessing land use and change patterns. Land-use classification and change detection were carried out on remotely sensed images of the area for 2010 and 2013. Change detection identified areas that went from agricultural, forest, or bare-land pre-earthquake to newly developed and urbanized areas post-earthquake. Areas to be sampled for mosquito larvae were subsequently identified. Mosquito collections comprised five genera and ten species, with the most abundant species being Culex quinquefasciatus 35% (304/876), Aedes albopictus 27% (238/876), and Aedes aegypti 20% (174/876). All three species were more prevalent in urbanized and newly urbanized areas. Anopheles albimanus, the predominate malaria vector, accounted for less than 1% (8/876) of the collection. A set of spectral indices derived from the recently launched Landsat 8 satellite was used as covariates in a species distribution model. The indices were used to produce probability surfaces maps depicting the likelihood of presence of the three most abundant species within 30 m pixels. Our findings suggest that the rapid urbanization following the 2010 earthquake has increased the amount of area with suitable habitats for urban mosquitoes, likely influencing mosquito ecology and posing a major risk of introducing and establishing emerging vector-borne diseases.

Effective chemical control relies on reducing vector population size. However, insecticide selection pressure is often associated with the development of resistant populations that reduce control success. In treated areas, these resistant individuals present an adaptive advantage due to enhanced survival. Resistance can also lead to negative effects when the insecticide pressure ceases. In this study, the biological effects of deltamethrin resistance were assessed in the Chagas disease vector Triatoma infestans. The length of each developmental stage and complete life cycle, mating rate, and fecundity were evaluated. Susceptible and resistant insects presented similar mating rates. A reproductive cost of resistance was expressed as a lower fecundity in the resistant colony. Developmental costs in the resistant colony were in the form of a shortening of the second and third nymph stage duration and an extension of the fifth stage. A maternal effect of deltamethrin resistance is suggested as these effects were identified in resistant females and their progeny independently of the mated male´s deltamethrin response. Our results suggest the presence of pleiotropic effects of deltamethrin resistance. Possible associations of these characters to other traits such as developmental delays and behavioral resistance are discussed.

The biological parameters related to the life cycles of three populations of Meccus picturatus (Usinger) (Hemiptera: Reduviidae), one of the main vectors of Trypanosoma cruzi Chagas (Trypanosomatida: Trypanosomatidae), were evaluated. A cohort of each of three populations from geographically isolated localities in western Mexico was maintained under similar laboratory conditions, after which all three populations were compared to each other. In each cohort, 50.9 to 72.1% of nymphs completed the cycle. The average time from N1 to adult was approximately 175 days and different for the three studied cohorts. The number of blood meals between molts varied from one to three. The hatching rates as well as the fecundity per female per day were different among the three studied populations. Our results show that the three isolated populations are statistically different in some parameters from each other, which points to the importance of studying the biological characteristics of local populations of triatomines for estimating their capacity of transmitting T. cruzi to reservoir hosts.

The transmission of dengue, the most important arthropod-borne viral disease in Brazil, has been intensified over the past decades, along with the accompanying expansion and adaptation of its Aedes vectors. In the present study, we mapped dengue vectors in Ouro Preto and Ouro Branco, Minas Gerais, by installing ovitraps in 32 public schools. The traps were examined monthly between September, 2011 through July, 2012 and November, 2012 to April, 2013. The larvae were reared until the fourth stadium and identified according to species. The presence of dengue virus was detected by real time PCR and agarose gel electrophoresis. A total of 1,945 eggs was collected during the 17 months of the study. The Ovitrap Positivity Index (OPI) ranged from 0 to 28.13% and the Eggs Density Index (EDI) ranged from 0 to 59.9. The predominant species was Aedes aegypti, with 84.9% of the hatched larvae. Although the collection was low when compared to other ovitraps studies, vertical transmission could be detected. Of the 54 pools, dengue virus was detected in four Ae. aegypti pools.

To explore the potential role of Ixodes ricinus as the presumed vector of Bartonella henselae in eastern Poland, ticks collected in various geographic locations were examined for the presence of B. henselae, and the results were matched against the prevalence of anti-B. henselae antibodies in individuals occupationally exposed to tick bites. The presence of Bartonella DNA was investigated by PCR in a total of 1,603 unfed Ixodes ricinus ticks. The presence of IgG antibodies against B. henselae was investigated in serum samples from 332 people occupationally exposed to tick bites (94 farmers and 238 forestry workers). The total prevalence of B. henselae in ticks was 1.7%; the infection rates in males (3.1%) and females (2.7%) were nearly ten times greater than in nymphs (0.3%). The prevalence of seropositive results in the risk group (30.4%), farmers (27.7%) and forestry workers (31.5%), was significantly greater compared to the control group (8.9%). The results showed a weak positive correlation between the degree of infection of ticks and humans living in the same geographic region. The lack of a direct relationship indicates that exposure to tick bites is only one of the factors contributing to the significant preponderance of a seropositive response to B. henselae in the forestry workers and farmers over the control group. Other factors must be considered, such as contact with cats, which are popular domestic animals in Polish villages, and exposure to cat fleas.

Arthropod-borne diseases remain a pressing international public health concern. While progress has been made in the rapid detection of arthropod-borne pathogens via quantitative real-time (qPCR), or even hand-held detection devices, a simple and robust maceration and nucleic acid extraction method is necessary to implement biosurveillance capabilities. In this study, a comparison of maceration techniques using five types of beads followed by nucleic acid extraction and detection were tested using two morphologically disparate arthropods, the Aedes aegypti mosquito and Xenopsylla spp. flea, to detect the zoonotic diseases dengue virus serotype-1 and Yersinia pestis. Post-maceration nucleic acid extraction was carried out using the 1-2-3 Platinum-Path-Sample-Purification (PPSP) kit followed by qPCR detection using the Joint Biological Agent Identification and Diagnostic System (JBAIDS). We found that the 5mm stainless steel beads added to the beads provided in the PPSP kit were successful in macerating the exoskeleton for both Ae. aegypti and Xenopsylla spp. Replicates in the maceration/extraction/detection protocol were increased in a stepwise fashion until a final 128 replicates were obtained. For dengue virus detection there was a 99% positivity rate and for Y. pestis detection there was a 95% positive detection rate. In the examination of both pathogens, there were no significant differences between qPCR instruments, days ran, time of day ran, or operators.

We engaged in field studies of native mosquitoes in a Cambridgeshire Fen, investigating a) the habitat specificity and seasonal dynamics of our native fauna in an intensively managed wetland, b) the impact of water-level and ditch management, and c) their colonization of an arable reversion to flooded grassland wetland expansion project. Studies from April to October, 2010 collected 14,000 adult mosquitoes (15 species) over 292 trap-nights and ∼4,000 pre-imaginal mosquitoes (11 species). Open floodwater species (Aedes caspius and Aedes cinereus, 43.3%) and wet woodland species (Aedes cantans/annulipes and Aedes rusticus, 32.4%) dominated, highlighting the major impact of seasonal water-level management on mosquito populations in an intensively managed wetland. In permanent habitats, managing marginal ditch vegetation and ditch drying significantly affect densities of pre-imaginal anophelines and culicines, respectively. This study presents the first UK field evidence of the implications of wetland expansion through arable reversion on mosquito colonization. Understanding the heterogeneity of mosquito diversity, phenology, and abundance in intensively managed UK wetlands will be crucial to mitigating nuisance and vector species through habitat management and biocidal control.

The tick Ixodes ricinus (L.) is the most important vector of tick-borne zoonoses in Europe. Apart from factors related to human behavior, tick abundance is a major driver of the incidence of tick-borne diseases in a given area and related data represent critical information for promoting effective public health policies. The present study analyzed the relationship between different environmental factors and tick abundance in order to improve the understanding of I. ricinus autecology and develop spatial predictive models that can be implemented in tick-borne disease prevention strategies. Ticks were sampled in 27 sites over a four-year period and different environmental variables were studied. Five simple models were developed that explain a large part of variation in tick abundance. Precipitation seems to play the most important role, followed by temperature, woodland coverage, and solar radiation. Model equations obtained in this study may enable the spatial interpolation and extension of tick abundance predicted values to sites of the same area, in order to build regional predictive maps. They could also be useful for the validation of large-scale spatial predictive maps.

The degree of reproductive isolation between Triatoma recurva (Stål) (Hemiptera: Reduviidae: Triatominae) and the six species of the genus Meccus plus T. mexicana (Herrich-Schaeffer) (Hemiptera: Reduviidae) was examined. Fertility and the segregation of morphological characteristics were examined in two generations of hybrids from crosses between these species. The percentage of couples with offspring (fertile) was low in the vast majority of sets of crosses, with the exception of that between T. recurva female and M. phyllosomus male. In all studied sets of crosses, no first- (F1) or second- (F2) generation individuals were morphologically similar to T. recurva but instead shared the morphology of the other parental species. A similar phenomenon was observed in the three successful sets of backcrosses. These results indicated that different recorded levels of reproductive fitness among T. recurva and the species of Meccus involved in this study, plus T. mexicana, are present and that they were apparently influenced by differing mechanisms of isolation. The presence of some degree of reproductive compatibility between studied triatomines of distinct genera (Meccus spp. and Triatoma spp.) reinforces the need for generic revision of the tribe Triatomini.

We investigated the role of carbon dioxide and host temperature in host attraction in frog-biting midges (Corethrella spp). In these midges, females are known to use frog calls to localize their host, but the role of other host-emitted cues has yet not been investigated. We hypothesized that carbon dioxide acts as a supplemental cue to frog calls. To test this hypothesis, we determined the responses of the midges to carbon dioxide, frog calls, and both cues. A significantly lower number of midges are attracted to carbon dioxide and silent traps than to traps broadcasting frog calls. Adding carbon dioxide to the calls does not increase the attractiveness to the midges. Instead, carbon dioxide can have deterrent effects on frog-biting midges. Temperature of calling frogs is not a cue potentially available to the midges. Contrary to our hypothesis, there was no supplemental effect of carbon dioxide when presented in conjunction to calls. Midge host-seeking behavior strongly depends on the mating calls emitted by their anuran host. Overall, non-acoustic cues such as host body temperature and carbon dioxide are not important in long-distance host location by frog-biting midges.

Documented information on the ecology of larval habitats in Botswana is lacking but is critical for larval control programs. Therefore, this study determined the characteristics of these habitats and the influences of biotic and abiotic factors in Tubu village, Botswana. Eight water bodies were sampled between January and December, 2013. The aquatic vegetation and invertebrate species present were characterized. Water parameters measured were turbidity (NTU), conductivity (µS/cm), oxygen (mg/l), and pH. Larval densities of Anopheles arabiensis mosquitoes and their correlation with abiotic factors were determined. Larval breeding was associated with ‘short’ aquatic vegetation, a variety of habitats fed by both rainfall and flood waters and sites with predators and competitors. The monthly mean (± SE_mean) larval density was 8.16±1.33. The monthly mean (±SE_mean) pH, conductivity, oxygen, and turbidity were 7.65±0.13, 1152.834±69.171, 5.59±1.33, and 323.421±33.801, respectively. There was a significant negative correlation between larval density and conductivity (r = -0.839; p < 0.01), while a significant positive correlation occurred between turbidity and larval density (r = 0.685; p < 0.05). Oxygen (r = 0.140; p > 0.05) and pH (r = 0.252; p > 0.05) were not correlated with larval density. Floods and diversified breeding sites contributed to prolonged and prolific larval breeding. ‘Short’ aquatic vegetation and predator-infested waters offered suitable environments for larval breeding. Turbidity and conductivity were good indicators for potential breeding places and can be used as early warning indices for predicting larval production levels.

In Australia, dengue control combines source reduction with lethal ovitraps to reduce Aedes aegypti populations during outbreaks. Lethal ovitraps are considered a sustainable and environmentally friendly method of controlling container-inhabiting mosquitoes, however, to-date, this claim has not been quantified. This study assesses the potential impact of lethal ovitraps on non-target organisms when used to control Ae. aegypti in tropical Australia. For retention of specimens, we substituted standard sticky ovitraps for lethal ovitraps. We collected 988 Ae. aegypti and 44,132 non-target specimens over 13 months from 16 sites. Although Ae. aegypti comprised only 2.2% of the total collection, they were were the eighth most dominant taxa collected, on the 93rd percentile. Of the non-target organisms, Collembola were the dominant taxa, 44.2%, with 36.8% and 10.5% Diptera and Hymenoptera, respectively. Of the Dipterans, 61% were family Phoridae. Lethal ovitraps were visited by 90 insect or invertebrate families in total. Ovitraps are attractive to Collembola, Phoridae, Sciaridae, Formicidae, and Culicidae, with minimal attraction by Apidae and other commonly monitored non-target organisms. For container-inhabiting mosquitoes, LOs are cost effective operationally, requiring minimal staff resources for placement and retrieval.

The swallow bug (Oeciacus vicarius) is the only known vector for Buggy Creek virus (BCRV), an alphavirus that circulates in cliff swallows (Petrochelidon pyrrhonota) and house sparrows (Passer domesticus) in North America. We discovered ants (Crematogaster lineolata and Formica spp.) preying on swallow bugs at cliff swallow colonies in western Nebraska, U.S.A. Ants reduced the numbers of visible bugs on active swallow nests by 74–90%, relative to nests in the same colony without ants. Ant predation on bugs had no effect on the reproductive success of cliff swallows inhabiting the nests where ants foraged. Ants represent an effective and presumably benign way of controlling swallow bugs at nests in some colonies. They may constitute an alternative to insecticide use at sites where ecologists wish to remove the effects of swallow bugs on cliff swallows or house sparrows. By reducing bug numbers, ant presence may also lessen BCRV transmission at the spatial foci (bird colony sites) where epizootics occur. The effect of ants on swallow bugs should be accounted for in studying variation among sites in vector abundance.

We evaluate three trapping methods for their effectiveness at capturing Culex pipiens and Culex torrentium, both enzootic vectors of bird-associated viruses in Europe. The comparisons, performed in two regions in Sweden, were among CDC traps baited with carbon dioxide, gravid traps, and ovitraps baited with hay infusion. The proportions of the two Culex species in a catch differed between trap types, with CDC traps catching a lower proportion of Cx. torrentium than both gravid traps and ovitraps. Between gravid traps and ovitraps, there was no difference in the proportions of the two species. The results indicate that Cx. torrentium may go undetected or underestimated compared to Cx. pipiens when using carbon dioxide baited CDC traps. The new insight of trap bias presented here adds an important dimension to consider when investigating these vectors of bird-associated viruses in the field.

Several studies have examined how climatic variables such as temperature and precipitation may affect life history traits in mosquitoes that are important to disease transmission. Despite its importance as a seasonal cue in nature, studies investigating the influence of photoperiod on such traits are relatively few. This study aims to investigate how photoperiod alters life history traits, survival, and blood-feeding activity in Aedes albopictus (Skuse) and Aedes aegypti (Linnaeus). We performed three experiments that tested the effects of day length on female survival, development time, adult size, fecundity, adult life span, and propensity to blood feed in Ae. albopictus and Ae. aegypti. Each experiment had three photoperiod treatments: 1) short-day (10L:14D), 2) control (12L:12D), and 3) long-day (14L:10D). Aedes albopictus adult females were consistently larger in size when reared in short-day conditions. Aedes aegypti adult females from short-day treatments lived longer and were more likely to take a blood meal compared to other treatments. We discuss how species-specific responses may reflect alternative strategies evolved to increase survival during unfavorable conditions. We review the potential impacts of these responses on seasonal transmission patterns, such as potentially increasing vectorial capacity of Ae. aegypti during periods of shorter day lengths.

From 2003 to 2007, entomological surveys were conducted in Lobito town (Benguela Province, Angola) to determine which Anopheles species were present and to identify the vectors responsible for malaria transmission in areas where workers of the Sonamet Company live. Two types of surveys were conducted: (1) time and space surveys in the low and upper parts of Lobito during the rainy and dry periods; (2) a two-year longitudinal study in Sonamet workers' houses provided with long-lasting insecticide-treated nets (LLIN), “PermaNet,” along with the neighboring community. Both species, An. coluzzii (M molecular form) and An. gambiae (S molecular form), were collected. Anopheles coluzzii was predominant during the dry season in the low part of Lobito where larvae develop in natural ponds and temporary pools. However, during the rainy season, An. gambiae was found in higher proportions in the upper part of the town where larvae were collected in domestic water tanks built near houses. Anopheles melas and An. listeri were captured in higher numbers during the dry season and in the low part of Lobito where larvae develop in stagnant brackish water pools. The infectivity rates of An. gambiae s.l. varied from 0.90% to 3.41%.

Prevalence studies of Borrelia burgdorferi and Anaplasma phagocytophilum have been rare for ticks from southwestern Pennsylvania. We collected 325 Ixodes scapularis ticks between 2011 and 2012 from four counties in southwestern Pennsylvania. We tested for the presence of Borrelia burgdorferi and Anaplasma phagocytophilum using PCR. Of the ticks collected from Pennsylvania, B. burgdorferi (causative agent of Lyme disease) was present in 114/325 (35%) and Anaplasma phagocytophilum (causative agent of Human Granulocytic Anaplasmosis) was present in 48/325 (15%) as determined by PCR analysis.