We examined the possible effects of host body size and throat gland development on the abundance of bloodfeeding nycteribiid and streblid flies parasitizing a Malagasy fruit bat, Rousettus madagascariensis G. Grandidier, 1928. Data were collected in the Parc National d'Ankarana in northern Madagascar during four visits: September 2014, 2015 (dry season), and January 2015, 2016 (wet season). Two bat fly species were identified, Eucampsipoda madagascarensisTheodor, 1955 (Nycteribiidae) and Megastrebla wenzeli (Jobling, 1952) (Streblidae). A positive correlation was found between host body size and abundance of E. madagascarensis during the four visits, suggesting that larger hosts have more parasites, and for M. wenzeli, this relationship was identified only during the wet season visits. In male hosts, body size and throat gland development are correlated with variation in E. madagascarensis abundance during the two seasons; this relationship was not found for M. wenzeli. We present some explanations for the observed patterns of bat fly abundance associated with throat gland development: increased vascularization and easier access to bloodmeals, chemical properties of gland secretions acting as attractants or perhaps being consumed, and modification of hair around the gland providing protection from bat grooming.

In the genus Culex, oviposition is often generalized as laying floating egg rafts directly on the surface of the water, as reported for arbovirus vectors Culex pipiens L. and Culex tarsalis Coquillett, both members of subgenus Culex. The current work reports on investigations of exceptions to this strategy by species of Melanoconion, a diverse and mostly tropical subgenus of Culex, with many arbovirus vectors. Wild gravid females from Vero Beach, FL, were introduced into outdoor cages with open water, partially covered water, and emergent solid structures for oviposition. Three species, Culex cedecei Stone and Hair, Culex iolambdis Dyar, and Culex pilosus Dyar and Knab laid vast majorities (80–100%) of egg clusters attached to solid surfaces, although at different heights and angles, while Culex atratus Theobald laid floating egg clusters (100%) directly on the water. When offered mud, open water, and woody material, Cx. iolambdis laid 86% of clusters on nearly vertical surfaces of mangrove rootlets. Culex iolambdis eggs laid on solid surfaces hatched within 3–4 d or dried up without hatching. Results suggest that Melanoconion species utilize a variety of oviposition strategies, yet most species studied oviposit out of water. For some species, such as Cx. pilosus, this is a strategy for diapause and desiccation resistance, while for others, such as Cx. iolambdis, ovipositing above water may protect eggs from predators or being flushed from larval habitat. A summary of oviposition strategies in genus Culex shows that above-water oviposition is widespread and that the Culex egg raft paradigm needs reassessment.

The southern districts of Odisha State in east-central India have been highly endemic for falciparum malaria for many decades. However, there is no adequate information on the abundance of the vector species or their bionomics in relation to space and time in these districts. Therefore, a study was carried out on the entomological aspects of malaria transmission to generate such information. Collections of mosquitoes were made once during each of the three seasons in 128 villages selected from eight districts. Villages within the foot-hill ecotype had a significantly greater abundance of Anopheles fluviatilis James s. l., whereas the abundance of Anopheles culicifacies Giles s. l. was significantly greater in the plain ecotype. The abundance of An. fluviatilis was maximum during the cold season, whereas An. culicifacies abundance was highest during summer and rainy seasons. The maximum likelihood estimation of the malaria infection rate in An. fluviatilis was 1.78%, 6.05%, and 2.6% in Ganjam, Kalahandi, and Rayagada districts, respectively. The infection rate of An. culicifacies was 1.39% only in Kandhamal district; infected females were not detected elsewhere. Concurrently, the annual malaria parasite incidence (MPI) was significantly higher in hill-top (17.6) and foot-hill (14.4) villages compared to plain villages (4.1). The districts with more villages in hill-top and foot-hill ecotypes also had a greater abundance of An. fluviatilis, the major malaria vector, and exhibited a higher incidence of malaria than villages within the plain ecotype, where An. culicifacies was the most abundant vector.

Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata was described in 2002, based on specimens caught in the wild in the municipality of Gentio do Ouro, Bahia, Brazil. In 2009, nymphs and adults were detected inside homes and sylvatic specimens were positive for Trypanosoma cruzi (Chagas). No information on the bionomics of T. sherlocki exists, although such data are considered essential to estimate its vector and colonization potential in domestic environments. Herein, the biological cycle of T. sherlocki was studied based on 123 eggs, with nymphs and adults fed on Mus musculus (Linnaeus). Nymphal development time phases, number of meals consumed, and stage-specific mortality rates were analyzed. Survival time under starvation conditions was measured between ecdysis and death among 50 nymphs (first to fifth instar) and 50 male and female adults. The median development time from egg to adult was 621.0 (CI: 489–656) d. The number of meals consumed ranged from 1 to 20 for nymphs of the first to fifth instar. The fifth instar showed the greatest resistance to starvation, with amean of 156.5 d. The high number of meals consumed by T. sherlocki favored infection with and transmission of T. cruzi. The full development of this species under laboratory conditions with a lowmortality rate indicates that this vector presents biological characteristics that may contribute to its adaptation to artificial human ecotopes. Its high resistance to starvation emphasizes the importance of entomological surveillance for this species.

Because they may demonstrate characteristics of the environment where a body has been laying prior to the discovery, flies are insects of forensic interest. We investigated the fly abundance and the effect of location in Kingdom of Saudi Arabia on fly species diversity that attack decomposing human and animal remains. Using baited traps deployed in each location, we collected 3,697 flies of seven species belonging to three families. Chrysomya albiceps Wiedmann represented 60.86% of the collected flies, whereas Musca domestica L. represented 25.8%; the other species made up < 6% each. To facilitate species identification by DNA barcoding, we sequenced a 710-bp “Folmer region” of cytochrome oxidase subunit 1 (COI) gene for 22 samples from collection sites distributed through entire Saudi Arabia. The COI sequences from Musca albina Wiedmann, Musca lucidula Loew, Musca calleva Walker, Musca sorbens Wiedmann, and Physiphora alceae Preyssler were obtained for the first time. This primary study indicates that even when Folmer primers were widely used in DNA barcoding, the Folmer's region is not adequate when discriminating between Musca species, and sequencing the whole COI or other genes is required for forensic purpose.

Remote sensing, a powerful tool for analyzing landscape factors, is being used to explore the spatial ecology of vectors of several diseases. This study aims to explore the role of buffer size in identification and quantification of geo-environmental factors from multispatial resolution satellite data and its application along with microclimatic data to kala-azar vector abundance modeling.

Sand fly abundance and microclimatic data were collected from 210 sample sites during the premonsoon and postmonsoon season of 2014 from Muzaffarpur district of Bihar (India). Linear imaging self-scanning sensor (LISS-III; 23.5 m) and advanced wide field sensor (AWiFS; 56 m) imageries were used for generating environmental variables at 300- and 500-m buffer zones. Four analytical models of sand fly density were developed and evaluated for predictive accuracy.

A total of 33 geo-environmental and four microclimatic variables were tested for the prediction of sand fly density, of which the best four were maximum temperature, relative humidity, Euclidean nearest-neighbor distance of settlement area to mixed bush-grass land, and surface water body. Predictive accuracy of the LISS-III models was found to be higher than AWiFS models at all buffer sizes.

The results show that geo-environmental parameters and microclimatic data are the best predictors for sand fly density modeling. Buffer sizes play an important role in identifying the explanatory variables. Model parameters may be useful in identifying predisposing factors of sand fly habitat suitability at the micro level.

The majority of the Yucatán State, México, presents subtropical climate that is suitable for many species of mosquitoes that are known to be vectors of diseases, including those from the genera Aedes and Culex. The objective of this study is to identify the geographic distribution of five species from these two genera and estimate the human population at risk of coming in contact with them. We compiled distributional data for Aedes aegypti (L.), Aedes (Howardina) cozumelensis (Diaz Najera), Culex coronator Dyar and Knab, Culex quinquefasciatus Say, and Culex thriambus Dyar from several entomological studies in Yucatán between March 2010 and September 2014. Based on these data, we constructed ecological niche models to predict the spatial distribution of each species using the MaxEnt algorithm. Our models identified areas with suitable environments for Ae. aegypti in most of Yucatán. A similar percentage of urban (97.1%) and rural (96.5%) populations were contained in areas of highest suitability for Ae. aegypti, and no spatial pattern was found (Moran's I = 0.33, P = 0.38); however, we found an association of abundance of immature forms of this species with annual mean temperature (r = 0.19, P≤0.001) and annual precipitation (r = 0.21, P≤0.001). Aedes cozumelensis is also distributed in most areas of the Yucatán State; Cx. quinquefasciatus, Cx. coronator, and Cx. thriambus are restricted to the northwest. The information generated in this study can inform decision-making to address control measures in priority areas with presence of these vectors.

Lyme disease is emerging in Canada due to geographic range expansion of the tick vector Ixodes scapularis Say. Recent areas of emergence include parts of the southeastern Canadian Prairie region. We developed a map of potential risk areas for future I. scapularis establishment in the Canadian Prairie Provinces. Six I. scapularis risk algorithms were developed using different formulations of three indices for environmental suitability: temperature using annual cumulative degree-days > 0 °C (DD > 0 °C; obtained from Moderate Resolution Imaging Spectroradiometer satellite data as an index of conditions that allow I. scapularis to complete its life cycle), habitat as a combined geolayer of forest cover and agricultural land use, and rainfall. The relative performance of these risk algorithms was assessed using receiver-operating characteristic (ROC) area under the curve (AUC) analysis with data on presence–absence of I. scapularis obtained from recent field surveillance in the Prairie Provinces accumulated from a number of sources. The ROC AUC values for the risk algorithms were significantly different (P < 0.01). The algorithm with six categories of DD > 0 °C, habitat as a simple dichotomous variable of presence or absence of forest, and normalized rainfall had the highest AUC of 0.74, representing “fair to good” performance of the risk algorithm. This algorithm had good (>80%) sensitivity in predicting positive I. scapularis surveillance sites, but low (50%) specificity as expected in this region where not all environmentally suitable habitats are expected to be occupied. Further prospective studies are needed to validate and perhaps improve the risk algorithm.

Dynamic simulation models provide vector abundance estimates using only meteorological data. However, model outcomes may heavily depend on the assumptions used to parameterize them. We conducted a sensitivity analysis for a model of Aedes aegypti (L.) abundance using weather data from two locations where this vector is established, La Margarita, Puerto Rico and Tucson, Arizona. We tested the effect of simplifying temperature-dependent development and mortality rates and of changing development and mortality thresholds as compared with baselines estimated using biophysical models. The simplified development and mortality rates had limited effect on abundance estimates in either location. However, in Tucson, where the vector is established but has not transmitted viruses, a difference of 5 °C resulted in populations either surviving or collapsing in the hot Arizona mid-summer, depending on the temperature thresholds. We find three important implications of the observed sensitivity to temperature thresholds. First, this analysis indicates the need for better estimates of the temperature tolerance thresholds to refine entomologic risk mapping for disease vectors. Second, our results highlight the importance of extreme temperatures on vector survival at the marginal areas of this vector's distribution. Finally, the model suggests that adaptation to warmer temperatures may shift regions of pathogen transmission.

Aedes aegypti (L.) is the vector responsible for transmitting dengue, chikungunya, yellow fever, and Zika viruses, as well as other pathogens. Microbial larvicides based on Bacillus thuringiensis Berliner israelensis (Bti) and Saccharopolyspora spinosa Mertz and Yao, such as VectoBac 12AS and Natular 2EC, have been shown to be effective in reducing larval populations of Ae. aegypti. We examined the gene expression of two detoxification enzymes, glucosyl and glucuronosyl transferases (AaeGGT1 and AaeGGT2), through developmental stages and a time course study in response to larvicide exposure using qualitative real-time polymerase chain reaction (qPCR). AaeGGT1 and AaeGGT2 gene expressions were differentially regulated during development of the immature stages.We also found that male adults had higher expression than female adults after controlling for age effects. AaeGGT1 and AaeGGT2 gene expression were both upregulated in response to VectoBac 12AS and Natular 2EC treatments with the maximum level of expression occurring 24h after treatment applications. This information sheds light on larvicideinduced changes in the physiology of Ae. aegypti with implications for development of mosquito control strategies.

Accurate species identifications are the essential first step in understanding the medical, economic, and ecological importance of black flies. The utility of DNA barcoding based on cytochrome c oxidase subunit 1 (COI) sequences was evaluated for identifying six common species of Palearctic black flies in the subgenus Wilhelmia, including several that are virulent pests. Chromosomally identified larvae from Turkey and Germany and COI sequences in GenBank were analyzed. Intraspecific genetic divergence was 0.7–3.5% (mean 1.6%), whereas interspecific genetic divergence was 2.7–16.9%. On the basis of COI barcodes, the six nominal species of Simulium (Wilhelmia) were clustered in three distinct clades with high levels of genetic divergence, using maximum likelihood and Bayesian analyses. All specimens of Simulium equinum (L.), Simulium pseudequinum Séguy, and Simulium paraequinum Puri were correctly identified. However, >75% of identifications were ambiguous for Simulium lineatum (Meigen) and Simulium turgaicum Rubtsov (Meigen) because of overlapping intra- and interspecific divergence of the two species and Simulium balcanicum (Enderlein), all three of which are chromosomally similar and nearly isomorphic. Phylogenetic evaluation showed that S. balcanicum, S. equinum, S. pseudequinum, and S. paraequinum were monophyletic, with high bootstrap and posterior probability values, but it also showed that S. lineatum and S. turgaicum were paraphyletic, each clustering in two distinct groups, suggesting the presence of cryptic taxa. Although DNA barcoding provided a partialmeans of identification and indications of additional biodiversity, other molecularmarkers are needed to clarify the limits of all pest species of the subgenusWilhelmia.

Ticks transmit a variety of pathogenic organisms to vertebrates, especially mammals. The fossil record of such associations is extremely rare. An engorged nymphal tick of the genus Ambylomma in Dominican amber was surrounded by erythrocytes from its mammalian host. Some of the exposed erythrocytes contained developmental stages of a hemoprotozoan resembling members of the Order Piroplasmida. The fossil piroplasm is described, its stages compared with those of extant piroplasms, and reasons provided why the mammalian host could have been a primate. The parasites were also found in the gut epithelial cells and body cavity of the fossil tick. Aside from providing the first fossil mammalian red blood cells and the first fossil intraerythrocytic hemoparasites, the present discovery shows that tick–piroplasm associations were already well established in the Tertiary. This discovery provides a timescale that can be used in future studies on the evolution of the Piroplasmida.

Culex coronator Dyar and Knab and Culex usquatus Dyar belong to the Coronator Group of the subgenus Culex. Culex coronator and Cx. usquatus are widespread and sympatric throughout their distribution range, which includes Brazil. Morphological identification of these species is based primarily on the characteristics of the male genitalia; females are indistinguishable using the qualitative characteristics employed in identification keys. The primary objective of this study was to distinguish females of Cx. coronator from those of Cx. usquatus employing both wing geometric morphometrics, and DNA sequences (NADH5, COI, Hunchback, and CAD). Additionally, we employed the isolation with migration model (IMa) to evaluate: 1) the migration rates and 2) the divergence time, between Cx. coronator and Cx. usquatus. Specimens were captured in Pariquera-Açu and Cananéia south-eastern São Paulo, Ribeira Valley, Brazil. Canonical variate analysis (CVA) demonstrated two groups in the morphospace. The accuracy of species recognition was moderate (82.6%) for Cx. coronator and low (60.8%) for Cx. usquatus. Bayesian analyses of concatenated gene sequences recovered from specimens of Cx. coronator separated the species into three lineages (herein referred to as Culex coronator A, B, and C), whereas Cx. usquatus specimens clustered into a single lineage. Lineages A and B of Cx. coronator intermixed with specimens of Cx. usquatus, and one specimen of Cx. coronator clustered into the Cx. usquatus lineage. The IMa analysis indicated that the divergence of Cx. coronator and Cx. usquatus is a slow process, with some degree of gene flow between the two species.

The wild species Rhodnius brethesi (Matta, 1919) (Hemiptera: Reduviidae) is found in areas of piassabais in microregion of Rio Negro, Amazonas, Brazil. Its geographical distribution overlaps the areas of Leopoldinia piassaba palm. In areas where palm trees are found, transmission cycle of Trypanosoma cruzi is related to the extractive activity of the palm fiber, exposing workers to wild vector transmission of this parasite. The close association with the palm tree L. piassaba suggests that this wild triatomine has special features in its sensory system allowing specificity of ecotope. The objective of the study is to identify the antennal sensilla phenotype and morphologically characterize the size and shape of the wings of wild R. brethesi, and to compare with the phenotype present in individuals reared in the laboratory. From the samples taken in the field, the presence of the species R. brethesi was found on both banks of the Rio Negro. The techniques used to verify the morphological patterns are important resources for observations of the triatomine populations, be they in an artificial or natural habitat.

Two new Neotropical species of the genus Oxysarcodexia Townsend (Diptera: Sarcophagidae) from Chaco ecoregion of Argentina are described, O. berthet, sp. nov. and O. ibera, sp. nov. Both species were recovered from localities in the eastern part of the Chaco forest (Humid Chaco). The descriptions are based on the male external morphology and terminalia. Photographs and SEM images are provided to aid in the identification of the species.

The black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), is a relevant species in waste and pest management, but is also of forensic and medical importance. A scanning electron microscopy (SEM) investigation of the antennae of both sexes of H. illucens is presented here for the first time. The antenna is composed of three regions: the scape, the pedicel, and the flagellum. The first two regions are single segments, whereas the third region, the longest one, is composed of eight flagellomeres. The scape and pedicel have microtrichia, chaetic sensilla, and rounded perforations. The flagellum is covered by different microtrichia, the morphology of which is described in detail. Two types of sensory pit are found on flagellomeres 1 to 6. An oval depression with trichoid sensilla extends from flagellomeres 4 to 6. On both sides of flagellomere 8 is a lanceolate depression covered by hair-like microtrichia. Morphometric and morphological analyses revealed some sex-related differences. The results of the SEM investigations are compared with those obtained on other species of the family Stratiomyidae and other brachyceran Diptera. The possible role of sensilla in sensory perception is also discussed in comparison with nondipteran species.

The subfossil remains of 14 cimicids (Hemiptera: Cimicidae) were recovered during archaeological investigations of the Paisley Five Mile Point Cave site (35LK3400), an exceptionally well-dated (n = 229 radiocarbon dates) late Pleistocene–early Holocene rock shelter site in south-central Oregon. Nine of the specimens have been assigned to three modern species of Nearctic Cimicidae—Cimex antennatus Usinger & Ueshima, Cimex latipennis Usinger & Ueshima, and Cimex pilosellus (Horváth)—whereas the remaining five individuals were too fragmentary to positively identify. The chronology of the insect assemblage puts one specimen at circa 5,100 calibrated years before present (cal. yr BP), and the remaining 13 range in age from 9,400 to almost 11,000 cal. yr BP. Although fossil and subfossil cimicid remains have been recovered at other archaeological sites, the fossil record for bed bugs is largely undocumented. The Paisley Caves specimens thus far represent the oldest remains of the genus in probable contact with humans on record.

Simulium (Simulium) umphangense, a new human-biting species of black fly, is described based on females captured while attacking humans in western and central Thailand. The female of this new species is similar to those of S. (S.) indicum Becher, S. (S.) nigrogilvum Summers, and S. (S.) vanellum Huang et al. in the Simulium griseifrons species-group of the subgenus Simulium Latreille in having the frons densely covered with golden-yellow short hairs, a character rarely found in the subgenus Simulium. This new species is readily distinguished from the three related species by having a darkened fore tibia, a bare subcosta (or rarely with a few hairs), and an ovipositor valve not protruded posteriorly. Additional diagnostic characters of this new species are noted. This is the seventh human-biting species of black fly in Thailand.

Ticks expend energy while host-seeking and must consume blood to advance to the next life stage. The energy required for activity is derived from the tick's lipid reserves, a valuable resource that sustains the tick until it finds the next host and can take another bloodmeal. The amount of lipid reserves in an unfed tick has been proposed as an index of tick biological age. Two different methods for aging nymphal blacklegged ticks, Ixodes scapularis Say, were analyzed in this study. To study lipid usage, colony-raised nymphs were held in lab-controlled chambers at 21.0 °C and ≥95% relative humidity, with a photoperiod of 12:12 (L:D) h. Samples of ticks were frozen at –80 °C every 2–3 wk, starting at 12-wk postmolt and continuing until 38-wk postmolt. Lipid reserves were determined indirectly through measurements of “physiological age” that estimate the energy a tick has based on the evaluation of morphometric size ratios of the tick scutum and alloscutum, and quantified directly through chloroform extractions of lipid from individual ticks. Morphometric age ratios and lipid amounts were compared to determine if morphometric measurements accurately estimated a tick's physiological state. Although the morphometric age ratio did correlate significantly with total tick lipid content, the predictive value of the ratio was not reliable; chloroform extraction results showed that lipid amounts declined steadily through the study and more accurately characterized the physiological condition of nymphal I. scapularis. The study of physiological aging of blacklegged ticks may lead to a better understanding of how changing environmental conditions affect tick longevity.

Nepeta essential oil (Neo; catnip) and its major component, nepetalactone, have long been known to repel insects including mosquitoes. However, the neural mechanisms through which these repellents are detected by mosquitoes, including the yellow fever mosquito Aedes aegypti (L.), an important vector of Zika virus, were poorly understood. Here we show that Neo volatiles activate olfactory receptor neurons within the basiconic sensilla on the maxillary palps of female Ae. aegypti. A gustatory receptor neuron sensitive to the feeding deterrent quinine and housed within sensilla on the labella of females was activated by both Neo and nepetalactone. Activity of a second gustatory receptor neuron sensitive to the feeding stimulant sucrose was suppressed by both repellents. Our results provide neural pathways for the reported spatial repellency and feeding deterrence of these repellents. A better understanding of the neural input through which female mosquitoes make decisions to feed will facilitate design of new repellents and management strategies involving their use.

Insects associated with carrion are critical to the decomposition process and nutrient cycling in ecosystems. Yet the communities of insects associated with carrion vary between locations, and detailed case studies are necessary for identifying differences and similarities among contrasting habitats. In this study, we examined temporal changes in the crawling insect community collected from rabbit carcasses placed in contrasting grassland and tree habitats in southeastern Australia. We collected 18,400 adult insects, including 22 species of fly, 57 species of beetle, and 37 species of ant. We found significant effects of habitat type and time, but not their interaction, on the composition of the entire insect community. Several ant species showed early and rapid colonization and highest abundances during early stages of decay, including Iridomyrmex purpureus (Smith, 1858) under trees, and Iridomyrmex rufoniger (Lowne, 1865) and Rhytidoponera metallica (Smith, 1858) in grassland. We found that most fly species showed highest abundance during active decay, but Chrysomya varipes (Macquart 1851) was more abundant under trees than in grassland during this time. Beetles peaked during active or advanced decay stages, with Saprinus and Omorgus the most abundant genera. Our study demonstrates that strong replication of contrasting environmental treatments can reveal new information on habitat preferences of important carrion insect species. The numerical dominance of ants early in decomposition has implications for insect community structure via potential competitive interactions with flies, and should be more rigorously examined in future carrion studies.

The tropical bed bug is scientifically recognized as a significant public health problem. While there is an increased awareness about their resurgence by medical and life science committees, efficient bed bug management still remains unresolved. The solution may soon arise, as information about bed bugs' infestation dynamics and systematics are becoming more distinguishable. Recent developments in studies about bed bugs are based on molecular intervention by determining their genetic variation and phylogeography. The aim of this study is to assess the phylogenetic relationships and genetic diversity among the populations of tropical bed bugs inhabiting Malaysia. A molecular genotyping study was conducted with 22 tropical bed bug populations composed of three individuals per population. The mitochondrial (COI) gene was used as a marker. The data obtained were analyzed using the T-Coffee, ClustalX, MEGA 6.0, and PAUP software. The results showed one main monophyletic clade that consisted of two groups: Ch01 and Ch02. Ch02 consists of samples from the Bandar Hilir population, differing from the other populations studied by one singleton base. However, as there were no changes in the amino acid, this singleton genetic variation was considered to have no effect on genetic differentiation. Ch01 shows similarity with some sequence of Cimex hemipterus (F.) from Thailand, suggesting an international diversity connection. The disparity index apparently suggests that all isolates are homogeneous populations and are supported by the low value of the mean pairwise distance between isolates. This study will increase the knowledge about phylogeographic diversity of tropical bed bug in Malaysia.

Horn fly, Haematobia irritans (L.), population estimates often serve as a diagnostic tool prior to implementing managerial control options available to researchers and producers. Digital photographs taken of cattle infested with horn flies have been suggested to provide similarly accurate population estimates as compared with traditional visual assessments. The objective of this study was to compare visual and digital techniques used to estimate horn fly populations. Sixteen Angus × Hereford yearling heifers artificially infested with four levels of horn flies (Low = 0 flies; Medium = 250 flies; High = 500 flies; Extreme = 1,000 flies) were evaluated. Population estimates were taken visually by experienced (VE1) and inexperienced (VE2) technicians, as well as digitally, with photographs taken on both lateral sides of the south-facing animal. Horn flies were counted in both photographs and combined (CDC) for full body estimates. In addition, the highest photographed side population times two (DDC) was used for comparison. Estimations were made at 0700, 1200, and 1900 h the day following infestation. A time of observation × infestation level interaction (P < 0.01) was detected. On average, VE1 population estimates were greater (P<0.01) than any other counting method observed. Morning estimates were greater (P < 0.05) than those taken at noon or in the early evening regardless of counting method. Further research regarding the standardization of these techniques to ensure more accurate population estimates is needed before these methods can be incorporated into integrated pest management programs.

Plant-derived essential oils can be used as insecticides for vector control. However, to establish their safety, it is necessary to perform toxicological studies. Herein, we evaluated the chemical composition and insecticidal activity of the essential oil from the leaves of Croton argyrophyllus on the third- and fourth-instar larvae and adult Aedes aegypti (L., 1762). We also evaluated the acute toxicity of the essential oil in adult female Mus musculus. The lethal concentration 50 (LC₅₀) and 90 (LC₉₀) of C. argyrophyllus essential oil on larvae of Ae. aegypti were 0.31 and 0.70mg _ml^-1, respectively, and 5.92 and 8.94mg ml^-1, respectively, on Ae. aegypti adults. The major components of the essential oil were spathulenol (22.80%), (E)-caryophyllene (15.41%), α -pinene (14.07%), and bicyclogermacrene (10.43%). It also displayed acute toxicity in adults of Mus musculus; the intraperitoneal and oral lethal dose 50 (LD₅₀) were 2,000mg kg^{- 1} and 2,500mg kg^{- 1}, respectively. The results showed that the essential oil from C. argyrophyllus leaves has insecticidal activity on Ae. aegypti larvae and adults at an average lethal concentration below the median lethal dose needed to cause acute toxicity in the common mouse.

The swallow bug, Oeciacus vicarius Horvath, is a hematophagous ectoparasite of the cliff swallow, Petrochelidon pyrrhonota Vieillot, and is closely related to bed bugs (Cimex spp.). Evolution of insecticide resistance has been documented for bed bugs but not studied in Oeciacus. For periods of 17 and 32 yr, two cliff swallow colonies in western Nebraska were treated during the summer breeding season using the organophosphate insecticide Dibrom. Despite continual treatments, O. vicarius has been observed frequently within these colonies. We evaluated the efficacy of Dibrom 8 on O. vicarius during the 2016 season at two treated colonies and four that had never experienced treatment. Dibrom 8 was found to be effective in 100% of trials, with immobilization within minutes and death within 72 h, for individuals from all colonies. In control treatments (water), individuals collected from treated colonies exhibited greater survival than individuals from untreated colonies, and those from active colonies (bugs fed) had greater survival than those from inactive colonies (bugs unfed). A residual effect was observed in both lab and field trials: 100% mortality occurred in the lab after exposure to filter paper substrates treated both 5 and 10 d earlier, and in the field, nests treated once early in the season had O. vicarius counts 43 d later that were <1% of those from untreated nests within the same colony. We hypothesize that the lack of resistance results from the limited potential for resistance allele fixation due to outbreeding and frequent immigration of insecticide-naıuml;ve individuals.

The house fly, Musca domestica L. (Diptera: Muscidae), is a disease vector of mechanically transmitted pathogens including bacteria, viruses, and protozoans. Opportunities for pathogen transmission can increase as fly longevity increases. Dietary preferences play an important role in insect longevity; therefore, we investigated house fly preferences, sucrose availability, and caloric constraints on house fly longevity. Experimental goals were: 1) to test the effects of calorie restriction on survival of house flies by manipulating concentrations of erythritol (low caloric content) and sucrose (high caloric content), and comparing commercial sweeteners of differing calorie content, 2) to identify house fly preferences for either erythritol or sucrose, and 3) to evaluate the insecticidal activity or toxicity of erythritol on house flies. Our data show that house flies may prefer high calorie options when given a choice and that house fly longevity likely increases as calorie content increases. Additionally, no significant differences in longevity were observed between the water only control (zero calories) and erythritol treatments. This suggests that decreased survival rates and death could be the result of starvation rather than insecticidal activity. This research furthers our understanding of house fly survival and sugar-feeding behavior.

Insecticide decay rate on different wall surfaces is of importance to indoor residual spray (IRS) programs used as a malaria control intervention. Past IRS operations showed increasing populations of endophilic malaria vectors resting on indoor surfaces from various sites in Uganda following use of Ficam VC (bendiocarb) insecticide; variability of insecticide life was believed to be primarily due to wall surface type. Bendiocarb longevity was tested in the northern Uganda districts of Amuru, Apac, and Pader to assess its residual efficacy on three commonly encountered wall surfaces. Wall types included mud and wattle, plain brick, and painted plaster. A susceptible mosquito strain (Anopheles gambiae Kisumu) was used in all trials. Nine houses in each of the three districts were set with three test cones and one control cone per house, divided evenly among the three wall surfaces. Bioassays were run monthly through 6 mo. Painted plastered surfaces produced 100% mortality (at 24 h) through 6 mo. Plain brick surfaces killed 100% of test mosquitoes through 4 mo, while mud and wattle wall surfaces produced a 98% mortality rate at 3 mo post spray. The KD₆₀ (knockdown at 60 min) for painted plastered surfaces was 100% for 6 mo, plain brick surface KD₆₀ was 80% at 6 mo, and the mud and wattle surface KD₆₀ was >80% at 3 mo. There was a significant effect on Ficam VC longevity by wall type and evidence of a relationship between test period and wall type on the KD₆₀.

Piwi-interacting RNAs (piRNAs) are a newly identified class of small noncoding RNAs. They are associated with chromatin organization, messenger RNA stability, and genome structure. Although the overexpression of piRNA-3312 in deltamethrin-susceptible (DS) strain of Culex pipiens pallens (L.) was observed in our previous large-scale transcriptome data, the roles of piRNA in insecticide resistance have not been clearly defined. The aim of the present study was to investigate how piRNA-3312 is involved in insecticide resistance. The lower expression level of piRNA-3312 in deltamethrin-resistant (DR) strain of Cx. pipiens pallens was confirmed by quantitative real time polymerase chain reaction (qRT-PCR). Overexpression of piRNA-3312 in the DR strain made the mosquitoes more sensitive to deltamethrin, whereas inhibiting the expression of piRNA-3312 in the DS strain made the mosquitoes more resistant to deltamethrin. Piwi-interacting RNA-3312 was also found to bind 3′ UTR (Untranslated Regions) of gut esterase 1 gene and could induce its degradation. In addition, knockdown of gut esterase 1 gene increased the sensitivity of DR strain to deltamethrin. In conclusion, we found that piRNA-3312 targeted the gut esterase 1 gene to negatively regulate the insecticide resistance. This finding facilitates the understanding of various functions of piRNAs and their association with insecticide resistance.

We describe a 2-yr trial to evaluate the ability of SELECT Tick Control System (TCS) host-targeted bait boxes to reduce numbers of host-seeking Ixodes scapularis nymphs in a residential neighborhood. After four successive 9-wk deployments, nymphal and larval I. scapularis infestation prevalence and intensity were significantly reduced on target small mammals. In addition, these deployments resulted in 87.9% and 97.3% control of host-seeking nymphs in treatment sites at 1 yr and 2 yr postintervention, respectively. Installation of a protective metal cover around the SELECT TCS bait boxes eliminated nontarget wildlife damage to bait boxes that resulted in failure of previous bait box types. The results are discussed in the context of the residential environment and future research needs.

The head louse Pediculus humanus capitis (De Geer) is a hematophagous ectoparasite that inhabits the human scalp. Infestations by this insect are commonly known as pediculosis, which is more common in younger groups. These infestations are asymptomatic; however, skin irritation from scratching occasionally may cause secondary bacterial infections. In recent years, the prevalence of pediculosis has increased in children; this increase has been attributed to louse resistance to the insecticides used as a control measure for infestation. The aim of the present study was to determine the presence and frequency of the knockdown resistance mutation (kdr) T929I in 468 head lice collected from 32 elementary schools in the metropolitan area of Nuevo Leon (24) and Yucatan (8), Mexico. This is the first report of a knockdown resistance (kdr) mechanism in head lice from Mexico. The T929I mutation was present in all of the sampled schools, with variability observed in its allelic and genotypic frequencies.

To control malaria in Madagascar, two primary vector control interventions are being scaled up: insecticide-treated nets and indoor residual spraying of bendiocarb, which was implemented in the Malagasy Central Highlands in 2009. The current efficacy of bendiocarb against Anopheles species was evaluated in a small-scale field trial. An experimental hut trial comparing the effectiveness of bendiocarb sprayed on five substrates (cement, wood, tin, mud, and vegetative materials) was carried out against Anopheles species in two study sites located in the eastern foothills of Madagascar. No significant difference was detected in either exophily or blood-feeding rates between treated and untreated huts. The mortality rate was significantly greater in treated huts compared to untreated huts. Efficacy up to 80% was found for 5 mo posttreatment. Although effective, bendiocarb has been used for 7 yr, and therefore an alternative insecticide may be needed to avoid the emergence of resistance.

Rickettsia felis is a human pathogen transmitted by the cat flea, Ctenocephalides felis (Bouché) (str. LSU), as well as an obligate symbiont of the parthenogenic booklouse Liposcelis bostrychophila (Badonnel) (str. LSU-Lb). The influence of genetic variability in these two strains of R. felis on host specialization and fitness and possible resulting differences on infection and transmission kinetics in C. felis is unknown. Utilizing an artificial host system, cat fleas were exposed to a R. felis str. LSU-Lb-infected bloodmeal and monitored for infection at 7-d intervals for 28 d. Quantitative real-time PCR was used to determine rickettsial load and infection density in newly exposed cat fleas, and transmission frequency between cat fleas. The effect of persistent R. felis infection on cat flea F₁ progeny was also assessed. At 7 d postexposure 76.7% of the cat fleas successfully acquired R. felis str. LSU-Lb. In R. felis str. LSU-Lb-exposed cat fleas, the mean infection load (6.15 × 10⁶), infection density (0.76), and infection prevalence (91/114) were significantly greater than R. felis str. LSU infection load (3.09 × 10⁶), infection density (0.68), and infection prevalence (76/113). A persistent R. felis str. LSU-Lb infection was detected for 28 d in adult cat fleas but neither female:male ratio distortion nor vertical transmission was observed in F₁ progeny. While infection kinetics differed, with higher intensity associated with R. felis str. LSU-Lb, no distinct phenotype was observed in the F₁ progeny.

This study investigated possible transovarial and transstadial transmission of Hepatozoon canis by Rhipicephalus sanguineus (Latreille) ticks collected from naturally infected dogs in a municipal dog shelter and the grounds of the shelter. Four hundred sixty-five engorged nymphs were collected from 16 stray dogs that were found to be infected with H. canis by blood smear and PCR analyses and maintained in an incubator at 28 °C for moulting. Four hundred eighteen nymphs moulted to adults 14–16 d post collection. Unfed ticks from the shelter grounds comprised 1,500 larvae, 2,100 nymphs, and 85 adults; were sorted according to origin, developmental stage, and sex into 117 pools; and screened by 18S rRNA PCR for Hepatozoon infection. Of 60 adult tick pools examined, 51 were infected with H. canis. The overall maximum likelihood estimate (MLE) of infection rate was calculated as 21.0% (CI 15.80–28.21). Hepatozoon canis was detected in 31 out of 33 female pools (MLE 26.96%, CI 17.64–44.33) and 20 out of 27 male pools (MLE 14.82%, CI 20.15–46.41). Among 42 unfed nymph pools collected from the shelter, 26 were infected with H. canis, and MLE of infection was calculated as 1.9% (CI 1.25–2.77). No H. canis DNA was detected in any of the gDNA pools consisting of larva specimens. Partial sequences of the 18S rRNA gene shared 99–100% similarity with the corresponding H. canis isolates. Our results revealed the transstadial transmission of H. canis by R. sanguineus, both from larva to nymph and from nymph to adult, in field conditions. However, there were no evidence of transovarial transmission.

PCR-based identification of vertebrate host bloodmeals has been performed on several vectors species with success. In the present study, we used a previously published PCR protocol followed by DNA sequencing based on primers designed from multiple alignments of the mitochondrial cytochrome b gene used to identify avian and mammalian hosts of various hematophagous vectors. The amplification of a fragment encoding a 359 bp sequence of the Cyt b gene yielded recognized amplification products in 192 female sand flies (53%), from a total of 362 females analyzed. In the study area of Casa Branca, Brazil, blood-engorged female sand flies such as Lutzomyia longipalpis (Lutz & Neiva, 1912), Migonemyia migonei (França, 1924), and Nyssomyia whitmani (Antunes & Coutinho, 1939) were analyzed for bloodmeal sources. The PCR-based method identified human, dog, chicken, and domestic rat blood sources.

Measuring rates of acquisition of the Lyme disease pathogen, Borrelia burgdorferi sensu lato Johnson, Schmid, Hyde, Steigerwalt & Brenner, by the larval stage of Ixodes scapularis Say is a useful tool for xenodiagnoses of B. burgdorferi in vertebrate hosts. In the nymphal and adult stages of I. scapularis, the duration of attachment to hosts has been shown to predict both body engorgement during blood feeding and the timing of infection with B. burgdorferi. However, these relationships have not been established for the larval stage of I. scapularis. We sought to establish the relationship between body size during engorgement of larval I. scapularis placed on B. burgdorferi-infected, white-footed mice (Peromyscus leucopus Rafinesque) and the presence or absence of infection in larvae sampled from hosts over time. Body size, time, and their interaction were the best predictors of larval infection with B. burgdorferi. We found that infected larvae showed significantly greater engorgement than uninfected larvae as early as 24 h after placement on a host. These findings may suggest that infection with B. burgdorferi affects the larval feeding process. Alternatively, larvae that engorge more rapidly on hosts may acquire infections faster. Knowledge of these relationships can be applied to improve effective xenodiagnosis of B. burgdorferi in white-footed mice. Further, these findings shed light on vector–pathogen–host interactions during an understudied part of the Lyme disease transmission cycle.

Mepraia spinolai (Porter) is a vector of Trypanosoma cruzi that causes Chagas disease. Females are always wingless, but males may be winged or wingless. We determined by PCR the infection percentage with T. cruzi of M. spinolai adults and nymphs in domestic, peridomestic, and wild collections, in different regions of Chile. In all regions, winged males were more abundant than females and wingless males. Winged males collected inside houses were less parasitized than were those from peridomestic and wild environments. Although winged males of M. spinolai have comparatively low levels of infection, this segment may still represent the greatest vector threat in this species for transmission of T. cruzi to humans and other vertebrates in domestic, wild, and peridomestic habitats. Winged males represent the dispersive form of this species that invades human dwellings. Feeding deprivation resulting from the time required to find a food source and to search for reproductive females could explain the lower infection rates (negatives) of winged males collected from inside houses in comparison with winged males collected from peridomestic and wild habitats.

Although the head louse, Pediculus humanus capitis De Geer, and body louse, Pediculus humanus humanus L., both have a worldwide distribution, the occurrence of head louse pediculosis appears to be more prevalent in modern societies despite systematic use of various pediculicides. This study tested head lice collected in rural Georgia and body lice collected in Russia for the prevalence of a kdr-biomarker that is associated with permethrin resistance. This study also screened lice for the presence of DNA from Bartonella quintana and Acinetobacter species. The kdr-permethrin resistance biomarker for the T917I mutation was detected by RFLP and PCR in 99.9% of head lice tested from Georgia, whereas only 2.9% of body lice from Russia tested positive for this kdr biomarker. DNA of B. quintana was detected in 10.3% of head lice from Georgia, whereas 84.8% of body lice from Russia tested positive. Acinetobacter DNA was detected in 80.8% (95% CI, 68–89%) of head lice from Georgia and all body lice from Russia tested.

Among true bugs (Insecta: Hemiptera), only hematophagous species (families Reduviidae, Cimicidae) have high veterinary and medical significance. In addition, several predatory and plant-feeding bug species, which also have piercing–sucking mouthparts, are known to occasionally bite humans. The majority of such examples are known from the New World. Here, we report the first case concerning the human-biting potential of the western conifer seed bug, Leptoglossus occidentalis Heidemann (Hemiptera: Coreidae). This is a phytophagous bug species, which has become widespread in North America, and has also been introduced into Europe where it shows a rapidly expanding geographical range.

Fleas are known vectors of zoonotic agents. Thirty-five fleas, including 28 Ctenocephalides felis (Bouché), four Pulex irritans (L.), and three Echidnophaga gallinacea (Westwood) from 19 rural dogs from southwestern Uganda were analyzed for the presence of Rickettsia spp. (ompB, gltA, and 17 kDa fragment genes) and Bartonella spp. (rpoB and ITS genes) by PCR. Rickettsial DNA was detected in 27 out of 28 of Ct. felis and in two out of four P. irritans. None of the E. gallinacea specimens harbored Rickettsia DNA. Rickettsia felis was confirmed in 12 Ct. felis and in the two P. irritans specimens with positive PCR-results. In addition, the presence of Candidatus Rickettsia asemboensis was evidenced in 15 Ct. felis. Bartonella spp. was not amplified in any sample. Our survey indicates that R. felis, the agent of the flea-borne spotted fever, is present in the study area. Besides, this is the first description of Ca. R. asemboensis in Uganda.

The black-legged tick, Ixodes scapularis Say, is the primary vector of Borrelia burgdorferi, a spirochete that causes Lyme disease, in eastern North America. Lyme disease risk has generally been considered to be focused in the Northeast and the northern Midwest in the United States, yet the distribution of the vector extends considerably more broadly. A recent analysis of the distribution of the species using ecological niche modeling approaches painted an odd biogeographic picture, in which the species is distributed in a “rimming” distribution across the northern Midwest and Northeast, and along the Atlantic and Gulf coasts of the eastern United States, but not broadly in the interior of eastern North America. Here, we reanalyze the situation for this species, and demonstrate that the distribution estimated in the previous study was a consequence of assumptions about relative weights applied to different error types. A more appropriate error weighting scheme for niche modeling analyses, in which omission error is prioritized over commission error, shows a simpler distribution, in which the species ranges continuously across eastern North America; this distributional pattern is supported by independent occurrence data from the eastern Great Plains, in Kansas. We discuss implications for public health planning and intervention across the region, as well as for developing effective and predictive maps of vector distributions and pathogen transmission risk.

A major mosquito-borne viral disease outbreak caused by Zika virus (ZIKV) occurred in Bahia, Brazil, in 2015, largely due to transmission by the mosquito, Aedes aegypti (L.). Detecting ZIKV in field samples of Ae. aegypti has proven problematic in some locations, suggesting other mosquito species might be contributing to the spread of ZIKV. In this study, several (five) adult Aedes albopictus (Skuse) mosquitoes that emerged from a 2015 field collection of eggs from Camaçari, Bahia, Brazil, were positive for ZIKV RNA; however, attempts to isolate live virus were not successful. Results from this study suggest that field-collected Ae. albopictus eggs may contain ZIKV RNA that require further tests for infectious ZIKV. There is a need to investigate the role of Ae. albopictus in the ZIKV infection process in Brazil and to study the potential presence of vertical and sexual transmission of ZIKV in this species.

Borrelia mayonii is a newly described member of the Borrelia burgdorferi sensu lato complex that is vectored by the black-legged tick (Ixodes scapularis Say) and a cause of Lyme disease in Minnesota and Wisconsin. Vertebrate reservoir hosts involved in the enzootic maintenance of B. mayonii have not yet been identified. Here, we describe the first isolation of B. mayonii from naturally infected white-footed mice (Peromyscus leucopus Rafinesque) and an American red squirrel (Tamiasciurus hudsonicus Erxleben) from Minnesota, thus implicating these species as potential reservoir hosts for this newly described spirochete.

Flea-borne rickettsiosis occurs worldwide and includes a number of pathogens, namely, Rickettsia typhi and Rickettsia felis. Most studies in the United States have occurred in southern Texas and California where flea-borne rickettsiosis is endemic, resulting in a lack of information from other regions of the country. Between March and August 2016, 222 fleas were collected from 52 client-owned dogs and cats in two urban areas in Oklahoma. Fleas were identified using morphological characteristics then pooled and tested by polymerase chain reaction (PCR) using published primers for gltA, ompB, and 17-kDa. The majority (98.6%) of fleas collected were Ctenocephalides felis (Bouché) followed by Pulex irritans (L) (1.4%). Overall, fleas collected from 30.0% (6/20) cats and 43.8% (14/32) dogs were infected with R. felis. Three C. felis-pools collected from three dogs, two in the Enid area (central Oklahoma) and one in the Elk City area (western Oklahoma), were infected with R. typhi as well as R. felis. ‘Candidatus R. senegalensis' was detected in one pool of fleas taken from a cat in Oklahoma City. This is the first evidence that flea-borne Rickettsia species occur in fleas obtained from client-owned dogs and cats in the Great Plains region. The impact of these Rickettsia species on public health in the region needs further investigation.