Tick-borne diseases are a growing problem in many parts of the world, and their surveillance and control touch on challenging issues in medical entomology, agricultural health, veterinary medicine, and biosecurity. Spatial approaches can be used to synthesize the data generated by integrative One Health surveillance systems, and help stakeholders, managers, and medical geographers understand the current and future distribution of risk. Here, we performed a systematic review of over 8,000 studies and identified a total of 303 scientific publications that map tick-borne diseases using data on vectors, pathogens, and hosts (including wildlife, livestock, and human cases). We find that the field is growing rapidly, with the major Ixodes-borne diseases (Lyme disease and tick-borne encephalitis in particular) giving way to monitoring efforts that encompass a broader range of threats. We find a tremendous diversity of methods used to map tick-borne disease, but also find major gaps: data on the enzootic cycle of tick-borne pathogens is severely underutilized, and mapping efforts are mostly limited to Europe and North America. We suggest that future work can readily apply available methods to track the distributions of tick-borne diseases in Africa and Asia, following a One Health approach that combines medical and veterinary surveillance for maximum impact.

Tick-borne illnesses have been on the rise in the United States, with reported cases up sharply in the past two decades. In this literature review, we synthesize the available research on the relationship between vegetation and tick abundance for four tick species in the northeastern United States that are of potential medical importance to humans. The blacklegged tick (Ixodes scapularis) (Say; Acari: Ixodidae) is found to be positively associated with closed canopy forests and dense vegetation thickets, and negatively associated with open canopy environments, such as grasslands or old agricultural fields. The American dog tick (Dermacentor variabilis) (Say; Acari: Ixodidae) has little habitat overlap with I. scapularis, with abundance highest in grasses and open-canopy fields. The lone star tick (Amblyomma americanum) (Linnaeus; Acari: Ixodidae) is a habitat generalist without consistent associations with particular types of vegetation. The habitat associations of the recently introduced Asian longhorned tick (Haemaphysalis longicornis) (Neumann; Acari: Ixodidae) in the northeastern United States, and in other regions where it has invaded, are still unknown, although based on studies in its native range, it is likely to be found in grasslands and open-canopy habitats.

The common bed bug (Cimex lectularius L.) is a known pest and an obligate blood-feeding ectoparasite. Bed bugs can feed on warm-blooded animals including humans, bats, poultry, and rabbits, but no research has investigated the use of companion animals (canines and/or felines) as a blood source. This study investigates how long known host DNA could be detected in a bed bug and the prevalence of bed bugs feeding on companion animals. Laboratory-reared bed bugs were fed host blood to determine how long DNA of human, feline, canine, and rabbit blood could be detected up to 21 d postfeeding. Additionally, 228 bed bugs were collected from 12 apartments with pets (6: canine, 5: feline, and 1: canine and feline), characterized as engorged or unengorged, and then screened with host-specific primers to identify the bloodmeal. Host meals of human, rabbit, feline, and canine blood were detected up to 21 d after feeding laboratory strains. All bed bugs died after feeding on the canine blood, but DNA could be detected up to 21 d post feeding/death. Of the field-collected bed bugs analyzed, human DNA was amplified in 158 (69.3%) bed bugs, canine DNA amplified in 7 bed bugs (3.1%), and feline DNA amplified in 1 bed bug (0.4%). Results of this study suggest that bed bugs predominately feed on humans and rarely feed on companion animals when they cohabitate in low-income, high-rise apartments. Additionally, results from this study warrant future investigations into host use by bed bugs in different housing structures and socioeconomic environments.

With the aim of identifying insects with potential value as indicator species in postmortem interval (PMI), the location or season of death of human or monogastric animals, two medium-sized large white pigs were used for the study during cold and warm seasons. Five stages of decomposition and their durations were observed and described in the pig carcasses during both seasons. Carcasses were first colonized by flies from seven taxa during fresh stage. Chrysomya marginalis (Wiedemann) (Diptera: Calliphoridae), Chrysomya albiceps, (Wiedemann) (Diptera: Calliphoridae) and Musca domestica (Walker) (Diptera: Calliphoridae) were recorded as the overall dominant adult flies found on the carcasses in both seasons. Colonization by beetles began during the bloated stage in the warm season and active stage in the cold season. Dermestes maculatus (De Geer) (Coleoptera: Dermestidae) and Necrobia rufipes (De Geer) (Cleridae) were the most abundant beetles in both seasons. Flies and beetles were generally abundant during the warm seasons as compared to the cold season. However, the difference was only significant for beetles. The highest number of flies were recorded in the bloated stage for both seasons, however they were reduced in the active stage which coincided with the introduction of predatory beetles. The arrival time of the different arthropod species and their association with different stages of decomposition during both seasons pointed to their value in estimating the PMI in forensic investigations in the locality of KwaZulu-Natal, South Africa. Consequently, they can potentially be useful in the estimation of PMI and other cases of criminal investigations.

Mosquito-borne pathogens have spread throughout tropical regions of the Western Hemisphere causing increased burden of disease in the region. Outbreaks of dengue fever, yellow fever, chikungunya, West Nile, and Zika have occurred over the past several years. Mosquito blood-feeding patterns need to be assayed to assist in determining which vertebrates could act as hosts of these mosquito-borne pathogens and which mosquito species could act as vectors. We conducted bloodmeal analyses of mosquitoes collected at Lomas Barbudal Biological Reserve, a dry tropical forest reserve in Costa Rica. Mosquitoes were collected using backpack aspirators and light, gravid, and resting traps, and then identified morphologically. Blood-fed mosquitoes underwent DNA extraction, PCR amplification, and sequencing of the vertebrate cytochrome b and cytochrome c oxidase 1 genes to identify vertebrate bloodmeal hosts. Several mosquitoes known to vector pathogens were found including Culex (Melanoconion) erraticus Dyar & Knab (Diptera: Culicidae), Cx. (Mel.) pedroi Sirivanakarn & Belkin, Aedes (Stegomyia) albopictus Skuse, Ae. (Ochlerotatus) scapularis Rondani, Ae. (Och.) serratus Theobald, and Ae. (Och.) taeniorhynchus Wiedemann. The most common bloodmeal hosts were basilisk lizards (Basiliscus vittatus) Wiegmann (Squamata: Corytophanidae) in Culex (Linnaeus) species and white-tailed deer (Odocoileus virginianus) Zimmermann (Artiodactyla: Cervidae) in Aedes (Meigen) species. These results show the diversity of mosquito species in a tropical dry deciduous forest and identify associations between mosquito vectors and potential pathogen reservoir hosts. Our study highlights the importance of understanding interactions between vector species and their hosts that could serve as predictors for the potential emergence or resurgence of mosquito-borne pathogens in Costa Rica.

The prevalence of mites of the genus Demodex and their associations with host age, gender, workplace, and comorbid skin and ocular conditions were studied in participants in Antofagasta, Chile, which is in a region with an extreme environment. We examined 680 participants aged 18–88 yr using standardized surface skin biopsies. Among them, Demodex had a prevalence of 13.5 % (95% confidence interval, 10.88–16.17). A slightly higher prevalence was observed in males (51.1%; 95% confidence interval, 40.9–61.3; nonsignificant) and participants in the 69–88 yr age group (50.0%; 95% confidence interval, 23.8–76.2; P < 0.05). Regarding the species involved, Demodex folliculorum was found in 89.1% (CI 82.7–95.5) of cases, while D. brevis was found in the remaining 10.9% of cases. A higher prevalence (25.0% CI 16.1–33.91) was observed in participants who worked indoors in generally enclosed and dust-rich environments (such as theaters, libraries, and administrative offices). There was also a strong association between Demodex prevalence and conjunctival hyperemia, with 35.9% (95% confidence interval, 9.1–35; OR 17.9) of the Demodex positive participants having this pathology compared to 10.3% of the noninfested participants. In summary, the prevalence of Demodex in Antofagasta, Chile (13.5%) was toward the lower end of the range reported among other regions around the world. Environmental factors such as exposure to the sun (including ultraviolet rays) or environmental pollution may affect the mites. In addition, Demodex genetics (related to virulence) and the ocular or skin microbiota may positively or negatively influence infestation and pathology.

The western mosquitofish, Gambusia affinis (Baird & Girard), has been used worldwide for the control of larval mosquitoes for more than 100 yr. We found that the western encephalitis mosquito, Culex tarsalis Coquillett (Diptera: Culicidae), can detect the presence of G. affinis in oviposition sites based on associated chemicals, leading to a decrease in the number of egg rafts laid. Three volatile chemical compounds were identified in the headspace above the water where G. affinis had been held for 24 h. Oviposition bioassays conducted using standards of the volatile compounds identified (dimethyl disulfide [DMDS], dimethyl trisulfide [DMTS], and S-methyl methanethiosulphonate) found that females reduced oviposition only when low concentrations of DMTS were present, but this response was not consistent across all trials and concentrations tested. DMDS, DMTS, and S-methyl methanethiosulphonate are known bacterial metabolic waste products and may be the source of the compounds. Two nonvolatile compounds of interest were found to be present in the Gambusia-exudate water. After tasting Cx. tarsalis were deterred from ovipositing onto Gambusia-treated water from which the bacteria had been removed by filtration, indicating that the kairomone may consist of nonvolatile compound(s). One of the nonvolatile compounds isolated from the Gambusia-treated water has a benzene ring structure similar to that of cholesterol but the structure of the two nonvolatile deterrents remains to be fully characterized. Our research shows that three volatile compounds and two nonvolatile compounds are present in water associated with G. affinis (Poeciliidae: Gambusia) and affect the oviposition behavior of Cx. tarsalis in laboratory bioassays.

Container Aedes mosquitoes are the most important vectors of human arboviruses (i.e., dengue, chikungunya, Zika, or yellow fever). Invasive and native container Aedes spp. potentially utilize natural and artificial containers in specific environments for oviposition. Several container Aedes spp. display ‘skip-oviposition’ behavior, which describes the distribution of eggs among multiple containers during a single gonotrophic cycle. In this study, we compared individual skip-oviposition behavior using identical eight-cup testing arenas with three container Aedes species: Aedes aegypti (Linnaeus), Aedes albopictus (Skuse), and Aedes triseriatus (Say). We applied the index of dispersion, an aggregation statistic, to individual mosquitoes’ oviposition patterns to assess skip-oviposition behavior. Aedes aegypti and Ae. albopictus utilized more cups and distributed eggs more evenly among cups than Ae. triseriatus under nutritionally enriched oviposition media (oak leaf infusion) conditions. When presented with a nutritionally unenriched (tap water) oviposition media, both Ae. aegypti and Ae. albopictus increased egg spreading behavior. Aedes albopictus did not modify skip-oviposition behavior when reared and assessed under fall-like environmental conditions, which induce diapause egg production. This study indicates specific oviposition site conditions influence skip-oviposition behavior with ‘preferred’ sites receiving higher amounts of eggs from any given individual and ‘non-preferred’ sites receive a limited contribution of eggs. A further understanding of skip-oviposition behavior is needed to make the best use of autodissemination trap technology in which skip-ovipositing females spread a potent larvicide among oviposition sites within the environment.

Entomotoxicology is an emerging science in the forensic investigation of death; it now influences judicial activities. Briefly, carrion flies ingest various chemicals as they feed on a cadaver. In entomotoxicology, such toxicological compounds are qualitatively and quantitatively analyzed in insects recovered from carrion. Some of these chemicals can disrupt flies' cellular physiology, leading to acceleration or delay in their developmental rate. In this study, the effect of zolpidem-contaminated tissue on the dipteran flies Chrysomya megacephala (Fabricius 1794) and Chrysomya saffranea (Bigot 1877) was assessed. Zolpidem tartrate is a sedative-hypnotic compound commonly used to treat anxiety and sleeping disorders, but it is associated with death from suicide. The effects were tested by providing the flies with liver tissue treated with various concentrations of zolpidem tartrate. The morphological parameters and development rate for all life cycle stages of treated insects were determined and used to estimate postmortem intervals. Results showed that the morphological parameters, i.e., weight, width, and length and rate of development of C. megacephala and C. saffranea was negatively associated with zolpidem tartrate concentration of C. saffranea, were negatively associated with the concentration of zolpidem tartrate, indicating that arthropod evidence evaluation can help establish the cause and time of death.

Successful monitoring of physiological resistance of malaria vectors requires about 150 female mosquitoes for a single set of tests. In some situations, the sampling effort is insufficient due to the low number of field-caught mosquitoes. To address this challenge, we demonstrate the feasibility of using the forced oviposition method for producing F₁ from field-caught Anopheles mosquitoes. A total of 430 and 598 gravid Anopheles females from four laboratory strains and five field populations, respectively, were tested. After blood feeding, gravid mosquitoes were individually introduced into transparent plastic vials, containing moistened cotton balls topped with a 4 cm² piece of filter paper. The number of eggs, hatching larvae, pupation, and adult emergence were recorded daily. The mean number of eggs per female mosquito ranged from 39.3 for Anopheles cracens to 93.6 for Anopheles dirus in the laboratory strains, and from 36.3 for Anopheles harrisoni to 147.6 for Anopheles barbirostris s.l. in the field populations. A relatively high egg hatching rate was found in An. dirus (95.85%), Anopheles minimus (78.22%), and An. cracens (75.59%). Similarly, a relatively high pupation rate was found for almost all test species ranging from 66% for An. minimus to 98.7% for Anopheles maculatus, and lowest for An. harrisoni (43.9%). Highly successful adult emergence rate was observed among 85–100% of pupae that emerged in all tested mosquito populations. The in-tube forced oviposition method is a promising method for the production of sufficient F₁ progeny for molecular identification, vector competence, insecticide resistance, and bioassay studies.

Chagas disease is a very important vector-borne disease in México, and Triatoma dimidiata sensu stricto (Latreille) is one of the most important vectors of Trypanosoma cruzi Chagas, mainly in southern and central states. In the Pacific Coast states with the highest prevalence of human T. cruzi infection, T. dimidiata s. s. is considered as a secondary vector. However, the vectorial capacity of those populations has not been studied. Therefore, the vector characteristics of three populations of T. dimidiata s. s. in western México were evaluated in this study. The populations were maintained in the laboratory at 27 ± 1°C and 75% ± 5% RH with a 12:12 h (light:dark) regime, fed on rabbits in a fortnight basis. The development times were short (172–238 d), and the number of bloodmeals to molt was low (11). Mortality was moderate (36–45%), the onset of feeding was relatively rapid (0.5–1.7 min), and feedings were extended (>15 min). More than 40% of individuals in most instars defecated in one of three categories: <1 min when feeding (5–37.9%), immediately after feeding (9–28.6%), or in <1 min post feeding (7–25.8%). The median number of laid eggs was high (over 2.5) in the three populations, as were the egg eclosion rates (>86%). Thus, the T. dimidiata s. s. in the three populations are potentially efficient vectors of T. cruzi and could contribute to the high prevalence of infection in human populations in western México.

Graphical Abstract

This study examined the effects of various diets on the development time and survival of the carrion beetle Thanatophilus rugosus (Fabricius, 1775). We attempted to find the best diet for rearing the species in laboratory conditions for entomological research and forensic purposes, and to further understand its feeding habits. The larval stages were monitored while feeding on three types of meat under a constant temperature. We discovered that the shortest development time in total was achieved in larvae fed with pork liver, followed closely by pork muscle. The longest development and lowest survival rates were observed when introducing a chicken diet. We were also able to identify specific stages during which the diet significantly affected the development.

The present study aimed to investigate the impact of gunshot residue (GSR) components (Pb, Sb, and Ba) on the life history parameters of Lucilia sericata (Meigen, 1826) (Diptera: Calliphoridae). This experiment was carried out at the Zoology Department, the Ondokuz Mayıs University in 2020. About 50 larvae, respectively were exposed to the chicken liver with four different concentrations of GSR particles and life history parameters recorded. A two-way analysis of variance (Two way ANOVA) was used on the larval weight, and length, development time, percentage of pupal and larval survival. Pearson's correlation coefficient was used to assess the association between life-history parameters and the concentration of GSR. The total development times decreased with increasing Sb, Ba, and Pb concentrations in the diets 1–3 d when compared to the control The results showed that Ba and Sb significantly decreased larval and pupal survival and GSR decreased the pupa and adult weight as compared to the control. We concluded that life-history parameters of L. sericata are sensitive to GSR residue and heavy metal changes in the environment. The presence of gunshot residue in the corpse should be kept in mind in a criminal investigation.

Adult Calliphoridae flies, as well as their immature stages collected from carcasses, have been used as evidence in forensic investigations to estimate the postmortem interval (PMI), particularly those of the genus Chrysomya as it is one of the first genera to colonize a corpse. Chrysomya rufifacies (Macquart 1842), due to its appearance in cadaveric remains, plays a fundamental role in the study of forensic entomology. For this reason, we determined the biological cycle of C. rufifacies under semicontrolled laboratory conditions: uncontrolled average fluctuating temperature of 29. 76 ± 3. 22°C, uncontrolled average fluctuating humidity of 48. 91 ± 11.13%, and a controlled photoperiod of 12/12 (L/O). We established that the total development time from oviposition to adult emergence of C. rufifacies was 6. 5 d. The eggs took 12 h to hatch after oviposition. The complete larval stage took 60 h (instar 1 = 12 h, instar 2 = 12 h, instar 3 = 24 h, instar 3 post-feeding = 12 h). The pupa had a duration of 84 h. The species needed a total of 4642.8(±4.59) accumulated degree-hours (ADH) to complete its biological cycle.

In the world, the impact of environmental conditions on the number of scorpion events was evaluated in North Africa,Middle East, and the Amazonian region but not in Europe. In mainland France, scorpion species described are Buthus occitanus (Amoreux, 1789), Belisarius xambeui (Simon, 1879) and 4 Euscorpiidae: Euscorpius concinnus (Koch, 1837), Euscorpius italicus (Herbst, 1800), Euscorpius tergestinus (Koch, 1837), and Tetratrichobothrius flavicaudis (De Geer, 1778). We aimed to describe the impact of environmental conduction on the number of scorpion events. For this, a retrospective multi-center study was conducted with data from the French poison control centers files about scorpion events between 1 January 2011 and 31 December 2020. During the study period, 975 incoming calls for scorpion events were recorded and 574 were related to scorpions native to mainland France and Corsica: B. occitanus (n = 86), Euscorpiidae species (n = 222), B. xambeui (n = 1), and undetermined species (n = 265). Cases were mostly reported along the Mediterranean coast, along rivers, and in cities with a trading port. The number of scorpion events was linked to the rivers' water level, rivers' flow, temperature, sunshine, and pluviometry (P < 0.05 for all variables). B. occitanus need warmest and driest environment than Euscorpiidae spp. A link between the severity of the envenoming and climatic condition or seasonality was not demonstrated.

Borrelia miyamotoi is a hard tick-associated relapsing fever spirochete that is geographically widespread in Ixodes spp. (Acari: Ixodidae) ticks, but typically occurs at low prevalence. Genetic variability has been described among strains derived from Asia, Europe, and North America, and among tick species that carry the infection, but little variability has been described within foci or tick species. Capitalizing on access to B. miyamotoi nucleic acid extracted from host-seeking Ixodes scapularis Say or Ixodes pacificus Cooley & Kohls from 16 states, we explored genetic variability based on sequence analysis of four amplicons described herein. Consistent with previous studies, we detected significant genetic differences between strains derived from I. scapularis (eastern United States) and I. pacificus (western United States) and identified two distinct sequences in the western United States (Am-West-1 and Am-West-2). Unique to this study, we identified two distinct sequences in the eastern United States (Am-East-1 and Am-East-2). Based on the 161 samples we analyzed, Am-East-1 was the only type represented in 50 B. miyamotoi-infected ticks collected from the Northeast (Vermont, Maine, New York, Connecticut, and Rhode Island), whereas ticks collected from the North-Central and Mid-Atlantic states harbored B. miyamotoi comprised of both Am-East-1 and Am-East-2. Further studies are needed to better characterize the phylogeography of B. miyamotoi and to discern if there are biologically meaningful differences among sequence types. To facilitate further exploration, we developed a polymerase chain reaction (PCR) assay designed to differentiate Am-East-1, Am-East-2, and Am-West sequence types without having to sequence the amplicon.

Aedes aegypti mosquito is the principal dengue vector in the Kingdom of Saudi Arabia (KSA); however, no study has addressed its ecology and population structure yet. Therefore, we report on Ae. aegypti phylo- and population genetics using three DNA markers: COI, ND4, and rDNA-ITS2. Sampling the immature stages of Ae. aegypti revealed that water storage tanks (34.3% of habitats) were the most productive and contained 33% of immatures stages. Other important habitats included containers for wastewater drainage (including air-conditioning and water cooler trays) and containers associated with ornamentation. Shallow water leakage spots (2.7% of habitats, 8% of immatures) can be considered rare-but-epidemiologically-important containers. Neighbor-joining (NJ) phylogenetic analysis of Ae. aegypti identified 8, 14, and 9 haplotypes of COI, ND4, and ITS2, respectively, and revealed high levels of genetic variation in Ae. aegypti populations of KSA. Global distribution of haplotypes also indicated multiple gene introductions into these populations, with high levels of intra-population genetic variation and continuous gene exchange. The neutrality values indicated a deficiency of alleles and suggested that the KSA Ae. aegypti loci tested did not follow a neutral model of molecular evolution. Fst values and AMOVA indicated that most of the genetic variation in the KSA Ae. aegypti populations is due to intra- rather than inter-population differences. This is the first comprehensive report on the phylo- and population genetics of Ae. aegypti from the Arabian Peninsula. This information expands our understanding of the ecology and population dynamics of this important arboviral vector for informed control efforts.

Culex quinquefasciatus, a member of the Culex pipiens complex, is widespread in Saudi Arabia and other parts of the world. It is a vector for lymphatic filariasis, Rift Valley fever, and West Nile virus. Studies have shown the deleterious effect of RNA interference (RNAi)-mediated knockdown of various lethal genes in model and agricultural pest insects. RNAi was proposed as a tool for mosquito control with a focus on Aedes aegypti and Anopheles gambiae. In this study, we examined the effect of RNAi of selected target genes on both larval mortality and adult emergence of Cx. quinquefasciatus through two delivery methods: soaking and nanoparticles. Ten candidate genes were selected for RNAi based on their known lethal effect in other insects. Disruption of three genes, chitin synthase-1, inhibitor of apoptosis 1, and vacuolar adenosine triphosphatase, resulted in the highest mortality among the selected genes using the two treatment methods. Silencing the other seven genes resulted in a medium to low mortality in both assays. These three genes are also active against a wide range of insects and could be used for RNAi-based mosquito control in the future.

Wolbachia pipientis (Hertig) is an endosymbiotic microorganism widespread among arthropods and other invertebrate hosts, and employed in strategies to reduce the incidence of arthropod-borne diseases. Here, we used a PCR-based approach for 16S RNA and wsp genes to investigate the prevalence, geographical distribution, and strains of Wolbachia in sand flies (Diptera: Psychodidae: Phlebotominae), the main vectors of the causative agents of leishmaniasis, from three biomes in Brazil: Amazon, Cerrado, and Caatinga. We found that: 1) Wolbachia DNA is present in most (66.7%) of the sampled sand fly species, including vectors of Leishmania spp. (Ross, Trypanosomatida: Trypanosomatidae), 2) the prevalence of Wolbachia DNA varies among species and populations, 3) some strains of Wolbachia may have wider geographical and host range in South America, and 4) two phylogenetic distinct wsp sequences might represent two novel strains for Wolbachia in South America sand flies. Those findings increase the basic knowledge about Wolbachia in South American sand flies and might foster further researches on its use to reduce the transmission of sand fly-borne parasites.

Culex longitubus Somboon, Namgay & Harbach is described as a new species of the Mimeticus Subgroup of the subgenus Culex. The larva is most similar to the larva of Cx. tianpingensis Chen from China, but is distinguished by the length of the siphon and the anal papillae, the form of the comb scales and pecten spines, and the development of setae 7-P, 13-T, 1-X, and 4-X. The adults have wing markings and male genitalia similar to those of species of the Mimeticus Complex. Phylogenetic analysis of COI sequences revealed that the new species is closely related to Cx. murrelli Lien of the Mimulus Complex. The immature stages of the new species were found in stagnant pools and marshes at high altitudes in several districts of Bhutan.

Identification of species involved in cadaveric decomposition, such as scavenger Diptera, is a fundamental step for the use of entomological evidence in court. Identification based on morphology is widely used in forensic cases; however, taxonomic knowledge of scavenger fauna is poor for many groups and for many countries, particularly Neotropical ones. A number of studies have documented the utility of a DNA barcoding strategy to assist in the identification of poorly known and diverse groups, particularly in cases involving immature states or fragmented organisms. To provide baseline knowledge of the diversity of scavenger Diptera in the Valley of Mexico, we generated a DNA barcode collection comprised of sequences of the cytochrome c oxidase subunit 1 (COI) gene for all families sampled at a nature reserve located in this region. We collected and identified specimens on the basis of morphology and a species delimitation analysis. Our analyses of 339 individuals delineated 42 species distributed across nine families of Diptera. The richest families were Calliphoridae (9 species), Sarcophagidae (7 species), and Phoridae (6 species). We found many of the species previously recorded for the Valley of Mexico, plus 18 new records for the region. Our study highlights the utility of DNA barcoding as a first-step strategy to assess species richness of poorly studied scavenger fly taxa.

Dermacentor variabilis is the most widely distributed three-host tick in North America, and transmits a variety of pathogens. Within the United States, this species has a discontinuous distribution, widespread east of the Rocky Mountains and with a few populations west of the Rockies. Phylogenetic evidence based on individual markers or relatively small data sets has suggested that populations at both sides of this geographic barrier may correspond to two different species. In this study, we further explore this hypothesis using an integrative taxonomy framework. Both molecular (mitochondrial and nuclear markers) and morphological analyses of specimens collected from central-eastern and western states were performed to explore species delimitation in this taxon. Results from these analyses were consistent, and provide strong evidence that D. variabilis actually corresponds to two species. Herein, the western populations are described as a new species, Dermacentor similis n. sp. The usefulness of integrative taxonomy in the context of species delimitation is also discussed.

Carrion is a dynamic and nutrient-rich resource that attracts numerous insect species that undergo succession due to the rapid change in the carrion resource. Despite this process being well-understood, few studies have examined resource change as a driver of carrion insect succession, and instead have focused on the effects of time per se, or on coarse, qualitative measures such as decay stage. Here we report on three field succession experiments using pig carcasses and human cadavers encompassing two winters and one summer. We quantified the effects of resource change (measured as total body score, TBS), carrion type, initial carrion mass, ambient temperature, and season on insect species richness and community composition. We found that all variables had an effect on different taxonomic or trophic components of the insect community composition, with the exception of initial carrion mass which had no effect. We found significant positive effects of TBS on beetle species richness and composition, while fly species richness was not significantly affected by TBS, but was by ambient temperature. TBS had a significant positive effect on all trophic groups, while ambient temperature also had a significant positive effect on the necrophages and predator/parasitoids. Our study indicates that resource change, as indicated by TBS, is an important driver of carrion insect species turnover and succession on carrion, and that TBS can provide information about insect ecological patterns on carrion that other temporal measures of change cannot.

An ecological study to evaluate necrophagous species richness, abundance, and diversity was conducted in four contrasting ecoregions of Los Angeles County, California, United States: a highly anthropized area, two moderately populated areas, and a sparsely populated area. Our hypothesis states that there will be higher abundance of exotic species in greatly anthropized ecoregions and higher diversity of native calliphorid flies in sparsely populated sites. A total of 2,883 blow flies belonging to 12 species were captured from the four ecoregions. The exotic Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) was the most abundant species, representing 72.84% of all Calliphoridae collected. The highest abundance was in the highly anthropized ecoregion. Differences of the blow fly assemblages across environmentally and anthropically different ecoregions were found, and that the relative abundance of the exotic species increased with anthropization. Our results suggest that both anthropization and elevation alter the structure of blow fly assemblages, modifying the abundance and species richness of Calliphoridae. The results of this study support the hypothesis of biotic homogenization, since we found a higher proportional abundance of exotic species and lower diversity of native blow flies in ecoregions heavily modified by human action. Additionally, the hypothesis of intermediate disturbance was supported as the highest diversity was found in the ecoregion with moderate anthropization mixed with natural elements. This is the first study to conduct an intensive survey of the Calliphoridae assemblages in Los Angeles County, which can be potentially used as a tool in human and veterinary health and forensic sciences.

Graphical Abstract

To better understand the population dynamics and dispersal ability of insect species, it is often helpful to derive a life table containing fundamental demographic data. The aim of this study was to determine a life table for the predatory necrophagous species Synthesiomyia nudiseta (van der Wulp, 1883) on a pig liver diet and under controlled laboratory conditions (29.5 ± 2. 5°C, RH 50 ± 15%, and a photoperiod of 12:12). This species has medical and veterinary importance and its distribution extends in tropical and subtropical areas and now it has been established in the southwestern of Europe. The mean adult longevity was 36. 18 ± 2. 06 d and the net reproduction rate, R, was 27.65 offspring/female, the mean generation time, T, was 22. 09 d, the finite rate of increase, λ, was 1. 16 d^–1, and the intrinsic rate of increase, r, was 0. 15 d^–1. These results indicate that S. nudiseta cannot be considered an r-strategist as the most common synanthropic necrophagous blowflies due to its predatory behavior; however, its invasive and colonist abilities are discussed. This is the first life table study of this species from Palearctic region to analyze the effect of its dispersal ability.

The Asian longhorned tick, Haemaphysalis longicornis Neumann, is an invasive species in the United States. Since its earliest recorded presence in West Virginia in 2010, H. longicornis has been reported from 15 states. While its public health significance in the United States is unclear, globally it transmits pathogens that infect livestock and humans, causing economic losses and substantial morbidity. Management and control of H. longicornis requires knowledge of its biology, ecology, and distribution. Here, we address the need for effective collection methods for host-seeking H. longicornis as an important step for accurately assessing tick abundance and potential disease risk. The number of H. longicornis collected were compared across three collection methods (dragging, sweeping, CO₂ traps) and three tick check distances (5 m, 10 m, and 20 m) were compared for dragging and sweeping. Field collections were conducted from June through August 2019 in Westchester County, New York, and ticks were grouped by life stage to assess collection method efficiency. Results indicated that implementing shorter (5 m) tick check distance was ideal for adult and nymphal collections. The dragging method proved better than sweeping for adult collections; however, there was no significant difference between the methods for nymphal collections, at any tick check distance evaluated. CO₂ traps attracted H. longicornis, but additional research is necessary to devise an effective tick retaining method before the traps can be implemented in the field. The results are presented to inform and support H. longicornis surveillance and control programs across the nation.

This study presents the diversity of mosquitoes collected from communes, endemic with malaria and dengue, located in Khanh Hoa and Binh Phuoc Provinces, Vietnam. A total of 10,288 mosquitoes were collected in the village and forested sites using standard larval dippers, cow-baited traps, ultra-violet light traps, and mechanical aspirators. Mosquito taxa were identified morphologically and species complexes/groups were further characterized molecularly. Five genera of mosquitoes were morphologically identified: Anopheles Meigen (21 species), Aedes Meigen (2 species), Culex Linnaeus (5 species), Mansonia Blanchard sp., and Armigeres Theobald sp. The PCR-based identification methods allowed the distinction of members of Maculatus Group, Funestus Group, and Dirus Complex; and DNA barcodes enabled the further identification of the Barbirostris Complex. Data reported here include the first report of An. saeungae Taai & Harbach and An. wejchoochotei Taai & Harbach from Vietnam, and re-emphasizes the significance of using molecular data in an integrated systematic approach to identify cryptic species and better understand their role in disease transmission.

Mat and liquid-type electric insecticide vaporizers continuously emit insecticides into the surrounding air. Because both the target insects, mostly mosquitoes, and humans are exposed to those insecticides, it is crucial to understand and monitor their deposition and spatial distribution in treated areas. In the current study, we examined the evaporation of insecticides from seven commercial liquid and mat vaporizers and measured their knock-down and insecticidal activity against the adult females of the Asian tiger mosquito, Aedes albopictus (Skuse). Electric vaporizers from three manufactures had differences in their heaters and active ingredients. Most had continued evaporation during hourly and daily monitoring; however, some liquid vaporizers failed to continue emission to their designated end periods. Overall, mosquitoes located at the upper position in a Peet-Grady chamber and in a field-simulation room exhibited faster knock-down activity than did mosquitoes in other areas, indicating that the insecticides accumulated on the ceiling area. Although most of mat and liquid vaporizers had <60 min of average KT₉₀ values when tested in the Peet-Grady chamber (1.8 × 1.8 × 1.8 m), they failed to have any knock-down in 2 h of observation in the field-simulation room (6.8 × 3.4 × 2.7 m) but showed 72. 8 ± 11.7% and 56. 7 ± 7.3% knock-down in the mat and liquid vaporizers, respectively, in 3 h of operation. Further study will be required to examine whether this relatively limited efficacy can be compensated by other physiological and behavioral effects, including disruption in host-seeking or blood-sucking activities.

Aedes albopictus is the vector of arbovirus diseases including yellow fever, dengue, Zika virus, and chikungunya fever, and it poses an enormous threat to human health worldwide. Previous studies have revealed that haedoxan A (HA), which is an insecticidal sesquilignan from Phryma leptostachya L., is a highly effective natural insecticide for managing mosquitoes and houseflies; however, the mechanisms underlying the response of Ae. albopictus after treatment with sublethal concentrations of HA is not clear. Here, high-throughput sequencing was used to analyze the gene expression changes in Ae. albopictus larvae after treatment with the LC₃₀ of HA. In total, 416 differentially expressed genes (DEGs) were identified, including 328 upregulated genes and 88 downregulated genes. Identification and verification of related DEGs were performed by RT-qPCR. The results showed that two P450 unigenes (CYP4C21 and CYP304A1), one carboxylesterase, and one ABC transporter (ABCG1) were induced by HA, which indicated that these detoxifying enzyme genes might play a major role in the metabolic and detoxification processes of HA. Additionally, acetylcholine receptor subunit α2 (AChRα2), AChRα5, AChRα9, and the glutamate receptor ionotropic kainate 2 (GRIK2) were found to be upregulated in HA-treated larvae, suggesting that HA affected the conduction of action potentials and synaptic transmission by disrupting the function of neural receptors. These results provide a foundation for further elucidating the target of HA and the mechanism of detoxification metabolism in Ae. albopictus.

Resistance status of Aedes albopictus (Skuse) from 13 districts in Sarawak State, Malaysia, was evaluated against four major classes of adulticides, namely organochlorine, organophosphate, carbamate, and pyrethroid. Adult bioassays were performed according to the World Health Organization (WHO) standard protocols to assess knockdown and mortality rates of Ae. albopictus. Among the tested pyrethroids, only cyfluthrin was able to exhibit complete knockdown. On the other hand, different susceptibility and resistance patterns were observed in other adulticides. As for mortality rates, the mosquitoes were susceptible to cyfluthrin and dieldrin but exhibited various susceptibilities to other tested adulticides. Cross-resistance was discovered within and between tested insecticide classes. Significant correlations were found within pyrethroid and carbamate classes (i.e., bendiocab and propoxur, P = 0.036; etofenprox and permethrin, P = 0.000; deltamethrin and lambda-cyhalothrin, P = 0.822; deltamethrin and permethrin, P = 0.042). Additionally, insecticides belonging to different groups were also found significantly correlated (i.e., malathion and deltamethin, P = 0.019; malathion and bendiocarb, P = 0.008; malathion and propoxur, P = 0.007; and bendiocarb and deltamethrin, P = 0.031). In conclusion, cyfluthrin was effective for Aedes albopictus control in Sarawak State and these data may assist local authorities to improve future vector control operations.

Indoor residual spray with deltamethrin remains the most common tool for reducing malaria transmission in Thailand. Deltamethrin is commonly used to spray the entire inner surfaces of the walls to prevent mosquitoes from resting. This study compared the mosquito landing responses on humans inside three experimental huts treated with deltamethrin at three different extents of wall coverage (25%, 50%, and full coverage), with one clean/untreated hut serving as a control. There were no significant differences between the numbers of Anopheles mosquitoes landing in the 50% and full coverage huts, whereas, in comparison to both of these, there was a significantly greater number landing in the 25% coverage hut. This study demonstrates that varying the percent coverage of indoor surfaces with deltamethrin-treated netting influences the blood-feeding success of wild Anopheles, and our findings suggest that it may be possible to reduce the extent of insecticide surface treatment while maintaining equivalent mosquito avoidance action to that seen in fully treated structures.

Indoor residual spraying (IRS) is one of the key vector control tools with a long history of use in the world. Ethiopia has set a goal to eliminate malaria from selected districts mainly by applying IRS and the distribution of long-lasting insecticidal nets. IRS is applied in low malaria transmission districts which are epidemic prone and in districts with high malaria transmission. Ethiopia uses insecticides that are recommended by World Health Organization; these insecticides must also be registered in Ethiopia. The registration of new and potential products requires confirmatory, local efficacy trials to be performed. Actellic 300CS, now registered, is one of such potential product. Actellic 300CS showed average mortalities of 99.6%, 99.6%, and 99.0% on the sprayed surfaces in the experimental huts, the top, middle, and bottom sections, respectively during the first 6 mo of the study period. Beyond 6 mo, (7, 8, and 9 mo) follow-up, mortalities for the top, middle, and bottom sections were 85.2%, 86.3%, and 85.2%, respectively. The results showed that the residual efficacy of Actellic 300CS was up to 9 mo with the first 6 mo exhibiting mortalities of greater than 99% while the next 3 mo showed mortalities exceeding 85%. Actellic 300CS was effective against fully susceptible laboratory-reared Anopheles arabiensis on all four surface types (rough, smooth, dung, and painted surfaces) tested in this study and could be used as one of the chemical insecticides of choice for the ongoing IRS programs in Ethiopia.

The house fly, Musca domestica L. (Diptera: Muscidae), is a nuisance pest often associated with livestock production, and it can also mechanically transmit the causal agents of human and veterinary diseases. We found that a 0.5% concentration of p-anisaldehyde, produced by many plants consumed by humans, repelled adult M. domestica in static air olfactometer tubes under laboratory conditions for ≥24 h, but by 48 h the repellent activity had worn off. Repellency, however, was not observed in response to 0.5% p-anisaldehyde that had been exposed to sunlight radiation lamps for 2 h. When p-anisaldehyde was aged in darkness for 48 h, it showed strong initial repellency for <1 h. The repellent action of 0.5% p-anisaldehyde was sufficient to keep adult M. domestica from landing on three different food sources when the botanical substance was misted onto the food sources, and when it was placed in proximity to, but not in contact with, the food sources. Extension of p-anisaldehyde's repellent action using solvents other than acetone is discussed.

Knockdown resistance (kdr) is a common mechanism of insecticide resistance in head lice to the conventionally used pyrethroid pediculosis and can be the result of various amino acid substitutions within the voltage-sensitive sodium channel (VSSC). In this study, 54 sequences from varied specimens were investigated to monitor well-known resistance mutations and probable new mutations. The Pediculus humanus capitis de Geer specimens were collected from 13 provinces in Iran. The specimens were stored in 70% ethanol until DNA extraction and PCR amplification of ∼900-bp fragment of VSSC. The sequences were analyzed using different bioinformatics software for the detection of well-known kdr substitutions and additional mutations potentially associated with kdr resistance in head lice. There were six new and an old (haplotype I) kdr haplotypes within the Iranian head louse population. K794E, F815I, and N818D amino acid substitutions were reported for the first time. The P813H mutation was the most prevalent amino acid substitution in eight provinces. Among 53 sequences, 26 (49%) were homozygous susceptible, and 27 (51%) were heterozygotes. Thus, 51% of the head lice collected in Iran harbored only the P813H allele. The exact test for the Hardy–Weinberg (H–W) equilibrium showed that genotype frequencies differed significantly from the expectation in East-Azerbaijan and Tehran provinces. Moreover, these populations had an inbreeding coefficient (Fis) <0, indicating the excess of heterozygotes. This observation suggests that the populations of head lice from Iran are currently under active selective pressure. For the rest of the populations, H–W equilibrium and the expectations were significantly in harmony. The results of the current study highlight molecular techniques in the accurate detection of resistance genotypes before their establishment within the head louse population. Accurate detection of resistant genotypes seems to be helpful in decision-making on lice control programs and resistance monitoring and management.

Graphical Abstract

Eastern equine encephalitis virus (EEEV) is an arbovirus endemic to the eastern United States. Human cases are rare but can be serious. The primary enzootic vector is Culiseta melanura (Coquillett) (Diptera: Culicidae), an ornithophagic mosquito. We conducted an aerial application of a granular methoprene formulation in Hockomock Swamp (Massachusetts), which represents a focus of EEEV transmission. Water collected from inside and outside Cs. melanura crypts was evaluated in bioassays of early fourth instar Cs. melanura larvae using treated and untreated water. Adult eclosion rates were 36% significantly lower in treated compared with untreated water (P < 0.05). Eclosion rates for water collected from inside crypts were significantly higher (62%) than rates from outside crypts (30%) (P < 0.05), indicating higher efficacy outside crypts. We tested whether reduced methoprene efficacy inside the crypts was due to reduced chemical penetration into this habitat. Chemical water analyses confirmed that methoprene concentrations were lower inside the crypts (0.1 ± 0.05 ppb) compared to water from outside crypts (1.79 ± 0.41 ppb). The susceptibility of Cs. melanura to methoprene was also determined to allow for comparison against concentrations observed in water collected from the field (LC-95: 1.95 ± 0.5 ppb). Overall, methoprene-treated water prevented mosquito development for up to 4 wk, but with a reduction in efficacy between 4- and 6-wk post-application. Our results suggest that aerial methoprene applications can effectively treat open water in wetlands but may not provide efficacious control of Cs. melanura due to an inability to penetrate larval habitats.

The house fly (Musca domestica Linnaeus) is an important disease vector. Insecticide resistance is an obstacle to effective house fly control. Previous studies have demonstrated that point mutations in acetylcholinesterase (Ace), carboxylesterase (MdαE7) and voltage-sensitive sodium channel (Vssc), and over-expression of CYP6D1v1 confer insecticide resistance in the house fly. However, information about the status and underlying mechanisms of insecticide resistance in Kazakhstani house flies is lacking. In this study, we investigated the occurrence of genetic mutations associated with insecticide resistance in field house flies collected at six different locations in southern Kazakhstan. Four mutations (V260L, G342A/V, and F407Y) in Ace and three mutations (G137D and W251L/S) in MdαE7 were detected with appreciable frequencies. Notably, haplotypes carrying triple-loci mutations in Ace and double mutations in MdαE7 were found in Kazakhstan. The L1014H and L1014F mutations in Vssc, and CYP6D1v1 resistance allele were detected at a low frequency in some of the six investigated house fly populations. Phylogenetic analyses of haplotypes supported multiple origins of resistance mutations in Ace and MdαE7. These observations suggest that house flies in southern Kazakhstan may exhibit significant resistance to organophosphates and carbamates. Regular monitoring of insecticide resistance is recommended to achieve effective house fly control by chemical agents in southern Kazakhstan.

Graphical Abstract

Phlebotomus argentipes is an established vector for Visceral leishmaniasis prevalent in the Indian subcontinent. Insect Glutathione S-transferases (GST) enzyme plays a pivotal role in the metabolism of xenobiotics and chemical insecticides. We report herein the identification and characterization of a delta class GST from the sandfly, P. argentipes. The resulting clone (rParg-GSTδ) is successfully sequenced, which revealed 76.43% and 66.32% gene identity with GST from Phlebotomus papatasi (Scopoli; Diptera: Psychodidae) and Lutzomiya longipalpis (Lutz and Neiva; Diptera: Psychodidae), respectively. The identified rParg-GST amino acid Blast results revealed 82.6% homology to delta class GST of Phlebotomus papatasi and more than 50% homology to Lepidoptera which comprises butterflies and moths. The Phylogenetic analysis of Parg-GST with different classes of Insect GSTs further supported its classification as delta class. A functional recombinant Parg-GSTδ protein (rParg-GSTδ) was expressed in Escherichia coli (Migula; Enterobacterales: Enterobacteriaceae) cells in a soluble form, purified to homogeneity and found to be active against a substrate 1-chloro-2,4-dintrobenzene (CDNB) and lipid peroxidation by-product 4-Hydrxynonenal (4-HNE). Interestingly, rParg-GSTδ demonstrates high dehydrochlorination activity against dichlorodiphenyltrichloroethane (DDT) i.e., 16.27 nM/µg in high performance liquid chromatography (HPLC) assay. These results provide evidence of direct DDT metabolism property exhibited by P. argentipes GST and set the foundation to decipher the metabolic resistance mechanism in P. argentipes against insecticides.

As tick-borne disease incidence increases and pathogens expand into new areas, the need for effective tick management strategies is paramount. In this 5-yr study (2014–2018) conducted in south central Wisconsin, we assessed whether an integrated tick management approach, deployed during peak tick activity (May–August), was more effective at reducing black-legged ticks (Ixodes scapularis Say (Ixodida: Ixodidae)), than individual interventions. Using a factorial design, invasive vegetation removal (Amur honeysuckle, Lonicera maackii Ruprecht (Dipsacales: Caprifoliaceae) and common buckthorn, Rhamnus cathartica Linnaeus (Rosales: Rhamnaceae)) was coupled with deployments of permethrin-treated cotton nesting materials (tick tubes) that target the white-footed mouse (Peromyscus leucopus Rafinesque (Rodentia: Cricetidae)). Results show that the probability of encountering a larval tick by drag sampling was unaffected by treatments at the cumulative 5-yr level. However, vegetation removal significantly reduced larval encounters in 2014, 2015, and 2018, by 33%, 57%, and 61% respectively, and reduced the density of questing nymphal (DON) ticks by 45% in 2015 compared to controls. Despite the limited effect on DON, vegetation removal significantly reduced the cumulative 5-yr density of Borrelia burgdorferi sensu stricto infected nymphs (DIN) (70%) compared to controls as a result of decreased nymphal infection prevalence. Sites treated with tick tubes had lower DIN (66%) and DON (54%) across the study and nymphs were reduced every year following the initial year of deployment compared to controls. Combining treatments did not further reduce DIN or DONs. We conclude that long-term integration of tick tubes with invasive vegetation removal does not provide additional benefit over individual treatments alone.

This study assessed the perception of pregnant women on indoor residual spraying (IRS), documented acceptability, and factors that significantly dictate willingness to use IRS among the pregnant women attending antenatal clinic in Ibadan Nigeria. A structured questionnaire was used to obtain relevant information from 500 pregnant women. Descriptive and principal component analysis (PCA) were done at 5% level of significance. Majority of the pregnant women had between good and fair knowledge of IRS. Less than 70% of the respondents were willing to allow IRS in their homes. Our PCA revealed that major factors responsible for acceptance and willingness to use IRS were associated with perceived benefits, advantages, and disadvantages of IRS rather than overall knowledge of IRS among the respondents. The analysis revealed that these factors were responsible for at least 80% of the reasons for a pregnant woman to willingly accept IRS or not. The factors that positively influenced willingness to use IRS include its benefit in controlling mosquitoes and other insects, reducing malaria incidence, and prolonged effects of IRS which is an added advantage over continuous purchase of aerosols. One major factor that may negatively affect the acceptance of IRS is the fact that you may have to pack out of the house and wait for more than 3 d before entering. This current study has identified some specific factors that seem to promote and/or reduce willingness to accept IRS as a malaria control intervention among pregnant women in South Western Nigeria.

Species of the Anopheles barbirostris complex (Myzorhynchus Series of the subgenus Anopheles) are potential vectors of malaria and filariasis parasites. Owing to the lack of reliable identification methods, the biting activity and host preference of the species within this complex have not been previous described. In this study, the trophic behavior and host preferences of the species in the complex were determined in Thailand, and a map of their geographical distributions constructed. Adult female mosquitoes in Chanthaburi and Prachuap Khiri Khan provinces were collected using human landings (indoor and outdoor) and cow-bait. Morphological characters were used to initially identify the mosquitoes, and then, species were further separated by multiplex PCR. Four species were identified in the An. barbirostris complex: An. wejchoochotei Taai & Harbach (Diptera: Culicidae), An. saeungae Taai & Harbach (Diptera: Culicidae), An. dissidens Taai & Harbach (Diptera: Culicidae), and An. campestris Reid (Diptera: Culicidae). The species showed exophagic and zoophilic behavior, except An. wejchoochotei in Chanthaburi, which displayed slight endophilic behavior. Indoor and outdoor blood feeding behavior of An. wejchoochotei and An. saeungae peaked at 2000 h. An updated distribution map of the An. barbirostris complex in Thailand is presented. With a better understanding of the sibling vector species and their spatial distributions, more specific and effective control strategies for vectors of malaria and filariasis in Thailand can be achieved.

Eastern equine encephalitis virus (EEEV) is the most pathogenic arbovirus endemic to the United States. Studies have demonstrated Florida's role as a regional reservoir for the virus and its ability to support year-round transmission. Previous research has developed risk index models for mapping locations most at risk for EEEV transmission. We compared vector abundance, vector feeding behavior, potential host species, and fauna presence at high versus low–moderate risk sites during the winter and spring. Predicted high-risk sites had a significantly greater abundance of mosquitoes overall, including Culiseta melanura (Coquillett) (Diptera: Culicidae), the primary enzootic vector of EEEV. Twenty host species were identified from Cs. melanura bloodmeals, with the majority taken from avian species. Culiseta melanura largely fed upon the Northern Cardinal (Cardinalis cardinalis (Passeriformes: Cardinalidae)), which accounted for 20–24.4% of the bloodmeals obtained from this species in years 1 and 2, respectively. One EEEV-positive mosquito pool (Cs. melanura) and nine EEEV seropositive sentinel chickens were confirmed during winter-spring collections from high-risk sites; no seropositive chickens nor mosquito pools were found at the low–moderate risk sites. These results suggest that high-risk sites for EEEV activity are characterized by habitats that support populations of Cs. melanura and which may also provide ample opportunities to feed upon Northern Cardinals. The overall low level of mosquito populations during the winter also suggests that control of Cs. melanura populations in winter at high-risk sites may prove effective in reducing EEEV transmission during the peak summer season.

During September–December 2018, 25 live ticks were collected on-post at Fort Leavenworth, Kansas, in a home with a history of bat occupancy. Nine ticks were sent to the Army Public Health Center Tick-Borne Disease Laboratory and were identified as Carios kelleyi (Cooley and Kohls, 1941), a species that seldom bites humans but that may search for other sources of blood meals, including humans, when bats are removed from human dwellings. The ticks were tested for numerous agents of human disease. Rickettsia lusitaniae was identified by multilocus sequence typing to be present in two ticks, marking the first detection of this Rickettsia agent in the United States and in this species of tick. Two other Rickettsia spp. were also detected, including an endosymbiont previously associated with C. kelleyi and a possible novel Rickettsia species. The potential roles of C. kelleyi and bats in peridomestic Rickettsia transmission cycles warrant further investigation.

Diseases caused by flaviviruses are a major public health burden across the world. In the past decades, South America has suffered dengue epidemics, the re-emergence of yellow fever and St. Louis encephalitis viruses, and the introduction of West Nile and Zika viruses. Many insect-specific flaviviruses (ISFs) that cannot replicate in vertebrate cells have recently been described. In this study, we analyzed field-collected mosquito samples from six different ecoregions of Argentina to detect flaviviruses. We did not find any RNA belonging to pathogenic flaviviruses or ISFs in adults or immature stages. However, flaviviral-like DNA similar to flavivirus NS5 region was detected in 83–100% of Aedes aegypti (L.). Despite being previously described as an ancient element in the Ae. aegypti genome, the flaviviral-like DNA sequence was not detected in all Ae. aegypti samples and sequences obtained did not form a monophyletic group, possibly reflecting the genetic diversity of mosquito populations in Argentina.

Burgeoning cases of tick-borne disease present a significant public health problem in the United States. Passive tick surveillance gained traction as an effective way to collect epidemiologic data, and in particular, photograph-based tick surveillance can complement in-hand tick specimen identification to amass distribution data and related encounter demographics. We compared the Federal Information Processing Standards (FIPS) code of tick photos submitted to a free public identification service (TickSpotters) from 2014 to 2019 to published nationwide county reports for three tick species of medical concern: Ixodes scapularis Say (Ixodida: Ixodidae), Ixodes pacificus Cooley and Kohls (Ixodida: Ixodidae), and Amblyomma americanum Linneaus (Ixodida: Ixodidae). We tallied the number of TickSpotters submissions for each tick species according to “Reported” or “Established” criteria per county, and found that TickSpotters submissions represented more than half of the reported counties of documented occurrence, and potentially identified hundreds of new counties with the occurrence of these species. We detected the largest number of new county reports of I. scapularis presence in Michigan, North Carolina, and Texas. Tick image submissions revealed potentially nine new counties of occurrence for I. pacificus, and we documented the largest increase in new county reports of A. americanum in Kentucky, Illinois, Indiana, and Ohio. These findings demonstrate the utility of crowdsourced photograph-based tick surveillance as a complement to other tick surveillance strategies in documenting tick distributions on a nationwide scale, its potential for identifying new foci, and its ability to highlight at-risk localities that might benefit from tick-bite prevention education.

With global expansion of the two main vectors of dengue, Aedes aegypti (Linnaeus, Diptera: Culicidae) and Aedes albopictus (Skuse, Diptera: Culicidae), there is a need to further develop cost-effective and user-friendly surveillance tools to monitor the population dynamics of these species. The abundance of Ae. aegypti and Ae. Albopictus, and associated bycatch captured by Male Aedes Sound Traps (MASTs) and BG-Sentinel (BGS) traps that were unbaited or baited with BG-Lures were compared in Cairns, Australia and Madang, Papua New Guinea. Mean male Ae. aegypti and Ae. albopictus catch rates in MASTs did not significantly differ when deployed with BG-Lures. Similarly, males of both these species were not sampled at statistically different rates in BGS traps with or without BG-Lures. However, MASTs with BG-Lures caught significantly less male Ae. aegypti than BGS traps baited with BG-Lures in Cairns, and MASTs without BG-Lures caught significantly more male Ae. albopictus than BGS traps without BG-Lures in Madang. Additionally, BG-Lures significantly increased female Ae. aegypti catch rates in BGS traps in Cairns. Lastly, bycatch capture rates in BGS traps were not significantly influenced by the addition of the BG-Lures. While this study provides useful information regarding the surveillance of Ae. aegypti and Ae. albopictus in these locations, further development and investigation is required to successfully integrate an olfactory lure into the MAST system.

Aedes aegypti (Linnaeus, 1762) is a major vector responsible for dengue transmission. Insecticides are being used as the most effective tool to control vector populations in Lahore, Pakistan. Control of Ae. aegypti is threatened by the development of resistance against insecticides. The current status of insecticide resistance was evaluated against pyrethroids (deltamethrin, cypermethrin, and lambda-cyhalothrin) in different populations of Lahore (Model Town, Mishri Shah, Sadar Cantt, Walton, and Valencia). The susceptibility of the larval and adult populations was tested following the standard WHO guidelines. Moderate to high levels of resistance were found against pyrethroids in the larval (RR₅₀: 3.6–27.2 and RR₉₀: 5–90) and adult populations (percentage mortality < 98%). Biochemical assays revealed a statistically significant increase in the enzyme level in all field populations compared to the laboratory strain. The value of esterase was one-fold higher, monooxygenase was 3.9- to 4.7-fold higher, and glutathione S-transferases was 1.9- to 2.6-fold higher in field populations compared to the laboratory strain. These results depict the presence of resistance against deltamethrin, cypermethrin, and lambda-cyhalothrin in field populations of Lahore mediated by metabolic enzymes i.e. esterases, monooxygenases, and glutathione S-transferase.

Culicoides Latreille species (Diptera: Ceratopogonidae) are insects of significant medical and veterinary importance worldwide because their bites can cause major annoyance, allergic reactions, and/or pathogen transmission to vertebrates, including humans. In this study, we examined the effect of host bloodmeal source on the fecundity and subsequent larval development traits of the progeny of Culicoides furens Poey, a nuisance species in coastal Florida. Field-collected females were fed on two different classes of hosts: birds (chicken) or mammals (human). Fecundity outcomes of the females and larval developmental traits of the progeny were recorded and compared between the two groups. The percentage of females that developed eggs, percentage of gravid females that deposited eggs, number of eggs produced per female, number of eggs deposited per female, egg hatch rates, larval survival rates, time to pupation, pupal sex-ratios, and adult eclosion rates were not significantly different between the two groups. Our results demonstrate that the host bloodmeal source has no significant effect on the fecundity and subsequent larval developmental life-history traits of the progeny in C. furens. Previous studies reported C. furens to be a generalist feeder that shows no host preference for birds or mammals. Collectively, these findings suggest that C. furens is well adapted, not just behaviorally, but also physiologically to efficiently utilize blood meals from avian and mammalian hosts, a quality that offers a major evolutionary advantage to the success of this midge species as a generalist feeder.

A cue for long-range vision allows mosquitoes to identify hosts and differentiate the ecological niches (e.g., habitats). However, the visual factors involved in attracting mosquitoes to a host are complex and have not been fully understood. Therefore, we assessed color preference to Aedes albopictus (Skuse) and Culex pipiens (Conquillett) as diurnal and nocturnal species, respectively, using seven fundamental colors including black, white, red, yellow, green, blue, and purple with each trap at 100 lux in a laboratory. We used a binary behavioral assay using the Mosquito Preference Index (MPI) as a preference ratio with a range of 0–1. Our analyses showed that Ae. albopictus had a greater response to black (MPIs, 0.7), followed closely by red, blue, and purple (MPIs, 0.6). We also found that red, blue, and purple were significantly higher (P < 0.05) than those of green (MPI, 0.5), white (MPI, 0.3), and yellow (MPI, 0.2). Similarly, the MPIs for Cx. pipiens were significantly higher at black and red (MPIs, 0.7; P < 0.05) compare to those of white and yellow (MPIs, 0.3; P < 0.05). The color preference of Ae. albopictus showed significant correlation to luminous intensities (L-value) (r = –0.640; P = 0.000) and blue intensities (b-value) (r = –0.372; P = 0.000) for all seven colors. In addition, Cx. pipiens negatively correlated (r = –0.703; P = 0.000) between color preference and L-value. Our analyses provide a greater understanding of how color plays a role in visual sensory stimuli, and how that could potentially affect mosquito host-seeking behavior.

Human granulocytic anaplasmosis (HGA) and human babesiosis are tick-borne diseases spread by the blacklegged tick (Ixodes scapularis Say, Acari: Ixodidae) and are the result of infection with Anaplasma phagocytophilum and Babesia microti, respectively. In NewYork State (NYS), incidence rates of these diseases increased concordantly until around 2013, when rates of HGA began to increase more rapidly than human babesiosis, and the spatial extent of the diseases diverged. Surveillance data of tick-borne pathogens (2007 to 2018) and reported human cases of HGA (n = 4,297) and human babesiosis (n = 2,986) (2013–2018) from the NewYork State Department of Health (NYSDOH) showed a positive association between the presence/temporal emergence of each pathogen and rates of disease in surrounding areas. Incidence rates of HGA were higher than human babesiosis among White and non-Hispanic/non-Latino individuals, as well as all age and sex groups. Human babesiosis exhibited higher rates among non-White individuals. Climate, weather, and landscape data were used to build a spatially weighted zero-inflated negative binomial (ZINB) model to examine and compare associations between the environment and rates of HGA and human babesiosis. HGA and human babesiosis ZINB models indicated similar associations with forest cover, forest land cover change, and winter minimum temperature; and differing associations with elevation, urban land cover change, and winter precipitation. These results indicate that tick-borne disease ecology varies between pathogens spread by I. scapularis.

Aedes aegypti (Linnaeus) is the main vector of most arboviruses in tropical and subtropical urban areas. In West Africa, particularly in Senegal, domestic and wild populations have been described. Both Ae. aegypti aegypti (Aaa) and Ae. aegypti formosus (Aaf) were found in progenies of Ae. aegypti families from several localities of Senegal. However, nothing is known about their resting and trophic behavior, which are key data for vector control. To fill this gap, blood-fed mosquitoes were collected monthly indoors and outdoors with BackPack aspirators and BG-Sentinel 2 traps between July and November 2019 from four urban sites. The enzyme-linked immunosorbent assay technique was used to analyze blood-fed Aaa and Aaf specimens. Both forms were found resting in all investigated places with the highest proportions found in scrap metals (51.7% for Aaa and 44.1% for Aaf) and used tires (19.2% for Aaa and 26.1% for Aaf). Blood-fed Aaf females showed lower occupation of the indoors environment compared to Aaa. Overall, the percentages of single bloodmeals from human were 80.5% (916/1138) for Aaa and 71.1% (263/370) for Aaf. A low frequency of other domestic hosts, including bovine, ovine, and cat were detected for both forms. This study provides the first data on resting and trophic behavior of Aaa and Aaf in Senegal. Both forms showed differences in their resting behavior but fed primarily on human and highlight the risk of arboviruses transmission in urban areas.

Mepraia spinolai, (Porter) 1934, is a diurnal triatomine endemic to Chile and a wild vector of the protozoan Trypanosoma cruzi, (Chagas) 1909, which causes Chagas disease. Behavioral changes in M. spinolai induced by this parasite have been reported previously, which include detection of a potential host, defecation latency, and some life history traits. In this study we assessed changes in locomotor and daily activity due to infection with T. cruzi. No difference was detected in distance traveled between infected and uninfected individuals. However, the groups differed in their daily activity patterns; infected individuals showed significant reduction of movements during the light phase and concentrated their activity in the dark phase. Uninfected individuals showed no differences in locomotor activity between the phases. The results suggest that T. cruzi induces a displacement in the activity of M. spinolai toward the dark phase of the circadian cycle, which may improve its vector competence.

Hand sanitizers are developed as alcohol-based liquid gel formulations, generally used to decrease the amount of infectious agents on human hands. Verdegen, LLC proposed to prepare an arthropod repellent gel for public use when the recent outbreaks of Zika infection vectored through Aedes mosquitoes in the American continents prompted multi-faceted emergency measures. Four different gel formulations were developed, comprising two of the most efficacious commercial arthropod repellent active ingredients, N,N-diethyl-3-methyl benzamide (deet) and 2-(2-hydroxyethyl)-1-piperidinecarboxylic acid 1-methylpropyl ester (picaridin), each at different concentrations (20 and 33% deet, or 20 and 33% picaridin). Compliance with the use of topical arthropod repellents remains an issue among military personnel. One of the most common complaints by Soldiers is that they do not like how the repellents applied on their skin leave behind an oily or greasy residue. These new gel formulations offer a user-friendly alternative for commonly used arthropod repellents formulations for the military and civilian personnel. We tested the efficacy and protection time of these new gel formulations in comparison with the commercially available cream formulations of deet and picaridin at similar concentrations. Our data show that gel formulations have better topical attributes, and offer equal or better biting protection for up to 48 h against host-seeking Aedes aegypti (L.) (Diptera: Culicidae) female mosquitoes.

Limited evidence suggests that the cervid parasite, Babesia odocoilei, is transovarially transmitted from adult female Ixodes scapularis Say to offspring. The prevalence of B. odocoilei in unfed larval I. scapularis and whether vertical transmission is crucial to pathogen maintenance are currently unknown. To investigate these questions, 275 unfed larvae from two Wisconsin counties were tested for B. odocoilei genetic material. Sixteen of 29 pools were positive for the parasite. The maximum likelihood estimation for overall larval infection prevalence was 7.8% (95% confidence interval: 4.7–12). This vertically acquired infection appears to be sustained transstadially in nymphal ticks the following year; however, our relatively small sample and replicate size warrants additional evaluation. Our study revealed further evidence of vertical transmission, a low and consistent infection prevalence in larvae, and the potential importance of vertical transmission in B. odocoilei maintenance.

The transmission of pathogens that cause leishmaniases occurs by the bite of female sand flies (Diptera: Psychodidae) in their vertebrate hosts, which makes the identification of their bloodmeal sources an important step for the control and epidemiology of these diseases. In Brazil, the state of Roraima has a great diversity of sand flies, vertebrate hosts, and protozoan Leishmania, but little is known about the host blood-feeding preferences of sand flies. Thus, we evaluated the bloodmeal sources of sand flies collected from their sylvatic habitats in Parque Nacional do Viruá, Roraima. Fieldwork was carried-out between 13th and 18th August 2019 using CDC light traps. Sand flies were slide-mounted and morphologically identified using the head and last segments of the abdomen. Engorged females had their DNA extracted, followed by amplification and sequencing of the cytochrome b (cytb) molecular marker for vertebrates. Sequences were analyzed and compared with those from GenBank using the BLASTn search tool, in addition to the reconstruction of a phylogenetic tree to demonstrate the clustering pattern of these sequences. A total of 1,209 sand flies were identified, comprising 20 species, in which the most abundant were Psychodopygus ayrozai (Barretto and Coutinho) (42.10%) and Psychodopygus chagasi (Costa Lima) (26.22%). Bloodmeal source identification was successfully performed for 34 sand flies, that confirm four vertebrate species, being the most abundant the armadillo Dasypus novemcinctus Linnaeus, 1758 (Cingulata: Dasypodidae).

The impact of tick bites during holidays in tropical countries is seldom reported. Here, we describe long-term skin alterations and itching experienced by a German tourist following the bite of a nymph of Dermacentor auratus Supino, 1897 in Cambodia. Tick infestation may be neglected by travelers. Therefore, careful travel history is necessary when travelers report a tick bite after spending holiday in tropical and subtropical countries.

Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 (Diptera: Psychodidae) has been found in several European countries. In Spain, sporadic records were reported in the early'80s in Catalonia (Northeast Spain), and it was never detected again. Recent entomological surveys carried out between 2004 and 2020 revealed the presence of several specimens of P. mascittii in Spain. The species identification was confirmed by both morphological and molecular analyses.The analyzed specimens belonged to the haplotype (COI_2) defined by one polymorphic site compared to other European specimens. Phlebotomus mascittii was found in low population densities in rural areas associated with livestock farms and in an urban cemetery during the summer season. This study provides the first records of this species in various localities along the Cantabrian cornice (Northern Spain) and represents its westernmost observation in the Palearctic region. The implications of the finding of this uncommon species are discussed at different levels, with emphasis on its suspected role in the transmission of leishmaniosis.

Borrelia mayonii is a recently discovered bacterial spirochete that causes Lyme disease and is transmitted by the blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae). To date, B. mayonii has been isolated from two vertebrate host species in Minnesota: field-caught white-footed mice (Peromyscus leucopus Rafinesque; Rodentia: Cricetidae) and American red squirrel (Tamiasciurus hudsonicus Erxleben). Here, we describe the first detection of B. mayonii in field-caught eastern chipmunks (Tamias striatus L. (Rodentia: Cricetidae)) from northern Wisconsin. During our study, we captured 530 unique small mammals and found an infection prevalence of 23.50% in field-caught eastern chipmunks (4/17) and 1.19% in Peromyscus spp. (5/420). Mean larval and nymphal burdens were determined for captured Blarina brevicauda (0, 0), Glaucomys volans (0.29, 0.14), Myodes gapperi (0.27, 0), Napaeozapus insignis (0, 0.25), Peromyscus spp. (1.88, 0.11), T. striatus (1.06, 0.65), and Sorex cinereus (0.09, 0). The high B. mayonii infection prevalence in eastern chipmunks suggests that the species may be an important reservoir for B. mayonii in the Upper Midwest.

A number of invasive tick species capable of transmitting pathogens have been accidentally introduced into the U.S. in recent years. The invasion and further range expansion of these exotic ticks have been greatly facilitated by frequent global travel and trade as well as increases in legal and illegal importation of animals. We describe the discovery of the first established populations of Haemaphysalis longicornis Neumann and the first fully engorged human parasitizing specimen documented through passive tick surveillance in Fairfield County, Connecticut, U.S. We also report several individual specimens of this invasive arthropod and vector of multiple pathogens of medical and veterinary importance collected through active tick surveillance from three counties (Fairfield, New Haven, and New London). Considering the potential for invasive ticks to transmit numerous native and emerging pathogens, the implementation of comprehensive surveillance programs will aid in prompt interception of these ticks and reduce the risk of infection in humans and wildlife.

Haemaphysalis longicornis Neumann (Asian longhorned tick) is an exotic and invasive tick species presenting a health and economic threat to the United States (U.S.) cattle industry due to its ability to transmit pathogens and infest hosts in large numbers. The objective of this study was to evaluate available products at causing H. longicornis mortality in a laboratory bioassay. The efficacy of products was evaluated at label rates using H. longicornis nymphs collected from a cattle farm in eastern Tennessee in two different bioassays (spray or dip) against untreated controls. After exposure, ticks were transferred to clean petri dishes and checked for mortality at 0, 1, 2, 3, 4, 21, 24, and 48 h post exposure. No mortality occurred in the untreated controls, whereas all treated ticks were dead within 24 h of exposure (P < 0.0001). These findings support the hypothesis that currently available spray and pour-on products are effective at causing H. longicornis mortality. We conclude that these acaricides can be used as a component to prevent H. longicornis dispersal and for control in the U.S.

In 2017, a single adult Triatoma sanguisuga (LeConte) (Hemiptera: Reduviidae) was identified from Indian Cave State Park in Nemaha County, NE. The following summer, a single adult specimen was found in a 4-H insect collection at the Nebraska State Fair from Richardson County. A subsequent investigation found that these were collected from a residence in Richardson County and three more adults were collected in 2019. In 2020, the latter three kissing bugs were submitted to Public Health Command-Central's DoD Food Analysis and Diagnostic Laboratory (DOD FADL) for confirmatory species identification and diagnostic testing for Trypanosoma cruzi, the causative agent of Chagas disease. One specimen tested positive for T. cruzi using a real-time dual-target PCR screen followed by confirmatory dual-target traditional PCR. Based on these findings, a survey plan was developed for Richardson and surrounding counties in southeast Nebraska. In July of 2020, two adult and seven nymphs (multiple instars) of T. sanguisuga were collected in Richardson County, one of which tested positive for T. cruzi. This is the first record of an established population of T. sanguisuga and T. cruzi-infected kissing bugs in Nebraska.

Culiseta (Allotheobaldia) longiareolata (Macquart) (Diptera: Culicidae) is an ornithophilic mosquito species that occurs in the southern Palaearctic Region from the Azores to Central Asia, the Ethiopian Region, India, and Pakistan. Although it has a widespread distribution range, the species was only recently reported in Western and Central Europe. Between 2017 and 2020, larvae, pupae, and adults of Cs. longiareolata (n = 161) were found at 13 distinct locations in Belgium (n = 4) and The Netherlands (n = 9). Collected mosquitoes were morphologically identified and the identification was then validated by COI DNA barcoding. These are the first records of the species in the above-mentioned countries. The present results suggest that Cs. longiareolata could be increasing its distribution range in temperate regions, indicating a warming climate. As the species might be a potential vector of bird pathogens (e.g., West Nile virus), its spread in Western Europe is noteworthy.

Graphical Abstract

Transgenic strains of the mosquito disease vector Aedes aegypti (L.) are being developed for population suppression or modification. Transgenic mosquitoes are identified using fluorescent protein genes. Here we describe DsRed and ZsGreen marker genes driven by the constitutive Ae. aegypti heat shock protein 83 (hsp83) promoter in transgenic mosquitoes. Transgenic larvae and pupae show strong full body expression of the red and green fluorescent proteins. This greatly assists in screening for transgenic individuals while making new or maintaining already established lines. Transient marker gene expression after embryo microinjection was readily visible in developing larvae allowing the separation of individuals that are more likely to produce transgenic offspring. The strongly expressed marker genes developed in this study should facilitate the detection of transgenic Ae. aegypti larvae or pupae in the field.

We report a case of envenomation by Steatoda borealis (Hentz, 1850) in Connecticut in late spring 2021. The bitten subject was a 60-yr-old female Caucasian. The bite occurred upon disturbing some books in her basement. She reported a painful stinging bite on her finger followed by the formation of an erythematous macule at the site of the bite which was gone the next day. There was no sign of necrosis. The spider was recovered directly at that time. The victim reported that later that night she experienced heart palpitations. This is the first report of envenomation by S. borealis, a common spider in Connecticut.

Rhipicephalus microplus is the main blooding-sucking ectoparasite of bovines and is regarded as important vectors of animal diseases such as Babesiosis. Mining protective antigens of R. microplus to develop antitick vaccine is the most potential tick control strategy. In this study, the specific primers were designed according to the conserved nucleotide sequence of enolase gene in Haemaphysalis flava, Ixodes ricinus, and Ornithodoros moubata. The fragment of enolase gene was obtained by PCR using cDNA template from fully engorged female R. microplus. The full-length cDNA of enolase gene was amplified using rapid amplification of cDNA ends (RACE). Expression pattern of enolase gene in different tissues of R. microplus was analyzed by real-time quantitative PCR (qRT-PCR). Results showed that the full-length enolase cDNA containing 2052 bp was obtained successfully. The complete cDNA included an ORF of 1305 nucleotides encoding a protein of 434 amino acids. The enolase exhibited 85.0% amino acid identity to the enolase of H. flava, 81.1% to I. ricinus enolase, and 81.3% to O. moubata enolase. qRT-PCR analysis indicated that the enolase had the highest expression in the salivary gland of R. microplus.