The theme of the 2018 Entomological Society of America, Entomological Society of Canada, and Entomological Society of British Columbia Joint Annual Meeting was Crossing Borders: Entomology in a Changing World. Following the theme of the meeting, papers selected for the 2018 ‘Highlights of Medical Entomology’ included reports of expanded ranges for Aedes albopictus (Skuse) (Diptera: Culicidae), Aedes japonicus japonicus (Theobald) (Diptera: Culicidae), Culex coronator Dyar and Knab (Diptera: Culicidae), Mansonia titillans (Walker) (Diptera: Culicidae), and Wyeomyia mitchellii (Theobald) (Diptera: Culicidae); a first report of mosquito feeding on earthworms and leeches; Cache Valley virus detection in Ae. j. japonicus; surveillance in high-risk areas of Canada for Lyme Disease; and increasing lone-star tick detections in Connecticut. The selection of papers served as a reminder of the increasingly difficult challenge of sustaining vector surveillance programs. Fluctuating funding and a decline in vector taxonomists and identification expertise contribute to the erosion of vector surveillance.

The field of urban entomology is primarily associated with the study of ants, bed bugs, cockroaches, termites, and other occasional invader pests that are found within or near human-made structures. A wide array of peer-reviewed studies were published in 2018. The topics of these articles ranged from genomes and basic biology of urban insects to various applied aspects of pest management. Key findings of these papers are presented and discussed from the perspective of the contributions they make to the discipline of urban entomology. Additionally, potential future research opportunities that are evident from these publications have been outlined.

The 2018 annual meeting of the Entomological Society of America meeting theme ‘Crossing Boarders’ guided the veterinary highlight compilation of papers, focusing on the themes: 1) emerging and re-emerging pest threats; 2) alternative control methods; and 3) tools for future research. Here the papers presented are summarized to serve as a presentation archive.

Vector-borne diseases (VBDs) cause significant morbidity and mortality each year in the United States. Over the last 14 yr, over 700,000 cases of diseases carried by ticks, mosquitoes, and fleas have been reported from U.S. states and territories to the Centers for Disease Control and Prevention. The number of reported cases has been increasing annually with two major trends: a steady increase in tick-borne diseases and increasing intermittent outbreaks of mosquito-borne arboviruses. The factors that are driving VBD introduction and emergence vary among diseases but are not likely to disappear, indicating that current trends will continue and probably worsen in the absence of effective prevention and control tools and implementation capacity. There are a number of challenges to preventing VBDs, including the lack of vaccines and effective vector control tools, insecticide resistance, and eroding technical capacities in public health entomology at federal, state, and local levels. For these reasons, a national strategy is needed to address VBD threats and to reverse the alarming trend in morbidity and mortality associated with these diseases.

Human head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), are hematophagous parasites that infest human heads. They are extreme host specialists suggesting a strict selection behavior towards the human head by the parasites. Despite the public health relevance of P. humanus capitis, the role of chemical clues to select the human head is not well known. In the present study, we attempted to find out whether head lice recognize and select the odor of the head over the odor of other parts of the human body. Our results using a two-choice olfactometer demonstrated that head lice were highly attracted by the volatile compounds of the human odor air, but they did not show preference for the volatiles of head compared to the volatiles of other parts of the body (forearm or foot). Conversely, when head lice were exposed in an experimental arena to the whole human odor of different parts of the body, they showed a preferential response to the whole head odor compared with the whole foot or forearm odors. These results suggest that head lice can at short distance of the host, be oriented towards the head odor of the human host in the selection of the scalp environment.

We investigated bactericidal and fungicidal properties of chitosan extracted from adults and nymphs from both German cockroach, Blattella germanica (Blattodea: Blattellidae) and American cockroach, Periplaneta americana (Dictyoptera: Blattidae). The cuticle of adults and nymphs extracted from both cockroaches were dried and ground. The powders were demineralized and deproteinized followed by deacetylation using NaOH. Finally, the chitosan yields were examined for antibacterial and antifungal activities. The degree of deacetylation (DD) was different between adults and nymph stages. The antimicrobial effect of American cockroach chitosan (ACC) and German cockroach chitosan (GCC) was tested against four bacteria and four fungi. The extracted chitosans from American cockroach, Periplaneta americana and German Cockroach, Blattella germanica suppressed the growth of Gram-negative/positive bacteria except Micrococcus luteus. The growth of Aspergillus flavus and Aspergillus niger were notability inhibited by the extracted chitosans. The antimicrobial effect of the chitosan depended on the cockroach species, with chitosan of the American cockroach showing more inhibitory effect. This difference may be due to differences in the structure of chitin between the two cockroach species.

The age of insects found on corpses is often used to estimate the minimum postmortem interval by forensic entomologists. Insect development is affected by biotic and abiotic factors, and temperature is the most important environmental factor that determines the length of insect development. Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) is a globally distributed fly that is commonly found on corpses, and this study investigated the development of C. rufifacies from China at various constant temperatures. At 16, 19, 22, 25, 28, 31, and 34°C, the developmental time from egg to adult was 870.17 ± 11.50, 566.20 ± 8.79, 406.38 ± 10.98, 291.14 ± 4.71, 232.59 ± 5.96, 192.47 ± 3.45, and 160.48 ± 7.15 h, respectively. We established three developmental models for C. rufifacies to estimate the age of the developing insect: the isomorphen diagram model, the isomegalen diagram model and the thermal summation model. Regression analysis was conducted to obtain the relationship between body length and development time of the larvae from hatching to wandering. The developmental threshold temperatures of the egg, 1st instar, 2nd instar, 3rd instar, and wandering larvae, and intra-puparial stages were 12.28 ± 0.30, 11.74 ± 0.95, 12.70 ± 0.55, 11.68 ± 0.96, 10.53 ± 1.53, and 12.51 ± 0.41°C, respectively. The developmental threshold temperature, and the thermal summation constant during the entire developmental period were 3759.95 ± 170.80 degree hours and 11.96 ± 0.38°C, respectively. This study provides an improved dataset to estimate the postmortem interval of corpses using C. rufifacies.

Musca domestica Linnaeus is an important public health pest with the ability to adapt to diverse climates. Assessment of variations in biology and life-history traits of insects along geographical gradients is important for a successful management plan in different regions. We investigated life-history traits and life table parameters of M. domestica from six different geographical regions of Punjab, Pakistan: Rahim Yar Khan (RYK), Bahawalpur (BWP), Multan (MTN), Lahore (LHR), Gujrat (GJT), and Murree (MRE). Overall, M. domestica from localities of lower latitude and elevations with higher mean temperatures completed their development faster than those from localities of higher latitude and elevations with lower mean temperatures. The immature developmental time was the longest for the MRE population that was collected from higher latitude and elevation with cooler climate, whereas the shortest for the RYK population from lower latitude with warmer climate. Pupal weights were heavier for the RYK, BWP, and MTN populations, all were from the lowest latitude and elevations with warmer climate, compared with rest of the field populations. Similarly, rate of adult eclosion, fecundity, egg hatching, longevity, and life table parameters such as intrinsic rate of population increase, mean relative growth rate, net reproductive rate, and biotic potential were significantly higher for the RYK, BWP, and MTN populations compared with the GJT, LHR, and MRE populations of M. domestica. The current results will probably be of importance when planning management of M. domestica in different geographical regions of Pakistan.

The seasonal activity of adults of eight forensically important blow fly species [Calliphora croceipalpis (Diptera: Calliphoridae), Jaennicke, Lucilia sericata (Meigen) (Diptera: Calliphoridae), L. cuprina (Wiedemann) (Diptera: Calliphoridae), Chrysomya chloropyga (Wiedemann) (Diptera: Calliphoridae), Ch. albiceps (Wiedemann) (Diptera: Calliphoridae), Ch. marginalis (Wiedemann) (Diptera: Calliphoridae), Ch. putoria (Wiedemann) (Diptera: Calliphoridae), Ch. megacephala (Fabricius) (Diptera: Calliphoridae)] was quantified from collections in Makhanda, South Africa. Flies were collected in traps baited with chicken liver and placed in the field at eight locations for four consecutive days each fortnight for 14 mo. The seasonal distribution of each species compared well to published seasonal distributions of these blow flies elsewhere in South Africa, with evidence of year-to-year variation within seasons that might be explained by weather. This information is important for determining when and where certain species are likely to occur and will be of use in forensic investigations and myiasis management plans.

The Anopheles mosquito Hyrcanus Group is widely distributed geographically across both Palearctic and Oriental regions and comprises 26 valid species. Although the species Anopheles sinensis Wiedemann (1828) is the most common in China and has a low potential vector rank, it has nevertheless long been thought to be an important natural malaria vector within the middle and lower reaches of the Yangtze River. A number of previous research studies have found evidence to support the occurrence of natural hybridization between An. sinensis and Anopheles kleiniRueda, 2005 (a competent malaria vector). We, therefore, collected a sample series of An. sinensis and morphologically similar species across China and undertook ribosomal and mitochondrial DNA analyses in order to assess genetic differentiation (Fst) and gene flow (Nm) amongst different groups. This enabled us to evaluate divergence times between morphologically similar species using the cytochrome oxidase I (COI) gene. The results of this study reveal significant genetic similarities between An. sinensis, An. kleini, and Anopheles belenraeRueda, 2005 and therefore imply that correct molecular identifications will require additional molecular markers. As results also reveal the presence of gene flow between these three species, their taxonomic status will require further work. Data suggest that An. kleini is the most basal of the three species, while An. sinensis and An. belenrae share the closest genetic relationship.

Forensically important flesh flies (Diptera: Sarcophagidae) often are not morphologically distinguishable, especially at the immature stage. In addition, female flies are quite similar in general morphology, making accurate identifications difficult. DNA-based technologies, particularly mitochondrial DNA (mtDNA), have been used for species-level identification. The cytochrome oxidase subunits I and II (COI-COII) sequences of Iranian Sarcophagidae are still unavailable in GenBank. In this study as many as 648 (540 males and 106 females) fly specimens from family Sarcophagidae, representing 10 sarcophagid species, including eight forensically important species were collected from seven locations in five Iranian provinces. Of these, 150 male specimens were identified based on both morphology of male genitalia and DNA sequencing analysis. Sequence data from the COI-COII regions for 10 flesh fly species collected in Iran were generated for the first time. Digestion of COI-COII region by restriction enzymes RsaI, EcoRV, and HinfI provided distinct restriction fragment length polymorphism profiles among the species and can serve as molecular markers for species determination. Phylogenetic analysis represented that the COI-COII sequences are helpful for delimitation of sarcophagid species and implementation in forensic entomology. However, the application of the COI-COII fragment as a species identifier requires great caution and additional species and markers should be studied to ensure accurate species identification in the future.

The genus Belminus Stål, 1859 is distinguished by markedly entomophagous species. This genus is a poorly studied group of the Triatominae (Jeannel, 1919), which includes hematophagous species that are vectors of Trypanosoma cruzi (Chagas, 1909), the etiological agent of Chagas disease. This study reports for the first time the description of the antennal sensilla of Belminus corredori (Galvão & Angulo, 2003), Belminus herreri (Lent & Wygodzinsky, 1979), and Belminus ferroae (Sandoval, Pabón, Jurberg & Galvão, 2007) nymphs and adults throughout scanning electron microscopy and optical microscopy. Nine morphological types of sensilla were identified in the three species, including bristles I and II, trichobothria, tapered hairs, thin-walled trichoid, thick-walled trichoid, basiconic, campaniform, and coeloconica sensilla. The analysis of the most abundant types of sensilla and their distribution in nymphs and adults shows similarities with the antennal phenotypes described in other Triatominae. Quantitative differences showed the important phenotypic plasticity of this morphological trait in the genus and allowed for the separation of the species in both sexes. These differences were associated with sexual dimorphism and wing condition (macropterous and brachypterous). The role of the antennal phenotype in sexual behavior, its usefulness as a taxonomic tool, and its relationship with the dispersal capacity of these species are discussed.

The Chiang Mai (CM) form of the genus Lutzia, subgenus Metalutzia, in Thailand is diagnosed and formally named Lt. chiangmaiensis Somboon & Harbach, n. sp. The species is contrasted with related species, and information is provided on its bionomics and distribution.

Two new species and a new subspecies of fleas belonging to the genus Ctenophthalmus Kolenati (Siphonaptera: Ctenophthalmidae) were described from Turkey. Ctenophthalmus (Spalacoctenophthalmus) beyzanurae n. sp. was collected from Nannospalax leucodon Nordmann (Mammalia: Rodentia) in Tekirdağ province while Ctenophthalmus (Euctenophthalmus) kefelioglui n. sp. and Ctenophthalmus (Euctenophthalmus) teres anatolicus n. ssp. were collected from Microtus guentheri (Danford & Alston) and Apodemus uralensis (Pallas) (Mammalia: Rodentia) in the Aksaray and Kars provinces of Turkey, respectively. Detailed illustrations of new taxa described were also provided.

Monitoring and collection of fly taxa (Diptera: Calliphoridae; Muscidae; Sarcophagidae) of medical, veterinary, and agricultural importance is often routine practice, providing data on target species presence, distribution and abundance. Collection practices currently involve baited trapping and while an inherent bias accompanying the choice of bait is acknowledged, there is little consistency in bait choice between studies and insufficient assessment of trapping success rates for bait types in current use. This study aimed to examine the effect of bait choice on trapping results for six commonly used bait types; a commercial bait (Envirosafe Fly Attractant, Envirosafe Products) and a combination of mixtures of liver, horse manure and 5% sodium sulfide (Na₂S). Trapping success was compared under different seasonal conditions (Summer, Autumn, and Spring) to determine the most attractive bait for calliphorid species, with a secondary comparison of kangaroo versus ox liver occurring under summer conditions. Baits containing Na₂S were the most successful in captures of all target taxa, with the addition of manure desirable, yielding increased diversity of taxa. Kangaroo liver baits attracted high numbers of Chrysomya spp. (Robineau-Desvoidy, Diptera: Calliphoridae), while Lucilia spp. (Robineau-Desvoidy, Diptera: Calliphoridae) were comparatively underrepresented in traps using liver alone. The use of a combined ox liver/manure/Na₂S bait is recommended as the gold standard for generic screening of necrophagous flies. Where more specific target fly taxa are desired, such as Chrysomya spp. or Lucilia spp. only, alternate baits such as kangaroo or lamb may prove more attractive/successful.

Aedes vigilax (Skuse) is a pest and vector species associated with coastal wetlands and the abundance of this mosquito has been identified as contributing to increased risk of mosquito-borne disease outbreaks. As urban development continues to encroach on these coastal wetlands, pest and public health impacts are becoming of increasing concern and in the absence of broadscale mosquito control. Urban planners are looking to buffer zones and other land use planning options to minimize contact between mosquitoes and humans but gaps in the understanding of dispersal ranges of mosquitoes hamper the adoption of these strategies. A mark-release-recapture experiment was conducted to measure the dispersal of this mosquito from an urban estuarine wetland in Sydney, Australia. An estimated total of over 150,000 wild caught female mosquitoes were marked with fluorescent dust and then released. A network of 38 traps was then operated for 5 d within an area of 28 km². A total of 280 marked mosquitoes was recaptured, representing less than 1% of the estimate 250,000 marked mosquitoes released. Marked mosquitoes were recaptured up to 3 km from the release point, providing an insight into the dispersal range of these mosquitoes. The mean distance traveled by marked mosquitoes was 0.83 km, a result reflecting the greater proportion of marked mosquitoes recaptured near release point. The findings of this study indicate that effective buffer zones between estuarine wetlands and high-density urban developments would be an impractical approach to minimizing pest and public health impacts associated with this mosquito.

As part of the response to autochthonous Zika transmission in the United States, the City of South Miami implemented a 6-mo period in which Wolbachia-infected WB1 Aedes aegypti (L.) males were released into an ∼170-acre area. Intracellular Wolbachia bacteria infections in Ae. aegypti cause early embryonic arrest (known as cytoplasmic incompatibility [CI]) and egg hatch failure, and inundative introductions have been suggested as a potential control tool. Throughout the release period, the Ae. aegypti population was monitored within both the release area and an equivalent area that did not receive WB1 male releases. The results show a significant reduction in egg hatch at the area receiving WB1 males, which is consistent with expectations for CI. Similarly, the number of Ae. aegypti was significantly reduced at the area receiving WB1 males, relative to the untreated area. The observed population reduction and results encourage additional work and replication of the Wolbachia biopesticide approach against Ae. aegypti, as an additional tool to be integrated with existing control tools for the control of this medically important vector and nuisance pest.

The use of insecticides such as deltamethrin, permethrin, and temephos has been a primary tool to manage Aedes aegypti (Linnaeus) in Punjab province, Pakistan; however, recent reports of control failure necessitate monitoring insecticide resistance. For this reason, we evaluated 12 field strains of Ae. aegypti from Punjab for resistance against deltamethrin, permethrin, and temephos along with underlying resistance mechanisms. For deltamethrin, high level of resistance was observed in Rawalpindi, Faisalabad, Sheikhupura, Lahore, Pattoki, and Kasur strains (RR_LC50 > 10-fold); moderate level of resistance in Sargodha, Gujranwala, and Sialkot strains (RR_LC50 = 5- to 10-fold), and low level of resistance in Okara, Multan, and Sahiwal strains (RR_LC50 < 5-fold). In the case of permethrin, high level of resistance was found in all the field strains, except the Okara strain that exhibited moderate resistance. For temephos, five field strains, viz. Faisalabad, Rawalpindi, Kasur, Lahore, and Gujranwala, showed high level of resistance; five strains, viz. Sheikhupura, Sialkot, Pattoki, Sahiwal, and Okara, showed moderate resistance, and two strains from Multan and Sargodha showed low resistance to temephos. Synergism bioassays implementing piperonyl butoxide and S,S,S-tributylphosphorotrithioate exhibited a nonsignificant effect on synergizing toxicity of deltamethrin and permethrin in all field strains except the Lahore strain, suggesting the possible role of target-site resistance mechanism. However, both synergists had a significant effect on synergizing toxicity of temephos in all field strains, suggesting the possibility of metabolic-based mechanism of insecticide resistance. In conclusion, the study confirmed the presence of resistance to deltamethrin, permethrin, and temephos in the studied field strains of Ae. aegypti from Punjab, Pakistan.

Background: Large distribution of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) contributed to a significant decrease in malarial mortality. Unfortunately, large insecticide resistance in malaria vectors occurred and is a threat to the future use of these control approaches. The purpose of this study was to explore a new approach for vector control. Patches containing Bacillus thuringiensis var. israelensis (Bti) solubilized Cry toxins mixed with sugar were developed and tested in the laboratory with pyrethroid-resistant Anopheles gambiae s.l. using tunnel tests.

Methods: Mosquitoes were released at 6:00 p.m. into a large tunnel separated by a bed net, perforated with nine holes, from a smaller chamber with a guinea pig. Nine Bti sugar patches (BSPs) were attached to the bed net between the nine holes. Fourteen hours later (8:00 a.m.), mosquitoes were collected from the tunnel and the guinea pig chamber. Live females were kept in cups and were fed a sugar solution (5%) for 72 h and delayed mortality was followed. The results were reported as passing, blood fed and mortality rates.

Results: Mosquito populations that are resistant to the insecticides in the bed net, exhibited high mortality (60%) in the presence of the BSPs. Untreated bed nets with patches in the tunnel test killed 66–95% of the mosquitoes that landed and untreated bed nets were superior to treated bed nets.

Conclusion: BSPs efficiently kill resistant mosquitoes that land on treated and untreated bed nets and thus could ultimately reduce the number of vector-borne malarial mosquitoes.

The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), transmits bovine babesiosis and anaplasmosis, and is endemic to Mexico, Latin and South America. Rhipicephalus (B.) microplus infestations within the United States are a continuing threat to U.S. cattle producers. An importation barrier between Texas and Mexico keeps the ticks from re-entering the United States. All cattle imported into the United States are dipped in an organophosphate (OP) acaricide and hand inspected for presence of ticks. Tick resistance has developed to most available acaricides, including coumaphos, the OP used in the cattle dip vats. OP-resistance can result from one or more mutations in the gene encoding the enzyme, acetylcholinesterase (AChE), resulting in production of an altered AChE resistant to OP inhibition. Previous research reported a large number of BmAChE1 mutations associated with OP resistance. We report baculovirus expression of recombinant tick BmAChE1 (rBmAChE) enzymes containing a single resistance-associated mutation, to assess their contribution to OP inhibition resistance. Surprisingly, of the naturally occurring BmAChE1 resistance-associated mutations, only D188G resulted in markedly reduced sensitivity to OP-inhibition suggesting that OP-insensitivity in BmAChE1 may result from the D188G mutation, or may possibly result from multiple mutations, each contributing a small decrease in OP sensitivity. Furthermore, an OP-insensitivity mutation (G119S) found in mosquitoes was expressed in rBmAChE1, resulting in 500-2000-fold decreased sensitivity to OP inhibition. Recombinant BmAChE1 with the G119S mutation demonstrated the lack of any structural prohibition to broad and high-level OP-insensitivity, suggesting potential increases in tick OP-resistance that would threaten the U.S. importation barrier to ticks.

Bed bugs have become a common urban pest with consequences on human health and economic costs to the hotel and tourism sectors. Insecticide resistance is considered an important factor in the current bed bug resurgence, and multiple resistance mechanisms could be working in the resistant bed bug populations. In the present study, we determined the resistance profile to four insecticides with a different mode of action in Cimex lectularius L. (Heteroptera: Cimicidae) field-collected colonies from Argentina. Furthermore, the synergism effect of piperonyl butoxide (PBO) with deltamethrin was investigated to explore the contribution of detoxification metabolism to resistance. Our results showed that most of the field-collected colonies are extremely resistant to deltamethrin and propoxur, much more than to azametiphos and imidacloprid. The differences in resistance ratios among field-collected colonies could be associated with different modes of action of insecticides used in control pest and the mechanisms involved in the resistance. PBO pretreatment led to a significantly decreased RR in pyrethroid-resistant colonies, suggesting an upturn of monooxygenase activity for deltamethrin detoxification. However, the high RR detected could involve other mechanisms as part of the whole resistant phenotype in colonies of C. lectularius resistant to pyrethroids.

A cluster-randomized controlled trial quantified the entomological efficacy of aerial ultra-low volume (AULV) applications of the insecticide chlorpyrifos against Aedes aegypti in Puerto Vallarta, México, during November–October 2017. The trial involved 16 large (1 × 1 km) clusters distributed between treatment-control arms. Primary endpoint was the abundance of Ae. aegypti indoors (total adults, females, and blood-fed females) collected using Prokopack aspirators. After four consecutive weekly cycles of AULV, all adult Ae. aegypti infestation indices were significantly lower in the treatment arm (OR and IRR ≤ 0.28). Efficacy in reducing indoor Ae. aegypti increased with each weekly application cycle from 30 to 73% (total adults), 33 to 76% (females), and 45.5 to 89% (blood-fed females). Entomological indices remained significantly lower in the treatment arm up to 2 wk after the fourth spraying round. Performing AULV spraying can have significant and lasting entomological impact on Ae. aegypti as long as multiple (ideally four) spray cycles are implemented using an effective insecticide.

Ixodids are globally distributed pests that transmit many disease agents. Increasing resistance to conventional acaricides raises the need for alternative tactics. Novaluron and pyriproxyfen are insect growth regulators (IGRs) that have variable potencies against acarines. We conducted in vitro and in vivo experiments to assess novaluron + pyriproxyfen (marketed as Tekko Pro) against four ixodid species. Laboratory assays on the brown dog tick, Rhipicephalussanguineus(Latreille) (Ixodida: Ixodidae), and the lone star tick, Amblyommaamericanum(L.) (Ixodida: Ixodidae), reduced metabolic activity in larvae and nymphs. Concentrations of novaluron + pyriproxyfen dried on filter paper impeded molting of larval R. sanguineus (less effective against nymphs). Molting A. americanum larvae were reduced by >95% using 4 and 8 µg/cm² eliminated molting; nymphal molting was reduced but not halted even at 16 µg/cm². On calves, novaluron + pyriproxyfen stopped larval A. americanum metabolic function 1 d post-treatment and larvae did not molt. When larvae were released 30 d after treatment, metabolic activity was reduced by 95% and molting was reduced by 94%. Southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae), larvae released 1 d after treatment on calves were 99% prevented from reaching adulthood. The treatment did not interfere with larval development when larvae were released 52 d after treatment. The cattle fever tick, Rhipicephalus (Boophilus) annulatus (Say) (Ixodida: Ixodidae), failed to reach adulthood when larvae were released on calves a day after treatment (residual activity was not assessed for R. annulatus). These IGRs, and possibly others, offer an alternative to conventional acaricides for ixodid control on cattle.

Plant essential oils (EOs) have been considered as spatial repellents to help disrupt the pathogen transmission cycle of mosquitoes. Our objective was to assess spatial repellency effects of EOs on the tropical yellow fever mosquito, Aedes aegypti (L.) (Diptera: Culicidae) and on local mosquito populations in coastal British Columbia (Canada). In laboratory experiments using protocols of the World Health Organization, three of the solitary EOs tested proved repellent to Ae. aegypti: cinnamon bark, lemongrass, and rosemary. Binary combinations of select EOs enhanced the repellent effect of single EOs through synergistic interactions. The EO blend of geranium and peppermint lowered the RD₅₀ (the dose required to obtain 50% repellency) of each solitary EO by >1,000-fold. Compared with binary EO blends, ternary EO blends were typically less repellent to mosquitoes, possibly due to a dilution effect of the most effective EO constituent(s) in the blend. In field experiments, the EO blend of lemongrass and cinnamon bark expressed spatial repellency towards the cool weather mosquito, Culiseta incidens (Thomson) (Diptera: Culicidae), even when this blend was disseminated from devices as much as 1 m away from a sentinel trap releasing attractive vertebrate host odorants and CO₂. Deployment of EOs as spatial repellents in small outdoor gatherings could help protect humans from mosquito-borne diseases, particularly when this tactic is coupled with other tools of mosquito management.

The first breeding populations of Aedes aegypti (Linnaeus) were identified in California in 2013, and have since been detected in 13 counties. Recent studies suggest two introductions likely occurred, with genetically distinct populations in the central and southern regions of the state. Given the threat of dengue, chikungunya, and Zika virus transmission, it is imperative to understand if these populations harbor genes that could confer resistance to pyrethrin-based insecticides, known as pyrethroids, the most commonly used class of adulticides in the state. In 2017, the California Department of Public Health initiated a pesticide resistance screening program for Ae. aegypti to assess the presence of specific mutations on the sodium channel gene (V1016I and F1534C) associated with knockdown resistance to pyrethroids. Mosquitoes collected between 2015 and 2017 from 11 counties were screened for mutations using real-time polymerase chain reaction assays. Results revealed distinctly different resistance profiles between the central and southern regions. The central population displayed nearly fixed resistant mutations at both loci, whereas the southern population was more variable. The relative proportion of resistant alleles observed in sampled mosquitoes collected in southern California increased each year from 2015 through 2017, indicating potential increases in resistance across this region. The presence of these mutations indicates that these mosquitoes may be predisposed to surviving pyrethroid treatments. Additional biological and biochemical assays will help better elucidate the mechanisms underlying insecticide resistance in California Ae. aegypti and prompt the use of pesticides that are most effective at controlling these mosquitoes.

Attractive toxic sugar baits (ATSBs) can be an effective vector control tool, especially in areas where aerial or aquatic applications of pesticides are undesirable or impractical. In general, there is a need to develop novel or alternative insecticides for vector control, and there is a demand from consumers for more ‘natural' pest control products. Sodium ascorbate (SA) is a naturally occurring antioxidant compound, found in fruits and vegetables, and is available commercially in the United States as a food additive and supplement. In this study, we continuously exposed groups of adult Aedes aegypti (L.), Anopheles stephensi Liston (Diptera: Culicidae), Phlebotomus papatasi Scopoli, and Lutzomyia longipalpis (Lutz & Neiva; Diptera: Psychodidae) to ATSBs containing SA in concentrations of 6, 8, 10, and 20%, and tracked their mortality over 10 d. We also exposed insects to a 20% SA–ATSB on a single day to determine the effect of a single exposure to the bait on mortality. Concentrations of ≥8% SA significantly reduced survival of both mosquito species over 10 d compared with sugar-fed controls. Sand fly mortality was inconsistent. A single exposure to 20% SA significantly reduced the survival of An. stephensi. Mosquitoes exposed to SA exhibited elevated catalase levels and cell death. The use of SA in ATSBs may be most effective in areas where sugar sources are scarce, and where mosquito species frequently sugar-feed. SA sugar baits may be a particularly attractive option for the general public looking to control mosquito populations using ‘natural’ alternatives to synthetic insecticides.

Among neglected tropical diseases, visceral leishmaniasis (VL) shows great relevance in global terms and is a serious public health concern due to the possibility of severe and lethal forms in humans. In this study, we evaluate entomological factors such as diversity and abundance of phlebotomine sand flies (Diptera:Psychodidae) and the Leishmania species circulating in these species in possible association with VL transmission in the Brazilian town Itaúna. The entomological collections were performed during three consecutive nights, always in the third week of each month, within a period of 12 mo. A total of 1,786 sand fly specimens were collected, from which 20% were collected inside houses. The influence of three local climatic variables (temperature, rainfall, relative humidity) on the population sizes of these insects was evaluated. Temperature was the most influential factor, with a significant positive correlation with the local population size of phlebotomine sand flies collected per month. Lutzomyia longipalpis (Lutz & Neiva, 1912) was the predominant species in the study area. Leishmania DNA was detected in nine out of 133 pools of sand fly females, using nested/ PCR, which resulted in a minimal natural infection rate of 2.91%. DNA from Leishmania infantum Nicolle, 1908 (Kinetoplastida: Trypanosomatida), was detected in Evandromyia cortelezzii (Bréthes, 1923), Ev. evandroi (Costa, Lima & Antunes, 1936), Ev. lenti (Mangabeira, 1938), and Ev. termitophila (Martins, Falcão & Silva, 1964), besides Lu. longipalpis. Our study indicates favorable conditions for VL spreading in Itaúna due to the presence of Lu. longipalpis and Le. infantum-infected phlebotomine sand flies.

The present study aimed to update the list of Aedes mosquito species occurring in Tunisia and to test the vector competence of Aedes (Ochlerotatus) caspius (Pallas) and Ae. (Ochlerotatus) detritus (Haliday), the locally most abundant and widespread species, to transmit Zika virus (ZIKV). In 2017–2018, mosquito larvae were collected from 39 different larval habitats in seven bioclimatic zones of Tunisia. The salinity and pH of each breeding site were measured. The survey revealed the presence of 10 Aedes species in Tunisia: Ae. (Stegomyia) albopictus (Skuse), Ae. (Ochlerotatus) berlandi (Séguy), Ae. caspius, Ae. detritus, Ae. (Finlaya) echinus (Edwards), Ae. (Finlaya) geniculatus (Olivier), Ae. (Acartomyia) mariae (Sergent and Sergent), Ae. (Ochlerotatus) pulcritarsis (Rondani), Ae. (Aedimorphus) vexans (Meigen), and Ae. (Fredwardsius) vittatus (Bigot). Of these 10 species, Ae. caspius and Ae. detritus were the most abundant in Tunisia. Aedes detritus and Ae. caspius larvae were reared until the imago stage under insectary conditions to test autogeny. The study showed that Ae. detritus is autogenous and stenogamous and Ae. caspius, anautogenous and eurygamous. Finally, the collected strains of these two species were experimentally infected with the Asian genotype of ZIKV, originally isolated from a patient in April 2014 in New Caledonia, to test their vector competence. Neither of these species was able to transmit ZIKV at 7 and 14 d postexposure. Further investigations are needed to test the competence of other Tunisian mosquito species that may be associated with ZIKV transmission.

The etiologic agent of Chagas disease, Trypanosoma cruzi, is transmitted by hematophagous insect vectors that subsist on repeated blood meals over their lives separated by periods of fasting. Using naturally infected Mepraia spinolai, we measured the influence of parasite infection on this host vector's mortality during regular feeding and after fasting. After their capture, the insects were fed twice with uninfected mice to evaluate parasitic infection in their fecal samples by microscopic observation and PCR. Then the insects were subjected to a fasting period, followed by a third (final) feeding. After each feeding, a fecal sample was obtained to evaluate T. cruzi infection. To determine its progress through ontogeny, mortality and ecdysis of the infected and uninfected nymphs and adults were recorded on three occasions, over 140 d, and analyzed. Detections of infection by T. cruzi between the two first feedings increased, but this detection level was generally reduced after final feeding unless reinfected. For nymphs (stages III–V), their mortality was highest when infected after the fasting period, whereas adults were equally resistant to death after fasting when infected with T. cruzi. Metacyclic trypomastigotes were principally excreted in the fecal samples. Our results confirm that T. cruzi is pathogenic to its invertebrate hosts under nutritional stress conditions, when nymphs' mortality is higher while infected than uninfected when they were hungry. These results are epidemiologically important because T. cruzi harms the fasting vector M. spinolai, reducing its lifespan and competence as a disease vector, and thereby its rates of parasite transmission.

This study records the clinical findings in nine hunting dogs showing systemic illness associated with trombiculids and identifies the mite species involved. In fall, coinciding with the seasonality of mites, all dogs were infested with mites and had been in the risk area (Sierra Cebollera Natural Park, La Rioja, Spain) a few hours before the onset of symptoms. The symptoms included vomiting, anorexia, weakness and lethargy, diarrhea, and even stupor. The clinical picture was fast-acting and potentially fatal. The infestations varied from low to severe. Molecular analysis of mites that fed on the dogs confirmed that they were larvae of Neotrombicula inopinata (Oudemans, Acari, Trombiculidae). This is the first time that N. inopinata has been identified as feeding on dogs and implicated in canine systemic illness associated with trombiculids. In contrast to other chiggers, N. inopinata does not seem to cause dermatitis. Likewise, the clinical and epidemiological similarity between the clinical symptoms we describe herein and the occurrence of seasonal canine illness (SCI) led us to suspect that this illness may be caused by infestation with these mites. The condition could be the consequence of severe infestation from large numbers of feeding mites, especially N. inopinata. Whether or not the cases were due to a severe allergic host response to salivary proteins or the result of the transmission of a new or emerging trombiculid-borne pathogen is not known.

Vector-borne diseases in the United States have recently increased as a result of the changing nature of vectors, hosts, reservoirs, pathogens, and the ecological and environmental conditions. Current information on vector habitats and how mosquito community composition varies across space and time is vital to successful vector-borne disease management. This study characterizes mosquito communities in urban areas of Oklahoma, United States, an ecologically diverse region in the southern Great Plains. Between May and September 2016, 11,996 female mosquitoes of 34 species were collected over 798 trap nights using three different trap types in six Oklahoma cities. The most abundant species trapped were Culex pipiens L. complex (32.4%) and Aedes albopictus (Skuse) (Diptera: Culicidae) (12.0%). Significant differences among mosquito communities were detected using analysis of similarities (ANOSIM) between the early (May–July) and late (August–September) season. Canonical correlation analysis (CCA) further highlighted the cities of Altus and Idabel as relatively unique mosquito communities, mostly due to the presence of Aedes aegypti (L.) and salt-marsh species and absence of Aedes triseriatus (Say) in Altus and an abundance of Ae. albopictus in Idabel. These data underscore the importance of assessing mosquito communities in urban environments found in multiple ecoregions of Oklahoma to allow customized vector management targeting the unique assemblage of species found in each city.

Vector-borne Chagas disease is endemic to the Americas and imposes significant economic and social burdens on public health. In a previous contribution, we presented an automated identification system that was able to discriminate among 12 Mexican and 39 Brazilian triatomine (Hemiptera: Reduviidae) species from digital images. To explore the same data more deeply using machine-learning approaches, hoping for improvements in classification, we employed TensorFlow, an open-source software platform for a deep learning algorithm. We trained the algorithm based on 405 images for Mexican triatomine species and 1,584 images for Brazilian triatomine species. Our system achieved 83.0 and 86.7% correct identification rates across all Mexican and Brazilian species, respectively, an improvement over comparable rates from statistical classifiers (80.3 and 83.9%, respectively). Incorporating distributional information to reduce numbers of species in analyses improved identification rates to 95.8% for Mexican species and 98.9% for Brazilian species. Given the ‘taxonomic impediment’ and difficulties in providing entomological expertise necessary to control such diseases, automating the identification process offers a potential partial solution to crucial challenges.

Tick surveillance provides essential information on distributions and encounter frequencies; it is a component of operational activities in public health practice. Our research objectives were a proof-of-concept for collaborative surveillance, which involved establishing an academic and government partnership to enhance tick surveillance efforts. The University of Tennessee (UT) collaborated with United States Department of Agriculture Forest Service, Southern Research Station Forest Inventory Analysis (FIA) in an Occupational Health and Safety partnership. UT provided FIA crews in the southeastern United States with vials containing 80% ethanol (July 2014–November 2017). Crew members were instructed to put all encountered ticks into the vials and return them to FIA headquarters. UT identified all submitted ticks to species and life stage, and screened Amblyomma americanum (L.) for Ehrlichia bacteria using a nested-PCR assay. From the 198 returned vials, 1,180 ticks were submitted, including A. americanum (90.51%; 202 larvae, 503 nymphs, and 363 adults), Dermacentor variabilis Say (7.12%; 1 nymph, 83 adults), Ixodes scapularis (Say) (1.61%; 19 adults), Amblyomma maculatum Koch (0.59%; 1 nymph, 6 adults), and Amblyomma cajennense (Fabricius) (0.17%; 1 nymph, 1 adult). FIA crews encountered A. americanum with Ehrlichia and collection information was used to generate baseline occurrence data of tick encounters. Results indicate that this collaborative-tick surveillance can be improved and used to generate useful data including pathogen detection, and because crews revisit these sites, changes in tick encounters can be monitored.

Public health authorities recommend a range of nonchemical measures to control blacklegged ticks Ixodes scapularis Say, 1821 (Ixodida: Ixodidae) in residential yards. Here we enumerate these recommendations and assess their relationship to larval tick abundance in 143 yards in Dutchess County, New York, an area with high Lyme disease incidence. We examined the relationship between larval tick abundance and eight property features related to recommendations from public health agencies: presence or absence of outdoor cats, wood piles, trash, stone walls, wood chip barriers separating lawn from adjacent forest, bird feeders, fencing, and prevalence of Japanese barberry (Berberis thunbergii DC [Ranunculales: Berberidaceae]). We assessed abundance of larval ticks using two methods, flagging for questing ticks and visual examination of ticks on white-footed mice Peromyscus leucopus Rafinesque, 1818 (Rodentia: Cricetidae). More questing larvae were found in yards where trash or stone walls were present. These effects were less pronounced as forest area increased within the yard. Counts of larvae per mouse were lower in properties with >75% of the yard fenced than in properties with less fencing. We find partial support for recommendations regarding trash, stone walls, and fencing. We did not detect effects of outdoor cats, bird feeders, barriers, wood piles, or Japanese barberry. There was low statistical power to detect effects of ground barriers (gravel, mulch, or woodchip), which were present in only two properties.

Understandings of natural enemy efficacy are reliant on robust quantifications of interaction strengths under context-dependencies. For medically important mosquitoes, rapid growth during aquatic larval stages could impede natural enemy impacts through size refuge effects. The identification of biocontrol agents which are unimpeded by ontogenic size variability of prey is therefore vital. We use functional response and prey preference experiments to examine the interaction strengths and selectivity traits of larvae of the cohabiting predatory midge Chaoborus flavicans (Meigen 1830) (Diptera: Chaoboridae) towards larval stages of the Culex pipiens (Diptera: Culicidae) mosquito complex. Moreover, we examine the influence of search area variation on selectivity traits, given its importance in consumer-resource interactions. Chaoborids were able to capture and consume mosquito prey across their larval ontogeny. When prey types were available individually, a destabilizing Type II functional response was exhibited towards late instar mosquito prey, whereas a more stabilizing Type III functional response was displayed towards early instars. Accordingly, search efficiencies were lowest towards early instar prey, whereas, conversely, maximum feeding rates were highest towards this smaller prey type. However, when the prey types were present simultaneously, C. flavicans exhibited a significant positive preference for late instar prey, irrespective of water volume. Our results identify larval chaoborids as efficacious natural enemies of mosquito prey, with which they frequently coexist in aquatic environments. In particular, an ability to prey on mosquitoes across their larval stages, coupled with a preference for late instar prey, could enable high population-level offtake rates and negate compensatory reductions in intraspecific competition through size refuge.

Aedes (Stegomya) albopictus (Skuse 1984), the Asian tiger mosquito, represents the most invasive and one of the medically most important mosquito vectors. Although native to South East Asia, the species has recently spread globally, and was registered in the city of Novi Sad (Serbia, Central Balkans) in August 2018. We characterized the invasive population using phenotypic (wing size and shape) and molecular (nuclear, internal transcribed spacer 2- ITS2, and mitochondrial, cytochrome c oxidase subunit I- COI) markers. The results of phenotypic analyses indicated that the Serbian population could be differentiated from the native (Thailand) and invasive (Hawaii and Florida) populations due to restricted gene flow, founder effect, and supposed different strain origin. The Serbian population showed genetic homogeneity, indicative of a small founder number (bottleneck invasion model). Despite the incorporation of ITS2 GenBank sequences into the data set, neither spatial (Geneland) nor nonspatial (BAPS) genetic structuring analyses helped infer the Serbian population origin. However, the comparison of the retrieved COI haplotype with previously characterized mitogenomes indicated a temperate strain origin, capable of overwintering. Such findings suggest that the newly registered Ae. albopictus population could be able to establish itself since previous studies outlined Novi Sad as a suitable area.

The synergistic effect of light-emitting diodes (LEDs) and kairomones on the attraction of sand flies to light traps was evaluated. Octenol and lactic acid were used as chemical attractants. Green LEDs and the incandescent lamps were used as light attractants. Five CDC-type light traps with the respective combination of attractants (incandescent lamp, incandescent lamp + chemical attractant, green LED, green LED + chemical attractant, and chemical attractant alone [without light]) were set between 18:00 and 06:00 following a Latin square design. A total of 6,536 sand flies and 16 species were collected. The most frequent species collected was Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera, Psychodidae) accounting for 43.21% of all individuals. Order of success (mean, SD) of lactic acid attractant fly capture was as follows: LED + lactic acid (36.83 ± 4.74), LED alone (34.87 ± 4.61), incandescent lamp + lactic acid (22.80 ± 3.19), incandescent lamp alone (12.67 ± 2.03), and lactic acid (0.46 ± 0.13). Order of success of octenol attractant fly capture was as follows: LED + octenol (37.23 ± 5.61), LED alone (35.77 ± 5.69), incandescent lamp + octenol (18.63 ± 3.28), incandescent lamp alone (14.67 ± 2.86), and octenol alone (1.80 ± 0.65). With exception of lactic acid + incandescent light, chemical synergists played no part in significantly increasing light trap capture of phlebotomine sand flies. However, the use of LEDs, with or without such attractants, provided significantly higher capture compared to the incandescent lamp with or without such chemicals, showing that LEDs are suitable and efficient light sources for surveillance and monitoring of phlebotomine sand flies in Brazil.