Initial data on the prevalence of storage mites in dry-stored food products and estimates of the presence of mites in human stool in the city of Minia, Egypt are provided. In total, 847 samples were collected randomly from houses and retail stores between March 2017 and February 2018. In addition, 1,000 human stool samples were collected for the detection of the presence of mites. Mites were extracted from 285 of 840 (33.9%) samples, and mite contamination was found to be most prevalent in wheat flour (73.3%). In total, 11 mite species belonging to six families were identified, with the pest species Tyrophagus putrescentiae (Schrank) (Acari: Acaridae) (TP) being the most prevalent (91.2% of samples). The seasonal density distribution showed the highest storage mite density in March–April, followed by October, and the lowest in January. In addition, mites were detected in 87 (8.7%) human stool samples, with significant associations between certain occupations and some personal characteristics. Therefore, more attention needs to be paid to intestinal acariasis arising from mite infestation of dry-stored food products.

Human head lice Pediculus humanus capitis (De Geer) (Phthiraptera: Pediculidae) are insect parasites closely associated with humans, feeding on the blood of their hosts and causing them skin irritation and probable secondary infections. Despite being a severe nuisance, very few studies have reported on intraspecific chemical communication in head lice. Here, we evaluated the attractive response of head lice to the volatile compounds and solvent extracts from their feces. We also chemically analyzed the main volatile components of these feces and those of the feces' extracts. Head lice were attracted to the methanol extract of their feces but not to the hexane or dichloromethane extracts, suggesting the polar nature of bioactive chemicals present in head louse feces. Follow-up chemical identifications, in fact, showed the presence of hypoxanthine, uric acid, and another purine tentatively identified as either guanine or iso-guanine. Additionally, head lice were significantly attracted by volatiles emitted from samples containing feces. The volatiles emanated from feces alone contained 19 identified substances: 2-pentanone, hexanal, heptanal, 3-methyl-3-buten-1-ol, octanal, sulcatone, nonanal, acetic acid, 2-ethyl-1-hexanol, decanal, 1-octanol, butyric acid, 1-nonanol, hexanoic acid, octanoic acid, 2,6-dimethyl-7-octen-2-ol, 2-undecanone, geranylacetone, and hexadecane. The major compounds found were decanal, nonanal, hexanal, and acetic acid, together representing approximately 60% of the identified compounds. This work represents the first chemical evidence of intraspecies communication among head lice. The results support the existence of active substances present in the feces of P. humanus capitis that may be involved in its aggregation behavior.

Considerable previous research has focused on predator-associated semiochemicals and how they affect mosquito oviposition behavior. However, most of this work has been done without taking into consideration either the natural density of the predators or how other semiochemicals in aquatic environments might affect the responses of gravid mosquitoes. The influence of mosquitofish density, source water (tap vs pond), presence of freshly laid egg rafts, and removal of a putative source of semiochemicals (bacteria) on oviposition by Culex tarsalis Coquillett (Diptera: Culicidae) was studied in laboratory bioassays. Culex tarsalis females were deterred from laying egg rafts on water that contained semiochemicals associated with Gambusia affinis (Baird & Girard), but this deterrence was not strongly associated with the density of fish used to condition aged tap water. The number of egg rafts laid onto Gambusia-exudate water made with either tap water (density < 1 fish per liter) or pond water was typically ≥ 50% of that onto water that did not house mosquitofish. Gravid mosquitoes tested individually did not reduce oviposition onto Gambusia-exudate water as compared to controls. Likewise, oviposition by females with ablated wings did not differ significantly between Gambusia-exudate water and controls. Oviposition onto filter-sterilized Gambusia-exudate water was reduced relative to unfiltered water, suggesting that semiochemicals deterring egg-laying were still present after bacteria were removed. Taken together, these findings suggest that the responses of gravid Cx. tarsalis to chemicals from habitats containing mosquitofish are complex and the origin of the semiochemicals present in the Gambusia-exudate water needs to be elucidated.

Electropenetrography (EPG) has been used for many years to visualize unseen stylet probing behaviors of plant-feeding piercing–sucking insects, primarily hemipterans. Yet, EPG has not been extensively used with blood-feeding insects. In this study, an AC–DC electropenetrograph with variable input resistors (Ri), i.e., amplifier sensitivities, was used to construct a waveform library for the mosquito arbovirus vector, Aedes aegypti (Linneaus), while feeding on human hands. EPG waveforms representing feeding activities were: 1) electrically characterized, 2) defined by visual observation of biological activities, 3) analyzed for differences in appearance by Ri level and type of applied signal (AC or DC), and 4) quantified. Electrical origins of waveforms were identified from five different Ri levels and AC versus DC. Mosquitoes produced short stylet probes (‘bites') that typically contained five waveform families. Behaviors occurred in the following order: surface salivation (waveform family J), stylet penetration through the outer skin (K), penetration of deeper tissues and location of blood vessels/pathway activities (L), active ingestion with engorgement (M), and an unknown behavior that terminated the probe (N). Only K, L, and M were performed by every insect. A kinetogram of conditional probabilities for waveform performance demonstrated plasticity among individuals in L and M, which were alternated. Now that EPG waveforms for mosquito feeding have been defined, EPG can be used as a tool for improved biological understanding of mosquito-borne diseases.

A study was performed to evaluate the implication of Protophormia terraenovae larvae as a surgical therapy for wounded skin. Three groups of sheep (n = 25) were considered based on larval doses. Groups 1 and 2 were artificially infested with low and high concentrations of L1 stage P. terraenovae, respectively, and group 0 served as a control. Skin biopsies were taken at 4 and 14 d postinfestation (D.P.If). A histopathological study was carried out to evaluate the lesions with a score, numbers of eosinophils and mast cells, and an immunohistochemical analysis of CD3, CD79α, and CD68 as T lymphocytes, B lymphosytes, and macrophages, respectively. The results indicated that higher larval doses led to faster regeneration by 14 D.P.If. Furthermore, the higher larval doses showed a high number of the CD68 marker and eosinophils and a low number of CD3 and CD79α markers and mast cells. In addition, the number of mast cells, T lymphocytes, and macrophage markers increased when the lesion progressed; however, a low number of immunolabeled CD79α cells and eosinophils were observed. The results indicate a possible positive effect of larvae in the healing of certain wounds.

There have been some investigations carried out about necrophagous flies and their biodiversity indices in Iran. In the present study, the species composition of flies, their richness, and biodiversity were investigated in central Iran. In a cross-sectional study, specimens were collected monthly in three different climate areas of Qom Province. Species diversity, species richness, and species evenness were calculated to estimate the species biodiversity of flies. In total, 7,291 flies were collected and identified using valid identification keys that classified flies into four families and 15 different species. The largest collection of flies was obtained in the mountainous areas comprising 36.2% of specimens. Moreover, five forensically important species of fly were collected in this region with the potential to cause human and animal myiasis: Calliphora vicina, Lucilia sericata, Musca domestica, Wohlfahrtia nuba, and Chrysomya albiceps. There were slight differences in species richness and diversity indices in the areas. In mountainous areas, the Shannon–Wiener index and richness were maximum (H' = 1.87, S = 13). The evenness based on the Shannon evenness index in the mountainous areas was 0.728 and 0.719 in desert areas, whereas it was the highest (0.764) in the semidesert areas. Moreover, high values of the Jaccard similarity index were observed in captured necrophagous fly species in all the areas. The necrophagous fly fauna and species diversity indices were high because of an increase in the evenness. In general, some well-known forensically important species of flies were collected that proved the potential of myiasis occurrence in this area.

Rickettsia typhi and Rickettsia felis (Rickettsiales: Rickettsiaceae) are flea-transmitted pathogens. They are important causes of acute febrile illness throughout the world. We, therefore, sought to identify the rickettsial species present in the fleas of dogs and cats in the department of Cauca, Colombia. In this study, we collected 1,242 fleas from 132 dogs and 43 fleas from 11 cats. All fleas were morphologically identified as Ctenocephalides felis (Bouché) adults and organized in pools for DNA extraction (234 pools from dogs and 11 from cats). The gltA gene from rickettsiae was targeted for screening amplification using conventional PCR. In total, 144 of the 245 pools (58.7%) were positive. The positive samples were then processed for the amplification of the 17kDa antigen gene (144/144; 100% positive) and sca5 gene (140/144; 97.2% positive). In addition, restriction enzyme length polymorphism analysis using NlaIV on the amplified product of the sca5 gene demonstrated several organisms: 21/140 (15%) were R. felis, 118/140 (84.3%) were Rickettsia asemboensis, and 1/140 (0.7%) were Candidatus Rickettsia senegalensis. Subsequent sequencing confirmed Candidatus Rickettsia senegalensis in C. felis collected from dogs the first reported from Colombia.

The widespread nominal black fly Simulium (Simulium) rufibasis Brunetti was reexamined morphologically, chromosomally, and molecularly to determine the status of populations in Japan and Korea with respect to S. rufibasis from the type locality in India and to all other known species in the S. (S.) tuberosum species-group. Morphological comparisons established that the species previously known as S. rufibasis in Japan and Korea is distinct from all other species. Consequently, it was described and illustrated as a new species, Simulium (S.) yamatoense. Simulium yokotense Shiraki, formerly a synonym of S. rufibasis, was morphologically reevaluated and considered a species unplaced to species-group in the subgenus Simulium. Chromosomal analyses of S. yamatoense sp. nov. demonstrated that it is unique among all cytologically known species of the S. tuberosum group and is the sister species of the Taiwanese species tentatively known as S. (S.) arisanum Shiraki. Populations of S. yamatoense sp. nov. included two cytoforms, based on the sex chromosomes. Cytoform A, including topotypical representatives, was found in Kyushu, Japan, whereas cytoform B was found in Korea and Honshu, Japan. Molecular analysis based on the COI mitochondrial gene generally corroborated morphological and chromosomal data that S. yamatoense sp. nov. is a distinct species and, like the chromosomal data, indicate that it is most closely related to S. arisanum, with interspecific genetic distance of 2.92–4.63%.

Derived from exhaustive search of mites of the genus Parichoronyssus associated with South American Bats, we found two new species associated with Phyllostomid, Emballonurid, and Noctiniolid bats: Parichoronyssus alexanderfaini n. sp. associated with Rhinophylla pumilio Peters; Parichoronyssus gettingeri n. sp. associated with Rhynchonycteris naso Wied-Newied, and Noctilio leporinus Linneo. Herein we give the description of those new species, and additionally is included several new records for Parichoronyssus from the region, as well as providing a key to the 11 species of Parichoronyssus.

The discovery of a new species, Periglischrus empheresotrichus, was determined through a review of museum collections, as well as a field survey of ectoparasites of island bats. This new species parasitizes on two bat species of the genus Monophyllus Leach, the Greater Antillean Long-tongued bat Monophyllus redmani Leach and the Lesser Antillean Long-tongued bat Monophyllus plethodon Miller. The female, male, deuthonymphs, and protonymph are described and illustrated. P. empheresotrichus n. sp. has an insular distribution, we evaluated the morphological variation of the adult populations, and concluded that intra-specific variation is correlated both with host species and locality (island) in the West Indies.

The assembly of species that colonize animal organic matter, their relative abundance, and dynamics are affected by the environmental and biogeographical conditions to which these resources are exposed. Baited trap studies are essential for research on the diversity, seasonality, distribution and population dynamics of necrophagous flies. Decomposing baits provide the necessary stimulus for flies to aggregate on them. In this study, three types of bait of animal origin with different organic chemical composition were compared in terms of the diversity, richness, abundance, and species composition of saprophagous flies species that were attracted to them. Bone-meal (BM), cow liver (CL), and rotten chicken viscera (CV) were used as bait to collect flies. In total, 3,387 Calliphoridae, Sarcophagidae, and Muscidae adult flies were collected. The most abundant species were Lucilia ochricornis (Wiedemann 1830), Chrysomya albiceps (Wiedemann 1819) (Diptera: Calliphoridae), Peckia (Sarcodexia) lambens (Wiedemann 1830) (Diptera: Sarcophagidae), and Ophyra capensis (Wiedemann 1818) (Diptera: Muscidae). The type of bait had significant effects on both the total richness (F_2,18 = 57.08; P < 0.0001) and the effective number of species (F_2,18 = 12.81; P = 0.0003) per trap. The average richness was higher in traps baited with chicken viscera, followed by cow liver and finally by bone-meal. The composition of cow liver and bone meal species constitute subsets of the species collected with chicken viscera, thus using the three baits would not increase the number of species detected. These results indicate that chicken viscera is the most efficient bait for testing or assessing necrophagous fly diversity.

Urban landscape features play an important role in the distribution and population spread of mosquito vectors. Furthermore, current insecticide and novel rear-and-release strategies for urban mosquito management rarely consider the spatial structure of the landscape when applying control practices. Here, we undertake a mark-recapture experiment to examine how urban features influence the movement and distribution of Australian container-inhabiting Aedes vectors. We pay attention to the role of semipermanent water storage containers, called rainwater tanks, and the influence of movement barriers, such as roads, on the spread and distribution of vector populations. Results suggest that Aedes aegypti (Linnaeus) (Diptera: Culicidae) were more likely to be captured around rainwater tanks, and that released males travel throughout residential blocks but do not cross roads. Conversely, female Aedes notoscriptus (Skuse) (Diptera: Culicidae) movement was uninhibited by roads and rainwater tanks did not influence female distribution or oviposition behavior. Using an isotropic Gaussian kernel framework, we show that vector movement is likely to be greater when applying a temporal effect, than when estimated by traditional methods. We conclude that a greater understanding on the role of urban features on vector movement will be important in the new age of rear-and-release mosquito control strategies, particularly those where estimations of movement are important for ensuring efficacy of application.

Dengue, yellow fever, Zika, and chikungunya arboviruses are endemic in tropical countries and are transmitted by Aedes aegypti. Resistant populations of this mosquito against chemical insecticides are spreading worldwide. This study aimed to evaluate the biological effects of exposure of pesticide-sensitive Ae. aegypti larvae (Rockefeller) to conidia of the entomopathogen, Metarhizium brunneum, laboratory strains ARSEF 4556 and V275, and any synergistic activity of phenylthiourea (PTU). In addition, to investigate the nature of any cross-resistance mechanisms, these M. brunneum strains were tested against the Rockefeller larvae and two temephos- and deltamethrin-resistant wild mosquito populations from Rio de Janeiro.Treatment of Rockefeller larvae with 10⁶ conidia/ml of ARSEF 4556 and V275 fungal strains resulted in significant decreased survival rates to 40 and 53.33%, respectively (P < 0.0001), compared with untreated controls. In contrast, exposure to 10⁴ or 10⁵ conidia/ml showed no such significant survival differences. However, the addition of PTU to the conidia in the bioassays significantly increased mortalities in all groups and induced a molt block. Experiments also showed no differences in Ae. aegypti mortalities between the fungal treated, wild pesticide-resistant populations and the Rockefeller sensitive strain. The results show the efficacy of M. brunneum in controlling Ae. aegypti larvae and the synergistic role of PTU in this process. Importantly, there was no indication of any cross-resistance mechanisms between Ae. aegypti sensitive or resistant to pesticides following treatment with the fungi. These results further support using M. brunneum as an alternative biological control agent against mosquito populations resistant to chemical insecticides.

Skin irritation has been reported to be the main adverse effect of excessive use of N,N-diethyl-m-toluamide (DEET) and ethyl 3-acetyl(butyl)amino (IR3535) commercial repellents. Therefore, there is an interest in alternatives of natural origin such as essential oils (EOs) and major compounds, which have repellent effects but have no contraindications. The main purpose of the present study was to identify the repellent effect of selected terpenes on Aedes aegypti Linnaeus, 1762 (Diptera: Culicidae) by in silico analysis based on their affinity with the odorant protein AaegOBP1. The protein-metabolite interactions in 20 terpenes were analyzed using the SwissDock tool. Terpenes presenting the highest affinity compared with commercial repellents were selected to evaluate repellent activity at concentrations 0.1, 10, and 25% against Ae. aegypti. Different periods (0–2, 2–15, 15–60 min) were evaluated with DEET as a positive control. The toxicity of terpenes was verified through Osiris and Molinspiration Cheminformatics Software, and cytotoxicity assays in Vero and HepaRG cells were performed using the MTT method. Two formulations were prepared with polyethylene glycol to evaluate skin long-lasting in vivo assay. The results showed four terpenes: geranyl acetate, nerolidol, α-bisabolol, and nerol, with affinity to AaegOBP1 comparable with DEET and IR3535. Geranyl acetate, nerolidol, and their mixtures showed no cytotoxicity and protection percentages close to 100% during the test at concentrations 10 and 25%. Long-lasting assays with geranyl acetate and nerolidol formulate showed 3 h as maximum protection time with 100% protection percentage. These metabolites and their mixtures are candidates to repellent formulations with times and protection percentages similar to DEET.

Mosquitoes of the Aedes genus are vectors for dengue, chikungunya, Zika, and yellow fever viruses. Mosquito repellents are an effective way to prevent mosquito bites and reduce the spread of mosquito-borne diseases. In the early 90s, the U.S. Environmental Protection Agency (EPA) published a list of active ingredients that pose minimum risk to human health that can be used as pesticides or repellents without passing the EPA registration process. The present study examined the efficacy of 21 of the active ingredients listed by the EPA 25 (B) exempt list and five commercially available sprays that only contained active ingredients from the EPA 25(B) list in repelling female Aedes aegypti (L.) females. We performed choice bioassays in a controlled laboratory environment, using aY-tube olfactometer to determine attraction rates of humans to female Ae. aegypti in the presence of one of the 21 active ingredients and five commercially available repellent sprays. We found that cinnamon oil, peppermint oil, spearmint oil, lemongrass oil, and garlic oil reduced mosquito attraction to human odor. Of the five commercial repellent sprays, only one reduced mosquito attraction for up to 30 min in our assay. The EPA 25 (B) list contains active ingredients that under the conditions of our assay repel Ae. aegypti.

Ixodid ticks vector pathogens of humans, livestock, and wildlife, and occur in tropical and temperate regions worldwide. We used the lone star tick, Amblyomma americanum (L.) (Ixodida: Ixodidae), as a model for other ixodid species, including the southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae). We assessed the lethality of CimeXa, an inert silica-based desiccant dust, and Drione, a silica-based desiccant dust with pyrethrin and piperonyl butoxide (PBO), against A. americanum larvae and nymphs on calves. CimeXa was highly lethal to larvae released onto shaved calf skin 72–96 h previously, and moderately toxic to nymphs. It did not affect immature life stages treated after they began feeding because desiccation was offset by ingestion of blood. Being inert, CimeXa can protect cattle from infestation before feeding occurs for a potentially indefinite period of time. Drione killed immatures within 24 h when released on treated skin and when applied to feeding ixodids because of the pyrethrin and PBO (PBO will also reduce resistance to pyrethrin if it develops). When CimeXa was applied to calves with the hair intact, ≈90% and ≈70% of A. americanum larvae and nymphs, respectively, were killed within 24 h and Drione killed 100% of each life stage. Drione also prevented and drastically reduced egg production and hatching. While CimeXa protects against infestation by immatures, Drione will also control ticks that are feeding. When pyrethrin and PBO in Drione eventually degrade, the silica aerogel carrier can continue to protect against infestation indefinitely.

Mosquitoes transmit serious diseases, which threaten humans and severely affect livestock. The half-lethal concentration (LC₅₀) was calculated by log probit analysis. The LC₅₀ and LC₉₀ values of larvicidal activity of Cassia fistula Linn. hexane-methanol soluble fraction (HMSF) after 24 h of exposure were 21.04 and 34.68 µg/ ml, respectively. The LC₅₀ values after 24 h of exposure were 84.09 µg/ml and 108.08 µg/ml for chloroform–methanol soluble fraction (CMSF) and ethyl acetate-methanol soluble fraction (EMSF) respectively. The percent hatchability of eggs exposed to the hexane extract was 90 ± 5.0, 68.33 ± 7.6, 46.6 ± 11.5, 10 ± 0.0, and 0 ± 0.0% at 10, 20, 40, 60, and 80 ppm, respectively. The pupicidal activity of the hexane extract at 40 µg/ml was 0.0%. The LC₅₀ value of adulticidal activity of the hexane extract was 12.8 mg/test tube. The biosafety of the hexane extract was assessed in nontarget organisms, i.e., zebrafish (Danio rerio) embryos and normal lung cells (BEAS-2B). The hexane extract of C. fistula was well tolerated by zebrafish embryos, and no mortality or toxicity was found in the embryos exposed to the highest tested concentration of 300 µg/ml. Similarly, all the concentrations tested against the normal lung cells (BEAS-2B) showed more than 95% survival. The gas chromatography–mass spectroscopy analysis identified 12 compounds, and 2-methyl hexanoic acid and 2-methyl butanoic acid were the major compounds identified in the hexane extract. The larvicidal activity of C. fistula extracts will help in the development of natural substitutes for vector management of mosquito populations.

Fitting long-lasting insecticidal nets (LLIN) as screens on doors/windows has a significant impact on indoor-adult Aedes aegypti (L.), with entomological reductions measured in a previous study being significant for up to 2 yr post-installation, even in the presence of pyrethroid-resistant Aedes populations. To better understand the mode of LLIN protection, bioassays were performed to evaluate the effects of field deployment (0, 6, and 12 mo) and damage type (none, central, lateral, and multiple) on LLIN efficacy. Contact bioassays confirmed that LLIN residual activity (median knockdown time, in minutes, or MKDT) decreased significantly over time: 6.95 (95% confidence interval [CI]: 5.32–8.58) to 9.24 (95% CI: 8.69–9.79) MKDT at 0- and 12-mo age, respectively, using a pyrethroid-susceptible Aedes strain. Tunnel tests (exposing human forearm for 40 min as attractant) showed that deployment time affected negatively Aedes passage inhibition from 54.9% (95% CI: 43.5–66.2) at 0 mo to 35.7% (95% CI: 16.3–55.1) at 12 mo and blood-feeding inhibition from 65.2% (95% CI: 54.2–76.2) to 48.9% (95% CI: 26.4–71.3), respectively; both the passage/blood-feeding inhibition increased by a factor of 1.8–2.9 on LLINs with multiple and central damages compared with nets with lateral damage. Mosquito mortality was 74.6% (95% CI: 65.3–83.9) at 0 mo, 72.3% (95% CI: 64.1–80.5) at 6 mo, and 59% (95% CI: 46.7–71.3) at 12 mo. Despite the LLIN physical integrity could be compromised over time, we demonstrate that the remaining chemical effect after field conditions would still contribute to killing/repelling mosquitoes.

Use of insecticidal baits risks the evolution of resistance to the feeding stimulant in the bait, not just to the active ingredient (toxicant). Sucrose-based baits are widely used against house flies, Musca domestica L. The baits are applied as dry granules, but readily liquefy. The proboscis extension reflex (PER) and consumption of alternative sweeteners, dry or in solution, were examined. Fructose, glucose, and xylitol merit further study as alternatives to sucrose. Dry, fructose, glucose, and xylitol elicited PER much more than sucrose, although not when in solution. Furthermore, dry or in solution, females and males ate as much or more fructose as sucrose. In solution, flies ate as much glucose as sucrose; although when dry, consumption was much less for glucose than sucrose. Dry, xylitol elicited as much consumption as sucrose for females, though less for males. In solution, for both sexes, xylitol elicited less consumption than sucrose did. Acesulfame potassium, sodium cyclamate, and sucralose do not look promising as they did not often elicit PER, whether dry or in solution. Erythritol also does not look promising. Erythritol elicited PER no more than sucrose did when dry and elicited PER much less than sucrose when in solution. Flies ate much less erythritol than sucrose whether dry or in solution.

Dermacentor variabilis is the predominant tick species in Nebraska and is presumed to be the primary vector of Rickettsia rickettsii associated with cases of Rocky Mountain spotted fever (RMSF). Interestingly, RMSF cases in Nebraska have increased on a year-to-year basis, yet the prevalence of R. rickettsii in D. variabilis ticks has not been established for Nebraska. Here we sought to set a baseline for the prevalence of R. rickettsii and other spotted fever group (SFG) rickettsiae harbored by D. variabilis ticks. Over a 3-yr period, D. variabilis were collected along the Platte River in south central Nebraska. Individual tick DNA was analyzed using endpoint PCR to identify ticks carrying SFG rickettsiae. In total, 927 D. variabilis were analyzed by PCR and 38 (4.1%) ticks tested positive for SFG rickettsiae. Presumptive positives were sequenced to identify the Rickettsia species, of which 29 (76%) were R. montanensis, 5 (13%) were R. amblyommatis, 4 (11%) were R. bellii, and R. rickettsii was not detected. These data indicate that R. rickettsii is likely at a low prevalence in south central Nebraska and spillover of R. amblyommatis into D. variabilis is likely occurring due to the invasive lone star tick (Amblyomma americanum). In addition, our data suggest that R. montanensis and R. amblyommatis could be associated with the increase in SFG rickettsiae infections in Nebraska.This information will be of value to clinicians and the general public for evaluating diagnosis of disease- and risk-associated environmental exposure, respectively.

Cimex hemipterus resistance to residual insecticides is a growing public health concern worldwide. Herein, we report the susceptibility status of C. hemipterus against pyrethroid (permethrin) and organophosphate (dichlorvos) insecticides in Dar es Salaam. The knockdown efficacy of dichlorvos (99%) was greater than that of permethrin (43%), and mortality at the end of 24 h was also greater for dichlorvos than for permethrin. Mortality in recommended concentration of permethrin was lower than manufacturer's expectations. In contrast, similar concentrations in dichlorvos resulted into mortality of 100% at the end of 24 h. LC_{50 and} LC_{95 values} for permethrin were 3.36 (2.356–4.364) and 2,887.39 (2,886.37–2,888.39), respectively, whereas those of dichlorvos were 7.91 (3.85–11.97) and 44.39 (40.33–48.45). Dar es Salaam city has high coverage and usage of long-lasting insecticidal nets (LLINs) for prevent malaria. The resistance in bedbugs to permethrin may be related to the widespread use of the insecticide in LLINs. We recommend of the change from pyrethroids dichlorvos for the control of bedbugs.

Cutaneous leishmaniasis (CL) by Leishmania braziliensis Vianna (Kinetoplastida: Trypanosomatidae) is a widespread disease in the western hemisphere and Brazil is the country reporting the majority of the human cases each year. Several phlebotomine sand flies have been regarded as putative or proven vectors of L. braziliensis and multiple vectors may be involved in the transmission of this parasite in the same endemic foci. In this study, we tested a subset of female phlebotomine sand flies collected in the frame of a previous study conducted in a military training camp, where outbreaks of CL by L. braziliensis have sporadically been reported. In total, 1,807 female phlebotomine sand flies were tested (individually or in pools) by real-time PCR and a minimum infection rate of 0.9% (95% CI: 0.6–1.5%) was detected. Positive females belonged to the species Psychodopygus complexus (Mangabeira) or Psychodopygus wellcomei Fraiha, Shaw & Lainson (females of these species are morphologically indistinguishable), Trichopygomyia longispina (Mangabeira), and Sciopemyia sordellii (Shannon & Del Ponte) (Diptera: Psychodidae). By restriction enzyme analysis, 13 samples (nine of Ty. longispina, two of Ps. complexus/wellcomei, and two of Sc. sordellii) presented a HaeIII restriction profile identical to L. braziliensis. The results of this study reinforce the notion that multiple vectors may be involved in the transmission of L. braziliensis in a single focus, ultimately making the epidemiological picture more complex than currently recognized.

Surveillance for diurnal container-inhabiting mosquitoes such as Aedes albopictus (Skuse), Aedes japonicus japonicus (Theobald), and Aedes triseriatus (Say) have routinely relied on the deployment of multiple trap types, including CO₂-baited light traps, gravid traps, oviposition traps, and BG-Sentinel. These trap configurations have met with varying degrees of effectiveness and in many instances likely under-sample these key mosquito vectors. Most recently, the BG-Sentinel trap used in conjunction with the human-scent lure has been largely accepted as the gold-standard for monitoring Ae. albopictus. However, its ability to attract other container-inhabiting Aedes species has not been fully evaluated. During 2018, we tested new scent lures, Trap Tech Lure-A and Lure-H (Bedoukian Research, Inc.), using BG-Sentinel traps with CO₂ in two regions of Connecticut, Stamford and Hamden, against the BG-Lure. Pooled mosquitoes were additionally screened for arbovirus infection. A total of 47,734 mosquitoes representing 8 genera and 32 species were captured during the study, with the Stamford site deriving on average three times as many mosquitoes per trap, adjusting for sampling effort. Lure-A and Lure-H outperformed the BG-Lure in terms of total numbers, diversity evenness, and the proportion of both Ae. j. japonicus and Ae. triseriatus. There were no significant differences among lures in capturing Ae. albopictus, and in terms of species richness. Fifty-seven isolates of virus (West Nile, Jamestown Canyon, and La Crosse viruses) were obtained during the study, with no significant difference between trap-lure. We highlight both novel lures as effective attractants for use in mosquito surveillance=, which either outperform, or equal, BG-Lure.

Here we conducted a systematic review and meta-analysis to reach a consensus on whether infected and uninfected mosquitoes respond differently to repellents. After screening 2,316 published studies, theses, and conference abstracts, we identified 18 studies that tested whether infection status modulated the effectiveness of repellents. Thirteen of these studies had outcomes available for meta-analysis, and overall, seven repellents were tested (typically DEET with 62% of outcomes), six mosquito species had repellence behaviors measured (typically Aedes aegypti (L.) (Diptera: Culicidae) mosquitoes with 71% of outcomes), and a broad diversity of infections were tested including Sindbis virus (Togaviridae: Alphavirus) (33% of outcomes), Dengue (Flaviviridae: Flavivirus) (31%), malaria (Plasmodium berghei Vincke & Lips (Haemospororida: Plasmodiidae) or P. falciparum Welch (Haemospororida: Plasmodiidae); 25%), Zika (Flaviviridae: Flavivirus) (7%), and microsporidia (4%). Pooling all outcomes with meta-analysis, we found that repellents were less effective against infected mosquitoes—marking an average 62% reduction in protective efficacy relative to uninfected mosquitoes (pooled odds ratio = 0.38, 95% confidence interval = 0.22–0.66; k = 96). Older infected mosquitoes were also more likely to show altered responses and loss of sensitivity to repellents, emphasizing the challenge of distinguishing between age or incubation period effects. Plasmodium- or Dengue-infected mosquitoes also did not show altered responses to repellents; however, Dengue–mosquito systems used inoculation practices that can introduce variability in repellency responses. Given our findings that repellents offer less protection against infected mosquitoes and that these vectors are the most dangerous in terms of disease transmission, then trials on repellent effectiveness should incorporate infected mosquitoes to improve predictability in blocking vector–human contact.

Q fever is a worldwide zoonosis caused by Coxiella burnetii (Derrick) Philip. It is a major cause of abortion among sheep and may be responsible for reproductive losses in red deer in Spain. Airborne transmission is the most widespread; however, some studies suggested that ticks may play a role, but little is known about their actual involvement in the C. burnetii cycle.The aim of this study was to determine the role that Hyalomma lusitanicum (Koch) tick plays in the maintenance of this agent among wildlife in the meso-Mediterranean areas. We processed by PCR 53 swabs from wild rabbits, 21 liver samples from red deer, and 236 ticks collected at different stages. Coxiella burnetii DNA was detected in 43.40% of wild rabbits and 38.09% of red deer, supporting the hypothesis that these animals are quite likely to serve as a reservoir in the field. We also found a high prevalence of C. burnetii in ticks (55.66%). It is worth noting that 50.45% of positive ticks were collected from negative hosts, suggesting that the pathogen probably was acquired at a previous tick stage. Our results suggest transstadial transmission, and the presence of bacterial DNA in the offspring of positive female ticks is the first evidence of the transovarial transmission of C. burnetii by H. lusitanicum.Thus, this tick species seems to play an important role as a bridge of infection in the wildlife cycle, although further studies are needed to confirm vector competence.

The analysis of vertebrate blood meals serves as an integral component of vector incrimination studies where feeding preferences and host associations influence vector-borne disease transmission. Diagnostic polymerase chain reaction (PCR)-based techniques have been widely used to determine host associations, yet applications for Culex (Diptera: Culicidae), which feed primarily on bird populations, have been limited by multistep PCR techniques that approach each potential host species singly. As a result, we have developed a multiplexed primer set targeting mitochondrial cytochrome b sequences that can distinguish human, bird, and mammalian host blood meals in a single PCR reaction, an improvement over previous analyses relying on single primers or other multiplex primer approaches through the inclusion of avian primers. To validate this new methodology, we demonstrate its application on blood samples as well as field-collected Culex samples. Although designed for applications with mosquito vectors, this multiplex PCR assay is not mosquito-specific, and should serve as a valuable tool for identifying the blood meals of other blood-feeding arthropods, contributing greatly to the study of vector-borne disease.

Avian influenza virus (AIV) H9N2 emerged in the 1990s as an economically important disease in poultry and occasionally infects humans and other mammals.The aim of this study was to evaluate the acquisition and retention of H9N2 AIV on and within the house fly, Musca domestica (Linnaeus 1758), under laboratory conditions. In first experiment, 100 adult house flies were divided into control and treatment groups equally.Treatment group was fed with a meal containing H9N2 virus, while control group was supplied with an identical meal without virus. Fifteen minutes after exposure in each group, flies were washed twice to remove surface particles, disinfected and then homogenized for testing.The two external body surface washes and the homogenate samples were tested for H9N2 to distinguish exterior from interior viral load. Second experiment was performed likewise but five flies from each group were taken at 0, 6, 24, 48, 72, 96, and 120 h post-exposure. All samples were subjected to real-time reverse-transcription polymerase chain reaction (RRT-PCR) for detecting H9-Specific viral RNA. Results of the first experiment showed that viral RNA was detectable in both of external surface and homogenates samples. Second experiment revealed that persistence of H9N2 AIVs on external body surface and within the body of M. domestica were 24 and 96 h, respectively. Moreover, viral RNAs concentration declined during the time after exposure to AIV H9N2 either outside or within house flies. Overall, house fly was able to acquire and preserve H9N2 AIV experimentally, which may contribute the spread of virus among poultry farms.

A model is developed of malaria (Plasmodium falciparum) transmission in vector (Anopheles gambiae) and human populations that include the capacity for both clinical and parasite suppressing immunity.This model is coupled with a population model for Anopheles gambiae that varies seasonal with temperature and larval habitat availability. At steady state, the model clearly distinguishes uns hypoendemic transmission patterns from stable hyperendemic and holoendemic patterns of transmission.The model further distinguishes hyperendemic from holoendemic disease based on seasonality of infection. For hyperendemic and holoendemic transmission, the model produces the relationship between entomological inoculation rate and disease prevalence observed in the field. It further produces expected rates of immunity and prevalence across all three endemic patterns. The model does not produce mesoendemic transmission patterns at steady state for any parameter choices, leading to the conclusion that mesoendemic patterns occur during transient states or as a result of factors not included in this study.The model shows that coupling the effect of varying larval habitat availability with the effects of clinical and parasite-suppressing immunity is enough to produce known patterns of malaria transmission.

It has been reported that starving ticks do not transmit spotted fever group Rickettsia immediately upon attachment because pathogenic bacteria exist in a dormant, uninfectious state and require time for ‘reactivation’ before transmission to a susceptible host. To clarify the length of reactivation period, we exposed guinea pigs to bites of Rickettsia rickettsii-infected Dermacentor variabilis (Say) and allowed ticks to remain attached for predetermined time periods from 0 to 48 h. Following removal of attached ticks, salivary glands were immediately tested by PCR, while guinea pigs were observed for 10–12 d post-exposure. Guinea pigs in a control group were subcutaneously inoculated with salivary glands from unfed D. variabilis from the same cohort. In a parallel experiment, skin at the location of tick bite was also excised at the time of tick removal to ascertain dissemination of pathogen from the inoculation site. Animals in every exposure group developed clinical and pathological signs of infection. The severity of rickettsial infection in animals increased with the length of tick attachment, but even attachments for less than 8 h resulted in clinically identifiable infection in some guinea pigs. Guinea pigs inoculated with salivary glands from unfed ticks also became severely ill. Results of our study indicate that R. rickettsii residing in salivary glands of unfed questing ticks does not necessarily require a period of reactivation to precede the salivary transmission and ticks can transmit infectious Rickettsia virtually as soon as they attach to the host.

Fleas transmit a variety of pathogens to humans but are relatively understudied in comparison to mosquitoes and ticks, including in Taiwan, where fleas in rural lowlands have never been systematically surveyed. In total, 700 fleas of four species were collected from 1,260 shrews and rodents at nine counties across lowland Taiwan. Nosopsyllus nicanus Jordan (Siphonaptera: Ceratophyllidae) and Xenopsylla cheopis Rothschild (Siphonaptera: Pulicidae) were the most abundant flea species (79.0 and 14.6% of total fleas, respectively); the former was largely limited to the islets, while the latter was restricted to the Taiwan main island. Rattus losea Swinhoe (Rodentia: Muridae) was the most common small mammal species (49.3% of total) and hosted the majority of fleas (88.3% of total). Five Rickettsia spp., including Rickettsia conorii Brumpt (Rickettsiales: Rickettsiaceae), Rickettsia felis Bouyer et al. Rickettsia japonica Uchida, Rickettsia raoultii Mediannikov, and Rickettsia rickettsii Brumpt or closely related species, were identified from 67 individually assayed fleas based on ompB and gltA genes. Rickettsia felis, mainly transmitted by fleas, was detected in one X. cheopis in southern Taiwan where a confirmed human case of infection with R. felis has been reported. The presence of R. felis, along with the other four tick-borne Rickettsia spp., demonstrates that a variety of rickettsiae circulate in rural lowland Taiwan and could pose risks to human health.

Sand flies (Diptera: Psychodidae) are natural vectors of Leishmania. For the initiation of sand fly experimental infections either Leishmania amastigotes or promastigotes can be used. In order to obtain comparable results, it is necessary to adjust and standardize procedures. During this study, we conducted promastigote- and amastigote-initiated infections of Leishmania infantum Nicolle, 1908 parasites in Phlebotomus (Larroussius) perniciosus Newstead, 1911 in two laboratories with different levels of biosafety protection. Protocol originally designed for a biosafety level 2 facility was modified for biosafety level 3 facility and infection parameters were compared. Particularly, specially designed plastic containers were used for blood feeding; feeders were placed outside the sand fly cage, on the top of the mesh; feeding was performed inside the climatic chamber; separation of engorged females was done in Petri dishes kept on ice; engorged females were kept in the cardboard containers until dissection. All experiments, conducted in both laboratories, resulted in fully developed late stage infections with high number of parasites and colonization of the stomodeal valve. We demonstrated that protocol originally designed for biosafety level 2 facilities can be successfully modified for other biosafety facilities, depending on the special requirements of the individual institution/laboratory.

The dog heartworm, Dirofilaria immitis (Leidy) (Spirurida: Onchocercidae), is a devastating parasite of domestic and wild canines vectored by a multitude of mosquito species. Although many species are implicated as vectors, not all contribute equally to disease transmission, with demonstrated variation in vector efficiency between and within species. We investigated the vector efficiency of mosquitoes derived from wild-caught North Carolina populations of two known heartworm vectors: a native species, Aedes triseriatus (Say) (Diptera: Culicidae), and an invasive species, Aedes albopictus (Skuse). We compared the parasite developmental times within the mosquito, mosquito longevity and fecundity, and the vector efficiency index between the two species. We found that the tested composite North Carolina population of Ae. triseriatus was an efficient vector of D. immitis under laboratory conditions, whereas the local composite population of Ae. albopictus was a competent but relatively poor vector. Compared with Ae. triseriatus, Ae. albopictus showed a longer time for parasite development, lower infection rates, and lower vector efficiency. Additionally, Ae. albopictus was the sole species to exhibit significant parasite-induced mortality. These results are in contrast to prior studies of populations of Ae. albopictus from locations outside of North Carolina, which have implicated the species as a highly competent heartworm vector.The variation seen for different strains of the same species emphasizes the heritable nature of D. immitis vector competence and highlights the need for local infection studies for accurate transmission risk assessment in a particular locale.

Mosquitoes pose an increasing risk in urban landscapes, where spatial heterogeneity in juvenile habitat can influence fine-scale differences in mosquito density and biting activity. We examine how differences in juvenile mosquito habitat along a spectrum of urban infrastructure abandonment can influence the adult body size of the invasive tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae). Adult Ae. albopictus were collected across 3 yr (2015–2017) from residential blocks in Baltimore, MD, that varied in abandonment level, defined by the proportion of houses with boarded-up doors. We show that female Ae. albopictus collected from sites with higher abandonment were significantly larger than those collected from higher income, low abandonment blocks. Heterogeneity in mosquito body size, including wing length, has been shown to reflect differences in important traits, including longevity and vector competence. The present work demonstrates that heterogeneity in female size may reflect juvenile habitat variability across the spatial scales most relevant to adult Aedes dispersal and human exposure risk in urban landscapes. Previous work has shown that failure to manage abandonment and waste issues in impoverished neighborhoods supports greater mosquito production, and this study suggests that mosquitoes in these same neighborhoods could live longer, produce more eggs, and have different vector potential.

A Musca domestica L. strain collected from Pakistan has recently been shown to be resistant to spinosad; however, there is scarce information about the mechanism of resistance. For this reason, we explored whether a metabolic-based mechanism was responsible by analyzing the activities of the metabolic detoxifying enzymes, carboxylesterases, glutathione S-transferases, and mixed-function oxidases, in both a spinosad-selected (Spin-SEL) strain of M. domestica and a susceptible counterpart (Lab-susceptible). The results revealed that both strains were statistically at par in terms of enzyme activities. The activity of carboxylesterases in the Lab-susceptible strain was 78.17 ± 3.06 in comparison to 79.16 ± 3.31 nmol min^–1 mg^–1 in the Spin-SEL strain. The activity of mixed-function oxidases was 51.58 ± 4.20 in the Lab-susceptible strain, whereas 54.33 ± 4.08 pmol min^–1 mg^–1 was recorded in the Spin-SEL strain. The activity of glutathione S-transferases was 86.50 ± 4.59 (Lab-susceptible) and 90.33 ± 2.81 nmol min^–1 mg^–1 (Spin-SEL). These results revealed that the studied enzymes might not be responsible for spinosad resistance in the studied strain of M. domestica. Therefore, studies should be extended to find out other possible mechanisms of spinosad resistance.

Drag-cloth sampling is the most commonly used method to sample for ticks. A cloth is dragged along the ground and checked for ticks at regular intervals to count ticks before they drop off.The distance between drag-cloth checks differs between studies, which could result in lower density estimates for studies with greater distances between checks. Here, we measured this effect by 1) calculating the rate at which nymphal and adult Ixodes scapularis Say ticks drop off the cloth per meter dragged and 2) measuring tick density by drag-cloth sampling with three different drag-cloth check interval distances. We found a higher drop-off rate for adult ticks, 0.083/m, than nymphal ticks, 0.047/m.The estimated density of ticks decreased with increasing check interval distance. Our results not only highlight the importance of accounting for check interval distance when estimating tick density, but also provide the first estimate of nymphal I. scapularis drop-off rate.

Eugenol is a major component of the essential oils in cloves and other aromatic plants. In insects, it produces toxic effects and repellency, and there is evidence that its site of action is the octopamine receptor. The objective of the present study was to explore whether the octopamine receptor is involved in the hyperactivity produced by eugenol in the blood-sucking bug Triatoma infestans (Klug). This insect is the main vector of Chagas disease in Latin America. Four treatments were topically applied on third instar nymphs: 1) octopamine, 2) eugenol, 3) phentolamine hydrochloride (an antagonist of the octopamine receptor) followed by octopamine, and 4) phentolamine hydrochloride followed by eugenol. Both octopamine and eugenol hyperactivated the nymphs. However, pretreatment with phentolamine hydrochloride inhibited the hyperactivating effect of both compounds. These results are in agreement with previous works on Drosophila melanogaster (Meigen) (Diptera: Drosophilidae) and the American cockroach. They suggest that the octopamine receptor is a possible site of action for eugenol.

Theileria spp. are tick-transmitted, intracellular apicomplexan protozoan parasites that infect a wide range of animals and, as such, can cause significant economic losses. The aim of the present study was to detect and analyze apicomplexan parasites from two different ectoparasites that were collected from Xinjiang Uygur Autonomous Region, China. The PCR-based detection of 18S rRNA indicated that Ornithodoros lahorensis specimens from Kashgar, Xinjiang, and Aksu were positive for Theileria spp., as were Melophagus ovinus specimens from Aksu. Meanwhile, phylogenetic analysis, based on the 18S rRNA gene sequences, revealed that the four amplified Theileria sequences could be attributed to T. ovis. To the best of our knowledge, this is the first study to report the detection of T. ovis DNA in M. ovinus and the first molecular identification study to confirm the detection of T. ovis in O. lahorensis in China. Accordingly, the present study extends the known distribution of T. ovis.

The adulticidal activity of six fungal strains of Metarhizium anisopliae (Metschn.) Sorokin s.l. (Hypocreales: Clavicipitaceae) against Aedes aegypti (L.) (Diptera: Culicidae) was assessed. These strains (CEP 085, CEP 087, CEP 120, CEP 350, CEP 404, and CEP 423) were isolated from soil samples or nondipteran hosts collected from areas in Argentina where Ae. aegypti is distributed. Bioassays were performed with four conidial concentrations plus a control of each fungal strain to determine the lethal concentrations (LC₅₀/LC₉₀), the median survival times (ST₅₀), the mean percentage of the surviving individuals, and the mean percentage of mycotized cadavers. The strains were able to infect and kill adult Ae. aegypti. Significant differences were found among the LC₅₀ values, with CEP 423 as the most virulent strain with the lowest LC₅₀ (2.4 × 10⁶ conidia/ml). At 1 × 10⁷ conidia/ml: no significant differences were found in the Kaplan–Meier survival functions among the strains; the ST₅₀ ranged from 5 (CEP 085) to 8 d (CEP 350); and the mean percentage of the surviving adults was between 13.3 (CEP 085, CEP 350 and CEP 423) and 46.7% (CEP 087). Significant differences were also found among strains in the mean percentage of cadavers with fungal sporulation. Strain CEP 423 produced the highest percentage of mycotized adults (70%). The concentration of the inoculum significantly affected the survival of individuals and the percentage of mycotized cadavers within each strain. Metarhizium anisopliae s.l. CEP 423 was selected as the most promising candidate for further research aiming to develop a mycoinsecticide against Ae. aegypti.

The age of a blow fly larval specimen found on a body can be used to estimate a minimum postmortem interval by comparing a physical feature of a larva (e.g., length) to a reference growth curve created under similar conditions. A better understanding of factors known to influence growth rates would lead to more precise estimates of larval age. A factor known to influence insect development, but almost always lacking in blow fly larval growth curves, is sex. We wanted to understand how sex might affect larval growth rates and therefore lead to more precise age estimates, in the forensically important blow fly Chrysomya megacephala (F.). We examined sex for an entire age cohort at 70 h and found on average, males were longer than females. To assess whether this difference would cause an investigator to interpret the results differently if comparing a larval specimen of one sex based on reference data from the other sex, we made sex-specific 95% prediction intervals. We did not find a difference when comparing a male specimen with the female prediction interval, but did find more females outside the interval than expected using the male interval. All females outside the male interval were shorter. Investigators commonly remove the largest individuals from a body to estimate age, so these females are less likely to be chosen. These results do not support the generation of sex-specific growth curves for this species, though this study's narrow scope means more information is required before making a final conclusion.

Culex pipiens Linnaeus and Culex quinquefasciatus Say are the primary vectors of West Nile and St. Louis encephalitis viruses in California. Pyrethrins and pyrethroids (synthetic pyrethrins) are the most widely used insecticides to control adult stage mosquitoes to prevent disease transmission. The most abundant and widespread mutation associated with pyrethroid resistance is the L1014F mutation of the voltage-sensitive sodium channel gene. Statewide, based on the testing of almost 2,000 mosquitoes from 14 counties, the resistant allele frequency was 71%. Although the L1014F mutation was found in all counties assessed, the resistance allele profiles differed between regions of California. The highest resistant allele frequency occurred in the Central region and lowest frequencies were from the Northern and Southern regions. Resistance allele frequencies observed in 2014–2016 are nearly 1.5 times higher than those from pre-2012, indicating that resistance profiles can change over time. Regular monitoring of the L1014F kdr mutation will help aid in operational decisions.

The use of stable isotope enrichment to mark mosquitoes has provided a tool to study the biology of vector species. In this study, we evaluated isotopic marking of Aedes aegypti (L.) (Diptera: Culicidae) in a laboratory setting. We determined the optimal dosage for marking adult Ae. aegypti mosquitoes with ¹³C and ¹⁵N. Additionally, Ae. aegypti mosquitoes were single and dually marked with ¹³C and ¹⁵N for up to 60 d postemergence without changes to adult body size or transgenerational marking. This report adds to the growing literature that explores the use of alternative marking methods for ecological and vector biology studies.