The introduction of West Nile virus (WNV) to North America in 1999 and its subsequent rapid spread across the Americas demonstrated the potential impact of arboviral introductions to new regions, and this was reinforced by the subsequent introductions of chikungunya and Zika viruses. Extensive studies of host–pathogen–vector–environment interactions over the past two decades have illuminated many aspects of the ecology and evolution of WNV and other arboviruses, including the potential for pathogen adaptation to hosts and vectors, the influence of climate, land use and host immunity on transmission ecology, and the difficulty in preventing the establishment of a zoonotic pathogen with abundant wildlife reservoirs. Here, we focus on outstanding questions concerning the introduction, spread, and establishment of WNV in the Americas, and what it can teach us about the future of arboviral introductions. Key gaps in our knowledge include the following: viral adaptation and coevolution of hosts, vectors and the virus; the mechanisms and species involved in the large-scale spatial spread of WNV; how weather modulates WNV transmission; the drivers of large-scale variation in enzootic transmission; the ecology of WNV transmission in Latin America; and the relative roles of each component of host–virus–vector interactions in spatial and temporal variation in WNV transmission. Integrative studies that examine multiple factors and mechanisms simultaneously are needed to advance our knowledge of mechanisms driving transmission.

Since West Nile virus (WNV) emerged in the United States in 1999, 22,999 neuroinvasive disease cases in humans were reported through 2017. These cases have arisen from an estimated seven million human infections. Population incidence is geographically heterogeneous and is highest in the West and Midwest. Upwards of 2% of the population in some jurisdictions may become infected during outbreaks. Before universal screening of the United States blood supply, this high infection incidence and that approximately 75% of those infected remain asymptomatic translated into a considerable risk of WNV transfusion transmission despite the short duration of viremia following infection. Universal blood donor screening has nearly eliminated the risk of WNV transfusion transmission, but at enormous cost. WNV transmission via transplanted organs carries extremely high morbidity and mortality. Improved vector surveillance and timely and effective response to surveillance data can reduce the impact of WNV and should remain public health priorities.

West Nile virus (WNV) (Flaviviridae: Flavivirus) was discovered in Africa more than 80 yr ago and became recognized as an avian pathogen and a cause of neurologic disease in horses largely during periodic incursions into Europe. Introduction of WNV into North America stimulated great anxiety, particularly in the equine industry, but also for pet owners and livestock producers concerned about the effect of WNV on other domestic animals. Numerous subsequent studies of naturally occurring and experimentally induced disease greatly expanded our understanding of the host range and clinical consequences of WNV infection in diverse species and led to rapid development and deployment of efficacious vaccines for horses. In addition to humans, horses are clearly the animals most frequently affected by serious, sometimes lethal disease following infection with WNV, but are dead-end hosts due to the low-magnitude viremia they develop. Dogs, cats, and livestock species including chickens are readily infected with WNV, but only occasionally develop clinical disease and are considered dead-end hosts for the virus.

West Nile virus (WNV) was first identified in North America almost 20 yr ago. In that time, WNV has crossed the continent and established enzootic transmission cycles, resulting in intermittent outbreaks of human disease that have largely been linked with climatic variables and waning avian seroprevalence. During the transcontinental dissemination of WNV, the original genotype has been displaced by two principal extant genotypes which contain an envelope mutation that has been associated with enhanced vector competence by Culex pipiens L. (Diptera: Culicidae) and Culex tarsalis Coquillett vectors. Analyses of retrospective avian host competence data generated using the founding NY99 genotype strain have demonstrated a steady reduction in viremias of house sparrows over time. Reciprocally, the current genotype strains WN02 and SW03 have demonstrated an inverse correlation between house sparrow viremia magnitude and the time since isolation. These data collectively indicate that WNV has evolved for increased avian viremia while house sparrows have evolved resistance to the virus such that the relative host competence has remained constant. Intrahost analyses of WNV evolution demonstrate that selection pressures are avian species-specific and purifying selection is greater in individual birds compared with individual mosquitoes, suggesting that the avian adaptive and/or innate immune response may impose a selection pressure on WNV. Phylogenomic, experimental evolutionary systems, and models that link viral evolution with climate, host, and vector competence studies will be needed to identify the relative effect of different selective and stochastic mechanisms on viral phenotypes and the capacity of newly evolved WNV genotypes for transmission in continuously changing landscapes.

In North America, the geographic distribution, ecology, and vectorial capacity of a diverse assemblage of mosquito species belonging to the genus Culex determine patterns of West Nile virus transmission and disease risk. East of the Mississippi River, mostly ornithophagic Culex pipiens L. complex mosquitoes drive intense enzootic transmission with relatively small numbers of human cases. Westward, the presence of highly competent Culex tarsalis (Coquillett) under arid climate and hot summers defines the regions with the highest human risk. West Nile virus human risk distribution is not uniform geographically or temporally within all regions. Notable geographic ‘hotspots' persist with occasional severe outbreaks. Despite two decades of comprehensive research, several questions remain unresolved, such as the role of non-Culex bridge vectors, which are not involved in the enzootic cycle, but may be involved in virus transmission to humans. The absence of bridge vectors also may help to explain the frequent lack of West Nile virus ‘spillover’ into human populations despite very intense enzootic amplification in the eastern United States. This article examines vectorial capacity and the eco-epidemiology of West Nile virus mosquito vectors in four geographic regions of North America and presents some of the unresolved questions.

The introduction of West Nile virus to North America in 1999 had profound impacts on human and wildlife health. Here, we review studies of WNV impacts on bird populations and find that overall impacts have been less than initially anticipated, with few species showing sustained changes in population size or demographic rates across multiple regions. This raises four questions: 1) What is the evidence for WNV impact on bird populations and how can we strengthen future analyses? We argue that future studies of WNV impacts should explicitly incorporate temporal variation in WNV transmission intensity, integrate field data with laboratory experimental infection studies, and correct for multiple comparisons. 2) What mechanisms might explain the relatively modest impact of WNV on most bird populations? We suggest that spatial and temporal variation in WNV transmission moderates WNV impacts on species that occur in multiple habitats, some of which provide refugia from infection. 3) Have species recovered from the initial invasion of WNV? We find evidence that many species and populations have recovered from initial WNV impact, but a few have not. 4) Did WNV cause cascading effects on other species and ecosystems? Unfortunately, few studies have examined the cascading effects of WNV population declines, but evidence suggests that some species may have been released from predation or competition. We close by discussing potentially overlooked groups of birds that may have been affected by WNV, and one highlight species, the yellow-billed magpie (Pica nutalli Audubon, 1837 [Passeriformes: Corvidae]), that appears to have suffered the largest range-wide impact from WNV.

The establishment of a tropical virus such as West Nile (WNV; Flaviviridae: Flavivirus) within the temperate latitudes of the continental United States was unexpected and perhaps contingent, in part, upon the ability of this invasive virus to persist during winter when temperatures become too cold for replication and vector mosquito gonotrophic activity. Our Forum article reviews research examining possible overwintering mechanisms that include consistent reintroduction and local persistence in vector mosquitoes and avian hosts, mostly using examples from research conducted in California. We conclude that the transmission of WNV involves so many vectors and hosts within different landscapes that multiple overwintering pathways are possible and collectively may be necessary to allow this virus to overwinter consistently within the United States.

Over the past 20 yr, many models have been developed to predict risk for West Nile virus (WNV; Flaviviridae: Flavivirus) disease in the human population. These models have aided our understanding of the meteorological and land-use variables that drive spatial and temporal patterns of human disease risk. During the same period, electronic data systems have been adopted by surveillance programs across much of the United States, including a growing interest in integrated data services that preserve the autonomy and attribution of credit to originating agencies but facilitate data sharing, analysis, and visualization at local, state, and national scales. At present, nearly all predictive models have been limited to the scientific literature, with few having been implemented for use by public-health and vector-control decision makers. The current article considers the development of models for spatial patterns, early warning, and early detection of WNV over the last 20 yr and considers some possible paths toward increasing the utility of these models for guiding interventions.

Over 50,000 human West Nile virus (WNV) (Flaviviridae: Flavivirus) clinical disease cases have been reported to the CDC during the 20 yr that the virus has been present in the United States. Despite the establishment and expansion of WNV-focused mosquito surveillance and control efforts and a renewed emphasis on applying integrated pest management (IPM) principles to WNV control, periodic local and regional WNV epidemics with case reports exceeding 2,000 cases per year have occurred during 13 of those 20 yr in the United States. In this article, we examine the scientific literature for evidence that mosquito control activities directed at either preventing WNV outbreaks or stopping those outbreaks once in progress reduce WNV human disease or have a measurable impact on entomological indicators of human WNV risk. We found that, despite a proliferation of research investigating larval and adult mosquito control effectiveness, few of these studies actually measure epidemiological outcomes or the entomological surrogates of WNV risk. Although many IPM principles (e.g., control decisions based on surveillance, use of multiple control methodologies appropriate for the ecosystem) have been implemented effectively, the use of action thresholds or meaningful public health outcome assessments have not been used routinely. Establishing thresholds for entomological indicators of human risk analogous to the economic injury level and economic thresholds utilized in crop IPM programs may result in more effective WNV prevention.

The purpose of this review is to elaborate the role of Periplaneta (P.) americana L. in modern and traditional Chinese medicine (TCM) and compare the use of the species in these two forms of medical treatments. From searches on Google Scholar, PubMed, and Web of Science databases, studies were identified involving TCMs with P. americana, which have a history of use over several thousand years, and demonstrate how extracts from this insect play a role in the treatment of diseases through antibacterial, antiviral, antitumor activity, and enhancement of immune function. Extracts from P. americana have not been fully developed for clinical use because the active components have not been completely purified or their molecular mechanisms thoroughly understood. The development of extraction technology in modern Chinese medicine has revealed that many extracts from P. americana are able to play an important role in the control of diseases such as cancer. Drugs such as ‘Kangfuxin Solution’ and ‘Xinmailong Injection’ are now widely used for gastrointestinal ulcers and chronic heart failure, having achieved beneficial curative effects in clinical studies. Based on this, the information from studies of P. americana in TCM and modern medicine should be combined and their respective advantages applied. This review provides an overview of the role of P. americana in modern and TCM and thus contributes to identification of further applications and area requiring drug development.

Locating and counting parasites on a host is a fundamental aspect of ecological research and basic veterinary and clinical practice. Variability in the biology and behavior of both hosts and parasites creates many methodological, logistical, and ethical considerations that must be made to collect this deceptively simple measurement. We identified methods that are used to count ticks (Acari: Ixodida, Leach 1815) on hosts by reviewing the methods sections of relevant published studies. Unfortunately, there is no best method agreed upon by scientists to collect ticks from hosts. In general, we suggest that studies focusing purely on counting ticks on hosts should use more sensitive methods to determine patterns of tick distribution on the surfaces of unconscious or deceased hosts in order to provide host body regions to target in future studies to maximize tick detection ability and limit the costs of research for researchers and the host animals involved. As ticks are counted on hosts for many different reasons, researchers must be goal oriented and chose methods that are appropriate for addressing their specific aims.

Cockroaches have the potential to disseminate bacteria in their environments and therefore a systematic review and meta-analysis was conducted to assess the state-of-the-art of our knowledge regarding bacterial contaminants of cockroaches. At least 78 bacterial species and 42 genera from 24 families and 11 orders of bacteria were reported to have contaminated cockroaches. At least 61, 42, 12, 13, 7, and 16 bacterial species have contaminated Blattella germanica (L.) (Blattaria: Ectobiidae), Periplaneta americana (L.) (Blattaria: Blattidae), Blatta orientalis (L.) (Blattaria: Blattidae), Diploptera punctata (Eschscholtz) (Blattaria: Blaberidae), Periplaneta fuliginosa (Serville) (Blattaria: Blattidae), and Supella longipalpa (F.) (Blattaria: Ectobiidae), respectively. Blattella germanica is the most commonly contaminated cockroach species, with the widest bacterial species diversity that threatens human health, followed by P. americana. Cockroach bacterial contaminants may result in the dissemination of opportunistic or pathogenic infections, particularly nosocomial and foodborne infections. One-way analysis of variance (ANOVA) revealed significant differences between bacterial contaminant species of cockroaches, species of cockroaches with bacterial contaminants, cockroach body part surfaces from the point of view bacterial contaminants and countries of cockroach infested with bacterial contaminants (P < 0.05). This statistical analysis indicates that the bacterial contaminants of the external cockroach body parts are potentially more harmful than from internal surfaces, and secondly, the bacterial contaminants of cockroaches in hospital environments are potentially more harmful than from other human environments. The survey indicated that the bacterial contaminant species of cockroaches appear to be mostly multiple drug resistant. The challenges of cockroaches as being potential vectors of pathogenic or opportunistic agents of human infections are discussed.

Ticks are widespread vectors for many important medical and veterinary infections, and a better understanding of the factors that regulate their population dynamics is needed to reduce risk for humans, wildlife, and domestic animals. Most ticks, and all non-nidicolous tick species, spend only a small fraction of their lives associated with vertebrate hosts, with the remainder spent in or on soils and other substrates. Ecological studies of tick-borne disease dynamics have emphasized tick–host interactions, including host associations, burdens, and efficiencies of pathogen transmission, while under emphasizing tick biology during off-host periods. Our ability to predict spatiotemporal trends in tick-borne diseases requires more knowledge of soil ecosystems and their effect on host and tick populations. In this review, we focus on tick species of medical and veterinary concern and describe: 1) the relationships between soil factors and tick densities; 2) biotic and abiotic factors within the soil ecosystem that directly affect tick survival; 3) potential indirect effects on ticks mediated by soil ecosystem influences on their vertebrate hosts; 4) the potential for tick-mediated effects on vertebrate host populations to affect ecosystems; and 5) possible nontarget impacts of tick management on the soil ecosystem. Soils are complex ecosystem components with enormous potential to affect the survival and behavior of ticks during their off-host periods. Hence, tick-borne disease systems present an excellent opportunity for soil ecologists and public health researchers to collaborate and improve understanding of these medically important and ecologically complex disease cycles.

Triatominae bugs (Hemiptera: Reduviidae) are usually associated with different vertebrate species, upon which many of them feed. Yet how these different blood meal sources influence key biological parameters is rarely investigated for triatomines. To fill this knowledge gap, this study sought to determine the effect of a domestic rat species (Rattus norvegicus Berkenhout (Rodentia: Muridae)), a domestic mice species (Mus musculus L. (Rodentia: Muridae)), and chickens (Gallus gallus domesticus L. (Galliformes: Phasianidae)), as blood meal sources upon several biological parameters (development time, number of required blood meals to moult and feeding and defecation behaviors) of the Mexican major vector Triatoma barberi Usinger. The three studied cohorts' development times were similar (325–338 d), but the number of required blood meals to moult (21), as well as the total mortality rate (26%), were both the highest in the cohort that fed on chickens. The longevity of females (186–190 d) was similar among the three studied cohorts, as was that of males. The median time elapsed between the presentation of a blood meal source and onset of feeding (10 min) was similar among the three studied cohorts, as were their feeding times and defecation patterns. Most of our studied parameters demonstrate how T. barberi can effectively take advantage of feeding on rodents as much as it does on hens. Those parameter results also show that T. barberi should be considered as a potential yet underappreciated vector in some areas, thus warranting a surveillance program of its current distribution area in Mexico.

Chrysomya megacephala Fabricius is one of the most common myiasis-causing and carrion-feeding fly species in the tropics. This species has a worldwide distribution due to globalization and therefore is of critical importance to the public health sector and forensic investigations. Development studies carried worldwide show that Ch. megacephala reared at similar temperatures may require different amounts of time to complete their cycle of growth. The present study determined for the first time the life-history information of this species in Sri Lanka. Chrysomya megacephala colonies were reared on three different food sources (bovine muscle, swine muscle, and swine liver) considering the most common forensic entomology rearing mediums and the capability of Ch. megacephala to cause myiasis in cattle. Furthermore, colonies were reared at four temperature regimes representing several specific development conditions of this blowfly: 1) 20°C representing the estimation of the lower developmental threshold for this species in Sri Lanka; 2) 25 and 27°C representing typical room/environmental temperatures in Sri Lanka; and 3) 38°C as this is the typical living animal temperature, representing myiasis conditions. Results show that temperature significantly affected larval length and width over time; however, tissue type was not a significant factor. The fastest development was recorded at 38°C for immature feeding on bovine muscle (162.15 h), followed by those fed swine liver (184.15 h) and swine muscle (208.00 h). The calculated minimum temperature threshold for Ch. megacephala was 13°C. Data generated will be crucial for future forensic investigations involving living or deceased individuals colonized by this species.

The impressive Sacristy of the Basilica of San Domenico Maggiore contains 38 wooden sarcophagi with the bodies of 10 Aragonese princes and other Neapolitan nobles, who died in the 15th and 16th centuries. To improve the knowledge about the entomofauna associated with bodies in archaeological contexts, herein we provide insights on the funerary practices and the insect community associated to Ferrante II King of Naples and other Italian Renaissance mummies of the Aragonese dynasty buried in the Basilica of St. Domenico Maggiore. We identified 842 insect specimens: 88% were Diptera (Muscidae, Fanniidae, and Phoridae), followed by 9% Lepidoptera (Tineidae) and 3% Coleoptera (Dermestidae and Ptinidae). Ninety-seven percent of the specimens were collected from the coffin of Francesco Ferdinando d'Avalos, which was the best preserved. A lack of fly species characterizing the first colonization waves of exposed bodies was noted.The most common fly was the later colonizing muscid Hydrotaea capensis (Wiedemann); only a few Fanniidae (Fannia spp.) were retrieved. The lack of blowflies, coupled with recording H. capensis as the dominant fly, supports our hypothesis that corpses have been kept indoors for a long time under confined environmental conditions. Other explanations include odorous oils/balms having been used in the embalming process, causing the delay or stopping the arrival of first colonizer flies. Hermetically sealing of the coffin with bitumen may also have played a role in preventing access to the corpses.This scenario describes a historical context characterized by a well-advanced knowledge of body preparation, with specific burial techniques adopted for nobles.

The decomposition of a living being involves a series of changes produced by a number of interacting abiotic and biotic factors. In this study, we analyzed the effect of the colonization of blowflies on the decomposition of chicken carcasses and on the emission of sulfur compounds.The loss of the mass of carcasses and the release rate of sulfur compounds were compared for 30 d in chicken carcasses with and without blowflies in field conditions.The tissue degradation was slower in the carcasses without insects compared to those colonized by blowflies.The decomposition stages of fresh, bloated, active decay, and advanced decay were observed in the carcasses without flies; while the decomposition stages of fresh, active decay, advanced decay, and dry remains were identified in carcasses with flies.Two sulfur compounds, dimethyl disulfide and dimethyl trisulfide, were present during the entire decomposition process.The emission of these compounds is not directly associated with the presence of the blowflies' immature stages during the whole decomposition process. However, in cadavers with insects, the highest emission of both compounds occurred in day 2, while in cadavers without insects, the peak of emission was observed in day 4. In addition, the presence of the larval stages I and II of Lucilia eximia (Wiedemann, 1819) (Diptera: Calliphoridae), Chrysomya rufifacies (Macquart, 1842) (Diptera: Calliphoridae), Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae), and Cochliomyia macellaria Fabricius, 1775 (Diptera: Calliphoridae) matched with the peak of emission of both compounds.

The inhibitory effect of Chrysomya rufifacies (Macquart) and Cochliomyia macellaria (Fabricius) larval excretions–secretions (ES) on Staphylococcus aureus was determined using a portable colorimetric method without the need for any dedicated spectral instrument. Polystyrene 96 well microplates were used and 100 µl of the bacterial inoculum (5 × 10⁵ CFU/ml) plus 100 µl of the dipteran exosecretions at different concentrations were added to each well. Subsequently, 50 µl of a 1% solution of the triphenyl tetrazolium chloride stain was added to each well to determine the bacterial viability. The color development in each well was measured with the ImageJ software S. aureus was exposed to different concentrations of the ES of both species individually. At a concentration of 800 ppm ES of C. rufifacies or Co. macellaria, bacterial growth was inhibited 97.45 ± 1.70% and 82.21 ± 1.88%, respectively. As expected, exposure to a lower concentration (i.e., 50 ppm) was less inhibitory (C. rufifacies ES, 77.65 ± 4.25% and Co. macellaria ES, 43.54 ± 4.63%). This study demonstrates for the first time the bactericidal activity of C. rufifacies and Co. macellaria ES against S. aureus. This finding is promising as it could result in the identification and synthesis of proteins capable of suppressing pathogen development in wounds. Additionally, the proposed method can simplify the use of expensive laboratory instruments for antimicrobial activity determination.

Babesia microti (Aconoidasida: Piroplasmida) (Franca, 1910) is an important tick-borne zoonotic parasite with rodents serving as reservoir hosts. In the present study, 536 rodents were captured from Burdur, Bartin, Giresun, and Yozgat provinces of Turkey between the years 2010 and 2012, and blood samples were examined for the presence of Babesia spp. using conventional PCR which targeted the 18S rRNA gene. The sequence analysis of PCR amplicons was tested for B. microti as well as for Hepatozoon spp., and Sarcocystis spp. Overall, 5.8% of the rodents were positive for B. microti: 41% in Myodes glareolus, 7.7% in Chionomys roberti, and 2% in Apodemus spp., whereas no Babesia DNA was detected in Mus macedonicus and Microtus spp. Six rodents were positive for Hepatozoon spp. and one rodent was positive for Sarcocystis spp. Overall, 14.9 and 4.5% of rodents captured from Bartin and Giresun provinces, respectively, were PCR positive for B. microti, whereas none of rodents captured in Burdur and Yozgat were positive for Babesia spp. The sequence data of B. microti from rodents revealed that all sequences belonged to the zoonotic genotype. Sequences of B. microti obtained from rodents of the Bartin province were genotypically closer to European isolates, whereas those obtained from rodents of the Giresun province were closer to Russian and Mongolian isolates.

Quill mites (Acariformes: Prostigmata: Syringophilidae) are a family of about 400 ectoparasite species known to infest birds for over 140 yr. All previously identified quill mites occur in the calamus, a cavity of the bird feather, where they spent their entire life except for a short dispersion period. In this study, we report the first record of quill mites dwelling in the quill walls during their entire life cycle. In short, we found all of the different life stages of the species Lawrencipicobia poicephali (Skoracki & Dabert 2002) (Acariformes: Syringophilidae) in the quill wall in 33 host individuals of six different host species from many different locations in Sub-Saharan Africa. Hosts were collected over a time period spanning more than 70 yr. Lastly, we report six new avian hosts for this ectoparasite species.

Forcipomyia (Forcipomyia) makanensis Hou sp. n. (Diptera: Ceratopogonidae) is described and illustrated based on male and female specimens from China. It is characterized by the male aedeagus triangular, with large cone-shaped process at apex, basal arch high, basal arm slender and curved, parameres separate narrower distance at base, cone-shaped apically, and the female subgenital plate pliers-shaped, without lateral process, with two spermathecae, oval, nearly equal. The new species is compared with the similar congener F. (Forcipomyia) lochmocola Zou and Yu, 1991. We provide separate keys for identification of the males and females of the species of subgenus F. (Forcipomyia) Meigen in China.

In forensic entomology, the minimum postmortem interval (PMImin) estimative is usually based on the oldest immature recovered from a local of death. The time spent by fly immatures in the intrapuparial period comprises more than 50% of their complete life cycles. An accurate estimate of the duration of this period will improve PMImin estimates. The blow fly Hemilucilia semidiaphana (Rondani 1850) was found in six criminal cases in the city of Curitiba. Even though there is data on the morphology of the larval instars and developmental rate of H. semidiaphana, the intrapuparial period has not been investigated. Here, we provide a detailed description of the intrapuparial morphological changes of H. semidiaphana, which might be useful to estimate minimum PMI. Samples of H. semidiaphana in the intrapuparial period were obtained from immatures reared on an artificial diet in incubators adjusted to 25°C or 20°C temperature regimes. Blow fly puparia of H. semidiaphana were fixed at intervals of 3 and 6 h until emergence of the adult. The external morphological traits of sampled immatures were analyzed using light microscopy. Our analysis of the intrapuparial period of H. semidiaphana provided 21 traits from which nine were age informative. These nine characteristics divide the developmental time (144 h at 25°C and 192 h at 20°C) into smaller sections. The developmental data provided, together with the time line allows a practical way to make interspecific comparisons as well as to estimate the age of H. semidiaphana based on the intrapuparial development.

The mosquito gut is divided into foregut, midgut, and hindgut. The midgut functions in storage and digestion of the bloodmeal. This study used light, scanning (SEM), and transmission (TEM) electron microscopy to analyze in detail the microanatomy and morphology of the midgut of nonblood-fed Anopheles aquasalis females. The midgut epithelium is a monolayer of columnar epithelial cells that is composed of two populations: microvillar epithelial cells and basal cells. The microvillar epithelial cells can be further subdivided into light and dark cells, based on their affinities to toluidine blue and their electron density. FITC-labeling of the anterior midgut and posterior midgut with lectins resulted in different fluorescence intensities, indicating differences in carbohydrate residues. SEM revealed a complex muscle network composed of circular and longitudinal fibers that surround the entire midgut. In summary, the use of a diverse set of morphological methods revealed the general microanatomy of the midgut and associated tissues of An. aquasalis, which is a major vector of Plasmodium spp. (Haemosporida: Plasmodiidae) in America.

Colonies of house flies (Musca domestica L. [Diptera: Muscidae]) and four species of parasitoids (Muscidifurax raptor Girault and Sanders, Muscidifurax zaraptor Kogan and Legner, Spalangia cameroni Perkins and Spalangia endius Walker) were established by making collections from dairy farms near Bell, FL, Beatrice, NE, Minneapolis, MN, and San Jacinto, CA. Colonies were assessed for heat tolerance by comparing life history parameters at 25–27°C and fluctuating hot (26.7–41.7°C) temperatures. Muscidifurax raptor, S. cameroni, and S. endius produced 24–28% as many progeny under hot conditions as at 25°C. Colonies of M. zaraptor were more heat-tolerant and produced an average 46.9% as many progeny under the hot regime compared with moderate conditions. There was little evidence for higher heat tolerance in parasitoid populations from historically hot locations (CA desert and FL). Colonies of M. raptor and S. endius that had been in culture for 24 yr were the least heat-tolerant with regard to progeny production. House flies collected from the same locations varied little in longevity, fecundity, or egg-to-adult survival under either hot or moderate regimes. Flies reared under hot conditions laid about half as many eggs (89/female) and had about half the egg–adult survival rate (47.3%) under hot compared with moderate conditions, indicating that heat stress had less effect on flies than on all of the parasitoids except M. zaraptor. An attempt to select for heat tolerance in flies by subjecting them to incremental increases in rearing temperatures for 20 generations resulted in little change in tolerance among the selected flies.

Most studies of the effects of low temperature on the development of immature stages of Aedes aegypti (L.) have been performed at constant temperatures in the laboratory, which may not accurately reflect the variable environmental conditions in the field. Thus, the aim of this study was to assess the effect of constant temperatures (CT) and fluctuating low temperatures (FT) on the fitness of Ae. aegypti of Buenos Aires, Argentina. Three CT treatments (12, 14, and 16°C) and three FT treatments (12, 14, and 16°C ± 4°C) were performed and then survival, development time, and size of adults analyzed for each treatment. The immature stages completed development in all the treatments, with an average survival of 88% at 16°C, 85% at 14°C, and 22% at 12°C, and showed no differences between the CT and FT treatments. Development times were similar between the CT and FT treatments at 16°C (average ± SD: 22.7 ± 2.0 d) and at 14°C (average ± SD: 30.5 ± 2.5 d), whereas at 12°C, they lasted longer under CT (average ± SD: 46.6 ± 5.1 d) than under FT (average ± SD: 37 ± 6.5 d). The sizes of the adults at 12 and 14°C were similar but larger than those at 16°C, and showed no differences between the CT and FT treatments. Compared to populations of other geographical regions assessed in previous studies, the shorter development times and the high survival at 14 and 16°C, and the ability to complete development at 12°C, a fact not previously reported, suggest that the Ae. aegypti population of Buenos Aires city has a higher tolerance to these conditions.

The feeding interactions between house dust mites (HDM) and microorganisms are key factors in the survival of mites in human environments. The suitability of different microbes for HDM is not known. Here, microbial species isolated from HDM cultures were offered to HDM in food preference tests under laboratory conditions. The microbial species were added to the rearing diet of mites to reach 7% of Saccharomyces cerevisiae and 10% of each tested microorganism. The suitability of each microbe-supplemented diet for Dermatophagoides pteronyssinus and Dermatophagoides farinae was compared in terms of mite population growth and mite preference in a cafeteria test. The effect of mite feeding on the respiration of microorganisms in the diet was observed in microcosms. HDM were able to feed and reproduce on some bacterial and fungal species, but the suitability of microbial species differed. Increasing the yeast Sa. cerevisiae in the diet from 7 to 17% appeared the most suitable for both mite species. Staphylococcus spp. bacteria were preferred for D. farinae and were suitable for reproduction. The population growth and feeding preferences of D. pteronyssinus and D. farinae with respect to microorganisms indicate that D. farinae can develop on a diet with bacterial (Staphylococcus nepalensis and Staphylococcus kloosii) additions, whereas D. pteronyssinus was successful on a diet with fungal (Aspergillus jensenii and Aspergillus ruber) additions. The bacteria Kocuria rhizophila and Bacillus cereus decreased population growth in D. pteronyssinus, whereas the yeasts Hyphopichia pseudoburtonii, Hyphopichia burtonii, and Candida ciferrii decreased population growth in D. farinae. These results indicate that some microorganisms are an important food source for HDM.

Despite awareness of the mutations conferring insecticide resistance in the bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), within the United States few studies address the distribution and frequency of these. Within the United States, studies have focused on collections made along the East Coast and Midwest, documenting the occurrence of two mutations (V419L and L925I) within the voltage-gated sodium channel α-subunit gene shown to be associated with knockdown resistance (kdr) to pyrethroids. Here, the distribution and frequency of the V419L and L925I site variants is reported from infestations sampled within Oklahoma and its immediately adjacent states. Additionally, the presence of a mutation previously undocumented in the United States (I935F) is noted. While novel in the United States, this mutation has previously been reported in Australian and Old World populations. No infestations were found to harbor wild-type individuals, and hence susceptible, at each of the three sites. Instead, ∼21% were found to possess the resistant mutation at the L925I site (haplotype B), ∼77% had mutations at both the V419L and L925I sites (haplotype C), and 2% possessed the mutation at the L936F site (haplotype A^b). The high frequency of haplotype C corresponds to previous studies in the United States, and contrasts dramatically with those of the Old World and Australia. The data presented here provide insight into the contemporary occurrence of kdr-associated insecticide resistance in the South Central United States, a region for which data have previously been absent. These data suggest that New World and Old World/Australian infestations are likely to have originated from different origins.

Insects are efficient vectors of bacteria and in the hospital environment may have a role in spreading nosocomial infections. This study sampled the flying insect populations of seven hospitals in the United Kingdom and characterized the associated culturome of Diptera, including the antibiotic resistance profile of bacterial isolates. Flying insects were collected in seven U.K. hospitals between the period March 2010 to August 2011. The bacteria carried by Diptera were isolated using culture-based techniques, identified and characterized by antimicrobial susceptibility testing. A total of 19,937 individual insects were collected with Diptera being the most abundant (73.6% of the total), followed by Hemiptera (13.9%), Hymenoptera (4.7%), Lepidoptera (2.9%), and Coleoptera (2%). From Diptera, 82 bacterial strains were identified. The majority of bacteria belonged to the Enterobacteriaceae (42%), followed by Bacillus spp. (24%) and Staphylococcus spp. (19%). Less abundant were bacteria of the genus Clostridium (6%), Streptococcus (5%), and Micrococcus (2%). A total of 68 bacterial strains were characterized for their antibiotic resistance profile; 52.9% demonstrated a resistant phenotype to at least one class of antibiotic. Staphylococcus spp. represented the highest proportion of resistant strains (83.3%), followed by Bacillus spp. (60%) and Enterobacteriaceae (31.3%). Diptera were the predominant flying insects present in the U.K. hospital environments sampled and found to harbor a variety of opportunistic human pathogens with associated antimicrobial resistance profiles. Given the ability of flies to act as mechanical vectors of bacteria, they present a potential to contribute to persistence and spread of antimicrobial-resistant pathogenic bacteria in the hospital environment.

The infestation with the human ectoparasite, Pediculus humanus capitis (De Geer), is a common public health problem affecting schoolchildren worldwide. In Chile, the main active ingredients present in the over-the-counter pediculicides contain pyrethroids. Despite the extended use of these products, there is no evidence of the insecticide resistance status of the head lice geographically located in Chile. The most extended resistant mechanism of pyrethroids consists of the target site insensitivity (Kdr) determined by the presence of mutations linked to insecticide-binding sites in the voltage-sensitive sodium channel.T917I is recognized as the main mutation in head lice, and detection is considered to be a biomarker of resistance.The goal of the present study was to detect the presence and distribution ofT917I mutation in five geographic locations of Chile. All five geographically selected louse populations had a frequency of pyrethroid resistance genes that ranged from 36 to 77%, and 94.9% of the collected head lice had one or twoT917I mutant alleles. Moreover, the frequency of the aggregate resistant alleles was 50.5%.This is the first evidence that head lice in Chile had the mutations commonly associated with the resistance to pyrethroids. Moreover, the overrepresentation of heterozygotes in the studied populations suggests that head lice in Chile are currently under active selective pressure.

Filth flies live in close proximity to humans and livestock and transmit pathogens. Current control relies on chemical insecticides, and flies can develop resistance to these insecticides. The public is also interested in natural and safer insecticides. Therefore, alternative pesticides compatible with the synanthropic nature of flies are needed. Four plant aliphatic methyl ketones were evaluated for control of adult house flies, Musca domestica L., blow flies, Cochliomyia macellaria (F.), and gray flesh flies, Sarcophaga bullata (Parker). In sealed petri dish assays, 2-heptanone, 2-octanone, 2-nonanone, and 2-undecanone exhibited fumigant activity against house flies with 24-h LC₅₀s of 6.9, 7.5, 8.0, and 9.2 µg/cm³, respectively. Further research focused on undecanone (a U.S. EPA-registered biopesticide). When tested in larger enclosures at 1.7, 2.3, and 2.8 µg/cm³, undecanone provided 60.4, 82.2, and 94.4% house fly mortality; 56.9, 75.6, and 92.5% flesh fly mortality; and 62.1, 84.5, and 97.9% blow fly mortality, respectively, after a 2-h exposure. In a two-choice behavioral assay with 194.6 µg/ cm² of the test compound on the treatment versus an untreated surface of the same area, the overall mean repellencies for blow flies were 84.7% for undecanone versus 87.6% for N,N-diethyl-meta-toluamide (DEET). For house flies, mean repellencies were 80.7% for undecanone and 84.9% for DEET. The house fly topical LD₅₀ for undecanone was 58.1 µg per fly. Undecanone was far less expensive for filth fly control than the gold standard for insect fumigation, methyl bromide.

Resistance status of Aedes albopictus (Diptera: Culicidae) collected from Sabah, East Malaysia, was evaluated against four major classes of adulticides, namely pyrethroid, carbamate, organochlorine, and organophosphate. Adult bioassays conforming to WHO standard protocols were conducted to assess knockdown and mortality rates of Ae. albopictus. Among tested pyrethroid adulticides, only cyfluthrin, lambda-cyaholthrin, and deltamethrin were able to inflict total knockdown. The other adulticide classes mostly failed to cause any knockdown; the highest knockdown rate was only 18.33% for propoxur. With regards to mortality rate, Ae. albopictus was unanimously susceptible toward all pyrethroids, dieldrin, and malathion, but exhibited resistance toward bendiocarb, propoxur, dichlorodiphenyltrichloroethane, and fenitrothion. Additionally, correlation analysis demonstrated cross-resistance between bendiocarb and propoxur, and malathion and propoxur. In conclusion, this study has disclosed that pyrethroids are still generally effective for Aedes control in Sabah, Malaysia. The susceptibility status of Ae. albopictus against pyrethroids in descending order was cyfluthrin > lambda-cyhalothrin > deltamethrin > etofenprox > permethrin.

Hermetia illucens (L.) (Diptera: Stratiomyidae), commonly known as black soldier fly is a dipterous species which can produce enteric pseudomyiasis in humans and domestic animals. Veterinary cases of intestinal myiasis caused by H. illucens in dogs have not been reported previously. We present this rare pseudomyiasis (= accidental myiasis) produced by infestation with several larvae of the intestinal tract in a dog inhabiting a city in the province of Buenos Aires, Argentina.

The summer of 2018 was very hot and dry with little rain nationwide in Japan. Particularly in July, temperature was 3°C higher in comparison with the normal year. We collected adults, pupae, third or fourth instar larvae of Aedes albopictus (Skuse) in Sendai city, Niigata city, and Kanazawa city in late July and early August 2018. Wild-collected adults were examined for reproductive capacity. On the other hand, wild-collected larvae and pupae were raised to adults in the laboratory and then examined for reproductive capacity. They did not lay eggs after single blood feeding. When they were supplied with bloodmeals twice or three times, they laid a small number of eggs, but these eggs did not hatch. When these adult individuals were maintained at 25°C for 3 wk and then fed with blood, they laid healthy eggs which developed to embryos. It is thought that extreme heat and/or extreme low humidity due to low precipitation reduced female reproductive capacity even if they could take bloodmeals.

Surveillance for blood-fed female mosquitoes was performed between August 2015 and February 2016 at sites along the periphery of the Aripo Savannas Environmentally Reserve (ASSR) located in northeastern Trinidad, West Indies. We collected engorged female mosquitoes representing 13 species. DNA extractions from dissected abdomens were subjected to PCR amplification with three primer pairs targeting the mitochondrial cytochrome oxidase I and cytochrome b gene sequences. High-quality sequence information and host identification were obtained for 42 specimens representing eight mosquito species with at least one primer combination. A broad range of vertebrates including humans were identified, but the majority were nonhuman mammals, both domestic and wild. Domestic dogs were the most common host and may represent potential sentinel species for monitoring local enzootic arbovirus activity in Trinidad. Culex declarator Dyer and Knab and Culex nigripalpus Theobald were the most common blood-fed mosquito species comprising 79.1% of the total number identified. These species obtained blood meals from birds, nonhuman mammals, and human hosts, and therefore pose significant risks as potential bridge vectors for epizootic arbovirus transmission in the ASSR area as well as other sylvan areas in Trinidad. These data represent the first such results for Trinidad.

Zika virus (ZIKV) has emerged as a globally important arbovirus and has been reported from all states of Brazil. The virus is primarily transmitted to humans through the bite of an infective Aedes aegypti (Linnaeus, 1762) or Aedes albopictus (Skuse, 1895). However, it is important to know if ZIKV transmission also occurs from Ae. aegypti through infected eggs to her offspring. Therefore, a ZIKV and dengue virus (DENV) free colony was established from eggs collected in Manaus and maintained until the third–fourth generation in order to conduct ZIKV vertical transmission (VT) experiments which used an infectious bloodmeal as the route of virus exposure. The eggs from ZIKV-infected females were allowed to hatch. The resulting F1 progeny (larvae, pupae, and adults) were quantitative polymerase chain reaction (qPCR) assayed for ZIKV. The viability of ZIKV vertically transmitted to F1 progeny was evaluated by cultivation in C6/36 cells. The effects of ZIKV on immature development of Ae. aegypti was assessed and compared with noninfected mosquitoes. Amazonian Ae. aegypti were highly susceptible to ZIKV infection (96.7%), and viable virus passed to their progeny via VT. Moreover, eggs from the ZIKV-infected mosquitoes had a significantly lower hatch rate and the slowest hatching. In addition, the larval development period was slower when compared to noninfected, control mosquitoes. This is the first study to illustrate VT initiated by oral infection of the parental population by using mosquitoes, which originated from the field and a ZIKV strain that is naturally circulating in-country. Additionally, this study suggests that ZIKV present in the Ae. aegypti can modify the mosquito life cycle. The data reported here suggest that VT of ZIKV to progeny from naturally infected females may have a critical epidemiological role in the dissemination and maintenance of the virus circulating in the vector.

Mosquito-borne diseases are on the rise globally, and have the potential to thrive along the Gulf Coast of the United States, where subtropical conditions may facilitate the introduction or movement of mosquito vectors. Despite surveillance efforts, Aedes aegypti (L.) had not been detected in the Gulf state of Alabama for nearly three decades. The detection of Ae. aegypti in Alabama may suggest remnant or reemergent populations of this vector. We conducted adult sampling between May and August of 2018 to capture mosquitoes during a time frame when all species should be active. This was to ensure no species were missed due to overwintering and to identify the distributions of Aedes mosquitoes of medical importance. No Ae. aegypti were detected in Alabama over the period of this study. We detected Aedes albopictus (Skuse) in 65 counties and the recently invasive Aedes japonicus japonicus (Theobald) in 30 counties across the state. These results indicate that while Ae. aegypti was recently reported from parts of Alabama, the state is not experiencing a major resurgence of the species, whereas Ae. albopictus remains ubiquitous. Further, results indicate that a third wave of Aedes invasion may be occurring, that of Ae. japonicus japonicus. All three of these species are medically important vectors and may pose threats to the public health of the Gulf Coast of the United States.