Variation in resource utilization plays a significant role in determining the success or failure of a species. Generalist species across numerous taxa have exhibited success in feeding strategies for a variety of reasons. This study investigated the colonization and oviposition habits of a generalist carrion-feeder, Hermetia illucens L. (Diptera: Stratiomyidae), to varying conditions of carrion decay and colonization. Oviposition treatments consisted of combinations of variably aged decaying rats, both uncolonized and colonized with a heterospecific carrion-feeder.The black soldier fly exhibited a greater preference for oviposition on aged carrion, regardless of carcass colonization status. However, when presented with the option to colonize a plant-based diet, there was a significantly greater (40.4%) oviposition response to this diet. Results of this study highlight the ability of the black soldier fly to colonize fresh or aged carrion with or without a primary colonizer present, further supporting the recognition of this species as a generalist.These data demonstrate potential inaccuracies in the utility of H. illucens as time of colonization indicator species for legal investigations, and, in some regard, can be considered an exploitative niche-specialist (i.e., relying on plant–carrion interface for immature development) when found in association with carrion.

The attraction of necrophagous insects, particularly blow flies, to corpses and carrion is of ecological, economic, and agricultural importance, although the mechanisms by which it occurs are not well understood. Much of the published research on blow fly attractants has focused on volatiles emitted from carrion surrogates, but little attention has been given to the possibility that blow fly eggs themselves may emit chemical cues that are responsible for conspecific and heterospecific insect attraction. In this study, the headspace volatiles emitted from eggs representing two aggregated oviposition events that were collected 1 mo apart from two species of the Calliphoridae family (Order: Diptera), Lucilia sericata (Meigen), and Phormia regina (Meigen) were analyzed via solid-phase microextraction-facilitated GC-MS.The volatiles' profiles were found to be consistent between samples representing the same species, but unique between the two species. Over 100 molecules covering a wide range of compound classes that included alcohols, aldehydes, esters, amines, ketones, and organosulfur compounds were identified.The profile of volatiles emitted from the L. sericata eggs contained several alkanes and aldehydes, whereas salient features of the P. regina headspace included numerous esters and ketones. Between the two species, 42 compounds were shared, several of which were carboxylic acids. Little overlap between the range of compounds detected and those reported to be emitted from decomposing remains was observed.

Graphical Abstract

On rabbit carcasses (Oryctolagus cuniculus L.) exposed in open and shaded habitats, the relative abundance of insects and species richness were observed. The decomposition process was classified into four stages: fresh, bloated, decayed, and dry. Except for the decayed stage, the elapsed time for each stage of decomposition was similar between open and shaded habitats, with all carcasses reaching dryness in 13 and 14 d, respectively. In total, 2009 insects were collected during the decomposition process with 1,863 flies belonging to seven families and 15 species, and 146 beetles belonging to six species and three families. Insect abundances rose between the fresh and decay stages. The dominant families of insects included Calliphoridae and Muscidae (80.9% of the collected insects) and accounting for 38.07% of the species richness, whereas Histeridae accounted for 4.3% of the collected insects and 14.29% of the species richness. The open habitat attracted 1,233 insects belonging to 18 families, including 1,142 flies and 91 beetles, whereas the shaded habitat attracted 776 insects belonging to 18 families, including 721 flies and 55 beetles. Diversity level and ratios of exclusive species are also reported for each habitat (open 61.4%; shaded 38.6%). Between habitats, there were substantially separate insect communities, however. In addition, there was a substantial difference in the insect number and species between decomposition stages. This study demonstrates that the exposure status needs to be evaluated and examined when estimating the time since death.

In Brazil, many studies on Forensic Entomology analyze the activity and succession of flies in animal models. Data on human corpses are always collected and evaluated in isolated cases. This study aimed to list the insect species associated with crime scenes investigated by the Technical-Scientific Institute of criminal expertise of the State of Rio Grande do Norte (ITEP-RN), in the Northeast of Brazil, a region exposed to high homicide rates. In total, 10 cases were investigated, of which 50% were in the initial stage of decomposition. The examined bodies were colonized by species of three orders of insects, Diptera, Coleoptera and Hymenoptera. The order Diptera represented 96% of the total insects, being represented by the following species: Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae), Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae), Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae), and Cochliomyia macellaria, (Fabricius) (Diptera: Calliphoridae); Musca domestica (Linnaeus) (Diptera: Muscidae); and unidentified females of the Sarcophagidae family. Among beetles, the occurrence of Dermestes maculatus (DeGeer) (Coleoptera: Dermestidae), Necrobia rufipes (De Geer) (Coleoptera: Cleridae), and Onthophagus sp. (Scarabaeidae), as well as unidentified specimens of the families Tenebrionidae and Histeridae were recorded. In addition, specimens of Ectatomma sp. (Formicidae) were also recorded. Considering that the Rio Grande do Norte state presents a high homicide rate and the last cadaver study was conducted over a decade ago, these records update the list of species associated with cadaveric decomposition and contribute to consolidate forensic entomology in the Northeast region.

A fine-toothed comb found in the Judean Desert and resembling an ancient louse comb was examined. Based on radiocarbon dating, it ranged between 1660 AD and 1950 AD. From the material accumulated between the teeth, an oribatid mite, a pseudoscorpion, exuviae of beetle larvae, a sucking louse (Polyplax brachyrrhyncha Cummings, 1915), as well as a fully engorged larva and a nymph of the ixodid tick Rhipicephalus turanicus Pomerantzev, 1936 were recorded. Additionally, the comb included numerous hairs of a spiny mouse (Acomys sp.). Although finding mites, beetle larvae, and a pseudoscorpion on a louse comb could be regarded as contamination, the findings of P. brachyrrhyncha, as well as of a larva and nymph of R. turanicus, are noteworthy. We hypothesize that the presence of animal lice and ticks could indicate some sort of pet grooming.

Given current and projected changes in the climate, the composition of mosquito species is predicted to shift geographically with implications for the transmission dynamics of vector-borne pathogens. Many mosquito species are rarely collected in Canada and their history is poorly understood; thus assessing their potential role as vectors for pathogenesis is difficult. Mosquitoes were collected from four trapping sites in Quebec Province, Canada, from June to September during 2018 and 2019 using BG sentinel traps. From all morphologically identified female mosquitoes, at least one specimen was selected for identification confirmation using the DNA-barcoding technique. Sequences were subjected to alignment and a Neighbor-Joining (NJ) tree was created using Geneious software. In total, 2,752 female mosquitoes belonging to 20 species over five genera: including Aedes (Ae.), Anopheles (An.), Culex (Cx.), Culiseta (Cu.), Coquillettidia (Cq.) were collected. The predominant mosquito was found to be Ae. cinereus. The highest number of mosquito species was captured in July, followed by August, September, and then June. Five genera were characterized by a distinctive set of cytochrome oxidase I (COI) sequences that formed well-supported clusters in the NJ-tree. The presence of Ae. japonicus in Quebec provides an initial look at the distribution of mosquito species in eastern Canada, which may put Canadians at risk of a wider range of arboviruses.

The carboxamide N, N-di-ethyl-meta-toluamide (DEET) is the most effective and widely used insect repellent today. However, drawbacks concerning the efficacy and the safety of the repellent have led to efforts to design new classes of insect repellents.Through quantitative structure–activity relationships, chemists have discovered two chemical groups of novel repellents: the acylpiperidines and the carboxamides, with the acylpiperidines generally more potent in biological assays. Although the exact mechanism of action of DEET and other repellents has not yet been thoroughly elucidated, previous research shows that the activity of insect odorant receptors are inhibited in the presence of repellents. The present electrophysiological study employs two-electrode voltage clamp with Xenopus laevis oocytes expressing AgOR2/AgOrco and AgOR8/AgOrco receptors to assess the effects of the novel repellents on Anopheles gambiae Giles (Insecta: Diptera: Culicidae) mosquito odorant receptors. The novel acylpiperidines and carboxamides reversibly inhibited (12–91%) odorant-evoked currents from both AgOR2/AgOrco and AgOR8/AgOrco receptors in a dose-dependent manner at all tested concentrations (30 µM to 1 mM). Furthermore, all the novel agents were more potent inhibitors of the receptors than DEET, with the acylpiperidines producing on average greater inhibition than the carboxamides. Interestingly, there was a correlation (r² = 0.72) between the percentage inhibition of AgOR2/AgOrco receptor currents and protection times of the acylpiperidines. Our results add to existing evidence that the repellency of a compound is linked to its ability to disrupt the insect olfactory system and that the acylpiperidines could represent a class of more effective alternatives to the current gold standard, DEET.

The role of aquatic arthropod diversity and community interactions of larval mosquitoes are important for understanding mosquito population dynamics. We tested the effects of aquatic macrophyte diversity and habitat structural complexity in shaping the predator and competitor invertebrate communities associated with mosquito larvae. Experimental mesocosms were planted with live aquatic macrophytes and allowed to be naturally colonized by local invertebrates. Results indicated a positive effect of macrophyte diversity on competitor diversity and a negative effect on predator diversity. In turn, predator diversity negatively impacted mosquito abundance through a direct effect, while competitor diversity showed an indirect negative effect on mosquito larval abundance through its positive effect on predator diversity. The enhancement of aquatic macrophyte diversity and structural complexity has practical applications for the reduction of mosquito populations in managed systems where complete source elimination is not possible.

The microbiota of mites is closely related to their growth, development, and pathogenicity. Therefore, it is necessary to study the bacteria in mites. Here, for the first time, based on 16s rRNA V3-V4 region, the microbiota of 45 samples of nine species in six families of medically important mites were analyzed using Illumina MiSeq sequencing technique.The results showed that, at the phylum level, Proteobacteria (56.20–86.40%) were the dominant, followed by Firmicutes (6.41–19.43%), Bacteroidetes (5.56–13.38%) and Actinobacteria (1.93–28.07%). But at the genera the microbiota of mites are different, showing four characteristics: 1) The microbiota is related to the parasitic host. Demodex folliculorum (Acariforms: Demodicidae) and D. brevis (Acariforms: Demodicidae), both parasitizing humans, showed similar microbial composition, as did D. canis (Acariforms: Demodicidae) and Sarcoptes scabiei canis (Acariforms: Sarcoptidae) parasitizing dogs, but D. caprae (Acariforms: Demodicidae) parasitizing sheep showed unique microbial community; 2) The microbiota is related to mite's species. Dermatophagoides farinae and Cheyletus malaccensis (Acariforms: Cheyletidae), both collecting from flour, show respective microbial composition; 3) The microbiota is related to the life stage. There were differences in microbiota between adults and larvae of D. farinae, but no differences observed in Psoroptes cuniculi (Acariforms: Psoroptidae); and 4) The microbiota is related to the blood-feeding state.The microbiota of blood-fed Ornithonyssus bacoti (Parasitiformes: Macronyssidae) adults was significantly higher than that of unfed adults. This indicates that the microbiota of mites is affected by mite species, parasitic host, growth stage and habitat.Therefore, understanding these influencing factors will have a very important guiding significance for the prevention and control of mite-borne diseases.

Aedes albopictus (Skuse, 1894) is a mosquito vector raising global health concerns owing to its transmission of dengue, Zika, and chikungunya viruses.This vector accounts for a large proportion of the Korean mosquito community; however, autochthonous clinical cases resulting from this species remain unreported in South Korea.This study aimed to examine the geographical genetic variations and sources of Ae. albopictus populations in South Korea exclusively on the basis of COI gene analysis from 292 samples collected from 37 localities in 2016–2018 and 290 reference sequences from GenBank. Thirty-eight haplotypes were identified among the 292 Ae. albopictus samples, with H1 (n = 190, 65.1%), H29 (n = 24, 8.22%), and H32 (n = 24, 8.22%) being the most common and widely distributed haplotypes in the mainland, southern coastal region, and Jeju Island, respectively. In general, high haplotype (≥0.5; 44.7%) and low nucleotide (≤0.00148 max.) diversity were observed in these populations. Based on eight regional groups, results of neutral tests and a mismatch analysis supported demographic expansions after bottlenecks. Furthermore, analysis of molecular variance, F_ST, and K2P distance showed that Gyeongsangnam-do, Jeju Island, and mainland groups were genetically differentiated. Haplotype network and phylogenetic analyses revealed that the sources of the main haplotypes are related to strains from other countries.The current findings need to be validated with additional sampling from heterogeneous habitats and different genetic markers. However, our results suggest that haplotype changes should be closely monitored for efficient vector surveillance and control.

Aedes aegypti (L.), the main vector of dengue and other arboviruses, was declared eradicated from Argentina in 1964; however, in 1987, it was detected again and nowadays it occurs in most of the country territory. To understand the transmission of vector-borne diseases, knowledge of the dispersal of vector populations is essential to evaluate the risk of pathogen transmission. We conducted a population genetic analysis of Ae. aegypti in 20 neighborhoods from Córdoba, the second largest city in Argentina, using 10 microsatellite loci. High genetic differentiation and the absence of an isolation by distance pattern was found using Weir and Cockerham's θ. Bayesian and multivariate clustering analyses showed that the studied sites included individuals with high membership coefficients (Q) in their populations, individuals with membership in another cluster, and admixed individuals. Individuals with high Q in clusters different from the population in which they were collected strongly suggests that passive transport is important in shaping the Ae. aegypti dispersal pattern in Córdoba city. Knowing the genetic structure of Ae. aegypti populations and their dispersal patterns would contribute to the implementation of vector control programs.

In recent years, bed bugs have experienced a remarkable resurgence on a near global scale. While reports have primarily focused on the common bed bug, Cimex lectularius (L.), which has resurged largely in temperate regions, in tropical regions the tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae), has reemerged as well. Recent reports of C. hemipterus introductions to subtropical and temperate regions, outside of the species natural distribution, suggest the potential for establishment and further spread. Establishment may be aided by insecticide resistance mechanisms, such as the presence of knockdown resistance (kdr)-associated mutations, which potentially confer resistance to pyrethroid, pyrethrin, and organochloride insecticides. Here, we present the first report of the detection and likely establishment of C. hemipterus in Honolulu, Hawaii, from samples collected in 2009 and 2019. Furthermore, through partial sequencing of the voltage-gated sodium channel, we report the presence of kdr-associated mutations in all samples. These findings have implications for the implementation of control strategies aimed at eradicating infestations.

Aedes albopictus (Skuse) was first reported in the Republic of Korea in 1941 (Suwon, Gyeonggi Province). Recently, a mosquito-monitoring program was established in response to the potential development of autochthonous infections of dengue, chikungunya, and Zika viruses, to identify the regional characteristics of Ae. albopictus in Suwon. Daily collections of adult mosquitoes were conducted using Blackhole UV-LED traps and BG-Sentinel traps baited with octenol, carbon dioxide or both at five sites in Suwon (Sungkyunkwan University, Manseok, Seoho, Olympic, and Jungang parks) from 1 July to 1 November 2016. Based on the ecological cycle of mosquitoes, an estimated association between the daily mean temperatures [2 wk prior to the collection date (lag 2 wk)] and relative population abundance were developed using a generalized linear model. The overall proportion of female mosquitoes collected from all traps was 88.3%. Higher female trap indices were observed for mosquitoes collected at BG-Sentinel traps. Additionally, the BG-sentinel trap was more effective collecting Ae. albopictus. Culex pipiens (Coquillett) was the most frequently collected mosquito using UV-LED traps, followed by Ae. vexans nipponii (Theobald), Ochlerotatus koreicus (Edwards), Cx. inatomi (Kamimura and Wada), and members of the Anopheles Hyrcanus Group. Within a certain temperature range, daily mean temperatures 2 wk prior to the collection period was significantly associated with a maximum abundance of Ae. albopictus (P < 0.05) at 26.2°C (95% confidence interval: 24.3–29.4°C).The BG-Sentinel trap is an effective trap for monitoring relative population abundance of Ae. albopictus, and trap catches are significantly correlated with daily mean temperatures.

The genus Otodectes Canestrini 1894, comprises an unique species, Otodectes cynotis (Hering, 1838), that is responsible for the ‘ear mange’ in terrestrial carnivores worldwide. Here we report its occurrence in four Brazilian host species from the southeastern region, based on records from Carnivora in the Acari Collection of the Butantan Institute, São Paulo state (IBSP). We also register a case report of a crab-eating fox, Cerdocyon thous L. 1758 found parasitized by this mite species.

Trehalose is a disaccharide that is the major sugar found in insect hemolymph fluid.Trehalose provides energy, and promotes growth, metamorphosis, stress recovery, chitin synthesis, and insect flight. The hydrolysis of trehalose is under the enzymatic control of the enzyme trehalase.Trehalase is critical to the role of trehalose in insect physiology, and is required for the regulation of metabolism and glucose generation. Trehalase inhibitors represent a novel class of insecticides that have not been fully developed. Here, we tested the ability of trehalose analogues to function as larvacides or adulticides in an important disease vector—Aedes aegypti. We show that validamycin A, but not 5-thiotrehalose, delays larval and pupal development and prevents flight of adult mosquitoes. Larval mosquitoes treated with validamycin A were hypoglycemic and pupae had increased levels of trehalose. Treatment also skewed the sex ratio toward male mosquitoes. These data reveal that validamycin A is a mosquito adulticide that can impair normal development of an important disease vector.

The Asian tiger mosquito, Aedes (Stegomyia) albopictus (Skuse), is a peridomestic, container-ovipositing mosquito commonly found throughout the southeastern United States. In the United States, Ae. albopictus is typically considered a nuisance pest; however, it is capable of transmitting multiple pathogens. Ae. albopictus is an important pest species and the target of numerous mosquito control efforts in the United States. Here, we evaluate the effectiveness and spatial extent of Ae. albopictus population reduction using a bifenthrin (AI Bifen IT, 7.9%) barrier spray and larval habitat management (LHM) in a temperate, suburban setting. Sixteen pairs of adjoining neighbors were randomly assigned to treatment groups with one neighbor receiving a treatment and the other monitored for evidence of a spillover effect of the treatments. Ae. albopictus populations in both yards were monitored for 33 d, with treatments occurring on the eighth day. Barrier sprays, both alone and combined with LHM, resulted in a significant reduction in Ae. albopictus abundance posttreatment. While LHM alone did not result in a significant reduction over the entire posttreatment period, Ae. albopictus populations were observed to be in decline during this period. No treatments were observed to have any reduction in efficacy 25 d posttreatment, with treatments involving LHM having a significantly increased efficacy. Yards neighboring treated yards were also observed to have reduced population sizes posttreatment, but these differences were rarely significant. These results provide insights into the population dynamics of Ae. albopictus following two common treatments and will be useful for integrated pest management plans.

Effective mosquito surveillance and management depend on a thorough understanding of the biology and feeding patterns unique to species and sex. Given that a propensity to sugar feed is necessary for some mosquito surveillance and newer control strategies, we sought to document the amount of total sugar in wild Aedes aegypti (L.) and Culex quinquefasciatus (Say) captured from five different locations in the Lower Rio Grande Valley (LRGV) of South Texas over 2 yr. We used Biogents Sentinel 2 (BGS2) traps in year 1 and aspirators, BGS2, and CDC resting traps in years 2 and 3 to collect adult mosquitoes.The hot anthrone test was used to quantify total sugar content in each mosquito. Additionally, the cold and hot anthrone tests were used to distinguish fructose content from total sugars for mosquitoes captured in 2019. Overall, Ae. aegypti females had significantly lower total sugar content than Ae. aegypti males as well as both sexes of Cx. quinquefasciatus. However, the percentage of Ae. aegypti positive for fructose consumption was four to eightfold higher than Ae. aegypti previously reported in other regions. The difference between locations was significant for males of both species, but not for females. Seasonality and trapping method also revealed significant differences in sugar content of captured mosquitoes. Our results reinforce that sugar feeding in female Ae. aegypti is less than Cx. quinquefasciatus, although not absent.This study provides necessary data to evaluate the potential effectiveness of sugar baits in surveillance and control of both Ae. aegypti and Cx. quinquefasciatus mosquitoes.

Effective suppression of container-inhabiting Asian Tiger [Aedes albopictus (Skuse)] (Diptera: Culicidae) and yellow fever [Aedes aegypti (L.)] (Diptera: Culicidae) mosquitoes presents one of the most intractable problems for modern mosquito control. Traditional tools often fail to control populations of these mosquito species, and are prohibitively expensive or have negative environmental impacts. Novel approaches and tools are urgently needed for integrated container-inhabiting mosquito management programs. One of the most promising techniques is autodissemination. We present the results of a long-term large-scale study conducted in a temperate urbanized environment representing typical Ae. albopictus habitats.Three treatment sites with autodissemination stations and three nearby reference sites were monitored for eggs, immature, and adult mosquitoes over a period of 3 yr from 2014 to 2016. Elevated larval and pupal mortality of 12–19% on average was the most notable outcome in sentinel cups of the treatment sites. The number of eggs in the treatment sites was significantly reduced in 2014, but not in 2015 or 2016. Adult populations remained similar in treatment and reference sites throughout the study.The impact of autodissemination on mosquito populations was lower than reported by previous investigations.Technical and logistical problems associated with wider coverage and working in multiple urban neighborhoods contributed to reduced efficacy. Incorporating autodissemination with routine mosquito control operations and commercializing this methodology for general public use will require further research on combining this tool with other novel or conventional technologies.

Bloodmeal feeding by the horn fly, Haematobia irritans (L.), is associated with reduced milk production and blood loss that ultimately prevents weight gain of calves and yearlings. Thus, blood feeding by H. irritans causes significant economic losses in several continents. As with other arthropods, resistance to the majority of commercialized insecticides reduces the efficacy of current control programs.Thus, innovative technologies and novel biochemical targets for horn fly control are needed. Salivary gland and Malpighian tubule function are critical for H. irritans survivorship as they drive bloodmeal acquisition and maintain ion- and fluid homeostasis during bloodmeal processing, respectively. Experiments were conducted to test the hypothesis that pharmacological modulation of H. irritans inward rectifier potassium (Kir) channels would preclude blood feeding and induce mortality by reducing the secretory activity of the salivary gland while simultaneously inducing Malpighian tubule failure. Experimental results clearly indicate structurally diverse Kir channel modulators reduce the secretory activity of the salivary gland by up to fivefold when compared to control and the reduced saliva secretion was highly correlated to a reduction in bloodmeal acquisition in adult flies. Furthermore, adult feeding on blood treated with Kir channel modulators resulted in significant mortality. In addition to validating the Kir channels of H. irritans as putative insecticide targets, the knowledge gained from this study could be applied to develop novel therapeutic technologies targeting salivary gland or Malpighian tubule function to reduce the economic burden of horn fly ectoparasitism on cattle health and production.

The invasive, human-biting Asian longhorned tick, Haemaphysalis longicornis Neumann, is establishing in the United States. This tick is a threat to public health in its native range in Asia, serving as a vector of severe fever with thrombocytopenia syndrome virus and Rickettsia japonica, the agent of Japanese spotted fever. However, there is a lack of published information specifically for H. longicornis concerning the efficacy of generally recommended personal tick bite prevention measures. We, therefore, evaluated permethrin-treated clothing and formulated human skin repellent products, representing the six repellent active ingredients generally recommended for tick bite prevention by the Centers for Disease Control and Prevention (CDC), against H. longicornis nymphs from a colony established with adult ticks collected in New York state. Reluctance of H. longicornis nymphs to stay in contact with nontreated human skin precluded the use of a human skin bioassay to optimally evaluate repellency. In a Petri dish choice bioassay, all tested product formulations were highly effective with estimated repellencies ranging from 93 to 97%. In addition, we observed strong contact irritancy of a summer-weight permethrin-treated garment against H. longicornis nymphs, with 96% of introduced ticks dislodging from the vertically oriented textile within 3 min. These preliminary studies indicate that personal tick bite prevention measures currently recommended by the CDC are effective against the invasive H. longicornis. However, additional studies are needed to explore the efficacy of the evaluated products against different life stages of H. longicornis, as well as ticks collected in the field rather than reared in the laboratory.

Aedes aegypti L. (Diptera: Culicidae) is one of the most medically important mosquito species, due to its ability to spread viruses of yellow fever, dengue fever, and Zika in humans. In this study, the insecticidal activity of 17 plant essential oils was evaluated via topical application against two strains of Ae. aegypti mosquito, Orlando (insecticide-susceptible) and Puerto Rico (pyrethroid-resistant). Initial screens with the Orlando strain showed that cucumber seed oil (2017 sample) was the most toxic, followed by sandalwood and thyme oil. When the essential oils were mixed with permethrin, they failed to show any significant synergism of insecticidal activity. Sandalwood and thyme oils displayed consistently high mortality against the resistant Puerto Rico strain, with low resistance ratios of 2.1 and 1.4, respectively. In contrast, cucumber seed oil showed significantly less activity against Puerto Rico mosquitoes, with a resistance ratio of 45. Bioactivity-guided fractionation of the 2017 sample of cucumber seed oil sample via flash column chromatography produced 11 fractions, and gas-chromatography/mass spectrometry analysis revealed that the three active fractions were contaminated with 0.33, 0.36, and 0.33% of chlorpyrifos-methyl, an organophosphorus insecticide, whereas inactive fractions did not show any trace of it.These results suggested that the insecticidal activity of cucumber seed oil was probably due to the presence of the insecticide, later confirmed with a clean batch of cucumber seed oil obtained in 2018, which showed negligible insecticidal activity.These findings demonstrate clearly the need for essential oil analysis to confirm purity before any claims are made about pesticidal potency.

Contact irritant (locomotor excitation) and noncontact spatial repellent avoidance behavior to deltamethrin and cypermethrin at dosages 0.025, 0.05, and 0.1 g/m² impregnated on papers were evaluated in the laboratory against deltamethrin- and cypermethrin-resistant field populations of female Aedes aegypti (L.) from Rayong and Chanthaburi Provinces,Thailand. Pyrethroid-resistant populations were compared with a susceptible laboratory strain (NIH-Thai) using an ‘excito-repellency’ (ER) test system. Both NIH-Thai and field mosquitoes had stronger contact irritancy responses compared to the relatively weak noncontact repellency effects. Contact assays with deltamethrin and cypermethrin at 0.1 g/m² showed high escape rates for Rayong (80.1 and 83.4%, respectively) and Chanthaburi (84.6 and 73.1%, respectively) mosquitoes. Cypermethrin produced significantly different (P < 0.05) percent escape responses in contact tests between NIH-Thai and field mosquitoes. Only deltamethrin contact at 0.05 g/m² produced a significant escape response (P < 0.001) between NIH-Thai and Rayong mosquitoes.These results suggest that there may not be an overall significant effect of background pyrethroid resistance on escape response, and the differences by comparisons may reflect inherent individual variation when using the ER bioassay system.The results show that pyrethroid resistance in Ae. aegypti does not appear to influence or reduce contact avoidance responses with the compounds tested. In particular, deltamethrin at 0.1 g/m² was an effective contact irritant and toxic compound against pyrethroid-resistant populations of Ae. aegypti.Therefore, 0.1 g/m² deltamethrin could be considered for residual applications of either fixed surfaces or materials (e.g., curtains) as a supplemental control measure against adult dengue vectors.

Vector control strategies recommended by the World Health Organization are threatened by resistance of Anopheles mosquitoes to insecticides. Information on the distribution of resistant genotypes of malaria vectors is increasingly needed to address the problem. Ten years of published and unpublished data on malaria vector susceptibility/resistance and resistance genes have been collected across Togo. Relationships between the spatial distribution of resistance status and environmental, socio-economic, and landscape features were tested using randomization tests, and calculating Spearman rank and Pearson correlation coefficients between mosquito mortality and different gridded values. Anopheles gambiae sensu lato was resistant to DDT, pyrethroids, and the majority of carbamates and organophosphates. Three sibling species were found (i.e., An. gambiae, Anopheles coluzzii, and Anopheles arabiensis) with four resistance genes, including kdr (L1014F, L1014S, and N1575Y) and ace1 (G119S). The most frequent resistance gene was L1014F. Overall, no association was found between the susceptibility/resistance status and environmental features, suggesting that evolution of resistance may be most closely related to extreme selection from local insecticide use. Nevertheless, further research is necessary for firm conclusions about this lack of association, and the potential role of landscape characteristics such as presence of crops and percentage of tree cover.

Insecticide resistance in Aedes aegypti mosquitoes poses a major threat to public health worldwide.There are two primary biological mechanisms that can lead to insecticide resistance, target site and metabolic resistance, both of which confer resistance to specific classes of insecticides. Due to the limited number of chemical compounds available for mosquito control, it is important to determine current enzymatic profiles among mosquito populations. This study assessed resistance profiles for three metabolic pathways, α-esterases, β-esterases, and mixed-function oxidases (MFOs), as well as insensitivity of the acetylcholinesterase (iAChE) enzyme in the presence of propoxur, among Ae. aegypti from the Central Valley and southern California. All field-collected Ae. aegypti demonstrated elevated MFOs and iAChE activity, indicating potential development of pyrethroid and organophosphate resistance, respectively. Although regional variations were found among α-esterase and β-esterase activity, levels were generally elevated, further suggesting additional mechanisms for developing organophosphate resistance. Furthermore, mosquito samples from southern California exhibited a higher expression level to all three metabolic enzymes and iAChE activity in comparison to mosquitoes from the central region.These results could help guide future mosquito control efforts, directing the effective use of insecticides while limiting the spread of resistance.

Ixodids are blood-feeding ectoparasitic vectors of many disease agents that infect humans, livestock, and wild animals. As ixodid resistance to conventional synthetic acaricides becomes increasingly problematic, natural products are receiving greater attention as possible alternative control tactics. Formic acid, produced by ants, is a commercially available product for fumigating varroa mites, Varroa destructor Anderson &Trueman, infesting honey bee, Apis mellifera L., hives, and it has been reported to repel ixodids. Lone star tick, Amblyomma americanum (L.), larvae and nymphs were used as a model ixodid to investigate deterrent, repellent, and lethal effects of formic acid as a fumigant and contact toxin in vitro in the laboratory. Although formic acid failed to deter or repel A. americanum, it was highly toxic as a fumigant to larvae at a 1% concentration even when exposure was limited to 5 min. Contact by crawling on wet, moist, and dry treated substrates under ventilated conditions causes >90% mortality to larvae in 5% formic acid concentration treatments within 30–120 min, and temporary immersion killed ≈60% of the larvae by 24 h after they were removed from the 5% formic acid treatment solution. Substantial nymphal mortality occurred after 1–1.5 h following exposure to substrate treated with the 10% concentration and immersion killed ≈45% of the nymphs. It appears that formic acid volatiles are more lethal to A. americanum immatures than direct contact with the external integument.

We examined whether routine fall yard maintenance, specifically depositing leaves removed from lawns and landscaping along forest margins, may increase densities of nymphal Ixodes scapularis Say and Amblyomma americanum (L.) ticks within these managed areas. Leaf blowing activities in fall 2017 and 2018 on residential properties in New Jersey, United States, significantly increased leaf litter depth in managed edge areas (range = 259.8–352.8 mm) compared to unmanaged edges (77.6–188.0 mm) and adjacent forest (39.4–166.2 mm). Drag sampling conducted on 20 and 30 dates in spring 2018 and 2019, respectively, yielded ≥3-fold more I. scapularis nymphs in managed edge plots compared to natural edge and forest plots in both years. In 2018, we collected more A. americanum nymphs from forest plots than from either natural or managed edge plots, but 2019 natural edge plots yielded the greatest number of the ticks. Nearly half of A. americanum adults were collected in forest plots in both years. Our data suggest that the acarological risk of human encounters with I. scapularis nymphs may be significantly greater in areas receiving an accumulation of leaves from leaf blowing or raking compared to adjacent unmanaged forest edges. This artificially elevated acarological risk can be mitigated if homeowners avail themselves of curbside leaf pickup or composting services offered by many municipalities or request that lawn/landscaping contractors remove collected leaves offsite, or at least to areas of less frequent use, rather than concentrating them along the lawn-forest edge.

In the last two decades, bed bugs (Cimex lectularius L. and Cimex hemipterus F.) have become perennial and difficult to control indoor pests. Current pest control options are severely constrained by high prevalence of insecticide resistance and availability and relatively high costs of alternative interventions. Among various measures to counter the drawbacks of insecticide resistance include efforts to diversify the modes of action of insecticides with residual applications of combinations of insecticides, which include a juvenile hormone analog (JHA). JHAs, such as hydroprene and methoprene, have a desirable safety profile and are effective against a variety of indoor pests. We evaluated the potential of hydroprene and methoprene to be incorporated into an ingestible bait, with dose–response studies on fifth-instar male and female bed bugs. Females were more susceptible than males to both JHAs, and methoprene was more effective by ingestion than hydroprene at inducing both lethal and sublethal effects. Ingestion of ≥10 µg/ml blood of either JHA by last instar nymphs reduced oviposition; untreated females that mated with males exposed to high concentrations of either JHA also exhibited lower oviposition. We suggest that methoprene could be incorporated into integrated pest management programs in liquid baits and residual sprays in combination with other active ingredients.

Aedes-borne virus disease control relies on insecticides to interrupt transmission. Temephos remains a key chemical for control of immature stage Aedes in Thailand and much of Southeast Asia. However, repeated use of insecticides may result in selection for resistance in vector populations, thus compromising operational intervention. Herein, the phenotypic response to temephos by Aedes aegypti (L.) and Aedes albopictus (Skuse) collected in Thailand and surrounding countries is presented. Data from 345 collection sites are included: 283 from literature review (244 sites with Ae. aegypti, 21 with Ae. albopictus, and 18 having both species sampled), plus 62 locations with Ae. aegypti in Thailand conducted between 2014 and 2018. Susceptibility assays followed WHO guidelines using the recommended discriminating dose of temephos (0.012 mg/liter) against late third to early fourth instar Ae. aegypti. Findings revealed 34 locations with susceptible Ae. aegypti, 13 with suspected resistance, and 15 indicating resistance. Published data between 1999 and 2019 in Thailand found Ae. aegypti resistant in 73 of 206 collection sites, whereas 3 locations from 11 sampled with low-level resistant in Ae. albopictus. From surrounding countries conducting temephos assays (Cambodia, Lao PDR, Myanmar, Malaysia, and Singapore), resistance is present in Ae. aegypti and Ae. albopictus from 27 of 56 and 19 of 28 locations, respectively. Routine insecticide susceptibility monitoring should be an operational requirement in vector control programs. Given the wide distribution and apparent increase in temephos-resistance, alternative larvicidal compounds must be considered if chemical control is to remain a viable vector control strategy.

Ticks can vector and transmit many pathogens and pose a serious human health threat throughout the world. After collection, many diagnostic laboratories must mechanically disrupt tick specimens for diagnostic testing and research purposes, but few studies have evaluated how well-commercial tissue homogenizers perform this task. We evaluated four commercially available tissue homogenizers: The Bead Ruptor 24 Elite, the Bullet Blender Storm, the gentleMACS Dissociator, and the Precellys 24. We quantitatively compared maceration level, nucleic acid quality, quantity, amplification, and DNA shearing to determine which machines performed the best. The Bead Ruptor 24 Elite had the highest overall score when disrupting a single, uninfected adult Amblyomma americanum (Linnaeus) (Ixodida: Ixodidae) and performed well in follow-on tests including disrupting individual juvenile samples and detecting pathogens from infected samples.

The Aedes aegypti mosquito is the primary vector of dengue, yellow fever, chikungunya, and Zika viruses. Infection with the dengue virus alone occurs in an estimated 400 million people each year. Likelihood of infection with a virus transmitted by Ae. aegypti is most commonly attributed to abundance of the mosquito. However, the Arizona-Sonora desert region has abundant Ae. aegypti in most urban areas, yet local transmission of these arboviruses has not been reported in many of these cities. Previous work examined the role of differential Ae. aegypti longevity as a potential explanation for these discrepancies in transmission. To determine factors that were associated with Ae. aegypti longevity in the region, we collected eggs from ovitraps in Tucson, AZ and reared them under multiple experimental conditions in the laboratory to examine the relative impact of temperature and crowding during development, body size, fecundity, and relative humidity during the adult stage. Of the variables studied, we found that the combination of temperature during development, relative humidity, and body size produced the best model to explain variation in age at death.

Aedes aegypti (L.) (Diptera: Culicidae) is a diurnal feeder that lives in close association with human populations. It is the principal vector of yellow fever, dengue fever and the Zika Virus. Issues of arboviral diseases have been on the ascendency in most countries including Ghana where Aedes mosquito is the main vector of yellow fever. A comparative study of the biting behavior of Ae. aegypti and the identification of subspecies were undertaken using molecular technique. Standard human landing technique was used to collect both indoor and outdoor biting mosquitoes at three zones located in the Upper East (Bolgatanga), Upper West (Nadowli), and Northern (Damongo) Regions of Ghana during the dry and rainy seasons between 0600 and 1800 Greenwich Mean Time (GMT). All collected mosquitoes were identified morphologically using taxonomic keys. random amplified polymorphic DNA polymerase chain reaction was used to categorize Ae. aegypti into subspecies. Adult female Aedes mosquitoes identified formed 62% (n = 1,206) of all female mosquitoes collected. Aedes aegypti 98% and Aedes vittatus 2% were the only Aedes species identified. Bolgatanga recorded the largest number of Ae. aegypti 42%, whereas Nadowli 22% recorded the least. Aedes vittatus was observed in Nadowli. Aedes aegypti exhibited a bimodal biting behavior peaking at 0600–0800 GMT and 1500–1600 h GMT. Molecular findings revealed 69% Ae. aegypti aegypti and 31% Ae. aegypti formosus as the two subspecies (n = 110). This information is important for implementing effective vector control programs in the three regions of the northern Ghana.

We conducted a molecular survey of Rickettsia in fleas collected from opossums, road-killed and live-trapped in peridomestic and rural settings, state parks, and from pet cats and dogs in Georgia, United States during 1992–2014. The cat flea, Ctenocephalides felis (Bouché) was the predominant species collected from cats and among the archival specimens from opossums found in peridomestic settings. Polygenis gwyni (Fox) was more prevalent on opossums and a single cotton rat trapped in sylvatic settings. Trapped animals were infested infrequently with the squirrel flea, Orchopeas howardi (Baker) and C. felis. TaqMan assays targeting the BioB gene of Rickettsia felis and the OmpB gene of Rickettsia typhi were used to test 291 flea DNAs for Rickettsia. A subset of 53 C. felis collected from a cat in 2011 was tested in 18 pools which were all bioB TaqMan positive (34% minimum infection prevalence). Of 238 fleas tested individually, 140 (58.8%, 95% confidence interval [CI]: 52.5–64.9%) DNAs were bioB positive. Detection of bioB was more prevalent in individual C. felis (91%) compared to P. gwyni (13.4%). Twenty-one (7.2%) were ompB TaqMan positive, including 18 C. felis (9.5%) and 3 P. gwyni (3.2%). Most of these fleas were also positive with bioB TaqMan; however, sequencing of gltA amplicons detected only DNA of Rickettsia asembonensis. Furthermore, only the R. asembonensis genotype was identified based on NlaIV restriction analysis of a larger ompB fragment. These findings contribute to understanding the diversity of Rickettsia associated with fleas in Georgia and emphasize the need for development of more specific molecular tools for detection and field research on rickettsial pathogens.

Studies done in Bouaké (Côte d'Ivoire) about 20-yr ago reported that Anopheles gambiae s.l. Giles was the major malaria vector. The present study aimed to update these data and to identify the main vectors. Mosquitoes were collected in Allokokro and Petessou villages between June 2014 and December 2015 using the human landing catching method. Potential breeding sites of An. gambiae s.l. were identified in August and October 2014 and mapped using GPS. Anopheles species were morphologically and molecularly [polymerase chain reaction (PCR)] identified. Ovaries of female were dissected to determine the parity and infection with Plasmodium was detected in head and thorax by quantitative PCR. In Allokokro, the biting rate of An. gambiae s.s was significantly greater than Anopheles coluzzii, whereas, in Petessou, biting rates of both species were comparable. Plasmodium falciparum (Haemosporida: Plasmodiidae), Plasmodium malariae (Haemosporida: Plasmodiidae), and Plasmodium ovale (Haemosporida: Plasmodiidae) identified in both villages. The infection rates of An. gambiae s.s. and An. coluzzii were not significantly different. The entomological inoculation rate (EIR) of An. gambiae s.s. for P. falciparum was 9-fold greater than that of An. coluzzii in Allokokro; however, in Petessou, the EIRs of both species were comparable. In both village, An. gambiae s.s was responsible for P. falciparum and P. ovale transmission whereas An. coluzzii transmitted all three Plasmodium species.

Culicoides midges vector numerous veterinary and human pathogens. Many of these diseases lack effective therapeutic treatments or vaccines to limit transmission. The only effective approach to limit disease transmission is vector control. However, current vector control for Culicoides midges is complicated by the biology of many Culicoides species and is not always effective at reducing midge populations and impacting disease transmission. The endosymbiont Wolbachia pipientis Hertig may offer an alternative control approach to limit disease transmission and affect Culicoides populations. Here the detection of Wolbachia infections in nine species of Culicoides midges is reported. Infections were detected at low densities using qPCR. Wolbachia infections were confirmed with the sequencing of a partial region of the 16S gene. Fluorescence in situ hybridization of Culicoides sonorensis Wirth and Jones adults and dissected ovaries confirm the presence of Wolbachia infections in an important vector of Bluetongue and Epizootic hemorrhagic disease viruses. The presence of Wolbachia in Culicoides populations in the United States suggests the need for further investigation of Wolbachia as a strategy to limit transmission of diseases vectored by Culicoides midges.

The importance of ticks in veterinary and medical science has received much attention. The dominant tick species in northeastern China, Ixodes persulcatus tick can transmit various pathogens to humans and animals and there are some studies on the microbiome composition of this tick. Our study characterized the bacterial communities in I. persulcatus by 16S amplicon pyrosequencing and described the differences of microorganisms in male and female tick and assessed the variation of microorganisms in the development stages in northeastern China. We mainly found the following bacteria genera: Pseudomonas (Pseudomonadales: Pseudomonadaceae), Citrobacter (Enterobacteriales: Enterobacteriaceae), and Rickettsia (Rickettsiales: Rickettsiaceae). Rickettsia is common and harmful pathogen transmitted by ticks. Meanwhile, we found there were differences between male and female tick of microbiomes, and the diversity of microbiome increased from engorged female ticks to eggs, but decreased when the eggs were molting into larvae. Our data showed that male ticks exhibited greater microbial diversity than female I. persulcatus tick and larvae presented with a different bacterial community compared to engorged female tick and hatched eggs. These findings may be useful for further understanding the interaction between I. persulcatus and microbiome biology.

Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) cause hemorrhagic disease (HD) in wild ruminants and bluetongue disease (BT) and epizootic hemorrhagic disease (EHD) in livestock. These viruses are transmitted by biting midges in the genus Culicoides (family Ceratopogonidae). Mortality from this disease can reach 90% in certain breeds of sheep and in white-tailed deer (Odocoileus virginianus). From January until December of 2012, we conducted a prospective study to determine the origin and routes of transmission of BTV and EHDV in captive deer and cattle. The objective was to determine the abundance of Culicoides spp. and BTV/EHDV infection prevalence in midges, cattle, and deer in an area experiencing an outbreak of BT and EHD. Agar gel immunodiffusion (AGID) tests to detect for EHDV and BTV antibodies were conducted on serum collected from cattle and deer, quantitative reverse transcriptase polymerase chain reaction (RT–qPCR) was utilized for BTV/EHDV RNA detection in tissues from dead deer, and CDC miniature black light traps baited with dry ice were deployed to capture insects. The AGID results showed 19 out of 29 cattle and 18 out of 58 white-tailed deer seroconverted for these viruses during the vector season. Tradition gel-based reverse transcriptase polymerase chain reaction was utilized to determine serotype. Sixteen cows were positive for EHDV-2, EHDV-6, or BTV-12 and 15 deer positive for EHDV-1, EHDV-6, or BTV-12. Specimens from 14 species of Culicoides (Dptera: Ceratopogonidae) (Culicoides arboricola Root and Hoffman, Culicoides biguttatus Coquillett, Culicoides crepuscularis Malloch, Culicoides debilipalpis Lutz, Culicoides furens Poey, Culicoides haematopotus Malloch, Culicoides hinmani Khalaf, Culicoides nanus Root and Hoffman, Culicoides neopulicaris Wirth, Culicoides paraensis Goeldi, Culicoides stellifer Coquillet, Culicoides variipennis Coquillet, Culicoides villosipennis Root and Hoffman, and Culicoides venustus Hoffman) were captured and tested for BTV and EHDV using RT-qPCR assays. BTV viral nucleic acid was detected in three pools from three different species of midges: C. crepuscularis, C. debilipalpis, and C. stellifer.

In view of recent cases of human and canine visceral leishmaniasis reported in Porto Alegre, Rio Grande do Sul, Brazil, we investigated the sand fly fauna inhabiting the neighborhoods of Morro Santana and Jardim Carvalho, Brazil, continuing a series of entomological surveys aimed to identify potential vectors of Leishmania (Ross, 1903) parasites. Sand flies were collected monthly from October 2016 to October 2017 using CDC light traps in the intradomiciliary and peridomiciliary environments of seven residences. Sand fly abundance was correlated to climatic variables. Females were pooled by species, location, and date for Leishmania DNA molecular screening using ITS1 and kDNA polymerase chain reaction. In total, 501 sand flies from five species were collected in which Lutzomyia gaminarai (Cordero, Vogelsang & Cossio, 1928) (Diptera: Psychodidae) (78%) was the most abundant species in the intradomiciliary sites while Migonemyia migonei (Franca, 1920) (Diptera: Psychodidae) (43.3%) was the most abundant in the peridomiciliary sites. A higher number of sand flies were collected during the warmest months, from December to March (Mann–Whitney statistical test – P < 0.001). Leishmania infantum DNA was detected in Lu. gaminarai (2), Pintomyia fischeri (Pinto, 1926) (1) and Mg. migonei (1). Leishmania braziliensis DNA was detected in Lu. gaminarai (1) and Pi. fischeri (1). Our results add support to the possible vector role of Pi. fischeri in the epidemiological cycle of Le. infantum in Brazil. Furthermore, the first documented detection of Leishmania DNA in Lu. gaminarai may be indicative of multiple vectors being involved in the Leishmania cycle within Porto Alegre.

A diversity of Borrelia burgdorferi sensu lato (Johnson, Schmid, Hyde, Steigerwalt & Brenner) (Spirochaetales: Spirochaetaceae) genomospecies, including the Lyme disease agent, Borrelia burgdorferi sensu stricto (s.s.), have been identified in the western United States. However, enzootic transmission of B. burgdorferi s.l. in small mammals and ticks is poorly characterized throughout much of the region. Here we report prevalence of B. burgdorferi s.l. in small mammal and tick communities in the understudied region of southern California. We found B. burgdorferi s.l. in 1.5% of Ixodes species ticks and 3.6% of small mammals. Infection was uncommon (∼0.3%) in Ixodes pacificus Cooley and Kohls (Acari: Ixodidae), the primary vector of the Lyme disease agent to humans in western North America, but a diversity of spirochetes—including Borrelia bissettiae, Borrelia californiensis, Borrelia americana, and B. burgdorferi s.s.—were identified circulating in Ixodes species ticks and their small mammal hosts. Infection with B. burgdorferi s.l. is more common in coastal habitats, where a greater diversity of Ixodes species ticks are found feeding on small mammal hosts (four species when compared with only I. pacificus in other sampled habitats). This provides some preliminary evidence that in southern California, wetter coastal areas might be more favorable for enzootic transmission than hotter and drier climates. Infection patterns confirm that human transmission risk of B. burgdorferi s.s. is low in this region. However, given evidence for local maintenance of B. burgdorferi s.l., more studies of enzootic transmission may be warranted, particularly in understudied regions where the tick vector of B. burgdorferi s.s. occurs.

Maintenance of laboratory colonies of insects and other arthropod pests offers significant research advantages. The availability, age, sex, housing conditions, nutrition, and relative uniformity over time of biological material for research facilitate comparison of results between experiments that would otherwise be difficult or impossible. A laboratory research colony of Phlebotomus papatasi (Scopoli), old world sand flies, was maintained with high-colony productivity for a number of years, but within a relatively short (4–6 mo) time period, colony productivity declined from over 10,000 flies per week to less than 100 per week. Mites and nematodes were both visible in the larval medium; however, the mites had been present throughout high productivity periods; therefore, it seemed reasonable to investigate the nematodes. PCR amplification of 18S rRNA yielded a clean cDNA sequence identified by BLAST search as Procephalobus sp. 1 WB-2008 (Rhabditida: Panagrolaimidae) small subunit ribosomal RNA gene, GenBank EU543179.1, with 475/477 nucleotide identities. Nematode samples were collected and identified as Tricephalobus steineri, (Andrássy, 1952) Rühm, 1956 (Rhabditida: Panagrolaimidae) based on morphological characteristics of the esophagus and the male copulatory apparatus. Mites (Tyrophagus putrescentiae [Acariformes: Acaridae]) may have played an additional predatory role in the loss of sand fly colony productivity. We hypothesized that the origin of the nematode infestation was rabbit dung from a local rabbitry used in preparation of the larval medium. Colony productivity was fully restored within 3 mo (two sand fly generational periods) by replacement of the rabbit dung from a clean source for use to prepare sand fly larval medium.

The potential for reinvasion of the United States by cattle fever ticks, Rhipicephalus (Boophilus) annulatus and Rhipicephalus microplus (Canestrini), which remain established in Mexico, threatens the viability of the domestic livestock industry because these ticks vector the causal agents (Babesia bovis and Babesia bigemina) of bovine babesiosis. The Cattle Fever Tick Eradication Program safeguards the health of the national cattle herd preventing the reemergence of bovine babesiosis by keeping the United States free of cattle fever ticks. Here, the collection of free-living southern cattle tick, R. microplus, larvae by sweeping flannel flags over vegetation in the wildlife corridor of Cameron and Willacy Counties, TX, is reported. Finding R. microplus larvae on vegetation complements reports of infestations in wildlife hosts inhabiting the southern Texas coastal plains. Land uses and environmental conditions have changed since cattle fever ticks were eradicated from the United States by 1943. These changes complicate efforts by the Cattle Fever Tick Eradication Program to keep cattle in the United States free of the cattle fever tick disease vectors. Current scientific research on technologies that could be used for area-wide management of fever tick larvae in south Texas and how this could be applied to integrated eradication efforts are discussed.

Since the last yellow fever (YF) outbreak was detected in Argentina in 2009, vector surveillance and studies of arbovirus infections are carried out intermittently specifically in areas where nonhuman primates of the Alouatta genus are present. We report in these areas of Corrientes province the detection of Haemagogus leucocelaenus (Dyar and Shannon) (Diptera: Culicidae) and Sabethes albiprivus (Theobald) (Diptera: Culicidae), both species involved in the forest YF cycle, and also the presence of Aedes albopictus (Skuse) in new areas in Argentina, which represents the southernmost citation for this species in South America. Aedes albopictus, a mosquito species native to Asia, was reported for the first time in Argentina in 1998, in Misiones province. Since then, no other report has indicated the extension of the distribution of this mosquito. This report shows the importance of performing continual entomological and arboviruses surveillance and highlights the impact that could result from the expansion of Ae. albopictus across Argentina.

The lone star tick, Amblyomma americanum Linnaeus, is ubiquitously present in the southeastern United States and will readily parasitize humans and companion animals. Fipronil is the active ingredient in several topically applied products used to manage ticks and fleas on companion animals. Reducing ectoparasite infestations on companion animals decreases the risk that pathogens vectored by these pests are transmitted to these animals and their owners. However, dependence on acaricides can promote the development of resistance. In this study, the Food and Agriculture Organization of the United Nations larval packet test was used to determine the lethal concentration (LC) values and discriminating concentration (DC) for fipronil in the lone star tick. The DC was calculated as 0.02080%. The low magnitude of the DC value suggests that fipronil is an effective active ingredient for A. americanum management. With the LC and DC values determined, emergent resistance can be monitored, potentially allowing for intervention should tolerance develop in A. americanum populations that are in frequent contact with humans and their companion animals.

The Oriental rat flea, Xenopsylla cheopis (Rothschild 1903), is a cosmopolitan flea usually found infesting domestic rats. This flea is a well-known major human plague vector in Madagascar. As part of field sampling, fleas and small mammals were collected in the village of South Andranofeno and the natural reserve of Sohisika, two sites of the district of Ankazobe, located in the Central Highlands of Madagascar. Rats inside houses and forest small mammals were trapped using Besancon Technical Services and pitfall traps, respectively. Their fleas were collected and preserved for laboratory works. Collected fleas from the village and forest belonged to five species, which were X. cheopis, Synopsyllus fonquerniei (Wagner and Roubaud 1932) (Siphonaptera: Pulicidae), Echidnophaga gallinacea (Westwood 1875) (Siphonaptera: Pulicidae), Ctenocephalides felis strongylus (Jordan 1925) (Siphonaptera: Pulicidae), Pulex irritans (Linnaeus 1758) (Siphonaptera: Pulicidae). After sampling in the forest zone, one specimen of X. cheopis was unexpectedly collected while infesting an endemic tenrec Setifer setosus (Schreber 1777) (Afrosoricida: Tenrecidae). Polymerase chain reaction (PCR) diagnosis on all collected fleas allowed detecting plague bacterium Yersinia pestis (Lehmann and Neumann 1896) (Enterobacterales:Yersiniaceae) on nine specimens of the endemic flea S. fonquerniei collected inside forest.The presence of the oriental rat flea in forest highlights the connection between human and wild environments due to animal movements and the fact that the rat flea can infest various hosts. As only one specimen of X. cheopis was collected on S. setosus, we hypothesize that flea was carried from the village to forest. Yersinia pestis infection of forest fleas outlines plague circulation in this sylvatic area.

Much of the bluetongue (BT) and epizootic hemorrhagic disease (EHD) research in North America focuses on white-tail deer and Culicoides sonorensis (Wirth & Jones) (Diptera: Ceratopogonidae), though several other biting midge species have been suggested as vectors. Culicoides stellifer (Coquillett) has been associated with hosts susceptible to hemorrhagic disease (HD), and more recently, specimens from Florida have tested positive for EHD and BT viral RNA. If C. stellifer is acting as a vector, this could have an impact on the distribution of HD in North America.To determine if gene flow is occurring across the range of C. stellifer within the southeast United States, a mitochondrial haplotype analysis was performed using the COI gene. Our haplotype network showed no population structure in C. stellifer from Florida,Texas, and South Carolina, as the overall genetic divergence between these sites was equal to the genetic divergence within each.We also compared these haplotypes to published sequences of C. stellifer collected in Ontario, Canada. Surprisingly, the genetic diversity of the flies from Ontario was two times greater than what was observed between the southeast U.S. collection sites.This considerable divergence could be evidence of a cryptic species. A better understanding of the connectivity between C. stellifer populations across all of North America will give insight into the distribution of HD. Our results show that gene flow is occurring between sites in the southeastern United States and potentially throughout the eastern distribution of the species.

An entomological survey was conducted in several localities of Rondônia State, and new records were obtained for seven sand fly species: Evandromyia apurinan Shimabukuro, Figueira & Silva, 2013, Evandromyia carmelinoi (Ryan, Fraiha, Lainson & Shaw, 1986), Micropygomyia echinatopharynxAndrade-Filho, Galati, Andrade & Falcão, 2004, Nyssomyia urbinattiiGalati & Galvis, 2012, Pintomyia duckeiOliveira, Alencar & Freitas, 2018, Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004), and Sciopemyia vattierae (Le Pont & Desjeux, 1992). Herein, we also describe the female of Pintomyia fiocruzi Pereira Júnior, Pessoa, Marivalva & Medeiros, 2019. The female has spermathecae with a detached apical ring similar to the spermathecae of Pintomyia serrana (Damasceno & Arouck, 1949), Pintomyia odax (Fairchild & Hertig, 1961), Pintomyia ottolinai (Ortiz & Scorza, 1963), and Pintomyia robusta (Galati, Cáceres & Le Pont, 1995). Here, we provide characters to separate Pi. fiocruzi from Pi. serrana and Pi. odax. Pintomyia ottolinai has a trans-Andean distribution and Pi. robusta has not been found in Brazil and in our studies, we associated the females found in this study with Pi. fiocruzi males found at the same collection sites. Our findings highlight the importance of entomological fauna surveys and demonstrate the need for taxonomists to study species diversity in Rondônia. This study increases the number of sand fly species recorded in Rondônia to 143.