Several arboviruses have emerged and/or reemerged in the New World in the past decades. Zika and chikungunya viruses, formerly restricted to Africa and perhaps Asia, invaded the continent, causing great concern. Dengue virus outbreaks have continued to occur in almost all countries, with millions of cases per year. West Nile virus rapidly invaded North America, and now cases have been found in Central and South America. Other arboviruses, such as Mayaro and Eastern equine encephalitis viruses, have increased their activity and have been found in new regions. Changes in pathogenicity have been documented for some viruses leading to unexpected disease. A diverse mosquito fauna, changing climate and vegetation, increased travel, and unplanned urbanization producing conditions for the proliferation of Aedes aegypti (L.), Culex quinquefasciatus Say, and other vectors have combined to strongly influence changes in the distribution and incidence of several arboviruses. The need for thorough studies of the mosquito fauna and modifications of environmental conditions, mostly in urban areas strongly influenced by social, political, and economic factors, is emphasized.

In 2011, a thriving population of Aedes albopictus (Skuse), the Asian tiger mosquito, was discovered within three cities in Los Angeles County over an estimated 52-km² urban area. Two years later in 2013, Aedes aegypti (L.), the yellow fever mosquito, was detected within several urban areas of Madera, Fresno, and San Mateo counties. State and local vector control agencies responded with an aggressive effort to eradicate or interrupt the spread of these two invasive mosquitoes; however, known populations continued to expand outward and new infestations were identified at an accelerated pace in central and southern California. By the end of 2015, one or both species had been detected within the jurisdictional boundaries of 85 cities and census-designated places in 12 counties. Herein we report on the discovery and widespread establishment of Ae. aegypti and Ae. albopictus in urban areas of coastal, central, and southern California between 2011 and 2015 and discuss the subsequent rapid changes to the activities and priorities of vector control agencies in response to this unprecedented invasion.

Leptotrombidium scutellare mites, the vector of Orientia tsutsugamushi, have rarely been reported to associate with Rickettsia species. Three hundred nineteen chiggers were collected from the ears of 32 rodents captured in Huangdao District of Qingdao City, China, in October 2015. The chigger samples were tested for Rickettsia, severe fever with thrombocytopenia syndrome virus, and hantavirus by PCR or RT-PCR amplification. All mites were classified morphologically and molecularly as L. scutellare chiggers. Rickettsial DNA sequences were amplified for four genes including 16S rRNA, ompB, gltA, and 17 kD protein genes. The minimum infection rate (MIR; number of positive pools/total specimens tested) of the Rickettsia species in the chiggers were 2.8% (9/319). Phylogenetic analysis indicated that individual genes were closely related to different Rickettsia species including R. felis (with 16S rRNA gene), R. australis (with gltA gene), an unnamed Rickettsia sp. TwKM02 (with ompB gene), and Rickettsia endosymbiont of soft tick Ornithodoros erraticus (with 17 kD protein gene). Phylogenic analysis of the concatenated sequence of 16S rRNA, gltA, ompB, and 17 kD protein genes indicated that the Rickettsia species from L. scutellare chigger was most closely related to R. australis and R. akari. These results indicated that the Rickettsia species in chiggers was unique; it was named Candidatus Rickettsia leptotrombidium. Severe fever with thrombocytopenia syndrome virus and hantavirus were not amplified from the chiggers, suggesting lack of infection of these pathogens in the chiggers. A unique Rickettsia species was detected in L. scutellare, which expanded the knowledge on the vector distribution of Rickettsia.

Avian movements are an important mechanism by which ticks and associated pathogens can colonize new locations. The temporal and geographic extent of tick and pathogen dispersal is dependent on tick phenology and host movements across a landscape. We evaluated bird-host traits and habitat characteristics that may influence tick infestation and subsequent dispersal in Illinois. To determine which characteristics of host species and habitats influence infestation, we examined captured birds in 22 forest patches across east-central Illinois during migratory periods in fall of 2012 and 2013, and spring of 2013 and 2014. We examined 1,028 birds of 78 species; 136 (13.2%) individuals representing 33 (42.3%) species were infested with at least one tick. We determined that infestation prevalence was greatest among birds that 1) forage primarily in the forest understory; 2) use large forest patches; 3) use patches with less invasive shrub cover. Furthermore, infestation intensity was greatest among birds captured during fall migration. These findings highlight the importance of avian and habitat traits that may influence tick infestation among passerine birds.

Culicoides biting midges transmit multiple ruminant viruses, including bluetongue virus and epizootic hemorrhagic disease virus, causing significant economic burden worldwide. To further enhance current control techniques, understanding vector–virus interactions within the midge is critical. We developed previously a double-stranded RNA (dsRNA) delivery method to induce RNA interference (RNAi) for targeted gene knockdown in adult Culicoides sonorensis Wirth & Jones. Here, we confirm the C. sonorensis inhibitor of apoptosis 1 (CsIAP1) as an anti-apoptotic functional ortholog of IAP1 in Drosophila, identify the ortholog of the Drosophila initiator caspase DRONC (CsDRONC), and demonstrate that injection of dsRNA into the hemocoel can be used for targeted knockdown in the midgut in C. sonorensis. We observed CsIAP1 transcript reduction in whole midges, with highest transcript reduction in midgut tissues. IAP1knockdown (kd) resulted in pro-apoptotic caspase activation in midgut tissues. In IAP1kd midges, midgut tissue integrity and size were severely compromised. This phenotype, as well as reduced longevity, was partially reverted by co-RNAi suppression of CsDRONC and CsIAP1. Therefore, RNAi can be directed to the midgut of C. sonorensis, the initial site of virus infection, using dsRNA injection into the hemocoel. In addition, we provide evidence that the core apoptosis pathway is conserved in C. sonorensis and can be experimentally activated in the midgut to reduce longevity in C. sonorensis. This study thus paves the way for future reverse genetic analyses of midgut–virus interactions in C. sonorensis, including the putative antiviral properties of RNAi and apoptosis pathways.

Lemurpediculus robbinsi sp. nov. is described from Crossley's dwarf lemur, Cheirogaleus crossleyi A. Grandidier, and Lemurpediculus claytoni sp. nov. is described from Sibree's dwarf lemur, Cheirogaleus sibreei Forsyth Major, from Madagascar. Both sexes of each new louse species are illustrated and distinguished from the two previously known species of Lemurpediculus: L. verruculosus (Ward) and L. petterorum Paulian. With the addition of two new species to the genus, an amended description of Lemurpediculus is provided. The two hosts of the new louse species are morphologically similar, endangered, obligately hibernating lemurs. These two species of lemurs are sometimes sympatric in rainforests in eastern Madagascar. Despite the morphological similarity of the two host species, their lice are morphologically distinct and are easiest to identify based on the shape of the subgenital plate of the female and the shape of the genitalia in the male. Both new species of lice should be considered to be endangered because their hosts are endangered. It is not known if either of the new species of lice are vectors of pathogens or parasites to their hosts.

Two new species, Simulium (Gomphostilbia) sunapii and S. (G.) rangatense, are described based on adults, pupae, and mature larvae from Flores, in the eastern part of the Sunda Archipelago, Indonesia. Simulium (G.) sunapii sp. nov. is placed in the S. asakoae species-group, representing the easternmost geographical record for the group in this archipelago. It is characterized by a small number of male upper-eye large facets in eight or nine vertical columns and 12 horizontal rows. Simulium (G.) rangatense sp. nov. is placed in the S. ceylonicum species-group and is characterized by the pupal gill with six filaments. This new species, together with two related species of the S. ceylonicum species-group in Flores, suggests the species radiation of this speciesgroup might have been accompanied by a reduction of the number of pupal gill filaments from eight to four through six. Taxonomic notes are provided to distinguish these two new species from related species.

Eleven species of myocoptid mites (Acariformes: Myocoptidae) are recorded from African rodents. Among them, three species are described as new for science: Myocoptes lophuromys sp. nov. from Lophuromys woosnami Thomas, 1906 (Rodentia: Muridae) from Uganda, Trichoecius hylomyscus sp. nov. from Hylomyscus anselli Bishop, 1979 (Rodentia: Muridae) from Tanzania (type host and locality) and Hylomyscus stella (Thomas, 1911) from Gabon, and Trichoecius faini sp. nov. from Hylomyscus arcimontanus Carlton and Stanley, 2005 from Tanzania. Gliricoptes graphiuriFain, 1970 syn. nov. is synonymized with Gliricoptes vulcanorumFain, 1970, and the female of Trichoecius angolensisFain, 1972 is described for the first time. New hosts and localities are provided for four species: Myocoptes grammomysFain, 1970 from Grammomys dryas (L., 1758) (Rodentia: Muridae) from Burundi and Uganda, Myocoptes spinulatusFain, 1970 from Dendromus mystacalis (Heuglin, 1863) (Rodentia: Muridae) from Tanzania, Crinicastor congolensisFain, 1970 from Grammomys macmillani (Wroughton, 1907) (Rodentia: Muridae) from Tanzania, and Trichoecius otomysFain, 1970 from Otomys typus (Heuglin, 1877) (Rodentia: Muridae) from Uganda. Three species are recorded from new localities: Gliricoptes vulcanorum from Tanzania, Trichoecius angolensis from South Africa, and Trichoecius lootensiFain, 1970 from Tanzania.

Little is known about the Anopheles fauna of Saint-Georges de l'Oyapock, a persistent malaria-endemic municipality in French Guiana. This study aimed to update the knowledge of local Anopheles diversity, and their ecology and role in malaria transmission. Sampling sessions were implemented between September 2013 and October 2014. Four species were identified from the 3,450 specimens collected: Anopheles darlingi Root, An. braziliensis, An. triannulatus s.l., and An. nuneztovari s.l. Anopheles darlingi was the predominant species. Its involvement in malaria transmission was suspected due to 1) its abundance, 2) the presence of a density peak during the malaria emergence period, and 3) a dynamic correlated with malaria cases observed two months later. Present and past studies show that the influence of environmental conditions on malaria vector dynamics is high, and may vary drastically according to the local context. This supports evidence that control strategies must be designed at fine scales.

Malaria is an important health issue in French Guiana. Its principal mosquito vector in this region is Anopheles darlingi Root. Knowledge of the spatial distribution of this species is still very incomplete due to the extent of French Guiana and the difficulty to access most of the territory. Species distribution modeling based on the maximal entropy procedure was used to predict the spatial distribution of An. darlingi using 39 presence sites. The resulting model provided significantly high prediction performances (mean 10-fold cross-validated partial area under the curve and continuous Boyce index equal to, respectively, 1.11—with a level of omission error of 20%—and 0.42). The model also provided a habitat suitability map and environmental response curves in accordance with the known entomological situation. Several environmental characteristics that had a positive correlation with the presence of An. darlingi were highlighted: nonpermanent anthropogenic changes of the natural environment, the presence of roads and tracks, and opening of the forest. Some geomorphological landforms and high altitude landscapes appear to be unsuitable for An. darlingi. The species distribution modeling was able to reliably predict the distribution of suitable habitats for An. darlingi in French Guiana. Results allowed completion of the knowledge of the spatial distribution of the principal malaria vector in this Amazonian region, and identification of the main factors that favor its presence. They should contribute to the definition of a necessary targeted vector control strategy in a malaria pre-elimination stage, and allow extrapolation of the acquired knowledge to other Amazonian or malaria-endemic contexts.

Aedes koreicus (Edwards) is an invasive mosquito species, like Aedes albopictus (Skuse) and Aedes japonicus japonicus (Theobald), that has already colonized a large part of northeastern Italy and other European countries. Despite its rapid expansion, information about adult distribution and trapping is lacking. Here, we conducted a 2-yr longitudinal survey using adult traps to investigate the spatiotemporal distribution of Ae. koreicus and evaluated the effectiveness of three trapping devices in Latin square experiments conducted in an urban site and a forested site. The following three different traps were compared: a CO₂-baited Biogents (BG) Sentinel trap, a CO₂-baited Centers for Disease Control and Prevention light trap (CDC trap), and a grass infusion-baited gravid trap.

In northern Italy, Ae. koreicus was collected from late April to early November, with peak of abundance observed in August. Aedes koreicus was more abundant in 2015 than in 2014 because of higher temperatures during summer. Unlike Ae. albopictus, the abundance of Ae. koreicus was not related to the altitude of the sampling locations in the range 241–660 m above sea level. The BG Sentinel and gravid traps collected significantly more Ae. koreicus than the CDC trap in the urban site, whereas there was no significant difference between the three traps in the forested site. In the urban site, the BG Sentinel trap and the gravid trap were the most effective for collecting Ae. albopictus and Culex pipiens L., respectively. In the forested site, Cx. pipiens was primarily collected by the CDC trap.

Repellents prevent mosquito bites and help reduce mosquito-borne disease, a global public health issue. Laboratory-based repellent bioassays predict the ability of compounds to deter mosquito feeding, but the variety of repellent bioassays and statistical analysis methods makes it difficult to compare results across methodologies. The most realistic data are collected when repellents are applied on the skin; however, this method exposes volunteers to chemicals and mosquito bites. Silicone membranes were investigated as an alternative to human skin in assays of repellent efficacy. Results from module system bioassays conducted in vitro with a silicone membrane were compared with in vivo bioassays conducted with N,N-diethyl-3-methylbenzamide (referred to as DEET), 1-piperidinecarboxylic acid 2-(2-hydroxyethyl)-1-methylpropylester (referred to as Picaridin), ethyl 3-[acetyl(butyl)amino]propanoate (referred to as IR3535), and para-menthane-3,8-diol (referred to as PMD) applied directly on the skin of the leg. No significant difference in mosquito feeding was found when comparing skin and volunteer-worn membrane controls using blood; however, feeding was significantly lower in unworn membrane controls using either 10% sucrose or blood, indicating that worn membranes are a possible surrogate for untreated human skin. Pooled data from six volunteers were used to generate dose–response curves of blood-feeding activity. Results from skin-applied repellents were modeled to determine if membranes could provide a predictive correlate for skin. Goodness-of-fit comparisons indicated that the nonlinear dose–response curves for the skin and membrane differed significantly for DEET and Picaridin, but did not differ significantly for IR3535 and PMD. With knowledge of the dose–response relationships and further modifications to this system, the membrane-based tests could be used for standardized repellent testing with infected vectors

Culex quinquefasciatus Say is an important disease vector throughout much of the world. Experiments were conducted to determine the effects of different larval habitat substrates on the fitness and biting efficiency of Cx. quinquefasciatus adults. Our findings indicate that the development time (egg to adult) of larvae reared in irrigation water was 8.63 d while that of larvae reared in distilled water was 17.10 d (Effect size = 0.95). However, the rate of adult emergence was similar for all the tested treatments. Furthermore, the mean weight of an egg raft varied between larval habitats: distilled water (1.83 mg), rainfall water (1.25 mg), irrigation water (1.52 mg), and sewerage water (2.52 mg) (Effect size = 0.91). But, the fecundity (eggs per female) and hatchability (%) were statistically similar in all the rearing mediums (Effect size = 0.79). Longevity of females in all the tested populations did not differ significantly (Effect size = 0.91). The mean relative growth rates of larvae reared in tap water (0.80) and distilled water (0.86) habitats were lower than growth rates in all other rearing habitats (Effect size = 0.96). The intrinsic rate of natural increase in tap water (0.27) and irrigation water (0.35) was significantly higher than that in distilled water (0.09) and sewerage water (0.16) (Effect size = 0.84). Adults reared in rain water had the highest biting efficiency among all the tested populations. These results provide useful information for the management of Cx. quinquefasciatus.

We investigated the dynamics and underlying causes of house (re)infestation with Triatoma infestans (Klug 1834) after a community-wide residual spraying with pyrethroids in a well-defined rural section of Pampa del Indio municipality (northeastern Argentina) over a 4-yr period. House infestation was assessed by timed manual searches, during insecticide applications, and by opportunistic householders' bug collections. All reinfested houses were selectively re-sprayed with insecticides. The resident population comprised Qom (66.6%) and Creole (33.4%) households, whose sociodemographic profiles differed substantially. The prevalence of house infestation dropped, less than expected, from 20.5% at baseline to 5.0% at 14 months postspraying (MPS), and then fluctuated between 0.8 and 4.2% over 21–51 MPS. Postspraying house infestation was positively and highly significantly associated with prespraying infestation. Most of the foci detected over 14–21 MPS were considered persistent (residual), some of which were moderately resistant to pyrethroids and were suppressed with malathion. Infestation patterns over 27–51 MPS suggested bug invasion from internal or external foci, but the sources of most findings were unaccounted for. Local spatial analysis identified two hotspots of postspraying house infestation. Using multimodel inference with model averaging, we corroborated that baseline domestic infestation was closely related to refuge availability, housing quality, and occurrence of peridomestic infestation. The diminished effectiveness of single pyrethroid treatments, partly attributable to moderate resistance compounded with rather insensitive vector detection methods and poor housing conditions, contributed to vector persistence. Improved control strategies combined with broad social participation are needed for the sustainable elimination of vector-borne human Chagas disease from the Gran Chaco.

The roots of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim., a well-known medicinal plant from Eastern Asia, are used worldwide for their known beneficial medicinal properties. Recently, the leaves have been used as an alternative to the roots. The present study was aimed at exploring the leaf essential oil as a potential source of compounds for mosquito management. Gas chromatography/mass spectrometry analysis of the leaf essential oil revealed 87 compounds, constituting 95.2% of the oil. a-Bisabolol (26.46%), β-caryophyllene (7.45%), germacrene D (6.87%), β-bisabolene (4.95%), and a-humulene (3.50%) were five of the major constituents. The essential oil was subjected to biting deterrence and repellent activity against mosquito Aedes aegypti. The biting deterrence of the oil produced a proportion not biting (PNB) value of 0.62 at 10 μg/cm² as compared with 0.86 of control DEET (N,N-diethyl-3-methylbenzamide) at a standard dose of 25 nmol/cm². Among individually selected compounds present in the oil (a-bisabolol, β-caryophyllene, a-humulene, and caryophyllene oxide), only a-bisabolol produced a PNB value of 0.80, equivalent to DEET at 25 nmol/cm², whereas the others were not repellent. The artificial mixture (AMES-1) of these four selected compounds produced a relatively high PNB value of 0.80. The repellent activity measured by minimum effective dosage (MED) for a-bisabolol and a-humulene produced MED values of 0.094 and 0.104 mg/cm², respectively, as compared with 0.023 mg/cm² of DEET. The leaf essential oil, the artificial mixture (AMES-1), and other binary and tertiary combinations of major compounds showed no repellent activity. In addition, morpho-anatomical features of the leaf are provided for correct identification of the species.

Dengue, chikungunya, and yellow fever are important vector-borne diseases transmitted by female mosquitoes when they feed on humans. The use of repellents based on natural products is an alternative for personal protection against these diseases. Application of chemicals with larvicidal activity is another strategy for controlling the mosquito population. The repellent and larvicidal activities of the essential oil from Eucalyptus nitens were tested against Aedes aegypti and Aedes albopictus, the main vectors of these arboviruses. The essential oil was extracted by hydrodistillation and then analyzed by gas chromatography–mass spectrometry. The main components of Eucalyptus nitens essential oil were found to be terpenes such as 1,8-cineole and p-cymene, followed by β-triketones and alkyl esters. The repellent activity of the essential oil against both species was significantly higher when compared with the main component, 1,8-cineole, alone. These results indicate that the repellent effect of E. nitens is not due only to the main component, 1,8-cineole, but also that other compounds may be responsible. Aedes aegypti was found to be more tolerant to the essential oil larvicidal effects than Ae. albopictus (Ae. aegypti LC₅₀ = 52.83 ppm, Ae. albopictus LC ₅₀ = 28.19 ppm). The repellent and larvicidal activity could be associated to the presence of cyclic β-triketones such as flavesone, leptospermone, and isoleptospermone.

Insecticide resistance is one of the primary threats to the recent gains in malaria control. This is especially true in Guinea, where long-lasting insecticidal nets are currently the primary vector control intervention. To better inform the national malaria control program on the current status of insecticide resistance in Guinea, resistance bioassays were conducted, using Anopheles gambiae s.l. Giles, in three sites. Molecular analyses were also done on An. gambiae s.l. to determine the species and find whether the target-site mutations kdr and Ace1^R were present. Susceptibility tests revealed resistance to DDT and pyrethroids, although mosquitoes were susceptible to deltamethrin in two of the three sites tested. Mosquitoes were susceptible to bendiocarb, except in Kissidougou, Guinea. The kdr-west mutation was widespread and the frequency was 60% or more in all sites. However, the Ace1^R mutation was present in low levels. Insecticide susceptibility should continue to be monitored in Guinea to ensure insecticide-based vector control methods remain effective.

Mosquitoes may develop resistance to insecticide active ingredients (AI). Thus, mosquitoes should be tested for resistance to confirm efficacy of insecticide-based control, inform management decisions, and protect public and environmental health. Our objectives were to determine a baseline of resistance for six AIs used in mosquito control in the United States to assess how resistance differs between mosquito collection location, AI, and mosquito species (container-ovipositing Aedes and Culex that may oviposit in containers or other sources). Field-collected eggs from 26 mosquito populations of five different species or hybrid species (Aedes albopictus Say, Aedes triseriatus Say, Culex pipiens L., Culex quinquefasciatus Say, Culex pipiens/quinquefasciatus) were obtained from four regions across the United States. Centers for Disease Control and Prevention bottle bioassays were used to determine baseline resistance and susceptibility status for six AIs (bifenthrin, deltamethrin, etofenprox, malathion, permethrin, and phenothrin). World Health Organization guidelines were used to classify mosquitoes as susceptible (98–100% mortality at diagnostic time [DT]), possibly resistant (80–97% mortality at DT), or resistant (< 80% mortality at DT). Aedes spp. mosquitoes were less likely to exhibit resistance, compared with Culex spp. mosquitoes. A high degree of resistance to etofenprox and malathion was observed (4–26-fold greater resistance to these two AIs compared with the other examined AIs). Baseline data on resistance and susceptibility for mosquitoes exposed to commonly used insecticides may help us evaluate resistance trends and highlight the importance of assessing local resistance trends before insecticide-based control measures are implemented.

Biorational insecticides are being increasingly used in integrated pest management programs. In laboratory bioassays, the pathogenicity of blastospores and conidia of the entomopathogenic fungus Metarhizium brunneum ARSEF 4556 was evaluated against larvae of three mosquito species. Three propagule concentrations (1 × 10⁶, 1 × 10⁷, and 1 × 10⁸ spores ml ^{– 1}) were used in the bioassays. Results showed that Aedes aegypti had lower survival rates when exposed to blastospores than when exposed to conidia, whereas the converse was true for Culex quinquefasciatus larvae. Anopheles stephensi larvae survival rates were similar when exposed to blastospores and conidia, except at the higher doses, where blastospores were more virulent. Several assays showed little difference in mortalities when using either 1 × 10⁷ or 1 × 10⁸ spores ml ^{– 1}, suggesting a threshold above which no higher control levels or economic benefit would be achieved. When tested at the lowest dose, the LT₅₀ of Cx. quinquefasciatus using blastospores, wet conidia, and dry conidia was 3.2, 1.9, and 4.4 d, respectively. The LT₅₀ of Ae. aegypti using blastospores, wet conidia, and dry conidia was 1.3, 3.3, and 6.2 d, respectively. The LT₅₀ of An. stephensi using blastospores, wet conidia, and dry conidia was 2.0, 1.9, and 2.1 d, respectively. These observations suggest that for optimized control, two different formulations of the fungus may be needed when treating areas where there are mixed populations of Aedes, Anopheles, and Culex.

Musca domestica L. were collected from cattle diagnosed with bovine ringworm to evaluate the potential of the house fly to disseminate Trichophyton verrucosum E. Bodin, a fungal dermatophyte that is the causative agent for ringworm in cattle. Fungal isolates were cultured from 45 individual flies on supplemented Sabouraud dextrose agar, and isolates were identified using morphological and microscopic approaches. Each isolate was identified further by PCR amplification of the ribosomal DNA locus with fungal-specific primers and subsequent amplicon sequencing. Trichophyton verrucosum was not identified using these approaches. However, 35 different fungal species representing 17 genera were cultured from collected flies, including several species that are allergenic and pathogenic to humans and animals. Several species within the fungal orders Hypocreales, Microascales, Onygenales, Saccharomycetales, Xylaniales, and Agaricales were observed for the first time on house flies. The most frequent fungus recovered was Cladosporium cladosporoides Fresen, which is known to be a ubiquitous, airborne allergen to humans.

Arthropod-borne diseases negatively affect humans worldwide. Understanding the biology of the arthropod vectors and the pathogens they harbor, the arthropods are moving targets as a result of climate change, ecosystem degradation, species introductions, and increased human travel. Viruses within the California serogroup of the genus Orthobunyavirus (family Bunyaviridae) are among the mosquito-borne viruses of concern owing to their zoonotic potential. Two of these, snowshoe hare virus (SSHV) and Jamestown Canyon virus, were shown, using a combination of serology and virus isolations, to circulate on the Island of Newfoundland, Canada, in the 1980s. More recently, serological analysis demonstrated that these two viruses continue to circulate on the Island in several domesticated and wild animals. Here, we detected the seroconversion to SSHV in wild snowshoe hares and in a single sentinel rabbit. The seroconversion in the sentinel rabbit occurred in early August (2011), which corresponded to the weeks of peak mosquito collections and the timing of the detection of SSHV in suspected mosquito vectors. A portion of the SSHV S segment sequence was generated from mosquito pools collected at sites near the sentinel rabbits and phylogenetically analyzed using the neighbor-joining method with other available California serogroup virus sequences. This analysis validated the SSHV identification but showed that the Newfoundland sequence fell outside the other SSHV sequences available, which originated from the United States between 1959 and 2005.

A cross-sectional study was conducted in the Eastern part of Zambia that previously reported a plague outbreak. The aim of the study was to evaluate the potential role of pigs, goats, and sheep as sero-surveillance hosts for monitoring plague, and to investigate the flea vectors and potential reservoir hosts to establish the current status of plague endemicity in the district. Serum samples were collected from 96 rodents, 10 shrews, 245 domestic pigs, 232 goats, and 31 sheep, whereas 106 organs were eviscerated from rodents and shrews. As for fleas, 1,064 Echidnophaga larina Jordan & Rothschild, 7 Xenopsylla cheopis (Rothschild), and 382 Echidnophaga gallinacea (Westwood) were collected from these animals in 34 villages. Enzyme-Linked Immunosorbent Assay (ELISA) and Polymerase Chain Reaction (PCR) tests were performed on serum, and organs and fleas to determine IgG antibodies against Fraction 1 antigen and pla gene of Yersinia pestis, respectively. ELISA results showed that 2.83% (95% CI = 0.59–8.05) rodents, 9.0% (95% CI = 5.71–13.28) domestic pigs, 4.7% (95% CI = 2.39–8.33) goats, and 3.2% (95% CI = 0.08–16.70) sheep were positive for IgG antibodies against Fra1 antigen of Y. pestis. On PCR, 8.4% (95% CI = 3.96–15.51) of the rodents were detected with Y. pestis pla gene, whereas all fleas were found negative. The common fleas identified were E. larina from pigs, whereas X. cheopis were the only fleas collected from rodents. The presence of sero-positive animals as well as the occurrence of X. cheopis on local rodents suggests that Y. pestis remains a risk in the district.

Cattle manure is one of the primary larval developmental habitats of house flies, Musca domestica (L.). Cattle serve as asymptomatic reservoirs of Shiga toxin-producing Escherichia coli (STEC), and bacteria are released into the environment in cattle feces. The USDA-FSIS declared seven STEC serogroups (O157, O26, O45, O103, O145, O121, and O111) as adulterants in beef products. In addition, the serogroup O104 was a culprit of a large outbreak in Germany in 2011. Our study aimed to assess the prevalence of seven non-O157 STEC (O26, O45, O145, O103, O121, O111, and O104) serogroups in adult house flies. Flies (n = 463) were collected from nine feedlots and three dairy farms in six states in the United States and individually processed. This involved a culturing approach with immunomagnetic separation followed by multiplex polymerase chain reactions for detection of individual serogroups and virulence traits. The concentration of bacteria on modified Possé agar ranged between 1.0 × 10¹ and 7.0 × 10⁷ (mean: 1.5 ± 0.3 × 10⁶) CFU/fly. Out of 463 house flies, 159 (34.3%) carried one or more of six E. coli serogroups of interest. However, STEC was found in 1.5% of house flies from feedlots only. These were E. coli O103 and O104 harboring stx1 and ehxA and E. coli O45 with stx1, eae, and ehxA. This is the first study reporting the isolation of non-O157 STEC in house flies from the confined cattle environment and indicating a potential role of this insect as a vector and reservoir of non-O157 STEC in confined beef cattle.

Efficient detection of enterohemorrhagic Escherichia coli O157:H7 is important to monitor the safety of food products obtained from cattle, and it has been primarily accomplished by analyzing manure samples by selective cultivation techniques, PCR, and ELISA. As each technique suffers from different biases, there may be value in using multiple methods and samples to increase detection efficiency. Difficulties associated with cattle manure sampling can be circumvented by isolation of E. coli O157:H7 from house flies, Musca domestica (L.), which present as an important vector for spreading diseases. Thus, isolation of pathogens directly from house flies provides information about the potential human health impact that house fly dispersal can have because of pathogen distribution. House flies can disperse from dairy farms, where E. coli O157:H7 endemically thrive in cattle, to restaurants where food is prepared and served. Here, we report that detecting E. coli O157:H7 in house flies was 2.7 times more frequent than in manure from nearby dairy farms. Flies appear to offer a promising alternative in efforts to detect E. coli O157:H7 in dairy farms, restaurants, processing plants, and other establishments.

Tick-borne pathogens transmitted by Ixodes scapularis Say (Acari: Ixodidae), also known as the deer tick or blacklegged tick, are increasing in incidence and geographic distribution in the United States. We examined the risk of tick-borne disease exposure in 9 national parks across six Northeastern and Mid-Atlantic States and the District of Columbia in 2014 and 2015. To assess the recreational risk to park visitors, we sampled for ticks along frequently used trails and calculated the density of I. scapularis nymphs (DON) and the density of infected nymphs (DIN). We determined the nymphal infection prevalence of I. scapularis with a suite of tick-borne pathogens including Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti. Ixodes scapularis nymphs were found in all national park units; DON ranged from 0.40 to 13.73 nymphs per 100 m². Borrelia burgdorferi, the causative agent of Lyme disease, was found at all sites where I. scapularis was documented; DIN with B. burgdorferi ranged from 0.06 to 5.71 nymphs per 100 m². Borrelia miyamotoi and A. phagocytophilum were documented at 60% and 70% of the parks, respectively, while Ba. microti occurred at just 20% of the parks. Ixodes scapularis is well established across much of the Northeastern and Mid-Atlantic States, and our results are generally consistent with previous studies conducted near the areas we sampled. Newly established I. scapularis populations were documented in two locations: Washington, D.C. (Rock Creek Park) and Greene County, Virginia (Shenandoah National Park). This research demonstrates the potential risk of tick-borne pathogen exposure in national parks and can be used to educate park visitors about the importance of preventative actions to minimize tick exposure.

Because of the increasing incidence of human ehrlichiosis in Wisconsin, we assessed reports of human infections by Ehrlichia chaffeensis and the distribution of its vector, the lone star tick (Amblyomma americanum (L.)). From 2008 through 2015, 158 probable and confirmed human cases of E. chaffeensis infections were reported to the Wisconsin Department of Health Services. Five cases without travel history outside of Wisconsin were confirmed as E. chaffeensis by polymerase chain reaction. Surveillance for the vector occurred from 2008 through 2015 and was based on active and passive methods, including examination of white-tailed deer, collections from live-trapped small mammals, submissions of ticks removed from wild and domestic animals through the Wisconsin Surveillance of Animals for Ticks (SWAT) program, digital or physical submissions by the public to the University of Wisconsin Insect Diagnostic or Medical Entomology laboratories, and active tick dragging. More than 50 lone star ticks (46 adults, 6 nymphs, and 1 larva) were identified. Lone star ticks were more commonly found in south central Wisconsin, particularly in Dane County, where discovery of more than one life stage in a single year indicates possible establishment.

Ceratopogonidae (Diptera) were collected on sheep farms in southern Ontario to establish whether Culicoides spp. pose a threat to the livestock industry. Specimens were collected in modified CO₂-baited Centers for Disease Control and Prevention light traps, returned to the laboratory, freeze-killed, and identified to species under a microscope. In addition to Culicoides variipennis (Coquillet), we found that Culicoides sonorensis Wirth & Jones occurred on a number of farms over a 2-yr period. These records represent a significant departure from C. sonorensis’ previously known geographical distribution. We present spatial and temporal distribution data for both species, with an emphasis on C. sonorensis. DNA sequence information is presented so that researchers lacking the necessary taxonomic skills can determine whether C. sonorensis is present in their collections. To differentiate C. sonorensis from C. variipennis, taxonomically reliable and informative traits were found in EF1a and, to a lesser extent, in ITS1, whereas the universal barcode region of cytochrome oxidase subunit 1 (CO1) was unsuitable.

The distribution of mosquito species in Rio Grande do Norte state, Brazil, was compiled from published data mid-2016 and a review of specimens deposited in the entomological collection of the Entomology Laboratory of the Federal University of Rio Grande do Norte. The existing records exist for 40 of the 167 municipalities in the state. The specimens in the Entomology Laboratory were collected using Shannon traps and by active search for immature individuals and from aquatic habitats using standard methods, in preserved Atlantic Forest and Caatinga remnants, located in urban and rural areas of the state. In total were recorded 76 species distributed into 25 subgenera, 15 genera, nine tribes, and two subfamilies, in addition to 15 new species records for the state.

In this study, ticks from pastoral communities in Kenya were tested for Rickettsia spp. infections in geographical regions where the presence of tick-borne arboviruses had previously been reported. Rickettsial and arbovirus infections have similar clinical features which makes differential diagnosis challenging when both diseases occur. The tick samples were tested for Rickettsia spp. by conventional PCR using three primer sets targeting the gltA, ompA, and ompB genes followed by amplicon sequencing. Of the tick pools screened, 25% (95/380) were positive for Rickettsia spp. DNA using the gltA primer set. Of the tick-positive pools, 60% were ticks collected from camels. Rickettsia aeschlimannii and R. africae were the main Rickettsia spp. detected in the tick pools sequenced. The findings of this study indicate that multiple Rickettsia species are circulating in ticks from pastoral communities in Kenya and could contribute to the etiology of febrile illness in these areas. Diagnosis and treatment of rickettsial infections should be a public health priority in these regions.

We report a case of urinary myiasis occurring in a 60-yr-old Iranian male patient with urinary tract problems and a history of travel to Thailand who was referred to Shafagh Medical Laboratory in Tehran (Iran). Larvae excreted in the patient's urine were confirmed by morphological identification key and DNA barcoding to belong to the species Megaselia scalaris Loew,which is known as the scuttle fly. Based on the patient's history, he was infectedwith M. scalaris in Thailand. To the best of our knowledge, this is the first report of urinary myiasis caused by M. scalaris in Thailand.

Insecticides based on botanical sources have taken on increased attention due to differing modes of action from current insecticides in use and the view that they may be environmentally friendly. Thymoquinone, a component in the essential oil of incense cedar heartwood, has been shown to have insecticidal action against adult mosquitoes. This study evaluated relative toxicities of thymoquinone, selected derivatives of thymoquinone, hydroquinone, and arbutin to determine if any had similar or better activity. The intrinsic toxicities of hydroquinone and thymohydroquinone were not significantly different from thymoquinone, while libocedrol and arbutin were significantly less toxic.

Aedeomyia squamipennis (Lynch Arribalzaga) is a tropical mosquito (Diptera: Culicidae) found throughout most of the American Tropics, from eastern Mexico through Argentina, including several Caribbean islands. Larvae are typically associated with bodies of water with dense growths of aquatic vegetation, particularly Pistia stratiotes L., water lettuce. Adult females feed predominantly on the blood of birds and seek hosts in forest canopies. Aedeomyia squamipennis is considered an important vector of Gamboa virus and avian malaria, and is also suspected of transmitting Venezuelan equine encephalitis virus. Collections from Florida City, FL, near the southern tip of the Florida peninsula, yielded larvae, adult males, and females of Ad. squamipennis, constituting a new genus and species record for Florida and the United States. The widespread availability of larval habitat and suitable hosts in Florida will likely lead to expansion of Ad. squamipennis in Florida, and perhaps into neighboring states. In South America, Ad. squamipennis is found as far south as Buenos Aires, Argentina, at ~35° S latitude, which is equivalent in N latitude to coastal North Carolina. The northern limit of expansion of Ad. squamipennis in North America will likely be limited by winter temperatures and availability of larval habitat.

Aedes aegypti (L.) is the primary vector of dengue, yellow fever, Zika, and chikungunya viruses, whereas Anopheles pseudopunctipennis (Theobald) is the principal vector for malaria in Latin America. The larval stage of these mosquitoes occurs in very different development habitats, and the study of their respective behaviors could give us valuable information to improve larval control. The aim of this study was to set up a bioassay to study basic larval behaviors using a video-tracking software. Larvae of An. pseudopunctipennis came from two localities in Salta Province, Argentina, while Ae. aegypti larvae were of the Rockefeller laboratory strain. Behaviors of individual fourth-instar larvae were documented in an experimental petri dish arena using EthoVision XT10.1 video-tracking software. The overall level of movement of larval An. pseudopunctipennis was lower than that for Ae. aegypti, and, while moving, larval An. pseudopunctipennis spent significantly more time swimming near the wall of the arena (thigmotaxis). This is the first study that analyzes the behavior of An. pseudopunctipennis larvae. The experimental system described here may be useful for future studies on the effect of physiological, toxicological, and chemosensory stimuli on larval behaviors.

This study evaluated the species composition of sand flies and identified potential vectors of Leishmania Ross species in rural areas of Porto Velho, Rondônia State, Brazil. American cutaneous leishmaniasis (ACL) is one of the gravest threats to public health in this state. Sand flies were collected over the course of 2014 and 2015 using HP light traps. Polymerase chain reaction was performed by targeting the Leishmania mkDNA region. In total, 2,344 sand flies were collected, from which 45 species, nine subgenera, and five species group were identified. The most abundant species were Lutzomyia antunesi (Coutinho) (n = 597, 25.47%), Lutzomyia ubiquitalis (Mangabeira) (n = 496, 21.16%), and Lutzomyia octavioi (Vargas) (n = 199, 8.49%). The greatest diversity occurred in the forest environment where the most abundant species were L. antunesi (n = 588, 25.07%), L. ubiquitalis (n = 493, 21.02%), L. octavioi (n = 199, 8.49%), and Lutzomyia flaviscutellata (Mangabeira) (n = 132, 5.63%). Two pools of L. ubiquitalis were positive for Leishmania DNA, which suggests that L. ubiquitalis is a putative vector of leishmaniasis in the municipality of Porto Velho.