Rickettsia parkeri Luckman (Rickettsiales: Rickettsiaceae) is a pathogenic spotted fever group Rickettsia transmitted by Amblyomma maculatum Koch (Acari: Ixodidae) in the United States. The acute innate immune response to this pathogen and the effect of tick feeding or salivary components on this response is largely unknown. We hypothesized that A. maculatum saliva enhances R. parkeri infection via downregulation of the acute cellular and cytokine immune response. C3H/HeN mice were intradermally inoculated with R. parkeri both with and without A. maculatum saliva. Flow cytometry and microscopic evaluation of inoculation site skin suspensions revealed that neutrophils and macrophages predominated at 6 and 24 h post R. parkeri inoculation, respectively. This cellular influx was significantly downregulated when A. maculatum saliva was inoculated along with R. parkeri. Inflammatory cytokines (interferon c and interleukins 6 and 10) were significantly elevated after R. parkeri inoculation. However, cytokine concentration and rickettsial load were not significantly modified by A. maculatum saliva during the acute phase of infection. These results revealed that tick saliva inhibits the cutaneous cellular influx during the acute phase of rickettsial infection. Further study is needed to determine the overall impact of this effect on the establishment of rickettsiosis in the host and development of disease.

Lutzomyia cruciata (Coquillet) is a vector of cutaneous leishmaniasis in Mexico and Central America. However, several aspects of its ecology and behavior are unknown, including whether a male pheromone partially mediates the sexual behavior of this sand fly. In this study, we evaluated the behavioral response of females to male abdominal extracts in a Y-tube olfactometer. The volatile compounds from male abdominal extracts were identified by gas chromatography–mass spectrometry and compared with those of female abdominal extracts. Finally, the disseminating structures of the putative sex pheromone were examined by scanning electron microscopy in the male abdomen. Females were more attracted to male abdominal extract than to the hexane control, suggesting the presence of male-produced sex pheromone. The male abdominal extracts were characterized by the presence of 12 sesquiterpene compounds. The major component, an unknown sesquiterpene with an abundance of 60%, had a mass spectrum with molecular ion of m/z 262. In contrast, the abdominal female extracts contained saturated fatty acids. Finally, we detected the presence of small “papules” with a mammiform morphology distributed on the abdominal surface of tergites IV-VII of male Lu. cruciata. These structures are not present in females. We conclude that Lu. cruciata males likely produce a pheromone involved in attracting or courting females.

This work was aimed at evaluating Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) life cycle, life tables, and reproductive and population parameters. This species is a necrophagous, synanthropic, cosmopolitan fly with broad distribution in the Neotropical region. Six successive generations from adult specimens collected in Bogotá, Colombia, were analyzed in laboratory conditions. The fly's life cycle was evaluated, using a natural diet of pig's liver (L) and two artificial diets—milk powder and egg (ME) and powdered liver (PL). The species' longest life cycle duration (in days), from egg to pupa, was obtained with the ME diet (24.6 ± 6.7) whereas the lowest value was recorded with the L diet (18.8 ± 3.6). Female adult forms achieved high longevity values, this being highest in ME (102.8 ± 3.6). The most efficient diets (L and ME) led to the following average reproductive and population values: 226.2 and 465.4 net reproductive rate (Ro), 22.2 and 47.1 cohort generation time (Tc), 0.24 and 0.13 intrinsic population growth rate (r_m), and 1.27 and 1.13 finite rate of increase (λ). The set of results (in terms of relatively high average values concerning the biological characteristics analyzed) showed the species' adaptability to the environmental and nutritional conditions established in a laboratory setting, thereby enabling their colonization.

Necrophagous species of insects provide useful complementary data to estimate the postmortem interval in forensic cases. Here, for the first time, we report on insect specimens collected from human corpses in Riyadh, Kingdom of Saudi Arabia. During the study, 14 beetle larvae were collected from the outdoor corpse (case report one) and five flies and seven beetles were collected from the indoor corpse (case report two). Sequencing was performed to study the mitochondrial DNA (mtDNA) as the prospective basis of an identification technique. The sequencing focused on a section of the cytochrome oxidase I encoding region of mtDNA. Two beetle species, Dermestes frischii (Kugelann) and Dermestes maculatus (De Geer) (Coleoptera: Dermestidae), and one fly species, Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae), were identified. These results will be instrumental in the implementation of a Saudi database of forensically relevant insects.

Forensic entomology is a science that uses insect fauna as a tool to assist in criminal investigations and civil proceedings. Although the most researched insects are the Diptera and Coleoptera, ants may be present in all stages of decomposition. The aim of this study was to evaluate the role of ants and their action on blow flies during the decomposition process. Experiments were performed in which four pig carcasses were exposed in the cold and dry season (November/2012 and March/2013) and four in the hot and wet season (May/2013 and August/2013). Flies were the first insects to detect and interact with the carcasses, and six species of the Calliphoridae family were identified. Ants (Hymenoptera: Formicidae) were the second group, with six subfamilies identified. Myrmycinae represented 42% of the species, followed by Formicinae (28%), Ectatominae and Ponerinae (both 10%), and Ecitoninae and Dolichoderinae (both 5%). The ants acted on the carcasses as predators of visiting species, omnivores, and necrophagous, in all cases significantly affecting the decomposition time, slowing it down when the ants preyed on adult and immature insects consuming the carcass, or accelerating it by consuming the carcass and creating holes that could serve as gateways for the action of other organisms. The ants also generated artifacts that could lead to forensic misinterpretation.

The castor bean tick Ixodes ricinus (L.) is the principal vector for a variety of viral, bacterial, and protozoan pathogens causing a growing public-health issue over the past decades. However, a national density map of I. ricinus is still missing. Here, I. ricinus nymphs in Germany were investigated by compiling a high-resolution map depicting the mean annually accumulated nymphal density, as observed by monthly flagging an area of 100 m². Input data comprise ticks collected at 69 sampling sites. The model domain covers an area of about 357,000 km² (regional scale). Two negative binomial regression models were fitted to the data to interpolate the tick densities to unsampled locations using bioclimatic variables and land cover, which were selected according to their significance by the Akaike information criterion (AIC). The default model was fitted to the complete dataset resulting in AIC = 842. An optimized model resulted in a significantly better value of AIC = 732. Tick densities are very low in urban (green) areas. Maximum annual densities up to 1,000 nymphs per 100 m² are observed in broad-leaved forests. The tick maps were verified by leave-one-out cross-validation. Root mean square errors of RMSE = 137 and RMSE = 126 nymphs per 100 m² were estimated for the two models, respectively. These errors are of the order of the interannual variation of the tick densities. The compilation of a high-resolution density map of unfed nymphal I. ricinus for Germany provides a novel, nationwide insight into the distribution of an important disease vector.

Lutzomyia fraihai (Martins, Falcão & Silva 1979), described from Três Braços municipality (Bahia state, Brazil), is closely related to Nyssomyia yuilli yuilli (Young & Porter, 1972), described from Rio Anori, Antioquia, Colombia. The former species, despite the morphological differences reported by its authors, was included as a junior synonym of Ny. yuilli yuilli, and this status has been accepted by sand fly-taxonomists. However, we had an opportunity to examine both taxa, and a clear difference was noted in their male genitalia. In this study, the morphometric/morphologic characters of specimens that had been identified as Ny. yuilli yuilli from different localities (Brazil, Peru, Colombia, Bolivia) and type material of both Ny. yuilli yuilli and Lu. fraihai were analyzed. The results revealed that males from Brazil (Bahia, Acre, Pará, and Rondônia states) and Peru present statistically significant differences in the ratio between paramere rectangular and digitiform areas as well as for the cercus length and width of the aedeagal ducts tips, when compared with the type specimens of Ny. yuilli yuilli, but are statistically similar to those characters of Lu. fraihai. However, females of the two species are indistinguishable. Thus, based on the male morphology, Ny. fraihaistat. rev. n. com. was resurrected from the synonymy of Ny. yuilli yuilli. The specimens from the Atlantic forest biome and the Amazon biome in Brazil, Peru, and Bolivia in fact belong to Ny. fraihai. This species presents a cis-Andean distribution, while Ny. yuilli yuilli has been found in both Andean and Trans-Andean areas.

The parasitic quill mites of the family Syringophilidae (Acariformes: Prostigmata) are recorded on ibises and spoonbills (Pelecaniformes: Threskiornithidae) for the first time. Four new species of the genus StibarokrisKethley, 1970 are described: 1) S. theristicus Skoracki, Zmudzinski & Unsoeld sp. nov. ex Theristicus caudatus (Boddaert, 1783) from Brazil, 2) S. geronticus Skoracki, Zmudzinski & Unsoeld sp. nov. ex Geronticus calvus (Boddaert, 1783) from South Africa, 3) S. brevisetosus Skoracki & Zmudzinski sp. nov. ex Plegadis falcinellus (L., 1766) from Turkey, and 4) S. plataleus Skoracki & Zmudzinski sp. nov. ex Platalea leucorodia L., 1758 from Austria. Additionally, a key to all described species in the genus is presented, and hypothesis of the Gondwanan origin of syringophilids associated with ibises and spoonbills is briefly discussed.

Seasonal, spatial, and habitat responses of carrion-associated Diptera assemblages can provide valuable information about the presence or absence of species and their relative abundance, and thereby enhance understanding of their responses to environmental variables and how this may have an impact on forensic investigations. Three different nature reserves (localities) within the Municipality of Tshwane, South Africa, were selected to determine whether species assemblages of carrion-feeding flies differ between seasons, localities, and habitat types. A total of 59,511 adult Diptera, identified to 35 species in eight different families, were collected using modified Redtop hanging traps, baited with liver and fish, during four seasons in three different habitat types. Species assemblages differed temporally, with season being the main factor determining species diversity and not locality or habitat. However, savanna and human-disturbed habitats supported a higher abundance and species richness than grassland habitats. Areas adjacent to the localities, such as large urban expanses in Dinokeng or agricultural holdings in Rietvlei, led to an increase in the abundance and mean species richness of carrion-associated Diptera, and in increased numbers of pest or invasive species such as Chrysomya megacephala (F.). Despite this, the overall species assemblages present in human-disturbed areas were very similar to those recorded in natural habitats.

The mosquito fauna (Culicidae) from remote northern areas of the State of Amazonas were sampled using Centers for Disease Control and Prevention, Shannon, Malaise, and Suspended traps, together with net sweeping and immature collections. One hundred and seven collections were performed in five localities along the Padauari River, State of Amazonas, Brazil, during June 2010. The 20,557 mosquitoes collected are distributed in 17 genera, representing 117 different species, of which four are new distributional records for the State of Amazonas. Furthermore, there are 10 morphospecies that may represent undescribed new taxa, eight of which are also new records for the State of Amazonas. The genus Culex had the highest number of species and the largest number of individuals. Aedes and Psorophora both represented 10% of the total sample and had the second highest number of species and individuals. The most abundant species was Culex (Melanoconion) gnomatos Sallum, Hutchings & Ferreira, followed by Aedes (Ochlerotatus) fulvus (Wiedemann), Culex (Melanoconion) vaxus Dyar, Culex (Melanoconion) portesi Senevet & Abonnenc, Psorophora (Janthinosoma) amazonica Cerqueira, Culex (Culex) mollis Dyar & Knab, Psorophora (Janthinosoma) albigenu (Peryassú), and Culex (Melanoconion) theobaldi Lutz. The epidemiological and ecological implications of mosquito species found are discussed and are compared with other mosquito inventories from the Amazon region. The results represent the most diverse standardized inventory of mosquitoes along the Padauari River, with the identification of 127 species-level taxa distributed in five localities, within two municipalities (Barcelos and Santa Isabel do Rio Negro).

The Lake Baringo and Lake Victoria regions of Kenya are associated with high seroprevalence of mosquito-transmitted arboviruses. However, molecular identification of potential mosquito vector species, including morphologically identified ones, remains scarce. To estimate the diversity, abundance, and distribution of mosquito vectors on the mainland shores and adjacent inhabited islands in these regions, we collected and morphologically identified adult and immature mosquitoes and obtained the corresponding sequence variation at cytochrome c oxidase 1 (COI) and internal transcribed spacer region 2 (ITS2) gene regions. A total of 63 species (including five subspecies) were collected from both study areas, 47 of which have previously been implicated as disease vectors. Fourteen species were found only on island sites, which are rarely included in mosquito diversity surveys. We collected more mosquitoes, yet with lower species composition, at Lake Baringo (40,229 mosquitoes, 32 species) than at Lake Victoria (22,393 mosquitoes, 54 species). Phylogenetic analysis of COI gene sequences revealed Culex perexiguus and Cx. tenagius that could not be distinguished morphologically. Most Culex species clustered into a heterogeneous clade with closely related sequences, while Culex pipiens clustered into two distinct COI and ITS2 clades. These data suggest limitations in current morphological identification keys. This is the first DNA barcode report of Kenyan mosquitoes. To improve mosquito species identification, morphological identifications should be supported by their molecular data, while diversity surveys should target both adults and immatures. The diversity of native mosquito disease vectors identified in this study impacts disease transmission risks to humans and livestock.

Culex pipiens L., the northern house mosquito, is the primary vector of West Nile virus to humans along the east coast of North America and thus the focus of much study. This species is an urban container-breeding mosquito whose close contact with humans and flexibility in host choice has led to its classification as a “bridge vector”; that is, it is thought to move zoonotic diseases to humans from vertebrate reservoirs. While this invasive species is now well documented in its established range, which expanded in 2001 to include Canada, the existence of populations of this species along the fringes of its range are less well known. Here we report, using morphological and genetic techniques, the existence of two locations where Cx. pipiens exists in Newfoundland in both expected and unexpected sites based on projected habitat suitability on the island.

The ability to mark individuals is a critical feature of many entomological investigations, including dispersal studies. Insect dispersal is generally investigated using mark–release–recapture techniques, whereby marked individuals are released at a known location and then captured at a measured distance. Ectoparasite dispersal has historically been challenging to study, in part because of the ethical concerns associated with releasing marked individuals. Here, we introduce the protein self-marking technique, whereby ectoparasites mark themselves in the field by feeding on the blood of an introduced host. We demonstrate the potential of this technique using laboratory-reared bed bugs (Cimex lectularius L.) that marked themselves by feeding on either rabbit or chicken blood. We then used enzyme-linked immunosorbent assays to detect host-specific blood serum proteins in bed bugs. We assessed these protein markers' ability to 1) distinctively identify marked individuals, 2) persist following multiple feedings on an alternate diet, 3) persist over time across a range of temperatures, and 4) transfer from marked to unmarked individuals. Protein markers were detectable in bed bugs before and after molting, remained detectible after multiple feedings on an alternate diet, persisted regardless of whether an individual was starved or fed on an alternate diet following original mark acquisition, and did not transfer between individuals. The duration of detectability depended on temperature. Our results suggest that protein self-marking is an effective technique for marking bed bugs and holds promise for use in dispersal studies of ectoparasitic insects.

Accumulating evidence suggests that enzootic transmission of pathogens such as West Nile virus (WNV) by mosquitoes is governed by host-bird interactions, including mosquito preferences for specific species and developmental stages of host birds, host bird availability, and host defensive behavior. Here, we examined how the attack rate of five mosquito species in southern California was influenced by the position of CO₂-baited traps in relation to distance from water and trap height. We identified 44,207 female mosquitoes representing five species: Aedes vexans Meigen, Anopheles franciscanus McCracken, Anopheles hermsi Barr & Guptavanij, and the two most abundant species which are also WNV vectors, Culex erythrothorax Dyar and Culex tarsalis Coquillett. Mosquito captures decreased markedly with trap height, and also decreased with distance from a riparian area but not with distance from an open water source lacking a vegetated border. The results of this study suggest that WNV-competent ornithophilic mosquitoes may amplify the virus especially in reservoir birds that roost or nest close to the ground and near riparian vegetation.

The spread of Aedes albopictus (Skuse) eastward in the mid-1980s from its initial establishment in Houston, TX, was associated with rapid declines and local disappearances of Aedes aegypti (L.) in Gulf Coast states and Florida where annual larval surveillance during the early 1990s described temporal and spatial patterns of competitive displacements in cemeteries and tire shops. Approximately 20 yr later in 2013–2014, we re-visited former collection sites and sampled aquatic immatures of these two species from tire shops in 10 cities on State Route 441 and from 9 cemeteries from Lakeland to Miami in southwest Florida. In the recent samples Ae. aegypti was recovered from three central Florida cities where it had not been detected in 1994, but its northern limit on Rte. 441, Apopka, did not change. Other evidence, such as trends at a few cemeteries, suggested a moderate resurgence of this species since 1994. Cage experiments that exposed female progeny of Ae. aegypti from recent Florida collection sites to interspecific mating by Ae. albopictus males showed that females from coexistence sites had evolved resistance to cross-mating, but Ae. aegypti from sites with no Ae. albopictus were relatively susceptible to satyrization. Habitat classifications of collection sites were reduced by principal component (PC) analysis to four variables that accounted for > 99% of variances; PCs with strong positive loadings for tree cover and ground vegetation were associated with collection sites yielding only Ae. albopictus. Within the coexistence range of the two species, the numbers of Ae. aegypti among total Aedes collected were strongly correlated in stepwise logistic regression models with two habitat-derived PCs, distance from the coast, and annual rainfall and mean maximum temperatures at the nearest weather station. Subtle increases in the range of Ae. aegypti since its previous displacements are interpreted in the context of the evolution of resistance to mating interference, realized versus fundamental niches of the two species, and persisting competition with Ae. albopictus.

During 2010–2016, tick specimens were solicited from veterinarians, biologists, and members of the public in Alaska. Eight species of ticks were recorded from domestic dogs. Some ticks were collected from dogs with recent travel histories to other countries or other U.S. states, which appears to explain records of ticks not native to Alaska such as Amblyomma americanum (L.) (lone star tick), Ixodes scapularis (Say) (blacklegged tick), and Ixodes ricinus (L.). However, we recorded Dermacentor variabilis (Say) (American dog tick) from dogs (and humans) both with and without travel history, suggesting that this nonindigenous tick could be establishing populations in Alaska. Other ticks commonly recorded from dogs included the indigenous Ixodes angustus Neumann and the invasive Rhipicephalus sanguineus (Latreille) (brown dog tick). Domestic cats were only parasitized by one tick species, the native I. angustus. Six species of ticks were recorded from humans: A. americanum (with and without travel history), Dermacentor andersoni Stiles (Rocky Mountain wood tick; travel associated), D. variabilis (with and without travel history), Haemaphysalis leporispalustris (Packard) (rabbit tick, native to Alaska), I. angustus, and R. sanguineus. Ixodes angustus predominated among tick collections from native mammals. Also, Ixodes texanus Banks (first record from Alaska) was collected from an American marten, Martes americana (Turton), H. leporispalustris was recorded from a snowshoe hare, Lepus americanus Erxleben, and Ixodes auritulus Neumann was collected from a Northwestern crow, Corvus caurinus Baird. The establishment of D. variabilis, D. andersoni, A. americanum, and/or I. scapularis in Alaska would have strong implications for animal and human health.

Entomopathogenic fungi are widely used to control arthropods not just in agricultural settings but also in Veterinary Medicine and Public Health. These products have been employed to control tick populations and tick-borne diseases. The effectiveness of these control measures not only depends on the fungi, but also on the tick species and environmental conditions. In Mesomediterranean areas, tick species are adapted to extreme climatic conditions and it is therefore especially important to develop suitable tick control strategies. The aim of this study was to evaluate the effectiveness of a new method of tick control which entails the application of a commercial strain of Beauveria bassiana (Balsamo, Vuillemin) on wild rabbit burrows under field conditions. Aqueous solutions of the product were applied using a mist blower sprayer into 1,717 burrows. Two trials were performed, one in spring and the other in summer. The parasitic index (PI) was calculated for 10 rabbits per treatment per time point on day +30, +60, and +90 posttreatment and efficiency was calculated by comparing the PI for ticks in treated and untreated rabbits. A total of 20,234 ixodid ticks were collected. Hyalomma lusitanicum Koch, 1844 was the most abundant tick feeding on rabbits. Treatment significantly reduced the PI in spring (by 78.63% and 63.28% on day +30 and +60, respectively; P<0.05), but appeared to be less effective in summer, with a marginally significant tick reduction of 35.72% on day +30 (P = 0.05). Results suggest that the efficacy of applications inside burrows could be temperature-dependent and that such applications could be an economic alternative to rabbit tick control during at least two months using a diluted solution of B. bassiana conidia.

Background: An insecticide susceptibility study was carried out on Anopheles gambiae sensu lato vectors in 11 districts in Ghana between October 2012 and January 2013. Methods:An. gambiae s.l. larvae were collected, bred under standard conditions, and 3–5 d postemerged females were used for bioassay. Between 22 and 25 female An. gambiae s.l. fed only 10% sugar were used for testing. Exposure was for 1 h (2 h for fenitrothion). An. gambiae that were knocked down were recorded every 10 min and mortalities recorded 24 h posttest. Eleven insecticides from four chemical classes were used: organochlorines, organophosphates, carbamates, and pyrethroids. Subsamples of An. gambiae were analyzed by polymerase chain reaction for species and knockdown resistance (kdr) allele determination. Results: Malathion was effective in killing An. gambiae in seven districts, fenitrothion in three districts, and propoxur in one district. The organophosphate and carbamate insecticides were effective in killing An. gambiae compared to pyrethroids and organochlorines. Of the limited samples analyzed, An. gambiae sensu stricto (39/110), An. coluzzii (66/110), and An. arabiensis (5/110) were identified. Few kdr (11/110) susceptible mosquitoes were detected. Homozygous kdr^RR (65/110) and heterozygous kdr^RS (8/110) genotypes were identified. Conclusions: An organophosphate insecticide is considered appropriate for indoor residual spraying (IRS) in the 11 districts earmarked for the IRS program in Ghana.

Permethrin resistance is widespread in Aedes aegypti (L.), the main dengue, zika, and chikungunya virus vector in Latin America and the Caribbean. A common mechanism of resistance to pyrethroids—knockdown resistance (kdr)—is conferred through mutations in the insect's voltage-dependent sodium channel. In this mosquito, around 10 replacement substitutions in the voltage-gated sodium channel gene (vgsc) have been reported in pyrethroid-resistant strains. Two of these mutations, named Ile1,016 and Cys1,534, are widespread in mosquito populations from Latin America and the Caribbean. This study assessed the levels of permethrin resistance and the frequency of two kdr mutations in eight Ae. aegypti populations collected in Puerto Rico in 2013. Permethrin resistance factors ranged from 33–214-fold relative to the New Orleans reference strain. The frequency of kdr mutation Ile1,016 ranged from 0.65 to fixation (1.0), and for Cys1,534 frequencies varied from 0.8 to fixation. Alarmingly, two populations—Carolina and Caguas—reached fixation at both loci. Our results suggest that permethrin effectiveness for Ae. aegypti control is compromised in these collections from Puerto Rico.

Invasive mosquito species can increase the transmission risk of native mosquito-borne diseases by acting as novel vectors. In this study, we examined the susceptibility of three exotic invasive mosquito species Aedes aegypti (L.), Ae. albopictus (Skuse), and Ochlerotatus japonicus (Theobald) to La Crosse virus (LACV) relative to the native primary vector Ochlerotatus triseriatus (Say). Adult females of the four mosquito species were orally challenged with LACV; incubated for 3, 5, 7, 9, or 11 d; and their midgut infection rates, dissemination rates, and effective vector competence were determined. Overall, Oc. japonicus (2.92) had the highest effective vector competence values, followed by Ae. albopictus (1.55), Ae. aegypti (0.88), and Oc. triseriatus (0.64). In addition, we assessed the relationship between mosquito size and LACV susceptibility for field-collected Oc. triseriatus and Oc. japonicus. We hypothesized that smaller adults would be more susceptible to LACV; however, our results did not support this hypothesis. Infected Oc. triseriatus tended to be larger than exposed but uninfected females, while infected and uninfected Oc. japonicus were similarly sized. These findings suggest that Oc. japonicus, Ae. albopictus, and Ae. aegypti have significant potential to transmit LACV and more research is needed to uncover their potential role in LACV epidemiology.

Anopheles mosquitoes may be incriminated as malaria vectors by observing sporozoites in their salivary glands and by testing heads or thoraces by an enzyme-linked immunosorbent assay (ELISA) to detect Plasmodium species-specific circumsporozoite proteins (CSP). This study tested Anopheles collected in Sungai Nyamuk Village for the presence of both Plasmodium falciparum and Plasmodium vivax CSP. The Anopheles spp. were collected by human landing collection indoors and outdoors and by indoor and outdoor resting catches in Sungai Nyamuk Village, Nunukan District, North Kalimantan Province from August 2010 to January 2012. Overall, 5,100 Anopheles spp. comprising 11 species were collected and 2,259 adult parous females were tested by ELISA. Of these, only one Anopheles peditaeniatus Leicester (3.8%, n = 26) and one Anopheles sundaicus sensu lato (0.6%, n = 157) that originated from outdoor biting catches tested positive for P. falciparum CSP. The remaining females from indoor biting, outdoor resting, and indoor resting catches were negative for P. falciparum and P. vivax proteins. Confirmation of these vectors biting outdoors indicated that P. falciparum transmission may be occurring outside of houses by An. peditaeniatus and An. sundaicus.

In total, 1,300 each of Anopheles kleini Rueda and Anopheles sinensis Wiedemann sensu stricto (s.s.) females (colonized from the Republic of Korea) and Anopheles dirus Peyton & Harrison (Thai strain) were allowed to feed on blood from Thai malaria patients naturally infected with Plasmodium vivax. The overall oocyst infection rates for An. dirus, An. kleini, and An. sinensis s.s. were 77.4, 46.1, and 45.9%, respectively. The mean number of oocysts was significantly higher for An. dirus (82.7) compared with An. kleini (6.1) and An. sinensis s.s. (8.6), whereas the mean number of oocysts for An. kleini and An. sinensis s.s. was similar. The overall sporozoite infection rates for An. dirus, An. kleini, and An. sinensis s.s. dissected on days 14-15, 21, and 28 days post-feed were significantly higher for An. dirus (90.0%) than An. kleini (5.4%), whereas An. kleini sporozoite rates were significantly higher than An. sinensis s.s. (<0.1%). The overall sporozoite indices for positive females with +3 (100-1,000 sporozoites) and +4 (>1,000 sporozoites) salivary gland indices were significantly higher for An. dirus (85.7%), compared with An. kleini (47.1%). Only one An. sinensis s.s. had sporozoites (+2; >10–100 sporozoites). These results indicate that An. kleini is a competent vector of vivax malaria. Although An. sinensis s.s. develops relatively high numbers of oocysts, it is considered a very poor vector of vivax malaria due to a salivary gland barrier.

Fleas and ticks are parasites of wild and domestic mammals, and can be vectors of several pathogens. In rural areas, domestic carnivores such as the domestic dog (Canis lupus familiaris L.), may act as a “bridge” between natural areas and human settlements where ectoparasites can be used as a metric of such link. The aim of this study was to identify fleas, ticks, and Rickettsia spp., collected from domestic and wild carnivores in a natural reserve and surrounding human settlements in Central Chile, using morphological keys and molecular analysis. We surveyed 170 households from which 107 dogs and eight cats were sampled. From the natural reserve, we sampled two chilla foxes (Pseudalopex griseus Gray), two lesser grison (Galictis cuja Molina), three kodkods (Leopardus guigna Molina), and four dogs. From dogs, we collected Ctenocephalides felis Bouché, Ctenocephalides canis Curtis, Pulex irritans L., and Rhipicephalus sanguineus s.l. Latreille; C. felis was the most frequent ectoparasite. Cats were infested only by C. felis and Rh. sanguineus s.l. From wild carnivores, we obtained C. canis and P. irritans, the latter being most frequent. Molecular analysis of P. irritans detected 10 haplotypes and two main clades, which tended to separate fleas from wild and domestic hosts. Molecular analysis of ompA and ompB genes confirmed the presence of Rickettsia felis in fleas collected from owned dogs and cats, which could represent a potential risk factor of R. felis transmission in the area.

Lutzomyia wellcomei (Fraiha, Shaw & Lainson) (Diptera: Psychodidae) can act as an important vector of Leishmania (Viannia) braziliensis. This study presents the results of collections carried out in a fragment of Atlantic Forest in a Conservation Unit of Rio Grande do Norte state. Collections occurred over 12 consecutive months using Shannon and CDC traps. A total of 777 sand flies from eight species were collected: Lutzomyia walkeri (Newstead), Lutzomyia evandroi (Costa Lima & Antunes), Lutzomyia wellcomei (Fraiha, Shaw & Lainson), Lutzomyia sordellii (Shannon & Del Ponte), Lutzomyia brasiliensis (Costa Lima), Lutzomyia lenti (Mangabeira), Lutzomyia longipalpis (Lutz & Neiva), and Lutzomyia abonnenci (Floch & Chassignet). Lutzomyia wellcomei was the most abundant species using the Shannon trap (97%) and L. walkeri in the CDC trap (81%). It is important to note the abundance of L. wellcomei in Shannon trap collections, which favors the capture of anthropophilic species. Lutzomyia wellcomei was only present in months where rainfall was above 100mm, confirming it as a species adapted to wetter months.

Zoonotic mosquito-borne viruses, such as the West Nile virus (WNV) and eastern equine encephalitis virus (EEEV), are major public health threats in the United States. Early detection of virus transmission and targeted vector management are critical to protect humans against these pathogens. Sentinel chickens and pool screening of mosquitoes, the most widely used methods of arbovirus early detection, have technical time-lags that compromise their early-detection value. The exploitation of sugar-feeding by trapped mosquitoes for arbovirus surveillance may represent a viable alternative to other methods. Here we compared effectiveness of sugar-impregnated nucleic-acid preserving substrates (SIPS) and sentinel chicken program for detecting WNV, EEEV, and St. Louis encephalitis virus in gravid traps, CO₂-baited light traps, and resting traps at 10 locations in two Florida counties. In St. Johns County, comparable numbers of EEEV detections were made by SIPS traps (18) and sentinel chickens (22), but fewer WNV detections were made using SIPS (1) than sentinel chickens (13). In Volusia County, seven arbovirus detections were made via the sentinel chicken program (one EEEV and six WNV), whereas only one arbovirus detection (WNV) was made using SIPS. CO₂-baited light traps captured >90% of total mosquitoes, yet yielded <30% of arbovirus detections. Resting traps and gravid traps captured a fraction of total mosquitoes, yet yielded roughly equivalent numbers of arbovirus detections, as did light traps. Challenges to successful deployment of SIPS include optimization of traps for collecting all vector species, increasing sugar-feeding rates of trapped vectors, and developing tractable methods for arbovirus detection.

Rickettsioses are re-emerging vector-borne zoonoses with a global distribution. Recently, Rickettsia sp. strain Atlantic rainforest has been associated with new human spotted-fever (SF) cases in Brazil, featuring particular clinical signs: eschar formation and lymphadenopathy. These cases have been associated with the tick species, Amblyomma ovale. From 2010 until 2015, the Brazilian Health Department confirmed 11 human SF cases in the Maciço de Baturité region, Ceará, Brazil. The present study reports the circulation of Rickettsia spp. in vectors from this entirely new endemic area for SF. A total of 1,727 ectoparasites were collected in this area from the environment, humans, and wild and domestic animals. Samples (n = 887) were screened by polymerase chain reaction (PCR), targeting the gltA and ompA rickettsial genes. Sequencing and phylogenetic analyses of gltA gene amplicons were carried out for 13 samples positive for both screening PCRs. Fragments of gltA and ompA from three samples were cloned, sequenced, and analyzed further. A. ovale and Rhipicephalus sanguineus specimens, collected from dogs, were found to be infected with Rickettsia sp. str. Atlantic rainforest, suggesting the importance of dogs in the epidemic cycle. Candidatus Rickettsia andeanae, Rickettsia felis, and Rickettsia bellii were also found infecting ticks and fleas in five municipalities, demonstrating the broad diversity of rickettsiae in circulation in the studied area. This study reports, for the first time, evidence of infection with Rickettsia sp. strain Atlantic rainforest in A. ovale and R. sanguineus in Ceará, and Ca. R. andeanae in an Atlantic rainforest environment of Brazil.

The Musca domestica antifungal peptide-1A (MAF-1A peptide) from housefly larvae was synthesized by solid-phase synthesis technique, and antiviral, antioxidant, and antifungal properties were evaluated in this study. Present results indicated that it could significantly inhibit the infection of influenza virus H₁N₁, Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), and Bombyx mori nuclear polyhydrosis virus (BmNPV), which displayed excellent virucidal activities. Antioxidant results demonstrated that the MAF-1A peptide had effective scavenging activity for hydroxyl and superoxide radicals, which were similar to that of ascorbic acid. Besides, antifungal results showed that it can also significantly inhibit the growth of four fungi, and the half inhibitory concentrations (IC₅₀) values were ∼59.3, 84.2, 144.9, and 48.5 μg/ml, respectively, highlighting an important role of MAF-1A peptide in the defense of M. domestica against pathogenic microorganisms. These results revealed that the MAF-1A peptide from housefly larvae has great potential as a natural ingredient for the exploitation of antiviral and antifungal therapeutic agents, avoiding abuse of chemical agents and environmental pollution.

California statewide West Nile virus (WNV) minimum infection rates in Culex thriambus Dyar mosquitoes are high; however, few specimens are submitted and tested each year, as their distribution seems limited to larval habitats along riparian systems. To evaluate the role of Cx. thriambus in the amplification, maintenance, and overwintering of WNV in Lake County, CA, the bionomics and vector potential of the species was investigated during 2014 and 2015. Culex thriambus was the most abundant mosquito species, with 1,153 adults and 7,624 immatures collected by vacuum aspiration and dip sampling, respectively, at the primary study site. Detection of WNV in four mosquito pools during September through November coincided with peak seasonality. Females entered and maintained a reproductive diapause during winter under field and seminatural conditions. Diapause was initiated in the majority of Cx. thriambus females by October and was terminated by 30 March. Some parous females (7.1%) and those in host-seeking arrest (7.1%) were collected throughout the winter period. An accrual of 679.51 degree-days (°D) was necessary for diapause termination under seminatural conditions. Culex thriambus females fed on 16 different avian species during spring and summer, and no mammalian feeds were detected. West Nile viral RNA was detected in four of 42 Cx. thriambus pools tested during June through November and infection rates ranged from 3.53–28.15/1,000 tested. In summary, WNV transmission may be increased along riparian corridors throughout California where Cx. thriambus mosquitoes remain relatively abundant.

Southern Zambia is the focus of strategies to create malaria-free zones. Interventions being rolled out include test and treat strategies and distribution of insecticide-treated bed nets that target vectors that host-seek indoors and late at night. In Macha, Choma District, collections of mosquitoes were made outdoors using barrier screens within homesteads or UV bulb light traps set next to goats, cattle, or chickens during the rainy season of 2015. Anopheline mosquitoes were identified to species using molecular methods and Plasmodium falciparum infectivity was determined by ELISA and real-time qPCR methods. More than 40% of specimens caught were identified as Anopheles squamosus Theobald, 1901 of which six were found harboring malaria parasites. A single sample, morphologically identified as Anopheles coustani Laveran, 1900, was also found to be infectious. All seven specimens were caught outdoors next to goat pens. Parasite-positive specimens as well as a subset of An. squamosus specimens from either the same study or archive collections from the same area underwent sequencing of the mitochondrial cytochrome oxidase subunit I gene. Maximum parsimony trees constructed from the aligned sequences indicated presence of at least two clades of An. squamosus with infectious specimens falling in each clade. The single infectious specimen identified morphologically as An. coustani could not be matched to reference sequences. This is the first report from Zambia of infections in An. squamosus, a species which is described in literature to display exophagic traits. The bionomic characteristics of this species needs to be studied further to fully evaluate the implications for indoor-targeted vector control.

Here, we present the results of a 2-yr sampling using malaise traps along the Atlantic Forest domain from the northeast to the south of Brazil. In total, 217 sand flies were collected, of which the most abundant species was Bichromomyia flaviscutellata (Mangabeira, 1942) (60.4%), followed by Psychodopygus ayrozai (Barretto & Coutinho, 1940) (11%) and Micropygomyia schreiberi (Martins, Falcão & Silva, 1975) (4.1%), and the remaining less abundant species comprised 10.1% of the total of sand flies collected. We report the occurrence for the first time of: 1) B. flaviscutellata, Pintomyia fischeri (Pinto, 1926), Ps. ayrozai, and Psychodopygus carreirai (Barretto, 1946) in the state of Alagoas; 2) Psychodopygus claustrei (Abonnenc, Lèger & Fauran,1979), Psychodopygus amazonensis (Root, 1934), and Sciopemyia sordellii (Shannon & del ponte, 1927) in the state of Bahia; 3) Nyssomyia anduzei (Rozeboom, 1942) in the state of Pernambuco; and 4) B. flaviscutellata, M. schreiberi, Ps. ayrozai, and Psychodopygus davisi (Root, 1934) in the state of Sergipe. Our results present novel records of sand flies collected with malaise traps in the Atlantic Forest domain demonstrating that different collecting methods such as malaise traps can provide new interesting data about these insects that are natural vectors of many pathogens.

Rickettsia bellii is a rickettsial species of unknown pathogenicity that infects argasid and ixodid ticks throughout the Americas. Many molecular assays used to detect spotted fever group (SFG) Rickettsia species do not detect R. bellii, so that infection with this bacterium may be concealed in tick populations when assays are used that screen specifically for SFG rickettsiae. We describe the development and validation of a R. bellii-specific, quantitative, real-time PCR TaqMan assay that targets a segment of the citrate synthase (gltA) gene. The specificity of this assay was validated against a panel of DNA samples that included 26 species of Rickettsia, Orientia, Ehrlichia, Anaplasma, and Bartonella, five samples of tick and human DNA, and DNA from 20 isolates of R. bellii, including 11 from North America and nine from South America. A R. bellii control plasmid was constructed, and serial dilutions of the plasmid were used to determine the limit of detection of the assay to be one copy per 4 μl of template DNA. This assay can be used to better determine the role of R. bellii in the epidemiology of tickborne rickettsioses in the Western Hemisphere.