Ontologies, which are made up by standardized and defined controlled vocabulary terms and their interrelationships, are comprehensive and readily searchable repositories for knowledge in a given domain. The Open Biomedical Ontologies (OBO) Foundry was initiated in 2001 with the aims of becoming an “umbrella” for life-science ontologies and promoting the use of ontology development best practices. A software application (OBO-Edit; *.obo file format) was developed to facilitate ontology development and editing. TheOBOFoundry now comprises over 100 ontologies and candidate ontologies, including the NCBI organismal classification ontology (NCBITaxon), the Mosquito Insecticide Resistance Ontology (MIRO), the Infectious Disease Ontology (IDO), theIDOMALmalaria ontology, and ontologies for mosquito gross anatomy and tick gross anatomy. We previously developed a disease data management system for dengue and malaria control programs, which incorporated a set of information trees built upon ontological principles, including a “term tree” to promote the use of standardized terms. In the course of doing so, we realized that there were substantial gaps in existing ontologies with regards to concepts, processes, and, especially, physical entities (e.g., vector species, pathogen species, and vector surveillance and management equipment) in the domain of surveillance and management of vectors and vector-borne pathogens. We therefore produced an ontology for vector surveillance and management, focusing on arthropod vectors and vector-borne pathogens with relevance to humans or domestic animals, and with special emphasis on content to support operational activities through inclusion in databases, data management systems, or decision support systems. The Vector Surveillance and Management Ontology (VSMO) includes >2,200 unique terms, of which the vast majority (>80%) were newly generated during the development of this ontology. One core feature of the VSMO is the linkage, through the has_vector relation, of arthropod species to the pathogenic microorganisms for which they serve as biological vectors. We also recognized and addressed a potential roadblock for use of the VSMO by the vector-borne disease community: the difficulty in extracting information from OBO-Edit ontology files (*.obo files) and exporting the information to other file formats. A novel ontology explorer tool was developed to facilitate extractionandexport of information fromtheVSMO*.obo file into lists of termsandtheirassociated unique IDs in *.txt or *.csv file formats. These lists can then be imported into a database or data management system for use as select lists with predefined terms. This is an important step to ensure that the knowledge contained in our ontology can be put into practical use.

Approximately 8% of calyptrate species diversity comes from the Calliphoridae, which includes flies of medical, veterinary, and forensic importance. The status of family Calliphoridae has for years been the central systematic problem of the superfamily Oestroidea, and phylogenetic relationships between the key groups of the Calliphoridae are unresolved and controversial. We reconstructed phylogenies of the Calliphoridae within the larger context of the other Oestroidea based on 5,189 bp of combined data from one mitochondrial (cytochrome oxidase subunit one) and three nuclear (carbamoylphosphate synthetase, elongation factor one alpha, and 28S ribosomal RNA) genes using maximum parsimony, maximum likelihood, and Bayesian methods. Trees obtained from the different phylogenetic methods were almost identical. Calliphoridae is polyphyletic, with the phylogenetic position of Mesembrinellinae still uncertain but clearly outside the lineage that includes other Calliphoridae and some noncalliphorids, and Polleniinae is the sister group of the family Tachinidae. Strong support for a sister group relationship between Rhiniinae and traditional calliphorid subfamilies conflicts with a recent proposal to give Rhiniinae family status. All calliphorid subfamilies (except Calliphorinae) for which we had more than one species were monophyletic. Melanomyinae was nested within Calliphorinae. Toxotarsinae was more closely related to Calliphorinae rather than, as indicated by morphology, to Chrysomyinae. Efforts to resolve the relationships of the Oestroid families were largely inconclusive, although the monophyly of the superfamily was strongly supported.

Knowledge of the fauna composition of anopheline mosquitoes, their ecological aspects and behavior, and influence of climatic variables on their population dynamics can help in understanding the transmission of Plasmodium parasites and thus develop more efficient strategies for the control of malaria. In the Central Atlantic Forest Biodiversity Corridor, southeastern Brazil, foci of introduced malaria have been reported among people returning from the Amazon region, north Brazil. Our objective was to evaluate and compare the anopheline fauna from a preserved environment and an adjacent peridomiciliary modified environment at the Central Atlantic Forest Biodiversity Corridor. We collected anopheline mosquitoes on a monthly basis from June 2004 to May 2006 from both these environments to understand the ecological aspects and their association with the occurrence of malaria. We captured 5,491 anopheline mosquitoes belonging to two subgenera and 11 species and studied the correlations between anopheline mosquito species and climatic variables. We considered Anopheles darlingi (Root) as the principal malaria vector and Anopheles albitarsis s. l. (Arribalzaga) as the secondary vector.

Even though house dust mites are one of the most important allergens, there have been few studies in China for their identification and diversity. In this study, we reported that Dermatophagoides siboney was found for the first time in Beijing, China, in a temperate zone and it was also the first reported in Asia. This survey of mite prevalence was carried out in several districts of Beijing, a city of thirty million people. House dust samples were collected from 38 homes of mite-allergic patients who visited our Allergy Department from December 2008 to January 2010. Out of 345 house dust samples collected, 64% contained mites. Dermatophagoides farinae was the predominant species in the mite population found and Dermatophagoides pteronyssinus was second, and Dermatophagoides siboney was the third. The positive rates of samples were higher in single-story homes and lower buildings. The seasonal density distribution of house dust mites showed the highest mite concentration in September through October, followed by May through July, December to next January; and lowest in March and November.

Wyeomyia (Wyeomyia) mitchellii (Theobald) was discovered in 2007 and 2008, respectively, in Moorea and Tahiti, two islands of the Society Islands (archipel de la Société) in French Polynesia. A few adult specimens were captured using a CDC backpack aspirator and BG-Sentinel mosquito traps. Wy. mitchellii larvae were found in water-impounding bromeliads and aroids at various survey sites around Tahiti. Imported bromeliads likely played a critical role in the introduction, establishment, and maintenance of Wy. mitchellii in Tahiti and Moorea. Bromeliads and aroids are common in residential areas, thereby increasing the exposure of human and domestic animals to Wy. mitchellii, which is of public health and veterinary concern. The establishment of Wy. mitchellii in French Polynesia requires further study.

Seasonal variation in dengue virus transmission in northwestern Thailand is inversely related to the magnitude of diurnal temperature fluctuations, although mean temperature does not vary significantly across seasons. We tested the hypothesis that diurnal temperature fluctuations negatively influence epidemiologically important life-history traits of the primary dengue vector, Aedes aegypti (L.), compared with a constant 26°C temperature. A large diurnal temperature range (DTR) (≈18°C daily swing) extended immature development time (>1 d), lowered larval survival (≈6%), and reduced adult female reproductive output by 25% 14 d after blood feeding, relative to the constant 26°C temperature. A small DTR (≈8°C daily swing) led to a negligible or slightly positive effect on the life history traits tested. Our results indicate that there is a negative impact of large DTR on mosquito biology and are consistent with the hypothesis that, in at least some locations, large temperature fluctuations contribute to seasonal reduction in dengue virus transmission.

The Amazon rainforest and the Brazilian Cerrado both possess high phlebotomine diversity. The fragmentation of these habitats has resulted in the appearance of human cases of cutaneous leishmaniasis. In one altered area of mixed primary vegetation (forest and Cerrado) and its adjacent settlement in the northeast state of Maranhão, Brazil, evidence exists for the active transmission of cutaneous leishmaniasis. Accordingly, an entomological investigation was performed in both the forest and the settlement to compare the phlebotomine vector fauna in each environment. The study was conducted from September 2009 to August 2010 in the municipality of Itapecuru Mirim in the state of Maranhão, Brazil. The phlebotomine species were captured using 24 light Center for Disease Control and Prevention traps that were placed in the forest and the settlement (peridomicile and intradomicile). The similarity between the phlebotomine compositions in the forest and those in the settlement was determined using a Principal Coordinate Analysis based on a dissimilarity matrix that was calculated using the Bray—Curtis index (relative abundance) and the Jaccard index (presence and absence of species). In total, 29 Lutzomyia species and one Brumptomyia species were collected. The phlebotomines were diverse and abundant in both the forest fragment (27 species, 4,606 specimens) and the settlement (22 species, 753 specimens). The most abundant species were L. infraspinosa (25%), L. davisi (21%), L. antunesi (21%), L. longipalpis (9%), L. saulensis (6%), L. flaviscutellata (5%), and L. wellcomei (4%). Some species were found strictly in the forest, other species were exclusive to the anthropic environment, and some species colonized both of the studied environments. The phlebotomines adaptation to these modified environments explains the autochthonous outbreak of cutaneous leishmaniasis.

Aedes aegypti (L.), the primary vector of dengue and dengue hemorrhagic fever, breeds and rests predominately inside human dwellings. With no current vaccine available, vector control remains the mainstay for dengue management and novel approaches continue to be needed to reduce virus transmission. This requires a full understanding of Ae. aegypti ecology to design effective strategies. One novel approach is the use of contact irritants at target resting sites inside homes to make the surface unacceptable and cause vectors to escape before biting. The objective of the current study was to observe indoor resting behavior patterns of female Ae. aegypti within experimental huts in response to two fabrics under consideration for insecticide treatment: cotton and polyester. Results indicate that fabric type, coverage ratio of dark to light fabric and placement configuration (vertical vs. horizontal) all influenced the resting pattern of mosquito cohorts. Findings from this study will guide evaluations of a push-pull strategy designed to exploit contact irritant behaviors and drive Ae. aegypti out of homes prefeeding.

Aedes (Finlaya) japonicus japonicus (Theobald) (Diptera: Culicidae) is recently invasive in North America and has expanded its range rapidly since 1998. Throughout its native and expanded range, Ae. j. japonicus larvae are commonly observed in many types of natural and artificial water-filled containers that vary in organic matter content and exposure to sunlight. Larvae are most often found in containers with decaying leaf material or algae, and we postulated that the added autocthonous primary production from algae could be both an important food source for larvae and an influential oviposition attractant to adult Ae. j. japonicus. We tested this hypothesis by placing plastic containers with varied levels of shading to manipulate algal density in the field, and then monitored oviposition by natural populations of Ae. j. japonicus. Over 99% of larvae hatching from eggs laid on the walls of our containers were Ae. j. japonicus, indicating that this species is a dominant colonizer of artificial containers in the study areas. Although full shading treatments effectively reduced algal biomass (significant reduction in chlorophyll a levels), at only one of three sites did this appear to affect Ae. j. japonicus oviposition. We conclude that algae in larval habitats are not a major factor in oviposition choices of adult Ae. j. japonicus females except when in situ primary production is high enough to substantially alter overall organic matter content cues.

Obtaining reliable gene expression data using real-time quantitative polymerase chain reaction (qPCR) is highly dependent on the choice of normalization method. We tested the expression stability of multiple candidate genes in the salivary glands (SG) and synganglia (SYN) of female Ixodes scapularis (Say) ticks in multiple blood-feeding phases. We found that the amount of total RNA in both the SG and SYN increases dramatically during tick feeding, with 34× and 5.8× increases from 62 and 7.1 ng of unfed tick, respectively. We tested candidate genes that were predicted from I. scapularis genome data to encode glyceraldehyde 3-phosphate dehydrogenase (gapdh), ribosomal protein L13A (l13a), TATA box-binding protein (tbp), ribosomal protein S4 (rps4), glucose 6-phosphate dehydrogenase (gpdh), and beta-glucuronidase (gusb). The geNorm and NormFinder algorithms were used to analyze data from different feeding phases (i.e., daily samples from unfed to fully engorged females over a 7-d period in three replicate experiments). We found that the rps4 and l13a genes showed highly stable expression patterns over the feeding duration in both the SG and SYN. Furthermore, the highly expressed rps4 gene makes it useful as a normalization factor when we perform studies using minute amounts of dissected tissue for qPCR. We conclude that rps4 and l13a, whether individually or as a pair, serve as suitable internal reference genes for qRT-PCR studies in the SG and SYN of I. scapularis.

Amblyomma variegatum F. are obligate hematophagous ectoparasites of livestock that serve as the vectors of Ehrlichia ruminantium (formerly known as Cowdria ruminantium), the causative agent of heartwater disease. In the light of the fact that they are blood-feeding, their salivary glands play prominent role in their acquisition of nutrients from the bloodmeal. Sialic acids are a major component of glycoprotein in mammalian blood fluid and cells. Sialome of hard ticks is still sparse. Here, for the first time, the possible expression of sialidase in A. variegatum was investigated. Our finding established the presence of type II sialidase-like activity in the three stages (larva, nymph, and adult) of the fed and unfed tick. There was no statistically significant difference in sialidase activity in the various stages of this ectoparasite (P > 0.05). The enzyme was purified by combination of salting out and ion exchange chromatography on DEAE—cellulose and hydroxylapatite columns. Characterization of the enzyme revealed that it is optimally active at 40°C and pH 5.5, and is activated by bivalent cations Zn² or Fe² . The enzyme has a K_m of 0.023 mM and V_max of 0.16 millimol/min with Fetuin as the substrate. To assess the susceptibility of some mammalian cells to the tick sialidase, we prepared erythrocyte ghost cells from different animals, which were incubated with the enzyme. Results revealed that the ruminant cells were better substrates. Our work and findings contribute to the preliminary characterization of the A. variegatum salivary proteome, and may pave way to the development of new acaricides.

Mating competitiveness trials have been conducted in large net-screened enclosures (8 by 5 by 2.8 m) built in a natural shaded environment, in the summers of 2006, 2007, 2008, and 2009 in northern Italy. Aedes albopictus (Skuse) males were radio-sterilized by applying gamma radiations at doses in the range 30–60 Gy. Gamma radiation was administered to aged pupae at the rate of 2.3 Gy/min. Reared radiated males (originally collected in Rimini, Forlì, Bologna, Matera, Pinerolo) and hybrid radiated males were tested against wild fertile males (originated from eggs collected in Rimini and Cesena) and reared fertile males, in multiple comparisons for mating competitiveness with reared or wild females. The ratio was kept constant at 100-100_100 (fertile males-radiated males_virgin females). Mating competitiveness was estimated through the calculation of the hatching rate of the eggs laid in oviposition traps positioned inside enclosures. No clear effect of the strains tested (reared, wild, or hybrid) was found. Results demonstrated that reducing the radiation dose from 60 to 30 Gy increases males' competitiveness. Laboratory investigations conducted after controversial results in the 2006 preliminary trials, showed that radiation induces precociousness in adult male emergence.

A nationwide investigation was carried out to determine the current susceptibility status of Culex quinquefasciatus Say populations against four active ingredients representing four major insecticide classes: DDT, propoxur, malathion, and permethrin. Across 14 study sites, both larval and adult bioassays exhibited dissimilar trends in susceptibility. A correlation between propoxur and malathion resistance and between propoxur and permethrin resistance in larval bioassays was found. The results obtained from this study provide baseline information for vector control programs conducted by local authorities. The susceptibility status of this mosquito should be monitored from time to time to ensure the effectiveness of current vector control operations in Malaysia.

Botanical insecticides offer novel chemistries and actions that may provide effective mosquito control. Toosendanin (TSN, 95% purity) is one such insecticide used to control crop pests in China, and in this study, it was evaluated for lethal and sublethal effects on larvae and females of the yellowfever mosquito, Aedes aegypti (L.). TSN was very toxic to first instar larvae after a 24 h exposure (LC₅₀ = 60.8 µg/ml) and to adult females up to 96 h after topical treatment (LD₅₀ = 4.3 µg/female) or ingestion in a sugar bait (LC₅₀ = 1.02 µg/µl). Treatment of first instars for 24 h with a range of sublethal doses (6.3–25 µg/ml) delayed development to pupae by 1 to 2 d. Egg production and larval hatching from eggs were dose dependently reduced (>45%) by TSN doses (1.25–10.0 µg) topically applied to females 24 h before or 1 h after a bloodmeal. Ingestion of TSN (0.031–0.25 µg/µl of sugar bait) by females 24 h before a bloodmeal also greatly reduced egg production and larval hatch; no eggs were oviposited by females ingesting the highest dose. Further studies revealed that topical or ingested TSN dose-dependently disrupted yolk deposition in oocytes, blood ingestion and digestion, and ovary ecdysteroid production in blood-fed females. Overall, our results indicate that TSN is an effective insecticide for Ae. aegypti larvae and adults, because of its overt toxicity at high doses and disruption of development and reproduction at sublethal doses.

The sand fly Phlebotomus papatasi Scopoli is the vector of Leishmania major (Yakimoff & Schokhor), which is maintained in populations of burrowing rodents. The purpose of this study was to conduct a laboratory study to determine the efficacy of oral treatment of rodents with fipronil for control of sand flies that feed on rodent feces as larvae or on rodent blood as adults. We determined through larval bioassays that fipronil was eliminated in feces of orally-treated hamsters at a level that was significantly toxic to sand fly larvae for 21 d after the hamsters had been withdrawn from a fipronil-treated diet. Through bloodfeeding bioassays, we also found that fipronil was present in the peripheral blood of hamsters at a concentration that was significantly toxic to bloodfeeding adult female sand flies for 49 d after the hamsters had been withdrawn from their treated diet. The results of this study suggest that fipronil acts as well as or better than feed-through or systemic insecticides that previously have been measured against sand flies, and is particularly promising because this single compound acts against both larvae and bloodfeeding adults. An area-wide approach using rodent baits containing a fipronil could suppress vector populations that originate in the vicinity of rodent reservoirs, and could be used to eliminate the most epidemiologically important part of the vector population: female sand flies that take bloodmeals on rodent reservoirs.

We tested the effectiveness of the rosemary oil-based insecticide, Eco-Exempt IC2, to control all stages of Ixodes scapularis (Say) in southern Maine. We selected plots in oak-pine forest where I. scapularis is endemic and recorded the abundance of ticks and nontarget arthropods before and after applications of IC2, bifenthrin (a synthetic pyrethroid), and water (reference treatment). Licensed applicators applied high-pressure spray treatments during the summer nymphal and fall adult seasonal peaks. Both acaricides sprayed during the summer nymphal season reduced nymphal I. scapularis/hour to zero. IC2 was as effective as bifenthrin in controlling nymphs through the rest of the nymphal season and also controlled adult ticks 9 mo postspray compared with 16 mo for bifenthrin, and both acaricides reduced larvae through 14 mo postspray. Both acaricides sprayed during the fall adult season reduced adult I. scapularis/hour to zero; IC2 controlled adult ticks 6 mo postspray compared with 1 yr for bifenthrin. Both fall-applied acaricides controlled nymphs 9 mo postspray and reduced larvae up to 10 mo postspray. Impacts on some nontarget arthropods was assessed. Colleoptera, Hymenoptera, and Collembola declined 1 wk postspray in acaricide-treated plots, and in IC2 plots all numbers rebounded by 20 d postspray. For bees and other flower-visiting insects there were no detectable reductions in nests produced, number emerged from nests, or number of foraging visits to flowering plants in IC2 or bifenthrin plots. IC2 was phytotoxic to the leafy portions of select understory plants that appeared to recover by the next growing season.

Seven commercial synthetic disinfectant and antibacterial soap products were evaluated as mosquito larvicides against Culex quinquefasciatus Say in the laboratory. Three aerosol disinfectant products, at 0.01% concentration resulted in 58–76% mortality of laboratory-reared fourth instar mosquito larvae at 24 h posttreatment. Four antibacterial soap products at 0.0001% concentration resulted in 88–100% larval mortality at 24 h posttreatment. The active ingredient of the antibacterial soap products, triclosan (0.1%) resulted in 74% larval mortality. One of the antibacterial soap products, Equate caused the highest mosquito larval mortality in the laboratory. Equate antibacterial soap at the application rate of 0.000053 ppm resulted in 90% mortality of the introduced fourth instar larvae of Cx. quinquesfasicatus in the outdoor pools. In laboratory and field bioassays, the antibacterial soap resulted in significant larval mosquito mortality.

A comprehensive 8-yr survey of acarine ectoparasites (ticks and mites) of bats was carried out in 18 localities from 2002 to 2009. Most of the surveys were conducted during 14 national biodiversity scientific expeditions throughout Malaysia. The objective was to identify acarines of known public health importance from bats and thus determine whether there is any potential public health risk in Malaysia. Trapping of bats was conducted using Harp traps and Mist nets. In total, 1,579 individuals comprising of 6 families and 52 species of bats were examined alive. In general, 25.6% of the bats were infested with acarines. Infestation rates of ticks, mesostigmatid mites, and chiggers on bats examined were 0.4, 10.4, and 14.7%, respectively. Their prevalence and mean intensity were tabulated. Genera of ticks extracted were Amblyomma, Dermacentor, Ixodes, and Ornithodoros. Of these genera, only two species can be identified to species level and they are Amblyomma cordiferum and Ixodes simplex. In total, 8 genera and 15 species of mesostigmatid mites were found; the species were Ancystropus eonycteris, Ancystropus zeleborii, Echinonysus nasutus, Laelaps aingworthae, Laelaps nuttalli, Laelaps sanguisugus, Laelaps sculpturatus, Longolaelaps longulus, Longolaelaps whartonii, Meristaspis lateralis, Meristaspis macroglossi, Paraperiglischrus rhinolophinus, Spinturnix acuminatus, Spinturnix americanus, and Spinturnix bakeri. Chiggers on bats were represented by 12 genera and 6 species; the species identified were Gahrliepia fletcheri, Riedlinia lipoxena, Trombigastia cadei, Walchiella impar, Walhiella oudemansi, and Whartonia caobangensis. The study produced an up-to-date list of acarine ectoparasites of bats in Malaysia where a total of 38 genera and 47 species of acarines were listed. Findings of the study demonstrated that 5 genera and 1 species of acarines that may pose potential health risks, can be found on bats.

Proteases are involved in the pathogenicity of allergy, increasing epithelial permeability and acting as adjuvants. Enzymatic activity is therefore important for the allergenicity of an extract and also affects its stability and safety. However, the enzymatic activity of extracts is not usually evaluated. The objective of this study was to evaluate the enzymatic activity of the most allergenic mite extracts and to investigate their allergenic properties. Extracts from nine allergenic mite species (Dermatophagoides pteronyssinus, Dermatophagoides farinae Hughes, Euroglyphus maynei, Lepidoglyphus destructor, Tyrophagus putrescentiae (Schrank), Glycyphagus domesticus (DeGeer), Acarus siro L., Chortoglyphus arcuatus, and Blomia tropicalis) were characterized. Protein and allergen profiles were characterized by sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE) and western-blot, respectively. Gelatinolytic activity was evaluated with a zymogram and the activity of other enzymes (cysteine, serine proteases, and esterases) was evaluated individually or with the API-ZYM system. The main differences in protease activity were found between house dust mites and storage mites. House dust mites presented higher cysteine protease activity while storage mites presented higher serine protease activity. These differences are in line with their trophic specialization. A wide range of different activities was found in all the extracts analyzed, reflecting the fact that the extracts preserve the activity of many enzymes, this being necessary for a correct diagnosis. However, enzymes may act as adjuvants and, therefore, could lead to undesirable effects in immunotherapies, making this activity not suitable for treatment products. Modified extracts with lower enzymatic activity could be more appropriate for immunotherapy.

The goal of this study was to determine the effect of vertically transmitted Arsenophonus and Rickettsia bacteria on locomotive ability of larvae of three eastern North American tick species: Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis. We conducted two different experiments on flat or inclined surfaces to measure tick motility. In each experiment, a moderately heated surface was used and placed at a selected incline. The individual's path across the surface was traced for a period of 2 min, or until the larval tick had moved off the surface. Following the microbial identification of Arsenophonus and Rickettsia, a mixed model analysis of variance (ANOVA) indicated that clutch microbial infection status had a significant effect on tick motility with Rickettsia increasing and Arsenophonus decreasing motility averaged over tick species and inclines. There was also a significant difference in motility among tick species and a highly significant effect of the Species*Incline interaction where Dermacentor had higher motility than Ixodes on the flat surface.

Anaplasma phagocytophilum (Dumler et al.) is the bacterial agent of human granulocytic anaplasmosis, an emerging infectious disease. The main vector of A. phagocytophilum in the United States is the blacklegged tick (Ixodes scapularis (Say)) and various small and medium-sized mammals are reservoirs. Previous studies indicate that birds are exposed to A. phagocytophilum; however, because no studies have directly investigated avian susceptibility, reservoir competence, and morbidity for A. phagocytophilum, uncertainty remains as to what role birds could play in its transmission ecology. In a controlled laboratory study, we tested whether two species, the American robin (Turdus migratorius (L.)) and the gray catbird (Dumetella carolinensis (L.)), can become infected with and then transmit A. phagocytophilum to feeding ticks, and whether exposed birds develop disease. Wild caught, seronegative birds (n = 10 per species) were exposed to A. phagocytophilum-infected I. scapularis nymphs (day 0). Transmission was assessed by xenodiagnosis on days 7, 14, 42, and 77; blood was assayed for bacteremia and serology. A. phagocytophilum was detected using quantitative polymerase chain reaction targeting the 16s rRNA gene. One robin infected 2 of 13 larval ticks (15%) on day 7; no other birds were found to infect feeding ticks at any time. Birds did not develop bacteremia, specific antibodies or significant illness because of exposure. Mouse controls became bacteremic, infected feeding ticks, and seroconverted. Our results suggest that these two avian species are unlikely to play a significant role in the maintenance of the agent of human granulocytic anaplasmosis and that avian serosurveys may not be a reliable indicator of A. phagocytophilum exposure.

House flies disseminate numerous species of bacteria acquired during feeding and breeding activities in microbe-rich habitats. Previous house fly surveys have detected the pathogen Staphylococcus aureus Rosenbach 1884, which causes cutaneous and septic infections in mammals, and enterotoxic food poisoning. We assessed the fate of GFP-expressing S. aureus (GFP-S. aureus) in the house fly alimentary canal with microscopy and by culture of whole flies and excreta. Furthermore, the concurrent expression of the antimicrobial peptide gene defensin was measured in the crop, proventriculus, midgut, and fat body. As soon as 4 h postingestion (PI), GFP-S. aureus were visualized as cocci or diplococci in the hindgut and rectum of flies fed ≈10⁵ colony forming units. Bacteria persisted up to 6 h PI but significantly decreased. Excretion of viable GFP-S. aureus peaked at 2 h PI and, although significantly less, continued up to 4 h PI. defensin was highly upregulated locally in the alimentary canal and systemically in fat body at 2, 4, and 6 h PI making this study the first to report, to our knowledge, an epithelial and systemic response to a bacterium with lysine-type peptidoglycan in flies exposed via feeding. While flies harbored S. aureus for up to 6 h PI, the highest probability of vectoring biologically relevant amounts of bacteria occurred 0–2 h PI. The combined effects of excretion, digestion and antimicrobial effectors likely contribute to loss of ingested bacteria. Nonetheless, house flies are relevant vectors for S. aureus up to 2 h PI and environmental reservoirs up to 6 h PI.

Mosquitoes vertically transmit many arthropod borne viruses, and as a consequence arboviruses are often present within the larval environment. We tested the hypothesis that Aedes aegypti (L.) and Aedes albopictus (Skuse) larvae were susceptible to dengue virus through two infection methods: exposure to dengue in the larval growth environment via viral supernatant, and exposure to infected tissue culture along with viral supernatant. In addition to investigating for the first time the susceptibility of larval Ae. albopictus to dengue virus, we analyzed the infection rate and viral titer of infected pools of Ae. aegypti when exposed to multiple serotypes of dengue. We found that both Ae. aegypti and Ae. albopictus larvae were susceptible to the three dengue virus serotypes to which they were exposed regardless of the exposure method and that there were significant differences between the serotypes in infection titer and infection rate. The finding that larval Ae. aegypti and Ae. albopictus are susceptible to dengue indicates that dengue might be able to spread among larvae within the larval habitat potentially contributing to the persistence of dengue in the environment.

Mosquitoes often are collected as part of an arbovirus surveillance program. However, trapping and processing of mosquitoes for arbovirus detection is often costly and difficult in remote areas. Most traps, such as the gold standard Center for Disease control light trap, require batteries that must be charged and changed overnight. To overcome this issue we have developed several passive traps for collection of mosquitoes that have no power requirements. The passive traps capture mosquitoes as they follow a CO₂ plume up a polyvinyl chloride pipe leading to a clear chamber consisting of a plastic crate. We believe the translucent, clear windows created by the crate inhibits escape. Once inside the crate mosquitoes readily feed on honey-treated Flinders Technology Associates cards that then can be processed by polymerase chain reaction for viral ribonucleic acid. Of the two designs tested, the box or crate-based passive trap (passive box trap, PBT) generally caught more mosquitoes than the cylinder trap. In Latin square field trials in Cairns and Florida, PBTs collected mosquitoes at rates of 50 to 200% of Center for Disease Control model 512 light traps. Mosquito collections by PBTs can be increased by splitting the CO₂ gas line so it services two traps, or by placing an octenol lure to the outside of the box. Very large collections can lead to crowding at honey-treated cards, reducing feeding rates. Addition of fipronil to the honey killed mosquitoes and did not impact feeding rates nor the ability to detect Kunjin viral ribonucleic acid by polymerase chain reaction; this could be used to minimize crowding affects on feeding caused by large collections. The passive traps we developed are made from inexpensive, commonly available materials. Passive traps may thus be suitable for collection of mosquitoes and potentially other hematophagous dipterans for pathogen surveillance.

This study looked at the influence of interannual variations in temperature and precipitation on seasonal mosquito abundances, the prevalence of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in the northeastern United States, and the capacity for local mosquito communities to maintain and transmit WNV, defined as vector community competence. Vector and virus surveillance took place within Middlesex County in New Jersey over two transmission seasons (2010 and 2011). Drought conditions during the 2010 season were associated with significant increases in the number of blood-fed Culex spp. mosquitoes collected per week, and significant increases in vector community competence, or the ability of local vector communities to transmit WNV, when compared with the wetter and milder 2011 season. These increases were associated with significantly higher weekly WNV infection rates in Culex spp. (i.e., Culex pipiens L. and Culex restuans L.) during the 2010 drought season. On a larger scale, the positive influence of drought on the amplification of WNV was also confirmed at the state level where early seasonal (June-July) increases in temperature and decreases in precipitation were strongly correlated with increases in yearly WNV infection rates over a 9-yr period (2003–2011). These data suggest that there may be clear temperature and precipitation thresholds beyond which epidemic levels of WNV transmission occur.

Dermatophagoides farinae Hughes, American house dust mite, is highly allergenic, producing symptoms in people worldwide. Identifying and cloning the allergens in this species may enable better diagnostic and therapeutic approaches. Here, we cloned, sequenced, and expressed the full-length cDNA encoding D. farinae group 10 allergen (Der f 10) isolated from dust mites in China. Bioinformatic analysis indicated that the 888 bp sequence encoded a cytoskeleton protein 295 amino acids long, with a molecular weight of ≈34 kDa. Sequence alignment with the group 10 allergens of Pyroglyphidae, Acaridae, and Glycyphagidae families revealed that the group 10 allergen from D. farinae is 95% similar to D. pteronyssinus Trouessart and Psoroptes ovis (Hering). These findings lay the groundwork for future studies, including large-scale production of recombinant Der f 10 allergen for diagnostic and therapeutic agents.

Myiases are caused by the presence of maggots in vertebrate tissues and organs. Myiases have been studied widely in humans, farm animals, and pets, whereas reports of myiasis in reptiles are scarce. We describe a case of myiasis caused by the Megaselia Scolaris (Loew) in an Indian python (Python molurus bivittatus, Kuhl) (Ophida: Boidae). The python, 15 yr old, born and reared in a terrarium in the mainland of Venice (Italy), was affected by diffuse, purulent pneumonia caused by Burkholderia cepacia. The severe infestation of maggots found in the lungs during an autopsy indicated at a myiasis.

Phlebotomus martini is a known vector of visceral leishmaniasis caused by Leishmania donovani in sub-Saharan Africa. The disease is known to be endemic in areas of north and south Sudan, Kenya, Ethiopia, Uganda, and Somalia but has not been reported from Tanzania. In this report we present the first documented collection of P. martini and P. vansomerenae in Tanzania. Sand flies were collected using standard dry-ice baited CDC light traps (John W. Hock Company, Gainesville, FL) from five sampling sites in the Arusha and Kilimanjaro regions from 14 to 20 July 2010. Phlebotomus martini was collected from all sites and represented 6.6% of the total identified sand flies. Phlebotomus martini ranged from 4.5 to 9.4% of the total identified catch from the four sites in the Kilimanjaro region and 17.9% of the total identified catch at the one collection site in the Arusha region. In addition, one male specimen of the sibling species, Phlebotomus vansomerenae, was found at Chemka Springs in the Kilimanjaro region. These data indicate the presence of an established population (s) of P. martini in northern Tanzania that could support L. donovani transmission in an area with no prior case history of visceral leishmaniasis.

An engorged female Amblyomma helvolum Koch tick was removed from an adult Varanus salvator Laurenti lizard during field collection in Thailand. After using polymerase chain reaction to amplify three genes (16S rDNA, gltA, and OmpA), we discovered the presence of a Rickettsia sp. of the Spotted Fever Group. Phylogenetic analysis revealed that this Rickettsia sp. is closely related to Rickettsia raoultii Mediannikov. Therefore, we report herein for the first time the detection of a novel Spotted Fever Group Rickettsia in an Amblyomma helvolum from a Varanus salvator in Thailand.