We describe and illustrate a new bat chigger, Whartonia (Fascutonia) natsumei Takahashi, Takahashi & Kikuchi n. sp., ex. Plecotus auritus (Linnaeus, 1758), from Japan, and summarize the host and distribution records of the chiggers of the genus Whartonia Ewing, 1944. With the description of a new species here, the number of Whartonia species increases to 40 composed of 22 species from Asia, 14 from North and South America in addition to Central America, and four from Africa.

Ochlerotatus (Finlaya) japonicus (Theobold) is newly established in Ontario, Canada. It was first discovered in 2001 during the province-wide West Nile virus mosquito surveillance program implemented by the Ontario Ministry of Health and Long Term Care. Although the numbers of adults trapped in CDC light traps were low in 2001 and 2002, they increased during the 2003 and 2004 surveillance seasons. Oc. japonicus larvae also have been collected in large numbers in the Niagara Peninsula in a variety of natural and artificial containers. The number of health units with records for Oc. japonicus has increased over the 4 yr of surveillance, illustrating the ability of this species to rapidly extend its range. As a potential arboviral bridge vector, its establishment in Ontario requires further study and should be considered a public health concern.

Flight dispersal of Triatoma infestans Klug is probably the most important mechanism for house reinfestation at a village scale after residual spraying with insecticides. The aim of the current study was to estimate the flight initiation probability of field-collected T. infestans and to assess how this probability was affected by sex, adult age, partial bloodmeal, and the presence of a host inaccessible for feeding. Four experimental series, each consisting of three to six consecutive nights and repeated measurements of flight initiation on each individually marked bug, were carried out in experimental huts inside closed cages under natural climatic conditions. We demonstrate that flight initiation probability of T. infestans is much higher than previously reported, responds to temperature in a sigmoid manner, and is higher in females than males, and that the frequency distribution of the number of flights per individual is highly aggregated in female and male bugs. The age of adults had strong effects on flight initiation, whereas the presence of an inaccessible host and a partial bloodmeal exerted no significant effects in models controlling for the effects of bug weight-to-length ratio. The high flight potential found is consistent with the rapid changes in reinfestation patterns observed in the field. The present estimates of flight probabilities and the identification of factors modifying them provide essential knowledge for modeling reinfestation patterns and for improving control strategies of T. infestans.

Most adult mosquito surveillance in Maryland is performed using dry ice-baited or unbaited Centers for Disease Control (CDC) miniature light traps suspended ≈1.5 m above the ground. However, standardized trapping methods may miss mosquito species involved in disease transmission cycles. During a 2-yr study, the effectiveness of the olfactory attractant 1-octen-3-ol alone and in combination with carbon dioxide was evaluated for collecting mosquito vector species. In addition, trap heights were examined to determine the optimal vertical placement to target various species. We evaluated the results during the second year by targeting selected species by using various habitat–height–bait combinations. Although Culex erraticus Dyar & Knab and Anopheles quadrimaculatus Say were not successfully targeted, Culex salinarius Coquillett, Aedes vexans Meigen, Anopheles bradleyi/crucians King, Coquillettidia perturbans Walker, Aedes sollicitans Walker, and Aedes taeniorhynchus Wiedemann were preferentially captured using targeted trapping schemes.

Prevalence of infection by Borrelia burgdorferi s.l. and spotted fever group (SFG) rickettsiae was estimated in host-seeking ticks in an area in Tuscany, central Italy, where Lyme borreliosis was reported in a forestry worker. B. burgdorferi s.l. was identified by polymerase chain reaction in 16.7% (95% CI = 10.3, 24.8) of Ixodes ricinus (L.) nymphs and 39.6% (95% CI = 26.5, 54.0) of adults. Borrelia lusitaniae accounted for 82.9% of positive samples, followed by Borrelia garinii (9.8%), Borrelia afzelii (2.4%), and Borrelia burgdorferi s.s. (2.4%). One Rhipicephalus spp. adult was infected with B. garinii (prevalence = 8.3%; 95% CI = 0.21, 38.5). Prevalence of infection by SFG rickettsiae was 38.5% (95% CI = 26.7, 51.4) in I. ricinus nymphs, 34.6% (95% CI = 22.0, 49.1) in I. ricinus adults, and 50% (95% CI = 21.1, 78.9) in Rhipicephalus spp. adults. Phylogenetic analysis showed the similarity of B. lusitaniae strains that were identified in this study and of a strain that was previously isolated from a human patient in Portugal. Results of this study confirm the dominance of B. lusitaniae in areas in the Mediterranean basin and the infection by SFG rickettsiae in I. ricinus.

The risk of Lyme disease for humans in the eastern United States is dependent on the density of host-seeking Ixodes scapularis Say nymphal stage ticks infected with Borrelia burgdorferi. Although many local and regional studies have estimated Lyme disease risk using these parameters, this is the first large-scale study using a standardized methodology. Density of host-seeking I. scapularis nymphs was measured by drag sampling of closed canopy deciduous forest habitats in 95 locations spaced among 2° quadrants covering the entire United States east of the 100th meridian. Sampling was done in five standardized transects at each site and repeated three to six times during the summer of 2004. The total number of adults and nymphs of the seven tick species collected was 17,972, with 1,405 nymphal I. scapularis collected in 31 of the 95 sites. Peak global spatial autocorrelation values were found at the smallest lag distance (300 km) and decreased significantly after 1,000 km. Local autocorrelation statistics identified two significant high-density clusters around endemic areas in the northeast and upper Midwest and a low-density cluster in sites south of the 39th parallel, where only 21 nymphs were collected. Peak nymphal host-seeking density occurred earlier in the southern than in the most northern sites. Spatiotemporal density patterns will be combined with Borrelia prevalence data as part of a 4-yr survey to generate a nationwide spatial risk model for I. scapularis-borne Borrelia, which will improve targeting of disease prevention efforts.

Trapping success, abundance, parity rate, and body size indices of Culex pipiens (L.) and Culex restuans Theobald, important vectors of West Nile virus (family Flaviviridae, genus Flavivirus, WNV), were determined for specimens captured from both ground level (≈1-m) and elevated (≈6- to 7-m) CO₂-baited CDC miniature light traps. Mosquitoes were collected from six study sites in Albany County, New York, from mid-May to mid-October 2004. There was no significant difference in abundance or mean parity for either Cx. pipiens or Cx. restuans between the elevated and the ground-level traps. Mosquitoes collected in elevated traps tended to be larger than mosquitoes collected at ground level. Elevated traps captured significantly fewer mosquitoes per trap-night than did ground traps, but a greater proportion of those captured were Culex spp. Therefore, elevated traps more efficiently captured Cx. pipiens and Cx. restuans with similar reproductive status and body size and reduced the time necessary to sort and identify collections. These findings may be beneficial to researchers or surveillance programs focusing on Cx. pipiens or Cx. restuans.

The development and survival of immatures of Culex vishnui (Diptera: Culicidae) complex, vectors of Japanese encephalitis virus, were studied in transplanted rice, Oryza savita L., fields during three crop growing seasons. The total duration of development from the first instar to adult emergence varied from 6 to 8 d. Survival rate estimates ranged from 0.003 to 0.524, but they generally were <0.1. Unusually high survival rates (0.192–0.524) were observed in summer and long-term monsoon crop seasons during 1993. A multiple regression method using backward elimination was used to analyze the factors responsible for these variations. The model identified nine parameters related with survival rates that explained 99% of the variance. Area of water surface and hardness were negatively related, whereas green algae, notonectid adults, anisopterans, dytiscids, salinity, water depth, and dissolved oxygen were positively associated with immature survival.

Seasonal abundance and life history of Fannia benjamini complex (Diptera: Muscidae) was studied in the coastal mountain community of La Habra Heights in Los Angeles County, California, with additional observations in drier, inland mountains in the Woodcrest area, Riverside County. The dominant species (>95% of fly collections) was Fannia conspicua Malloch, whereas Fannia benjamini Malloch also was present. Both species could be collected nearly year-round by netting adults (almost exclusively females) attracted to people, although F. benjamini was relatively more common in cooler weather (November–April). In La Habra Heights, adult activity peaked in June and July (both species), whereas peak activity in Woodcrest was late April through June for F. conspicua and February through April for F. benjamini. Field-collected adult females provided only water died within 2–4 d, whereas females provided with water plus a dry milk and sugar mixture survived up to 128–130 d in the laboratory. Males of F. conspicua (1–40 individuals) swarmed at heights of 0.5–4.0 m near Aptenia cordifolia (L.f.) Schwantes, an exotic, succulent, low-growing ground cover plant. Eggs of F. conspicua were deposited singly on older, decaying leaves of this plant, and adult F. conspicua emerged from Aptenia-covered areas in the field. In the laboratory, larvae of F. conspicua grazed on microbial surface films associated with Aptenia. Emergence of F. conspicua from field-collected soil–Aptenia mixtures (held in the laboratory) occurred from 1.5 to 5 mo after collection. Widespread Aptenia planting since the 1980s has probably resulted in F. conspicua becoming a severe human nuisance in some areas.

Studies were carried out at three sites in the highlands of western Kenya (Iguhu and Mbale in Kakamega and Vihiga districts, respectively, and Marani in Kisii district) and at one site in the western Kenya lowlands (Kombewa in Kisumu district) to determine the spatial–temporal dynamics of malaria vectors and intensity of malaria transmission from June 2003 to June 2004. At the highland sites, Anopheles gambiae Giles predominated, constituting >80% of the vector species, whereas An. funestus Giles made up <20%. In contrast, at the lowland site, An. funestus made up 68% of the vector species. The mean annual indoor resting densities of An. gambiae at Iguhu were 5.0 female mosquitoes per house per night, 14.2- and 26.3-fold greater than those at Mbale and Marani. During the main transmission season, the indoor resting densities of An. gambiae increased 4.1-, 10.1-, and 5.0-fold over the dry season period in Iguhu, Mbale, and Marani, respectively. The estimated annual entomological inoculation rate (EIR) at Iguhu was 16.6 infectious bites per person per year (ib/p/yr), 1.1 at Mbale, and 0.4 at Marani. This suggests high spatial variation in vector abundance and malaria transmission intensity. At the lowland site, Kombewa, the total annual EIR was 31.1 ib/p/yr and the indoor resting densities during the transmission season increased 7.1-fold in An. funestus and 18.5-fold in An. gambiae sensu lato over the dry season. The low level of transmission in the highlands suggests that it may be disrupted by vector control methods such as residual spraying.

The southern house mosquito, Culex quinquefasciatus Say (Diptera: Culicidae), is closely associated with humans and is a vector of filariasis. Use of semiochemicals for control of mosquitoes is a novel and potentially ecofriendly control approach. Human skin emanations are known to attract mosquitoes. In the current study, olfactory and behavioral responses of host-seeking female Cx. quinquefasciatus to various chemical components of human skin emanations (carboxylic acids, alcohols, and aldehydes) were evaluated separately at three doses (0.01, 0.1, and 1 μg) for electroantennogram (EAG) and three doses (0.1, 1, and 10 μg) for behavioral assay. Results of EAG studies indicated that all carboxylic acids elicited significant olfactory responses except tetradecanoic acid (C₁₄) and octadecanoic acid (C₁₈). In particular, hexanoic acid (C₆) elicited a maximum, eight-fold olfactory response compared with the solvent control. Ethylene glycol and benzyl alcohol exhibited significant EAG and behavioral responses, whereas among aldehydes, nonanal elicited high EAG and behavioral responses, but only at all three doses tested compared with control. Some carboxylic acids elicited significant behavioral responses, attracting ≈75% of females tested toward chemical stimuli in a Y-tube olfactometer.

Ovitraps baited with grass infusions were evaluated in the field as oviposition attractants or stimulants for Aedes (Stegomyia) mosquitoes. Significantly higher oviposition by Aedes aegypti (L.) was observed in areas with no vegetation, but intense human presence, whereas Aedes albopictus (Skuse) oviposition was significantly higher in areas with vegetation, regardless of human presence. Ovitraps baited with infusions of Panicum maximum (Jacq.) collected significantly more eggs than controls containing tap water. Moreover, the number of eggs deposited in ovitraps baited with infusion made from fresh, mature leaves was higher compared with dried leaves or fresh, immature leaves. Anaerobically fermented of grass infusions were more attractive than either aerobically fermented or sterilized infusions of P. maximum. The fermentation age of the grass infusion also affected the oviposition response of mosquitoes. Significantly more eggs were deposited in baited ovitraps containing a fresh leaf infusion of P. maximum fermented for 15 or 20 d, compared with the controls. This demonstrates that 15- or 20-d anaerobic fermentation made of fresh, fully mature leaves of P. maximum is the optimum infusion for ovitrap-based Aedes (Stegomyia) surveillance mosquitoes in Brazil. Neglecting to consider these differences in infusion attractiveness may introduce significant variation in surveillance results.

We examined whether predators and competitors influence selection of oviposition sites by Anopheles gambiae Giles. Mosquitoes in cages laid significantly fewer eggs in rainwater conditioned with a predator (backswimmers, Notonecta sp.) than in unconditioned rainwater. Rainwater conditioned with a putative competitor (tadpoles, Xenopus sp.) also had fewer eggs than unconditioned rainwater. Similarly, mosquitoes laid significantly fewer eggs in rainwater conditioned with five and 50 An. gambiae larvae than in unconditioned rainwater. When larvae were present, significantly more eggs were laid in containers with five larvae than in containers with higher densities, but the differences in number of eggs laid were not significant among the densities of 40, 70, and 100 larvae. This study demonstrated that caged An. gambiae females avoid oviposition in habitats with supposed competitors and predators.

Attraction of Culex quinquefasciatus Say, Culex tarsalis Coquillett, Culex nigripalpus Theobald, and Aedes aegypti (L.) to avian and other host odors was investigated in a dual-port olfactometer. Although attraction to a human arm was high for Ae. aegypti (>80%) and low for all Culex spp. (<25%), all species responded similarly to a chicken (55.3–73.6%). Responses of Ae. aegypti, Cx. quinquefasciatus, and Cx. nigripalpus to feathers were low (<20%) but greater than to controls. There was no difference in attraction of Cx. tarsalis to feathers or controls. Responses to CO₂ (5 ml/min) were low for all species (<15%) except Cx. tarsalis, which were moderate (24.5%). When feathers were combined with CO₂, the resulting attraction was additive or lower than responses to feather and CO₂ alone for all species except for Cx. tarsalis, which had responses that were three-fold greater than expected if responses were additive. The CO₂–feather treatments were less attractive than a chicken for all species. When olfactometer assays were extended from 3 to 20 min, responses by Ae. aegypti significantly increased to a chicken and CO₂ and attraction of Cx. quinquefasciatus significantly greater to chickens, CO₂, and feathers. None of the volatile compounds previously identified from feathers or uropygial glands tested were attractive. Both feather-rubbed cotton balls and hexane extracts of feathers were attractive and as attractive as feathers; however, ether extracts were not attractive. Feathers clearly contribute to the attraction of host-seeking Culex spp., and future studies will focus on identification of the attractant compounds.

Analyses of 11 isoenzyme loci of Ochlerotatus eatoni (Edwards, 1916), endemic to two Macaronesian Islands (Madeira and Tenerife, Canary Islands), revealed substantial genetic structure in the study populations. Samples from sites on the south and north of Madeira displayed a significant reduction of variability compared with those from central Madeira and Tenerife. The Tenerife population exhibited a severe deficit of heterozygosity with similar magnitude across all the loci examined. The complex pattern of variation in Oc. eatoni is because of interplay of breeding structure, genetic drift, and geographical and historical factors. From these findings, we concluded that island colonization by Oc. eatoni was not marked by founder effect.

Most single nucleotide polymorphism (SNP) detection requires expensive equipment and reagents. The oligonucleotide ligation assay (OLA) is an inexpensive SNP assay that detects ligation between a biotinylated “allele-specific detector” and a 3′ fluorescein-labeled “reporter” oligonucleotide. No ligation occurs unless the 3′ detector nucleotide is complementary to the SNP nucleotide. The original OLA used chemical denaturation and neutralization. Heated OLA (HOLA) instead uses a thermal stable ligase and cycles of denaturing and hybridization for ligation and SNP detection. The cost per genotype is ≈US$1.25 with two-allele SNPs or ≈US$1.75 with three-allele SNPs. We illustrate the development of HOLA for SNP detection in the Early Trypsin and Abundant Trypsin loci in the mosquito Aedes aegypti (L.) and at the α-glycerophosphate dehydrogenase locus in the mosquito Anopheles gambiae s.s.

In total, 320 phelebotomine sand flies from Alibozlu (Osmaniye), Kizyusuflu (Osmaniye), and Sanliurfa in southeastern Turkey were tested for the detection and identification of Leishmania in vector sand flies by enzyme-linked immunosorbent assay with species-specific monoclonal antibodies. We used monoclonal antibodies that recognize both Leishmania tropica and Leishmania major, and a monoclonal antibody specific only to L. tropica. Phosphate-buffered saline and monoclonal antibody M₂ recognizing Leishmania amazonensis were used as controls. Infection rates of sand flies were 0.9% in Alibozlu, 0% Kizyusuflu, and 3.6% in Sanliurfa. Positive sand flies were identified as Phlebotomus sergenti Parrot and Phlebotomus major syriacus Adler & Theodor.

Serotonin is an important neuromessenger used in a variety of signaling pathways throughout the animal kingdom. In insects, serotonin has been demonstrated to mediate feeding and feeding-related behaviors. In this study, serotonin antibody was localized in cells and processes throughout the central nervous system (CNS) and midgut of female horse fly Tabanus nigrovittatus Macquart. In the CNS, immunoreactivity was localized in cells and processes throughout the brain and ventral nerve cord. In the midgut, a fine network of immunoreactive processes was observed running along the outer surface of the midgut, with a decrease in innervation toward the posterior region of the midgut.

Ethanolic extracts from the Ginkgo biloba L. exocarp from the Chinese ginkgo were assayed against larvae of three strains of Culex pipiens pallens Coquillett. The chemical compositions were detected using a Hewlett–Packard 6890/5973 mass spectrometric detector. The larvicidal bioassay was carried out according to the recommendations of the World Health Organization. The analysis of the essential oil of ginkgo exocarp showed that its major components are ginkgo acid (85.3%) and ginkgo phenolic (5.69%). The larvicidal bioassay showed that extracts of ginkgo exocarp have LC₅₀ of 18.6, 12.7, and 25.0 mg/liter for deltamethrin-susceptible, deltamethrin-resistant, and field strains, respectively. The acute toxicity concentrations of the ginkgo extracts that killed 50% (LD₅₀) of Wistar rats within 2 wk and young carp within 96 h were 4947.2 mg/kg and 557.9 mg/liter, respectively. These results are promising in creating new, effective, and affordable approaches to mosquito control.

The effects of azadirachtin A added to the standard diet on the development, mortality, and metamorphosis of Lutzomyia longipalpis Lutz & Neiva, 1912 were studied. Concentrations of 0.1, 1.0, and 10.0 μg of azadirachtin/mg of diet significantly increased larval mortality in comparison with nontreated insects. Concentrations 0.1 and 1.0 μg blocked the molt of larvae, which remained as third instars until the end of the experiment. The 10 μg/mg concentration resulted in greater molt inhibition. In this group, all insects stopped their development as second instars. Simultaneous addition of ecdysone (1 μg/mg) to the standard diet containing azadirachtin counteracted the effects of azadirachtin on mortality and inhibition of ecdysis. These results indicate that azadirachtin is a potent growth inhibitor of L. longipalpis.

Malaria control in southern Mozambique is currently by indoor residual carbamate insecticide treatment, with pyrethroid-treated bed-nets distributed to pregnant women and children under five in northern Mozambique. The susceptibility of Anopheles funestus s.s. to pyrethroid, carbamate, organochlorine, and organophosphorus insecticides was determined by World Health Organization adult mosquito susceptibility tests at 19 localities in Mozambique, from March 2000 to July 2002. Biochemical assays were carried out on mosquitoes from the same families to detect shifts in the quantity or activity of enzyme families involved in insecticide detoxification. An. funestus from all localities remained fully susceptible to DDT and the organophosphorus insecticide malathion. A high level of pyrethroid resistance was detected in An. funestus populations in southern Mozambique. An. funestus outside Maputo province were still susceptible to pyrethroids. An. funestus from six localities also were resistant to carbamate insecticides propoxur and bendiocarb. Both pyrethroid and carbamate resistance occurred in five of these six localities. Mosquitoes from five of the localities with elevated p450 estimates, compared with the insecticide-susceptible Durban strain, were pyrethroid-resistant. The only exception to this trend was Mozal, which had elevated p450 estimates but full pyrethroid susceptibility by bioassay. The lack of cross-resistance between pyrethroids and DDT in Mozambican An. funestus suggests that a kdr-type target site resistance mechanism has not been selected. Low levels of insecticide-insensitive acetylcholinesterase, the target site for carbamates and organophosphates, were found in all populations tested. The high level of metabolically based pyrethroid resistance has implications for current malaria control programs in Mozambique.

Malaria control in the southern part of Mozambique is currently by indoor residual spraying with a carbamate insecticide and by pyrethroid-treated bed-nets distributed to pregnant women and children under five in northern Mozambique. The susceptibility of Anopheles gambiae s.s. and Anopheles arabiensis Patton to pyrethroid, carbamate, organochlorine, and organophosphorus insecticides was determined by World Health Organization adult mosquito susceptibility tests at 17 localities in Mozambique, from March 2000 to July 2002. Biochemical assays were carried out on mosquitoes from the same families to detect shifts in the quantity or activity of enzyme families involved in insecticide detoxification. An. gambiae s.s. from all localities remained fully susceptible to DDT and the organophosphorus insecticide malathion. A low level of pyrethroid resistance was detected in populations in southern Mozambique. Populations outside Maputo province were still susceptible to pyrethroids. Low level resistance to the carbamate propoxur also was detected in An. arabiensis from two localities. Mosquitoes from five of the localities had elevated p450 estimates, compared with the insecticide susceptible Durban strain. The lack of cross-resistance between pyrethroids and DDT in Mozambican populations suggests that a kdr-type target site resistance mechanism has not been selected. Increased frequencies of insecticide insensitive acetylcholinesterase, the target site for carbamates and organophosphates, were found in 16 of the populations tested. Although vector control with bendiocarb is not being compromised by the presence of the acetylcholinesterase mechanism alone, the high level of insensitive acetylcholinesterase unless sensibly managed may have long-term implications for malaria control programs in Mozambique.

We performed experiments to determine whether an extract of Sarcoptes scabiei (De Geer) influenced cytokine expression by human T-lymphocytes. Peripheral blood mononuclear cells from five sensitized donors and four donors without sensitization to scabies mites were challenged with a T-cell mitogen alone, with scabies extract (SS) alone, or with mitogen and SS together. Supernatants were analyzed for the cytokines interferon-γ (IFNγ), interleukin (IL)-2, IL-4, and IL-10. No IL-2 or IL-4 was produced in response to scabies extract. Cells from both naïve and sensitized donors produced large amounts of IFNγ and IL-10. The lack of IL-4 but high levels of IL-10 suggests that IL-10 was likely secreted by type 1 T-regulatory cells, which were activated by something in the scabies extract. IL-10 has anti-inflammatory and immune-suppressive effects. It may play a key role in depressing the inflammatory and immune responses in humans so that clinical symptoms are not seen until 4–6 wk after a person becomes infested with scabies mites.

Whether the house fly, Musca domestica L., gut is a permissive environment for horizontal transfer of antibiotic resistance and virulence genes between strains of Escherichia coli is not known. House flies were immobilized and force fed suspensions of defined, donor strains of E. coli containing chloramphenicol resistance genes on a plasmid, or lysogenic, bacteriophage-born Shiga toxin gene stx1 (bacteriophage H-19B::Ap1). Recipient strains were E. coli lacking these mobile elements and genes but having rifampicin as a selectable marker. Plasmid transfer occurred at rates of 10⁻² per donor cell in the fly midgut and 10⁻³ in the fly crop after 1 h of incubation postfeeding. Bacteriophage transfer rate was ≈10⁻⁶ per donor cell without induction, but induction with mitomycin C increased rates of transfer to 10⁻² per donor cell. These findings show that genes encoding antibiotic resistance or toxins will transfer horizontally among bacteria in the house fly gut via plasmid transfer or phage transduction. The house fly gut may provide a favorable environment for the evolution and emergence of pathogenic bacterial strains through acquisition of antibiotic resistance genes or virulence factors.

We evaluated the molestus form of Culex pipiens pipiens (L.) (hereafter referred to as “molestus”) captured near Tashkent, Uzbekistan, for their ability to transmit Japanese encephalitis (family Flaviviridae, genus Flavivirus, JEV) and West Nile (family Flaviviridae, genus Flavivirus, WNV) viruses under laboratory conditions. These molestus were highly competent laboratory vectors of WNV, with infection and dissemination rates of 96 and 81%, respectively. Approximately 75% of female molestus that fed after development of a disseminated infection transmitted virus by bite. Therefore, ≈60% of those molestus taking a second bloodmeal between 16 and 25 d after an infectious bloodmeal would be expected to transmit WNV by bite. In contrast, these molestus were less efficient vectors of JEV, with infection and dissemination rates of 51 and 25%, respectively. In addition, only 33% of individuals with a disseminated infection transmitted JEV by bite, indicating a significant salivary gland barrier. Therefore, only ≈8% of orally exposed individuals would be expected to transmit JEV by bite if they took a second bloodmeal 16–25 d later. These data indicate that the molestus form of Cx. p. pipiens should be considered a potentially important vector of WNV in Uzbekistan and may become involved in the transmission of JEV, should this virus be introduced into Uzbekistan.

The developmental cycle of the cyclically transmitted African trypanosome involves an obligatory passage through the tsetse fly, Glossina spp. This intricate relationship requires the presence of molecules within the insect vector, including a midgut lectin, that interact with the trypanosome. Recently, a gene encoding for a proteolytic lectin, with trypanosome-transforming activity, was isolated from a midgut cDNA library of Glossina fuscipes fuscipes Austen in our laboratory. Using the same approach, we have identified a similar gene from a midgut cDNA library of Glossina austeni (Newstead). The protein encoded by this gene was expressed in bacteria and a baculovirus-based expression system. The baculovirus-expressed lectin was found in the medium of baculovirus-infected Sf-21 cell cultures, indicating that the tsetse fly-derived signal peptide was recognized and cleaved by the Sf-21 cells. The baculovirus-expressed protein also was glycosylated despite the absence of classical O-linked and N-linked sugar attachment motifs. Both the baculovirus- and bacterium-expressed lectin proteins were shown to agglutinate trypanosomes and rabbit red blood cells in vitro. This agglutination was strongly inhibited by d-glucosamine. d-Glucosamine also inhibited the action of the authentic and recombinant lectins upon the chromogenic substrate Chromozym TRY. Interestingly, both baculovirus- and bacterium-expressed lectins showed no significant differences in terms of these activities, indicating that a sugar moiety is not essential for biological activity. Our results provide an important molecular tool for further characterization of Glossina proteolytic lectin.

Culex tarsalis Coquillett females were infected with the NY99 strain of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) and then incubated under constant temperatures of 10–30°C. At selected time intervals, transmission was attempted using an in vitro capillary tube assay. The median time from imbibing an infectious bloodmeal until infected females transmitted WNV (median extrinsic incubation period, EIP₅₀) was estimated by probit analysis. By regressing the EIP rate (inverse of EIP₅₀) as a function of temperature from 14 to 30°C, the EIP was estimated to require 109 degree-days (DD) and the point of zero virus development (x-intercept) was estimated to be 14.3°C. The resulting degree-day model showed that the NY99 WNV strain responded to temperature differently than a lineage II strain of WNV from South Africa and approximated our previous estimates for St. Louis encephalitis virus (family Flaviviridae, genus Flavivirus, SLEV). The invading NY99 WNV strain therefore required warm temperatures for efficient transmission. The time for completion of the EIP was estimated monthly from temperatures recorded at Coachella Valley, Los Angeles, and Kern County, California, during the 2004 epidemic year and related to the duration of the Cx. tarsalis gonotrophic cycle and measures of WNV activity. Enzootic WNV activity commenced after temperatures increased, the duration of the EIP decreased, and virus potentially was transmitted in two or less gonotrophic cycles. Temperatures in the United States during the epidemic summers of 2002–2004 indicated that WNV dispersal and resulting epicenters were linked closely to above-average summer temperatures.

Xanthurenic acid (XA) has been implicated as an inducer in vivo of exflagellation in Plasmodium spp. Consequently, the development of Plasmodium gallinaceum was assessed in a white-eye mosquito strain, kh^w, of Aedes aegypti (L.), which is deficient in XA because of a mutation of the gene encoding the enzyme kynurenine hydroxylase, and in a transformed line of kh^w mosquitoes that carry the wild-type cn gene of Drosophila melanogaster Meigen and express a functional enzyme necessary for XA production. Although XA was not detectable in kh^w mosquitoes by using high-pressure liquid chromatography with electrochemical detection, parasites were able to develop. Transformed kh^w mosquitoes failed to consistently support parasite development at higher prevalences and mean intensities than did the nontransformed kh^w lines, even though XA was detectable. These data suggest that factors other than XA may play a role in initiating Plasmodium development in vivo.

Laboratory-reared Culicoides sonorensis Wirth & Jones were infected with vesicular stomatitis virus serotype New Jersey (family Rhabdoviridae, genus Vesiculovirus, VSNJV) through intrathoracic inoculation. After 10-d incubation at 25°C, these insects were allowed to blood feed on four steers. Two other steers were exposed to VSNJV through intralingual inoculation with 10⁸ tissue culture infective dose₅₀ VSNJV. All six steers became seropositive for VSNJV. The results demonstrate the ability of C. sonorensis to transmit VSNJV to livestock. Only the animals intralingually inoculated with VSNJV showed clinical signs in the form of vesicles at the site of inoculation. Uninfected C. sonorensis allowed to feed on the exposed animals did not become infected with VSNJV. Animals infected by C. sonorensis showed a slower antibody response compared with intralingually inoculated animals. This is probably because of different amounts of virus received via insect transmission and syringe inoculation. A significant difference was found in the serum acute-phase protein α-1-acid glycoprotein in animals that received VSNJV through C. sonorensis transmission. These animals had previously been exposed to insect attack in the field compared with intralingually inoculated animals and C. sonorensis-infected animals that had been protected from insect attack. The failure to observe clinical signs of vesicular stomatitis through transmission of VSNJV by C. sonorensis may explain widespread subclinical infections during vesicular stomatitis epidemics.

Immature Aedes mosquitoes were found in domestic, peridomestic, and tree hole habitats within three socioeconomic strata of Jodhpur, a city within an arid area of Rajasthan, India, endemic for dengue. Peridomestic habitats served as a persistent source of Aedes vectors, especially those used for watering cows for religious reasons that were located within high socioeconomic areas. Domestic (indoor) containers within low socioeconomic strata showed a higher container index (27.0%) than periurban areas with cattle sheds (14.3%) or high socioeconomic areas (18.1%). Mosquitoes were collected in tree holes at zoos and gardens supporting several species of monkeys. Six of 67 Aedes albopictus Skuse reared from immatures collected in tree holes tested positive for dengue antigen acquired through vertical transmission, possibly indicating a persistence mechanism for dengue virus within an urban environment.

The wetness and greenness indices created using Landsat Thematic Mapper (TM) data from June 1995 and 1997 and July 2002 were tested for their ability to predict the location of sites with different levels of nymphal blacklegged tick, Ixodes scapularis Say, abundance in Rhode Island. In 1995, there were statistically significant differences in the mean of greenness and wetness indices between sites classified as low and moderate tick abundance areas (P = 0.005 and P = 0.041, respectively). In 1997, there also were statistically significant differences in the mean of the greenness and wetness indices, but these differences were between the grouping of low/moderate tick abundance and the high tick abundance category (P = 0.023 and P = 0.015, respectively). The same indices from the 2002 image were not significant predictors of tick abundance. It may be that Landsat TM-derived indices can be used to predict nymphal blacklegged tick abundance in years (e.g., 1995 and 1997) when tick abundance is lower than average but not in years when it is higher (e.g., 2002). Thus, it seems unlikely that these remotely sensed indices will be very useful for modeling nonperidomestic Lyme disease risk over a large region in Rhode Island.

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) invaded southern California during 2003, successfully overwintered, amplified to epidemic levels, and then dispersed to every county in the state. Although surveillance programs successfully tracked and measured these events, mechanisms that allowed the efficient overwintering and subsequent amplification of WNV have not been elucidated. Our current research provided evidence for three mechanisms whereby WNV may have persisted in southern California during the winters of 2003–2004 and 2004–2005: 1) continued enzootic transmission, 2) vertical transmission by Culex mosquitoes, and 3) chronic infection in birds. WNV was detected in 140 dead birds comprising 32 species, including 60 dead American crows, thereby verifying transmission during the November–March winter period. Dead American crows provide evidence of recent transmission because this species always succumbs rapidly after infection. However, WNV RNA was not detected concurrently in 43,043 reproductively active female mosquitoes comprising 11 species and tested in 1,258 pools or antibody in sera from 190 sentinel chickens maintained in 19 flocks. Although efficient vertical transmission by WNV was demonstrated experimentally for Culex tarsalis Coquillett infected per os, 369 females collected diapausing in Kern County and tested in 32 pools were negative for WNV. Vertical transmission was detected in Culex pipiens quinquefasciatus Say adults reared from field-collected immatures collected from Kern County and Los Angeles during the summer transmission period. Chronic infection was detected by finding WNV RNA in 34 of 82 birds that were inoculated with WNV experimentally, held for >6 wk after infection, and then necropsied. Frequent detection of WNV RNA in kidney tissue in experimentally infected birds >6 wk postinfection may explain, in part, the repeated detection of WNV RNA in dead birds recovered during winter, especially in species such as mourning doves that typically do not die after experimental infection. In summary, our study provides limited evidence to support multiple modes of WNV persistence in southern California. Continued transmission and vertical transmission by Culex p. quinquefasciatus Say seem likely candidates for further study.

The invasion of different southern California landscapes by West Nile virus (WNV) and its subsequent amplification to epidemic levels during 2004 enabled us to study the impact of differing corvid populations in three biomes: the hot Colorado desert with few corvids (Coachella Valley), the southern San Joaquin Valley (Kern County) with large western scrub-jay but small American crow populations, and the cool maritime coast (Los Angeles) with a large clustered American crow population. Similar surveillance programs in all three areas monitored infection rates in mosquitoes, seroconversion rates in sentinel chickens, seroprevalence in wild birds, numbers of dead birds reported by the public, and the occurrence of human cases. Infection rates in Culex tarsalis Coquillett and sentinel chicken seroconversion rates were statistically similar among all three areas, indicating that highly competent mosquito hosts were capable of maintaining enzootic WNV transmission among less competent and widely distributed avian hosts, most likely house sparrows and house finches. In contrast, infection rates in Culex pipiens quinquefasciatus Say were statistically higher in Kern and Los Angeles counties with elevated corvid populations than in Coachella Valley with few corvids. Spatial analyses of dead corvids showed significant clusters near known American crow roosts in Los Angeles that were congruent with clusters of human cases. In this area, the incidence of human and Cx. p. quinquefasciatus infection was significantly greater within corvid clusters than without, indicating their importance in virus amplification and as a risk factor for human infection. In contrast the uniform dispersion by territorial western scrub-jays resulted in a high, but evenly distributed, incidence of human disease in Kern County.

Birds from 45 species were sampled during three spring seasons from an isolated canyon on the Sutter Buttes in California for the presence of subadult stages of Ixodes pacificus Cooley & Kohls, and for infection with Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner. These birds were found to have an infestation prevalence of 45%, a density of 1.7 ticks per bird, and an intensity of 3.8 ticks per infested bird. There was a significant difference in the I. pacificus infestations between canopy and ground-dwelling birds. Birds also demonstrated an overall infection with B. burgdorferi of 6.4% with significant difference between bird species. Amplification and subsequent sequencing of the 23s-5s rRNA intergenic spacer region of the Borrelia genome from one bird, a hermit thrush, Catharus guttatus (Nuttall), showed that the infection in this bird was caused by B. burgdorferi sensu stricto; the first such finding in a bird from the far west. Our results suggest that birds play a role in the distribution and maintenance of I. pacificus, and possibly of B. burgdorferi, at the Sutter Buttes, CA.

This study aimed to identify the major mosquito vectors of Ross River virus (family Togaviridae, genus Alphavirus, RRV) and to explore the threshold of mosquito abundance necessary for RRV transmission in Brisbane, Australia. Data on the monthly counts of RRV cases by statistical local areas from the Queensland Health and the monthly mosquito abundance in Brisbane between November 1998 and December 2001 from the Brisbane City Council were used to assess the pairwise relationship between mosquito abundance and the incidence of RRV disease over a range of time lags using cross-correlations. We used time series Poisson regression models to identify major mosquito species associated with incidence of RRV after adjusting for overdispersion, maximum temperature, autocorrelation, and seasonality. Our results show that Aedes vigilax (Skuse) (relative risk [RR] = 1.32; 95% CI = 1.01–1.74 per 100 mosquitoes per trap) and Culex annulirostris (Skuse) (RR = 1.14, 95% CI = 1.04–1.24 per 100 mosquitoes per trap) were most strongly associated with RRV transmission at a lag of 1 mo. Classification and regression tree (CART) analyses indicate that the occurrence of RRV was associated with an average monthly mosquito abundance of Aedes vigilax above 72 and Cx. annulirostris above 52. The validation analyses indicate that the crude agreement between predicted values and actual observations was 76% (sensitivity, 61%; specificity, 80%). The results may have applications as a decision support tool in planning disease control and risk-management programs.

Previous studies have identified several anopheline species integral to the transmission of malaria in Belize. The highly efficient vector, Anopheles darlingi Root, is currently considered the most important. The preferred larval habitat of An. darlingi has been described as floating detritus patches, which are commonly associated with overhanging spiny bamboo, Guadua longifolia (E. Fourn.), along river margins. The objectives of this study were to use remote sensing and geographic information system (GIS) tools to 1) define the landscape features (i.e., river curvature, land cover, and house locations) associated with An. darlingi-positive breeding habitats and 2) determine the association between cleared land cover and the growth of spiny bamboo. A systematic survey was conducted in which all detritus patches of at least 1 m² were sampled, mapped using GPS, and characterized by cause of habitat lodging. Bamboo stretches growing along the river margins also were mapped. Spatial analyses of satellite imagery found no associations between river characteristics or land cover with positive An. darlingi habitats. In addition, there was no significant difference in cleared versus forested land cover in relation to the presence or absence of bamboo. Results indicate that the average distance from homes to negative habitats was significantly greater than from positive detritus mats. Based on the land cover and river characteristics used, our results do not support the use of remote sensing as a predictive tool to locate specific areas within rivers positive for An. darlingi habitats.

Numerous studies have investigated the role of weather on insect species. For mosquitoes, these studies have yielded mixed results. Although it is clear that weather impacts mosquito population dynamics, these investigations have failed to accurately characterize their fluctuations. We use a novel graphical method to examine large numbers of meteorological aggregations of varying lengths and lags simultaneously to establish relationships between these summary variables and mosquito counts, to gain a better understanding of the weather effects. Poisson regression models were developed to characterize the population dynamics of Aedes sollicitans (Walker) by using meteorological data and a 34-yr set of daily mosquito count data. The models accurately characterize mosquito dynamics over time and space. The aggregated meteorological variables included in the model were lowest minimum tides between days 27 and 14 before trapping, total precipitation between days 22 and 9, total precipitation on day 1 and the day of trapping, cooling degree-days on day 0, average minimum relative humidity between days 28 and 9, lowest stream flow from day 11 to day 0, and lowest minimum temperature between days 28 and 13.

In southeastern Canada, most populations of Ixodes scapularis Say, the Lyme disease vector, occur in Carolinian forests. Climate change projections suggest a northward range expansion of I. scapularis this century, but it is unclear whether more northerly habitats are suitable for I. scapularis survival. In this study, we assessed the suitability of woodlands of the Lower Great Lakes/St. Lawrence Plain region for I. scapularis by comparing tick egg survival in four different woodlands. Woodlands where I. scapularis are established, and sand dune where I. scapularis do not survive, served as positive and negative control sites, respectively. At two woodland sites, egg survival was the same as at the positive control site, but at two of the sites survival was significantly less than either the positive control site, or one of the other test sites. Egg survival in all woodland sites was significantly higher than in the sand dune site. Ground level habitat classification discriminated among woodlands in which tick survival differed. The likelihood that I. scapularis populations could persist in the different habitats, as deduced using a population model of I. scapularis, was significantly associated with variations in Landsat 7 ETM data (normalized difference vegetation index [NDVI] and Tasselled Cap indices). The NDVI index predicted habitat suitability at Long Point, Ontario, with high sensitivity but moderate specificity. Our study suggests that I. scapularis populations could establish in more northerly woodland types than those in which they currently exist. Suitable habitats may be detected by ground-level habitat classification, and remote-sensed data may assist this process.

We used drag sampling to examine the geographical distribution patterns of ixodid ticks engaging in open (non-nidicolous) host-seeking behavior in dense woodland habitats of the climatically and ecologically diverse Mendocino County in north coastal California. The findings based on this sampling methodology reflect risk of human exposure to host-seeking ticks rather than the true distribution of the ticks. Drag sampling in 78 sites yielded 7,860 nymphal or adult Ixodes pacificus Cooley & Kohls, 220 Dermacentor occidentalis Marx, 150 Ixodes spinipalpis Hadwen & Nuttall, 15 Hemaphysalis leporispalustris (Packard), 12 Ixodes angustus Neumann, 12 Ixodes auritulus Neumann, and a single Dermacentor variabilis (Say). I. pacificus, which is the primary vector of the Lyme disease spirochete Borrelia burgdorferi to humans in California, occurred in all 78 sites examined. D. occidentalis, another tick species commonly biting humans in California, and H. leporispalustris typically were encountered in oak-associated woodlands in the central or eastern parts of the county. In contrast, three species of Ixodes ticks (I. angustus, I. auritulus, and I. spinipalpis) most commonly were found questing openly in woodlands with redwood present in the western part of the county. I. angustus and I. spinipalpis are occasional human biters and known experimental vectors of B. burgdorferi. Our study represents the first collection of large numbers of openly host-seeking I. spinipalpis ticks. Univariate tests of associations between presence of ticks (D. occidentalis, H. leporispalustris, I. angustus, I. auritulus, or I. spinipalpis) and environmental geographical information systems–remote sensing (GIS/RS)-based data indicated that elevation, number of growing degree-days, and tasseled cap brightness, greenness, and wetness are especially useful predictors of presence of openly host-seeking ticks. Combinations of the above-mentioned GIS/RS-based data yielded significant logistic regression models for habitat suitability of host-seeking ticks for all five above-mentioned species. The model equations were used to create spatial surfaces of predicted presence of suitable habitat for openly host-seeking ticks in Mendocino County dense woodlands.

Insecticide-treated bednets (ITNs) significantly reduce malaria vector populations. Susceptibility to ITNs differs by vector species, and culicine mosquitoes have not been shown to be significantly affected by the use of ITNs. We examined the impact of 2–4 yr of ITN use on malaria vector species distribution and culicine mosquitoes. Routine entomological surveillance was conducted in adjacent areas with and without ITNs from November 1999 to January 2002. Use of ITNs reduced the proportion of Anopheles gambiae Giles relative to Anopheles arabiensis Giles. The number of culicines per house was significantly lower in the ITN area than in the neighboring area. Changes in the An. gambiae sibling species distribution may help to explain apparent mosquito behavioral changes attributed to ITNs. Reductions in culicines by ITNs may have implications for community perceptions of ITN effectiveness and for control of other diseases such as lymphatic filariasis.

Experimental approaches provide insight into the dynamics of species competition and coexistence, but studies of ectoparasitic arthropods are rare. Host choice experiments were undertaken in Amazonian Peru, by using obligate bat fly parasites of Seba’s short-tailed bat, Carollia perspicillata (L.). The fly species Trichobius joblingi Wenzel strongly preferred clean hosts to those previously infested with individuals of T. joblingi. Abundance thresholds may limit conspecific parasites on host individuals, and similar morphologies and habitat use may increase intraspecific competition among individuals of T. joblingi. T. joblingi did not distinguish between clean hosts and those previously infested with flies of another species, Speiseria ambigua Kessel. Results suggest that the presence of a morphologically dissimilar species does not preclude subsequent colonization. Differential microhabitat selection, aided by adaptive morphology, may facilitate species coexistence among ectoparasitic bat fly species.

The blacklegged tick, Ixodes scapularis Say, first reported in Indiana in 1987, has now been detected in more than half of Indiana’s counties. The first case of human granulocytic ehrlichiosis (human anaplasmosis) in Indiana was reported in 2002. We now report the detection of Anaplasma phagocytophilum and Babesia odocoilei (Emerson and Wright 1968) in I. scapularis ticks collected in northern Indiana. Using polymerase chain reaction analysis, 41 of 193 adult ticks (21.2%) collected from deer were positive for A. phagocytophylum, and 22 (11.4%) were positive for Babesia sp. Restriction fragment analysis of 12, and sequencing of another five of the amplified products identified these parasites as B. odocoilei. Five ticks (2.6%) were coinfected. Eight of 68 questing adult ticks (11.8%) were positive for A. phagocytophilum; seven (10.3%) were positive for Babesia sp. Six of the latter seven positive samples were determined to be B. odocoilei by restriction fragment analysis and sequencing of two samples. None of 39 pools of nymphs was positive for Babesia sp. Three of 15 ticks (20%) collected from a dog were positive for A. phagocytophilum and three ticks (20%) were positive for Babesia sp. One was confirmed as B. odocoilei. One tick was coinfected. This is the first report of the presence of these two agents in ticks in Indiana.