The sand fly Lutzomyia longipalpis (Lutz & Neiva) (Diptera: Psychodidae: Phlebotominae) is the main vector of American visceral leishmaniasis. Adult males produce a terpenoid sex pheromone that in some cases also acts as male aggregation pheromone. We have analyzed the correlation between male pheromone production levels and pheromone gland cell morphogenesis after adult emergence from pupae. The abdominal tergites of L. longipalpis males were dissected and fixed in glutaraldehyde for transmission electron microscopy, or the pheromone was extracted in analytical grade hexane. Pheromone chemical analysis was carried out at 3- to 6-h intervals during the first 24 h after emergence and continued daily until the seventh day. All extracts were analyzed by gas chromatography. For the morphological analysis, we used insects collected at 0–6, 9–12, 12–14, and 96 h after emergence. Ultrastructural data from 0- to 6-h-old adult males revealed smaller pheromone gland cells with small microvilli at the end apparatus. Lipid droplets and peroxisomes were absent or very rare, but a large number of mitochondria could be seen. Lipid droplets started to appear in the gland cells cytoplasm ≈9 h after adult emergence, and their number and size increased with age, together with the presence of several peroxisomes, suggesting a role for these organelles in pheromone biosynthesis. At 12–15 h after emergence, the lipid droplets were mainly distributed near the microvilli but were smaller than those in mature older males (4 d old). Pheromone biosynthesis started around 12 h after emergence and increased continuously during the first 3 d, stabilizing thereafter, coinciding with the period when males are more able to attract females.

The male accessory glands in Triatoma are tubular and produce substances with some functions related to production of the spermatophore. In the current study, the cytochemistry of male accessory glands was evaluated in starved Triatoma brasiliensis and adult Triatoma melanica. The storage of carbohydrates and proteins in T. melanica male accessory glands occurs earlier than in T. brasiliensis. In addition, the occurrence of eletron-lucent granules without carbohydrates and proteins suggests that other substances are released by these glands, which may be used for lubrication of the male genitalia. Male T. brasiliensis has more intense secretory activity in the fifth day of adult life, which may indicate a higher reproductive capacity. The analysis of lipid production in male accessory glands can contribute to the knowledge of spermatophore formation in these species.

In total, 167 individuals of 30 species of snakes belonging to 22 genera and five families were examined for tick infestation from November 2008 to March 2010. Only two species of snakes, Ptyas mucosa (L., 1758) (Indian rat snake) and Naja naja (L., 1758) (spectacled cobra), were found infested by ticks. All ticks collected were identified to be Amblyomma gervaisi [previously Aponomma gervaisi (Lucas, 1847) ]. The average prevalence of these ticks on Indian rat snakes (ra = 48) was 29.16%, with abundance of 7.02 ticks per individual; on spectacled cobras (ra = 20), average prevalence was 30.00%, with abundance of 6.9 ticks per individual. The nymphs and males were predominant. All the ticks were found on the dorsal aspect of the body of the snake, and no ticks were recorded on the head, tail, or ventral body. The rate of tick infestation was highest in scrubland and was lowest in evergreen forests. Female Indian rat snakes showed higher tick infestation rates than male Indian rat snakes. Using Mann—Whitney U test, we found that longer snakes of both species had significantly higher rate of tick infestation in both the species of snakes.

With the exception of an airplane equipped with nets, no method has been developed that successfully samples red imported fire ant, Solenopsis invicta Buren, sexuals in mating/dispersal flights throughout their potential altitudinal trajectories. We developed and tested a method for sampling queens and males during mating flights at altitudinal intervals reaching as high as ∼140 m. Our trapping system uses an electric winch and a 1.2-m spindle bolted to a swiveling platform. The winch dispenses up to 183 m of Kevlar-core, nylon rope and the spindle stores 10 panels (0.9 by 4.6 m each) of nylon tulle impregnated with Tangle-Trap. The panels can be attached to the rope at various intervals and hoisted into the air by using a 3-m-diameter, helium-filled balloon. Raising or lowering all 10 panels takes ∼15–20 min. This trap also should be useful for altitudinal sampling of other insects of medical importance.

Vegetation patterns and the presence of large numbers of nesting herons and egrets significantly altered the number of host-seeking Culex tarsalis Coquillett (Diptera: Culicidae) collected at dry ice-baited traps. The numbers of females collected per trap night at traps along the ecotone of Eucalyptus stands with and without a heron colony were always greater or equal to numbers collected at traps within or under canopy. No Cx. tarsalis were collected within or under Eucaplytus canopy during the peak heron nesting season, even though these birds frequently were infected with West Nile virus and large number of engorged females could be collected at resting boxes. These data indicate a diversion of host-seeking females from traps to nesting birds reducing sampling efficiency.

Phlebotomus papatasi (Diptera: Psychodidae) is the Old World sand fly vector of zoonotic cutaneous leishmaniasis caused by Leishmania major (Trypanosomatidae: Kinetoplastida), a debilitating and disfiguring protist parasitic disease prevalent throughout southern Mediterranean countries, the Middle East, as well as southern and eastern European countries, where it is regarded as a serious public health problem. Little is known of the mating ecology of P. papatasi, and, in particular, the role (if any) of pheromones is not known. In this laboratory- and field-based study, we have shown that a male-produced sex pheromone exists in P. papatasi. Young female P. papatasi are attracted to the headspace volatiles of small groups of males, males and females together, but not females alone. Males were not attracted to males, females, or mixed groups of males and females in the laboratory. Larger groups of males or males and females together were repellent in the laboratory study. Field experiments showed that Centers for Disease Control (CDC) light traps baited with small groups of males and females together were attractive to females, but not males. CDC traps baited with large groups of males and females together caught significantly fewer females and males than the control traps; however, the proportion of females caught compared with males overall was much higher than with CDC traps baited with small numbers of males and females. These results suggest that females may be attracted in preference to males to the vicinity of the baited traps and are highly sensitive to the concentration of male pheromone. It also suggests that P. papatasi mating behavior is fundamentally different from that of Lutzomyia longipalpis, where large mating aggregations of males and females occur.

Aquatic prey show behavioral modifications in the presence of predation-risk cues that alleviate their risk from predation. Aedes albopictus (Skuse), Aedes japonicus (Theobald), and Culex pipieos L. are invasive mosquitoes in North America, and their larvae are prey for the native mosquito predator, Toxorhynchites rutilus (Coquillett). Ae. albopictus and Ae. japonicus are recent invaders, whereas Cx. pipiens has been in the United States for >100 yr. In the presence of predation-risk cues from Tx. rutilus larvae, Cx. pipiens larvae increased the time spent resting at the surface (least risky behavior) more than the other prey species. Ae. japonicus larvae increased resting at the surface of the containers more than Ae. albopictus larvae in the presence of predation-risk cues. Cx. pipiens larvae spent more time motionless at the surface even in the absence of predation-risk cues when compared with the other species, indicating that Cx. pipiens larvae are the least vulnerable prey. As compared with the other prey species, Ae. albopictus larvae exhibited more high-risk behaviors both in the presence and absence of predation-risk cues, indicating that they are the most vulnerable prey. Ae. albopictus is the superior competitor; however, predation by Tx. rutilus larvae may prevent competitive exclusion by Ae. albopictus and promote coexistence among the three prey species.

The effects of colonization on host-seeking behavior of mosquitoes was examined by comparing attraction responses of newly colonized Aedes aegypti (L.) from field-collected eggs in Puerto Rico to that of the Gainesville (Florida) strain, originally from Orlando (Florida) and in colony since 1952. Females from the Orlando and the F₀ through F₁₀ generations of the Puerto Rico strain were evaluated using attractant odors in a triple-cage dual-port olfactometer. Two attractant sources were used: odors from the hand of a volunteer and a standard blend of L-lactic acid, acetone, and dimethyl disulfide. Convergence of the percentage of attraction responses occurred around the F₄–F₆ generations of the Puerto Rico strain. Both the Orlando and Puerto Rico strains exhibited similar responses for tests with the remaining F₇–F₁₀ generations. A temporal effect on mosquito responses was observed for both strains regardless of the attractant blend used in tests. This study indicates that Ae. aegypti host-seeking behavior changes significantly over the first four to six generations after introduction into the laboratory, whereas the field-collected strain increases in attraction response until it stabilizes at a new level.

In this study, we examined the effect of weather and moon illumination on sand fly activity, as measured by light trap collections made between 2 May 2003 and 25 October 2004 at Tallil Air Base, Iraq. Wind speed, temperature, dew point, percentage of sky cover, and moon illumination were entered into principal components analysis. The resulting principal components were entered into stepwise regression to develop a model of the impact of the weather on sand fly collections. Wind speed, percentage of sky cover, and moon illumination each had a strong inverse relationship with the number of sand flies collected, whereas temperature displayed a direct relationship to sand fly collections. Our data indicate that sand fly light trap catches at Tallil Air Base are highest on warm, clear nights with low wind speed and minimal illumination from the moon.

The cDNA of a trypsin gene from Aedes (Ochlerotatus) taeniorhynehus (Weidemann) was cloned and sequenced. The full-length mRNA sequence (890 bp) for trypsin from Ae. taeniorhynchus (AetTryp1) was obtained, which encodes an open reading frame of 765 bp (i.e., 255 amino acids). To detect whether AetTryp is developmentally regulated, a quantitative real-time polymerase chain reaction was used to examine AetTryp1 mRNA expression levels in different developmental stages of Ae. taeniorhynehus. AetTryp1 was expressed at low levels in egg, larval, and pupal stages, but was differentially expressed in adult Ae. taeniorhynchus, with highest levels found in 5-d-old female adults when compared with teneral adults. In addition, AetTryp1 mRNA expression differed between sexes, with expression levels much lower in males. However, in both males and females, there was a significant increase in AetTryp1 transcription levels as age increased and peaked in 5-d-old adults. AetTryp1 expressed in 5-d-old female Ae. taeniorhynchus significantly increased after 30 min postblood feeding compared with the control. The AetTryp1 mRNA expression in 5-d-old female Ae. taeniorhynchus was affected by different concentrations of permethrin.

This study assesses the potential of cladocerans as competitors for controlling the oviposition and larval abundances of mosquitoes. Control of mosquito larvae involving the use of antagonists has focused mostly on predators. We hypothesized that cladoceran competitors have a strong potential to control larval populations of some species of mosquitoes that can be early colonizers of newly-filled waterbodies, and should be less efficient competitors. To test this hypothesis, the establishment and development of larval populations of wild Culex pipiens were investigated in outdoor microcosms varying in terms of Daphnia magna populations. When the population was well established (i.e., high densities of D. magna), oviposition was fully inhibited and there was consequently no mosquito larvae. When the population was more recently established (i.e., lower densities of D. magna), oviposition and larval development of Cx. pipiens occurred. In the absence of D. magna, oviposition, larval biomass, and abundance of Cx. pipiens reached high values. In this situation, conspecifics were inhibiting further oviposition of Cx. pipiens. Based on these results, we suggest that competing Zooplankton species, such as D. magna, could be used for the control of mosquito species such as Cx. pipiens. This approach could be beneficial for the management of wetlands.

Glutathione S transferases (GSTs) are multifunctional enzymes involved in detoxification of xenobiotic compounds in majority of the insect groups. Significance of insect GSTs is their elevated level of activity in association with insecticide resistance. This investigation was to explore the metabolic status of GSTs in two Indian DDT-resistant malaria vectors, Anopheles culicifacies and Anopheles annularis, and one DDT-susceptible vector, Anopheles fluviatilis. Malkangiri and Koraput districts of Orissa State, endemic for falciparum malaria and having a long insecticide spraying history, were the study areas. F₁ progeny was raised from wild-caught females of the three vectors and used for biochemical assays to detect the GST-mediated DDT resistance mechanism. Results of the enzyme assay showed a significant 3-fold increase in GST activity in DDT-resistant An. annularis compared with its susceptible population. In DDT-resistant An. culicifacies, the median GST activity (71.8 µmol/min/mg) was almost the same as estimated in the DDT-resistant An. annularis (74.6 µmol/min/mg), suggesting that the GST activity estimated in An. culicifacies could be an elevated level for detoxification of DDT. Furthermore, the GST activity in DDT-resistant An. culicifacies and An. annularis was significantly higher than that in the DDT-susceptible An. fluviatilis, which had a GST activity of 20.0 µmol/min/mg. Also, the GST-mediated DDT detoxification was confirmed by comparing GST activity in wild-caught females with that in their F₁ progeny.

Spinosad is a naturally derived insecticide that has shown potential as a mosquito larvicide. To determine the activity of spinosad against blackflies, late-instar larvae from a community comprising Simulium triittatum (63.6%) and seven other species, including three known vectors of onchocerciasis in Mexico (S. metallicum, S. ochraceum, and S. callidum), were subjected to concentration-mortality laboratory bioassays following World Health Organization guidelines. Cephalic capsule measurements confirmed the relatively homogeneous distribution of experimental larvae. The 50% lethal concentration of spinosad was estimated at 1.48 ppm spinosad (95% confidence interval: 1.07–2.33) for a 10-min exposure period, whereas larvae treated with 0.05 ppm of the organophosphate temephos experienced 61% mortality. Immature aquatic insects were identified to genus and tested for their susceptibility to spinosad in the laboratory. After exposure to 12 ppm spinosad for 10 min, ephemeropterans, odonates, trichopterans, and hemipterans did not experience significantly increased mortality over that of untreated controls, whereas a significant increase in mortality was observed in spinosad-treated Plecoptera (P < 0.001). Tilapia and trout fry exposed to 12 ppm spinosad for 10 min did not experience increased mortality at 24-h postexposure over that of the controls. We conclude that spinosad is less toxic than temephos to these blackfly species, but is likely to have a low impact on nontarget members of the aquatic community.

Dengue and dengue hemorrhagic fever are mosquito-borne viral diseases that coincide with the distribution of Aedes aegypti (L.), the primary vector in the tropical and semitropical world. With no available vaccine, controlling the dengue vector is essential to avoid epidemics. This study evaluates the efficacy of a new smoke-generating formulation containing pyriproxyfen and permethrin in Puerto Libertad, Misiones, Argentina. A fumigant tablet (FT) was applied inside the houses by the community members and compared with a professional application. A treatment combining the application of fumigant tablets indoors and ultralow volume fumigation outdoors was also assessed. The community perceptions and practices about dengue disease and the acceptance of this new nonprofessional FT were evaluated through surveys. Results show >90% adult emergence inhibition and 100% adult mortality with these treatments. More than 80% of the residents applied the FT and preferred participating in a vector control program by using a nonprofessional mosquito control tool, instead of attending meetings and workshops promoting cultural changes.

We conducted a series of field experiments in 2003 and 2004 to evaluate the efficacy of a variety of insecticides and insecticide application technologies for the control of phlebotomine sand flies at Tallil Airbase, Iraq. During the experiments, 53,263 sand flies were collected. The experiments evaluated the following: 1) routine sand fly control operations using a variety of residual and area-wide insecticides; 2) a combination of five different insecticide application methods in and around tents; 3) residual application of lambda-cyhalothrin and ultra-low volume application of Pyrethrins in houses; 4) carbaryl and lambda-cyhalothrin applied as barrier sprays; 5) a deltamethrin-impregnated fence; 6) lambda-cyhalothrin applied as a residual spray in concrete manholes; 7) deltamethrin-treated flooring in tents; and 8) ultra-low volume-applied malathion. Although some of the experiments resulted in limited reductions in the number of sand flies collected in light traps, in no instance did we completely eliminate sand flies or reduce populations for a sustained period. The implications of these findings are discussed.

Several species of captive and wild birds have been found to be infected with various avian blood protozoa in Japan. We investigated the prevalence and transmission of avian malaria parasite and determined the bloodmeal hosts of mosquitoes collected in a zoological garden in Tokyo, Japan, by using the polymerase chain reaction. In total, 310 unfed and 140 blood-fed mosquitoes of seven species were collected by using sweep nets and CDC traps. Bloodmeal identification indicated that mosquitoes had fed on 17 avian and five mammalian species, including captive animals. The results of avian malaria parasite detection from mosquitoes with avian bloodmeals indicated that Culex pipiens pollens Coquillet is a main vector of avian Plasmodium in the current study site and that some captive and wild birds could be infected with avian malaria parasites. Furthermore, the distances between the collection site of blood-fed mosquitoes and the locations of their blood-source captive animals were estimated. Most females with fresh bloodmeals were found within 40 m of caged animals, whereas half-gravid and gravid females were found between 10 and 350 m from caged host animals. We demonstrated that blood-fed mosquitoes can provide useful information regarding the mosquito vector species of avian malaria parasites and allows for noninvasive detection of the presence of avian malaria parasites in bird populations.

Avian influenza caused by avian influenza virus (AIV) has a negative impact on poultry production. Low-pathogenic AIV (LPAIV) is naturally present in wild birds, and the introduction of the virus into domestic poultry is assumed to occur through contact with wild birds and by human activity, including the movement of live and dead poultry, and fomites such as clothing and vehicles. At present, the possible role of insects in the spread of AIV is dubious. The objective of the present work was to investigate the potential transmission of LPAIV by persistence of the virus in the alimentary tract of house flies, Musca domestica L. (Diptera: Muscidae). Flies were fed three virus concentrations of two AIV strains and then incubated at different temperatures for up to 24 h. The persistence of the two virus strains in the flies declined with increasing incubation temperatures and incubation periods. Similarly, increased virus uptake by the flies increased the persistence of virus. Persistence of infective AIV in flies differed significantly between the two virus strains. The laboratory experiments of the present study indicate that the house fly can be a potential carrier of AIV.

The focal distribution of tick-borne encephalitis virus (TBEV; Flaviviridae, Flavivirus) appears to depend mainly on cofeeding transmission between infected Ixodes ricinus L. nymphs and uninfected larvae. To better understand the role of cofeeding ticks in the transmission of TBEV, we investigated tick infestation of rodents and the influence of microclimate on the seasonality of questing I. ricinus ticks. A 3-yr study was carried out at four sites, including two confirmed TBEV foci. Free-living ticks and rodents were collected monthly, and microclimatic data were recorded. A decrease in questing nymph density was observed in 2007, associated with low relative humidity and high temperatures in spring. One site, Thun, did not show this decrease, probably because of microclimatic conditions in spring that favored the questing nymph population. During the same year, the proportion of rodents carrying cofeeding ticks was lower at sites where the questing nymph density decreased, although the proportion of infested hosts was similar among years. TBEV was detected in 0.1% of questing ticks, and in 8.6 and 50.0% of larval ticks feeding on two rodents. TBEV was detected at all but one site, where the proportion of hosts with cofeeding ticks was the lowest. The proportion of hosts with cofeeding ticks seemed to be one of the factors that distinguished a TBEV focus from a non-TBEV focus. The enzootic cycle of TBEV might be disrupted when dry and hot springs occur during consecutive years.

Rift Valley fever virus (RVFV) causes outbreaks of severe disease in domestic ungulates as well as humans in Africa. There is a logical concern that RVFV could be introduced into the Americas and cause significant health and economic damage based on the precedent of the introduction and spread of West Nile virus (WNV). Unfortunately, there are currently no licensed diagnostic assays available for RVFV in the Americas. In this work, we report on the ability of a novel dipstick assay, VectorTest RVFV antigen assay, modeled on the VecTest assay for WNV, to detect a RVFV-infected female within a pool of mosquitoes. The dipsticks provided results in <20 min, were easy to use, and did not require a laboratory with containment facilities. Although readily able to detect a mosquito with a disseminated RVFV infection, it only occasionally detected RVFV in a mosquito with a nondisseminated infection, and therefore may fail to detect some pools that actually contain one or more positive mosquitoes. The RVFV dipstick assay was highly specific and did not react with samples to which had been added yellow fever, West Nile, Venezuelan equine encephalitis, sandfly fever Naples, sandfly fever Sicilian, or sandfly fever Toscana viruses. The RVFV assay can provide a rapid, safe, easy-to-use assay to alert public health personnel to the presence of RVFV in mosquitoes. Results from this assay will allow a rapid threat assessment and the focusing of vector control measures in high-risk areas.

The presence or absence of the biting midge Culicoides sonorensis Wirth & Jones (Diptera: Ceratopogonidae), a primary vector of bluetongue viruses (genus Orbivirus, family Reoviridae, BTV) in North America, was assessed on ranches and farms across the Northern Great Plains region of the United States, specifically Nebraska, South Dakota, and North Dakota, as part of a 2-yr regional study of BTV exposure among cattle. Blacklight/suction trap samples totaling 280 2-night intervals were taken at 140 aquatic sites (potential larval habitat for C. sonorensis) on 82 livestock operations (ranches and farms) that span a south-to-north gradient of expected decreasing risk for exposure to BTV. In Nebraska, C. sonorensis populations were common and widespread, present at 15 of 18 operations. Of 32 operations sampled in South Dakota, seven of which were sampled in successive years, 18 were positive for C. sonorensis; 13 of 14 operations located west of the Missouri River were positive, whereas 13 of 18 operations east of the river were negative. Of 32 operations sampled in North Dakota, seven of which were sampled both years, 12 were positive for C. sonorensis. Six of eight operations located west and south of the Missouri River in North Dakota were positive, whereas 18 of 24 operations east and north of the river were negative for C. sonorensis. These data illustrate a well-defined pattern of C. sonorensis spatial distribution, with populations consistently present across Nebraska, western South Dakota, and western North Dakota; western South Dakota, and North Dakota encompass the Northwestern Plains Ecoregion where soils are nonglaciated and evaporation exceeds precipitation. In contrast, C. sonorensis populations were largely absent east of the Missouri River in South Dakota and North Dakota; this area comprises the Northwestern Glaciated Plains Ecoregion and Northern Glaciated Plains Ecoregion where surface soils reflect Wisconsinan glaciation and precipitation exceeds evaporation. In defining a well-demarcated pattern of population presence or absence on a regional scale, the data suggest that biogeographic factors regulate the distribution of C. sonorensis and in turn BTV exposure. These factors, ostensibly climate and soil type as they affect the suitability of larval habitat, may explain the absence of C. sonorensis, hence limited risk for exposure to BTV, across the eastern Northern Plains, upper Midwest, and possibly Northeast, regions of the United States.

During 2007–2010, we examined which container types in Mérida, México, are most productive for Aedes aegypti (L.) immatures. Surveys for mosquito immatures followed routine surveillance methodology and container type classifications used by Servicios de Salud de Yucatán. Our main findings were that 1) small and larger discarded containers that serve no particular purpose and therefore can be removed from the environment contribute strongly to larval and pupal production in Mérida, and 2) the importance of different container types can vary among sets of residential premises as well as between dry and wet periods. These results may help to guide future implementation in Mérida of control efforts that target the most productive container types for Ae. aegypti immatures. Furthermore, if the Patio Limpio cleanup campaign that currently is ongoing in Mérida proves successful in removing discarded containers as important immature development sites, then we should see dramatic changes in the most productive container types in the future as the mosquito is forced to switch to other container types, which perhaps also will be easier to include in highly targeted mosquito control interventions.

In total, 1,000 Ixodes ricinus L. ticks were collected from a small recreational forest area in central Germany (Thuringia) and investigated for the presence of Borrelia spp., Babesia spp., Anaplasma spp., Rickettsia spp., Coxiella burnetii, and Francisella tularensis. Overall, 43.6% of the ticks were infected with at least one pathogen. In 8.4% of ticks double infections were detected, and 1.6% harbored more than two pathogens. In this study, we present data on the coexistence of established and emerging pathogens in questing nymphs and adult ticks in a recreational area in central Germany, indicating the need for further studies for a reliable risk assessment.

In 2006–2007, stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), were suspected of being enzootic vectors of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) during a die-off of American white pelicans (Pelecanus erythrorhynchos Gmelin) (Pelecanidae) in Montana, USA. WNV-positive stable flies were observed feeding en masse on incapacitated, WNV-positive pelicans, arousing suspicions that the flies could have been involved in WNV transmission among pelicans, and perhaps to livestock and humans. We assessed biological transmission by infecting stable flies intrathoracically with WNV and testing them at 2-d intervals over 20 d. Infectious WNV was detected in fly bodies in decreasing amounts over time for only the first 6 d postinfection, an indication that WNV did not replicate within fly tissues and that stable flies cannot biologically transmit WNV. We assessed mechanical transmission using a novel technique. Specifically, we fed WNV-infected blood to individual flies by using a cotton swab (i.e., artificial donor), and at intervals of 1 min—24 h, we allowed flies to refeed on a different swab saturated with WNV-negative blood (i.e., artificial recipient). Flies mechanically transmitted viable WNV from donor to recipient swabs for up to 6 h postinfection, with the majority of the transmission events occurring within the first hour. Flies mechanically transmitted WNV RNA to recipient swabs for up to 24 h, mostly within the first 6 h. Given its predilection to feed multiple times when disturbed, these findings support the possibility that the stable fly could mechanically transmit WNV.

This study compared the spatial and temporal patterns of Culex tarsalis Coquillett and Aedes vexans Meigen populations and examined their relationships with land cover types and climatic variability in Sioux Falls, SD. Between 24 and 30 CDC CO₂-baited light traps were set annually in Sioux Falls from May to September 2005–2008. Land cover data were acquired from the 2001 National Land Cover Dataset and the percentages of selected land cover types were calculated within a 600-m buffer zone around each trap. Meteorological information was summarized from local weather stations. Cx. tarsalis exhibited stronger spatial autocorrelation than Ae. vexans. Land cover analysis indicated that Cx. tarsalis was positively correlated with grass/hay, and Ae. vexans was positively correlated with wetlands. No associations were identified between irrigation and the host-seeking population of each species. Higher temperature in the current week and 2 wk prior and higher precipitation 3–4 wk before collection of host-seeking adult mosquitoes had positive influences on Cx. tarsalis abundance. Temperature in the current week and rainfall 2–3 wk before sampling had positive influences on Ae. vexans abundance. This study revealed the different influences of weather and land cover on important mosquito species in the Northern Great Plains region, which can be used to improve local vector control strategies and West Nile virus prevention efforts.

A laboratory model of myiasis as a result of Dermatobia hominis (L.) larvae was developed using mice as hosts. Mice in three groups were each infested with one newly hatched larva and skin biopsies processed for histopathology at 4, 12, and 20 d postinfestation (dpi). Mice in three other groups were each subjected to implantation of one larva collected from an infested (donor) mouse at 4, 12, and 20 dpi. Skin lesions of these receptor mice were then assessed at 10, 14, and 6 d postimplantation (dpimp), respectively. The inflammatory process in infested mice at 4 dpi was discrete, consisting of a thin necrotic layer around the larva, edema, many neutrophils, few eosinophils, mast cells, and proliferation of fibroblasts. At 12 dpi, there was a thicker necrotic layer, edema, many neutrophils and eosinophils, few mast cells, neoformation of capillaries, proliferation of the endothelium and fibroblasts, and early stages of fibrosis. These histopathological characteristics together with fibrosis were observed over a large area of the lesion at 20 dpi. Mice submitted to larval implantations demonstrated similar skin histopathology to that seen in the infested rodents, 10 dpimp corresponding to 12 dpi and 6 or 14 dpimp to 20 dpi. In all mice, the progressive acute inflammatory process followed a sequence linked to factors such as size of larvae and presence of secretory-excretory products. Both infested mice and those implanted experimentally with D. hominis larvae were shown to be suitable models for the study of the parasite-host relationship in this important zoonotic myiasis.

Salivary glands, midgut, Malpighian tubules, and ovaries were dissected from infected, colony-derived Amblyomma maculatum (Gulf Coast ticks) injected as nymphs with either Rickettsia parkeri (a spotted fever group rickettsia [SFGR] ; treatment) or phosphate-buffered saline (negative control). For comparison, similar tissues were dissected from hemolymph-positive, field-collected ticks. Tissues were analyzed by indirect fluorescent antibody (IFA) tests. All phosphate-buffered saline-injected ticks were IFA negative, whereas SFGR were detected by IFA in 100% of the salivary glands and ovaries and 78 and 75% of midgut and Malpighian tubule samples, respectively, of R. parkeri-injected ticks. Nearly 22% (10/46) of the field-collected ticks were hemolymph positive. Of those, SFGR were detected by IFA in 80% of the salivary glands, 67% of the ovaries, and 60% in the midgut and Malpighian tubules. This is the first study to assess the distribution of SFGR in select tissues of A. maculatum ticks.

Although domestic animals may not be permissive for Plasmodium, they could nevertheless play a role in the epidemiology of malaria by attracting Anopheles away from humans. To investigate interactions between domestic animals and mosquitoes, we assayed immunoglobulin G (IgG) antibodies directed against the salivary proteins of Anopheles gambiae in domestic animals living in Senegalese villages where malaria is endemic. By Western blotting, sera from bovines (n = 6), ovines (n = 36), and caprines (n = 36) did not react with Anopheles whole saliva. In contrast, equine sera recognized proteins in both saliva and salivary gland extracts. Two of the major immunogens (32 and 72 kDa) were also reactive in extracts from other major mosquito genera (Aedes and Culex), but reactions to Anophehs-specific antigens were detected in 12 of 17 horses. These data suggest that horses strongly react to Anopheles bites, and further experiments on horses are warranted to investigate the impact of this domestic animal species on the transmission of human malaria.

The extensive use of insecticides to control vector populations has lead to the widespread development of different mechanisms of insecticide resistance. Mutations that confer insecticide resistance are often associated to fitness costs that prevent them from spreading to fixation. In vectors, such fitness costs include reductions in preimaginal survival, adult size, longevity, and fecundity. The most commonly invoked explanation for the nature of such pleiotropic effects of insecticide resistance is the existence of resource-based trade-offs. According to this hypothesis, insecticide resistance would deplete the energetic stores of vectors, reducing the energy available for other biological functions and generating trade-offs between insecticide resistance and key life history traits. Here we test this hypothesis by quantifying the energetic resources (lipids, glycogen, and glucose) of larvae and adult females of the mosquito Culex pipiens L. resistant to insecticides through two different mechanisms: esterase overproduction and acetylcholinesterase modification. We find that, as expected from trade-off theory, insecticide resistant mosquitoes through the overproduction of esterases contain on average 30% less energetic reserves than their susceptible counterparts. Acetylcholinesterase-modified mosquitoes, however, also showed a significant reduction in energetic resources (20% less). We suggest that, in acetylcholinesterase-modified mosquitoes, resource depletion may not be the result of resource-based trade-offs but a consequence of the hyperactivation of the nervous system. We argue that these results not only provide a mechanistic explanation for the negative pleiotropic effects of insecticide resistance on mosquito life history traits but also can have a direct effect on the development of parasites that depend on the vector's energetic reserves to fulfil their own metabolic needs.

Host bloodmeals of indigenous Caribbean mosquitoes have not been studied previously. We identified vertebrate DNA in 90 blood-engorged mosquitoes belonging to four genera (Aedes, Culex, Deinocerites, and Uranotaenia) and 12 species that were collected in Puerto Rico within a geographic and temporal focus of West Nile virus transmission in 2007. It was found that 62 (68.8%) bloodmeals were from reptiles, 18 (20.0%) from birds, and 10 (11.1%) from mammals. Only one bloodmeal of 18 derived from Culex (Culex) species was passerine, suggesting a preference for nonpasserine birds and other vertebrates (i.e., reptiles) among the candidate WNV vectors. We interpret the results with respect to vectorial capacity for West Nile virus, an emerging arbovirus throughout the Caribbean Basin.

Reproductive isolation between Triatoma mexicana (Herrich-Scaeffer) and the six species of the Phyllosoma complex, belonging to the genus Meccus, was examined by analyzing the fertility of experimental hybrids. The percentage of couples with offspring was highest in the set of crosses between Meccus phyllosomus (Burmeister) and T. mexicana, and lowest in those between Meccus pallidipennis (Stål) and Meccus bassolsae (Alejandre-Aguilar, Nogueda-Torres, Cortés-Jimé nez, Jurberg, Galvão, and Carcavallo) with respect to T. mexicana. Crosses between Meccus mazzottii (Usinger), Meccus picturatus (Usinger), and Meccus longipennis (Usinger) and T. mexicana had scarce offspring, whereas offspring from crosses between M. phyllosomus and T. mexicana were abundant. However, all F₁ nymphs died in different instars before reaching sexual maturity. The biological evidence in this study suggests that different degrees of reproductive isolation exist between T. mexicana and the different species of Meccus. Our data also support the current status of T. mexicana as a separate species.

The carbon dioxide-baited trap is the most common and effective method for sampling vector life-stage Amblyomma americanum (L.) (Acari: Ixodidae), although confounding environmental variables are rarely considered. A mark-recapture experiment was designed to compare recapture proportions of A. americanum nymphs and adults between two habitat types: old field and oak-hickory forest. Powdered fluorescent dye was used to mark A. americanum ticks released in 1-m increments from carbon dioxide-baited traps. Adults were recaptured in significantly higher proportion than nymphs, but habitat type had no significant effect on recapture proportions. Tick abundance is an important parameter in the estimation of human risk of exposure to tick-borne disease and the influence of life stage on capture rates should be considered when calculating entomological risk.

Container-dwelling mosquitoes use a wide variety of container habitats. The bottle cap is often cited as the smallest container habitat used by container species. When containers are small, the habitat conditions can greatly affect evaporation rates that in turn can affect the species dynamics within the container. An evaporation rate model was adapted to predict evaporation rates in mosquito container habitats. In both the laboratory and field, our model was able to predict actual evaporation rates. Examples of how the model may be applied are provided by examining the likelihood of Aedes albopictus (Skuse), Aedes aegypti (L.), and Culex pipiens pipiens (L.) completing their development within small-volume containers under typical environmental conditions and a range of temperatures. Our model suggests that under minimal direct sunlight exposure, both Ae. aegypti and Ae. albopictus could develop within a bottle cap before complete evaporation. Our model shows that under the environmental conditions when a plastic field container was sampled, neither Ae. albopictus or Cx. p. pipiens could complete development in that particular container before the water evaporated. Although rainfall could replenish the habitat, the effects of evaporation would increase larval density, which could in turn further decrease developmental rates.