Biting midges of the genus Culicoides Latreille have been incriminated in transmission of bluetongue. Since 1998, the disease has spread across Europe provoking the largest epidemic ever recorded with important economic loses. Some species of the subgenus Avaritia and Culicoides have been described as candidate vectors involved in these epizootics. Both subgenera contain groups of cryptic species that could differ in their vectorial capacity. For this reason, the correct identification of vector species is considered an essential issue in epidemiological programs. In the current study, the usefulness of wing form in differentiating morphologically similar species of the subgenus Culicoides by means of geometric morphometric techniques is assessed in specimens previously identified through molecular analyses based on cytochrome oxidase subunit I gene sequences. Significant differences between species were detected in the two components of form, i.e., size and shape. Although wing size was affected by temperature, wing shape showed a more stable specific variation, allowing the proper classification of a high percentage of specimens. In addition, the concordance between phylogenies inferred from molecular data and phenetic clusters suggests the existence of a phylogenetic signal in wing shape. These findings enhance the use of this complex phenotypic trait not only to infer genetic relationships among species of the subgenus Culicoides but also as a potentially powerful tool to differentiate cryptic species within the genus.

Eudusbabekia paralepidoseta new species, was recorded on the Hart's little fruit bat Enchistenes hartii (Thomas) in the southern part of Mexico. The female and male are described and illustrated. E. paralepidoseta n. sp. represents the 32nd species in the genus. From the 31 known species of Eudusbabekia known to infest phyllostomid and mormoopid bats, E. paralepidoseta n. sp. has some morphological features similar to Eudusbabekia lepidoseta Jameson, 1971, including shapes of almost all dorsal and ventral setae; the presence of a patch of supernumerary; mostly broad to thick, medium, and almost scale-like setae; and the absence of setae 2b. However, E. paralepidoseta n. sp. can be differentiated from E. lepidoseta, by the reduced number of supernumerary setae on the female venter (37–43). The close morphological and therefore evolutionary similarities between E. lepidoseta and E. paralepidoseta n. sp. suggest possible close evolutionary relationships between their hosts, Sturnira lilium (Geoffroyi and St.-Hilaire) and E. hartii, which belong to the monophyletic subfamily Stenodermatinae.

The female of the phlebotomine sand fly Lutzomyia reclusa Fernández & Rogers 1991 [=Pintomyia (Pifanomyia) reclusa (Fernández & Rogers) sensu Galati], is described for the first time, based on specimens collected in the Department of Cajamarca, in northern Peru. The female can be recognized from other species of the series pia, species group Verrucarum, by wing venation with beta shorter than half of alpha, labrum just shorter than head width but longer than flagellomere 1, palpomere 5 much longer than palpomere 3, arrangement of cibarial armature, and form of spermathecae and relative size of spermathecal ducts. Diagnostic characters and measurements of the male of Lu. reclusa are provided as well.

A morphometric and molecular study of adult male and female Lutzomyia shannoni (Dyar 1929) collected at seven different locations within the southeastern United States was conducted to assess the degree of divergence between the grouped specimens from each location. The collection locations were as follows: Fort Bragg, NC; Fort Campbell, KY; Fort Rucker, AL; Ossabaw Island, GA; Patuxent National Wildlife Research Refuge, MD; Suwannee National Wildlife Refuge, FL; and Baton Rouge, LA. Forty males and forty females from each location were analyzed morphometrically from 54 and 49 character measurements, respectively. In addition, the molecular markers consisting of the partial cytochrome c oxidase subunit I (from 105 sand flies: 15 specimens/collection site) and the partial internal transcribed spacer 2 (from 42 sand flies: six specimens/collection site) were compared. Multivariate analyses indicate that the low degree of variation between the grouped specimens from each collection site prevents the separation of any collection site into an entity that could be interpreted as a distinct population. The molecular analyses were in concordance with the morphometric study as no collection location grouped into a separate population based on the two partial markers. The grouped specimens from each collection site appear to be within the normal variance of the species, indicating a single population in the southeast United States. It is recommended that additional character analyses of L. shannoni based on more molecular markers, behavioral, ecological, and physiological characteristics, be conducted before ruling out the possibility of populations or a cryptic species complex within the southeastern United States.

Circadian patterns of flight activity in mosquitoes can influence pathogen transmission by regulating dispersal potential of vectors and contact rates between vectors and reservoir and/or dead-end hosts. We investigated circadian activity patterns of Culex erraticus (Dyar and Knab) at a wetland field site in central Alabama, by aspirating resting adults and questing females in the morning and evening hours, respectively. Mosquitoes were aspirated at regular time intervals to determine the time of day during which peak resting site-seeking and host-seeking activities occurred. Day-to-day variation in activity patterns due to wind, humidity, and temperature was examined using stepwise linear regression. We found a distinct peak in flight activity during the morning hours (2 h before and 2 h after sunrise) for females and males of Culex erraticus, the most commonly encountered species at the site. The exact time of the peak varied from day to day, and was largely a function of temperature. A less distinct peak in activity was observed for questing females in the evening, although flights generally commenced just after sunset and peaked 30–60 min after sunset. A significant amount of day-to-day variation in the number of questing females was attributable to relative humidity. Our study demonstrates predictable patterns of circadian activity for Cx. erraticus, a suspected bridge vector of eastern equine encephalitis virus. Moreover, these patterns are modulated by environmental conditions. This information may be used to develop vector control strategies and make predictions about factors that affect the spread of mosquito-vectored pathogens.

To study the impact of Amazonian forest fragmentation on the mosquito fauna, an inventory of Culicidae was conducted in the upland forest research areas of the Biological Dynamics of Forest Fragments Project located 60 km north of Manaus, Amazonas, Brazil. The culicid community was sampled monthly between February 2002 and May 2003. CDC light traps, flight interception traps, manual aspiration, and net sweeping were used to capture adult specimens along the edges and within forest fragments of different sizes (1, 10, and 100 ha), in second-growth areas surrounding the fragments and around camps. We collected 5,204 specimens, distributed in 18 genera and 160 species level taxa. A list of mosquito taxa is presented with 145 species found in the survey, including seven new records for Brazil, 16 new records for the state of Amazonas, along with the 15 morphotypes that probably represent undescribed species. No exotic species [Aedes aegypti (L.) and Aedes albopictus (Skuse)] were found within the sampled areas. Several species collected are potential vectors of Plasmodium causing human malaria and of various arboviruses. The epidemiological and ecological implications of mosquito species found are discussed, and the results are compared with other mosquito inventories from the Amazon region.

Adult Fannia conspicua Malloch were captured by sweep net at a southern California coastal mountain community to determine diel flight activity and crop/gut sugar (gut sugar) concentration. Male swarming activity was monitored by visual estimation of swarm numbers. Gut sugar content of captured flies was determined by cold anthrone assay. Peak host seeking by female flies generally occurred in early morning (0700–0800) and early evening (1900–2000). Variation in female host-seeking activity was significantly explained by the time elapsed since sunrise or time remaining until sunset, with temperature, humidity, and wind speed having small, but significant effects on activity. Male swarming activity occurred more generally throughout the day, with peaks in midmorning and mid-afternoon and reduced swarming during periods of highest female host-seeking activity. Male swarming behaviors were only minimally explained by environmental variables. Flies of both sexes commonly fed on fructose sugars with 99.94% of host-seeking female flies (n = 1,647), and 98.93% of swarming male flies (n = 1,398) with gut sugar levels exceeding those of starved flies. Host-seeking female flies had significantly higher overall sugar content than swarming male flies. Male flies had peak gut sugar levels at 0800–0900 and 2000. Female flies had broad peaks in gut sugar level from 0700 to 1200 and 1600 to 1900. Stepwise regression showed that variation in gut sugar level was poorly explained by environmental variables for both sexes.

Culiseta morsitans (Theobald) is a mosquito vector of eastern equine encephalitis virus. Gravid Cs. morsitans were collected from resting shelters during June–September of 2004 and 2005 associated with a natural breeding area known as Toad Harbor-Big Bay Wildlife Management Unit in Oswego County, New York. Specimens were held individually in water-filled containers to encourage egg laying. Conditions in captivity provided three site options for oviposition, as follows: on a paper filter above the water line, on the filter at the water line, or on the water surface. Oviposited eggs were counted, as were eggs found retained in abdomens of dissected females. Fecundity for 181 female Cs. morsitans was based on the total egg count for each female. There were 89 females that oviposited ≥ 1 egg and produced an average of 158.4 eggs ± 4.9 eggs with a range of 51–318 eggs. Of the 89, there were 63 captive females that oviposited all their eggs, producing a mean of 160.4 ± 4.9 eggs, range 81–254 eggs. There were 62 egg masses oviposited above the water line within a range of 2–68 mm (average = 24 mm), 12 egg rafts or oviposition of scattered eggs directly on the water surface, and no rafts oviposited on filter paper directly at the water line. Wing length was used as a measure of mosquito size. The mean wing length for 154 females was 4.37 ± 0.02 mm (range, 3.8–4.9 mm).

Male reproductive success is dependent on insemination success and reproductive output. During mating, male mosquitoes transfer not just sperm, but also seminal fluid proteins that may have profound effects on mated female biology and behavior. In this study, we investigated the role of male body size and mating history on semen depletion, female longevity, and reproductive success in Aedes aegypti L. Small and large males were mated in rapid succession with up to five females. Our results indicate that large males had greater mating capacity than small males. A reduction in fecundity by >50% was observed in females that were fourth to mate with small males in comparison with females that mated earlier in sequence. For females mated to large males, this reduction became evident for females that mated fifth in sequence. No loss of fertility (measured as hatch rate) was observed in females that were third-fifth in mating sequence compared with females mated to virgin males. When females were maintained on a low-quality (5% sucrose) diet, those mated to virgin males had a greater longevity compared with females mated third in sequence. We conclude that small males experience more rapid seminal depletion than large males, and discuss the role of semen depletion in the mated female. Our results contribute toward a better understanding of the complexity of Ae. aegypti mating biology and provide refined estimates of mating capacity for genetic control efforts.

Variation in cold hardiness was studied in the laboratory for the nondiapausing eggs of nine Aedes (Stegomyia) species from eastern Asia and Pacific islands, ranging from the tropics to the cool-temperate zone. Species included were Aedes aegypti (L.), Aedes albopictus (Skuse), Aedes flavopictus Yamada, Aedes galloisi Yamada, Aedes hensilli Farner, Aedes palauensis Bohart, Aedes paullusi Stone and Farner, Aedes riversi Bohart and Ingram, and Aedes scutellaris (Walker). For species with distributions covering two or three climatic zones, at least one population was included from each zone, except for Ae. aegypti (subtropical populations were unavailable). Some eggs of four species confined to the tropical zone survived short exposure to freezing temperature (≥-5°C) when humidity was high. There were inter- and intraspecies variations in cold hardiness of nondiapausing eggs. Cold hardiness in each climatic zone differed in that populations of species with more northerly distributions were more cold-hardy than species with southerly distributions. In Ae. albopictus and Ae. flavopictus that cover three climatic zones, populations from cooler regions were more cold-hardy than populations from warmer regions. A possibility of southern species without diapausing eggs to colonize temperate eastern Asia was discussed.

A temporal observational study on culicid entoniofauna was conducted in a region characterized as a fragment of the Atlantic Forest that forms the Tinguá Biological Reserve in the State of Rio de Janeiro. This investigation was performed with the aim of analyzing the influence of climatic factors (temperature and relative air humidity) on the activity levels at different times of the day among mosquito species within the ecosystems that form the Tinguá Biological Reserve. The abundance index and dominance coefficient were calculated in relation to 61 mosquito species that were caught at four sampling sites, in the mornings, afternoons, and evenings. The results revealed that culicid species were distributed with greater incidence during the two diurnal periods and that their preference for times of the day was directly influenced by the climatic variables analyzed. The latter acted as limiting factors for occurrences of mosquito species.

The Asian rock pool mosquito Aedes japonicus (Theobald) inhabits natural and artificial container habitats, some of which are occupied by the native treehole mosquito Aedes triseriatus (Say), a vector of LaCrosse encephalitis virus. A laboratory experiment was used to evaluate the effects of nutrient limitation and interspecific interactions between these species. The goal was to address two related hypotheses. First, interspecific interactions between these species show competitive asymmetry with the invasive mosquito Ae. japonicus being favored over Ae. triseriatus. Second, competitive stress at the larval stage alters adult longevity. There was minimal evidence for competitive asymmetry between these two species. Mosquito and population performance showed clear negative density-dependent effects with similar effects of intra- and interspecific interactions. Only Ae. japonicus development time showed competitive asymmetry over Ae. triseriatus, providing weak support for the first hypothesis. For both species, competition resulted in lower adult longevity compared with low competition, providing support for the second hypothesis. These results suggest both species are similarly affected by intra- and interspecific competition and underscore the importance of the effects of larval competition that continue into adulthood and alter parameters important to transmission of vector-borne diseases.

The chronic effects of exposure of Culex restuans (Theobald) and Aedes albopictus (Skuse) (Diptera: Culicidae) to low concentrations of malathion were examined by exposing larvae of the two species to four malathion doses at 20, 25, and 30°C and maintaining the resulting adults at 25°C. For both species, a significant temperature by malathion interaction on survival was found. Greater temperatures at the highest malathion dosage had significantly lower survivorship than in other treatments, but this effect was not observed at 20°C. These results suggest that low temperature may buffer against some of the negative effects of malathion. For both species, temperature but not malathion had significant effects on female developmental time to adulthood and adult longevity. Temperature also affected adult female size for Ae. albopictus but not Cx. restuans. Ae. albopictus females developed faster as temperature increased, lived longer when larvae were maintained at 30°C than at 20°C, and were larger when larvae were maintained at 25°C than at 20 or 30°C. Cx. restuans females developed faster at 25 and 30°C than at 20°C and lived longer at 25°C than at 20 or 30°C. The estimated finite rate of increase (λ′) for Cx. restuans was significantly lower at 20°C than at 25 and 30°C, whereas that of Ae. albopictus was significantly influenced by an interaction between temperature and malathion with significantly lower λ′ at 20°C than at 25 and 30°C for all malathion treatments except 0.014 mg liter^-1. Understanding how pesticides interact with abiotic environmental conditions will contribute to management decisions about vector control practices.

Responses of the northern fowl mite (NFM), Ornithonyssus sylviarum (Canestrini & Fanzago) (Acari: Macronyssidae), and the chicken body louse (CBL), Menacanthus stramineus (Nitzsch) (Phthiraptera: Menoponidae), to variation in temperature, light, and humidity were assessed in bioassays. The location on a continuous thermal gradient at which each ectoparasite arrested was recorded and analyzed. NFM adults arrested at an average temperature of 30.09 ± 0.34°C. Adult CBL and first-instar CBL nymphs arrested at 33.69 ± 0.20°C and 34.99 ± 0.26°C, respectively. Groups of each ectoparasite were placed into clear glass vials (n = 10/vial) with one half shaded, and vials were exposed to three light levels, as follows: high (200 µmolm ^-2s^-1) , low (4 µmolm ^-2s^-1), and nearly no light (0 µmolm ^-2s^-1). The vial cap edges provided an opportunity to assess the interactive effect of light with harborage. NFM avoided light and sought harborage. In low light, the harborage preference overrode the tendency to avoid light. CBL avoided the harborage and showed a minimal preference for light. A four-level humidity gradient was established in two separate experimental arenas for NFM and CBL. Trials were run in ambient light (4 µmolm ^-2s^-1) for the NFM and in nearly no light for the CBL. The NFM gradient used 38 ± 2%, 54 ± 7%, 73 ± 3%, and 90 ± 4% RH, whereas the CBL gradient used 42 ± 5%, 48 ± 7%, 63 ± 4%, and 73 ± 5% RH. NFM showed no humidity response in the walking bioassay, but the CBL settled at the lowest humidity level. Temperature and humidity on different hen body regions were related to the bioassay results and observed on-host ectoparasite distributions.

Triatoma dimidiata Latreille is a major vector of Chagas disease with an extensive geographic distribution from Central Mexico, through Central America, to northern South America. As a result of its variability in phenetic and genetic characters, disagreement concerning its taxonomic status has been raised. In this study, the cuticular hydrocarbon pattern of T. dimidiata populations from Mexico, Belize, Guatemala, Honduras, Costa Rica, and Colombia was analyzed by capillary gas chromatography coupled to mass spectrometry; linear discriminant analysis was used to help elucidate population structure. Vector populations segregated into five distinct groups; specimens from Yucatan Peninsula, together with those from Central Mexico, Central America, and Colombia corresponded to different T. dimidiata subspecies, a putative different species comprising insects from Belize, together with an isolated population collected at bat caves in Guatemala. The analysis revalidates the earlier division of T. dimidiata into three subspecies, T. d. maculipennis, T. d. dimidiata, and T. d. capitata; and an additional subspecies and a distinct species are proposed.

Species of the genus Culex Linnaeus have been incriminated as the main vectors of lymphatic filariases and are important vectors of arboviruses, including West Nile virus. Sequences corresponding to a fragment of 478 bp of the cytochrome c oxidase subunit I gene, which includes part of the barcode region, of 37 individuals of 17 species of genus Culex were generated to establish relationships among five subgenera, Culex, Phenacomyia, Melanoconion, Microculex, and Carrollia, and one species of the genus Lutzia that occurs in Brazil. Bayesian methods were employed for the phylogenetic analyses. Results of sequence comparisons showed that individuals identified as Culex dolosus, Culex mollis, and Culex imitator possess high intraspecific divergence (3.1, 2.3, and 3.5%, respectively) when using the Kimura two parameters model. These differences were associated either with distinct morphological characteristics of the male genitalia or larval and pupal stages, suggesting that these may represent species complexes. The Bayesian topology suggested that the genus and subgenus Culex are paraphyletic relative to Lutzia and Phenacomyia, respectively. The cytochrome c oxidase subunit I sequences may be a useful tool to both estimate phylogenetic relationships and identify morphologically similar species of the genus Culex.

The phylogeographical history of the pest fly screwworm, Cochliomyia hominivorax (Coquerel), was studied using partial mitochondrial DNA sequences of the control region, Cytochrome c oxidase (CO) subunit I and CO subunit II from 361 individuals collected across its current geographic range. Analyses showed marked genetic differentiation on a macrogeographic scale. The genetic diversity in the species is structured into four main “regional groups,” corresponding to Cuba, the Dominican Republic, and the North and South Amazon region. Results indicated that the distribution of screwworm genetic diversity was mainly shaped by historical events, i.e., colonization of Caribbean islands, vicariance in the Amazon region and population expansion. Demographic history analyses revealed that the population expansion started ≈20–25,000 yr ago and recently increased exponentially. We hypothesized that the initial period of expansion was probably associated with environmental amelioration in the late Pleistocene and the exponential increase with resource availability in recent times. The population expansion is probably responsible for the low divergence and the lack of genetic and geographic correlation in the South Amazon region but did not erase the genetic structure pattern on a continental scale. The screwworm is one of the most damaging livestock pests in South and Central America, and the pattern of genetic variability distribution reported here suggests that the Caribbean area and the North and South Amazon regions could be considered as independent units for future pest control programs.

Larvicidal activity of methanolic plant extracts of Lantana cramera (P1) root and Anacardium occidentale (P2) leaf was investigated against the larvae of the three mosquito species (Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti reared in the laboratory), and the respective glutathione S-transferase (GST) activity was analyzed as an index of protection against the extracts. The LC₅₀ (extract concentration that shows 50% mortality) values of Pl extract for An. stephensi, Ae. aegypti, and Cx. quinquefasciatus were 132.55,27.82, and 11.68 ppm, respectively, whereas those of P2 extract were 56.81, 912, and 10.79 ppm, respectively. In general, in the untreated groups, the level of GST activity was significantly higher in Ae. aegypti in comparison with An. stephesi and Cx. quinquefasciatus. However, the enzyme activity failed to show any response when treated with either of the plant extracts in Ae. aegypti. However, an increase in the GST activity was recorded in extract-treated larvae of both An. stephensi and Cx. quinquefasciatus. The results of the current study suggest that both the plant extracts show species-specific mosquitocidal potential. Induction of GST activities in survived An. stephensi and Cx. quinquefasciatus larvae suggests the role of this enzyme in conferring resistance to the plant extracts.

Anopheles arabiensis Patton (Diptera: Culicidae) larvae were reared from hatching to the adult stage in the laboratory under a range of diet and larval concentrations using a factorial design. The range circumscribed most of the larval densities and diet concentrations that would allow larval growth and survival using the particular diet formulation and water volume we tested. We determined how these variables affected three outcomes, as follows: larval development rate, survival, and wing length. As has been reported previously, negative density dependence of survival as a function of increased larval density was the prevalent effect on all outcomes when diet was limiting. When diet was not limiting, density dependence was not observed, and three cases of overcompensatory survival were seen. We discuss these results in the context of diet and larval densities for mass rearing and the effect of larval competition on control strategies.

Mosquito resistance to chemical insecticides is considered a serious threat for the sustainable use of contemporary malaria vector control methods. Fungal entomopathogens show potential as alternative biological control agents against (insecticide-resistant) anophelines. This study was designed to test whether the fungus, Beauveria bassiana, could be delivered to mosquitoes on netting materials that might be used in house screens, such as eave curtains. Tests were conducted to determine effects of formulation, application method, netting material, and nature of mosquito contact. Beauveria had a twice as high impact on Anopheles gambiae s.s. longevity when suspended in Shellsol solvent compared with Ondina oil (HR = 2.12, 95% confidence interval = 1.83–2.60, P < 0.001), and was significantly more infective when applied through spraying than dipping. Polyester and cotton bednets were the most effective substrates for mosquito infections, with highest spore viability on cotton nets. Whereas fungal impact was highest in mosquitoes that had passed through large-meshed impregnated nets, overall efficacy was equal between small- and large-meshed nets, with ≤30-min spore contact killing >90% of mosquitoes within 10 d. Results indicate that the use of fungal spores dissolved in Shellsol and sprayed on small-meshed cotton eave curtain nets would be the most promising option for field implementation. Biological control with fungus-impregnated eave curtains could provide a means to target host-seeking mosquitoes upon house entry, and has potential for use in integrated vector management strategies, in combination with chemical vector control measures, to supplement malaria control in areas with high levels of insecticide resistance.

The therapeutic and persistent efficacy of a single subcutaneous injection of a long-acting formulation of moxidectin at a concentration of 1 mg/kg body weight was determined against Rhipicephalus (Boophilus) microplus (Canestrini), along with the concentration-time blood sera profile in treated cattle. The therapeutic efficacy against ticks of all parasitic stages on cattle at the time of treatment was >99.9%, and the mean tick number, index of fecundity, engorgement weight, and egg mass weight of ticks recovered from treated animals were all significantly lower than ticks from untreated animals. The index of fecundity, engorgement weight of females, and egg mass weight of ticks recovered from treated animals infested at weekly (7-d) intervals between 14 and 63 d posttreatment were significantly lower than for ticks on untreated animals, whereas the number of ticks per animal recovered from treated cattle remained lower than that of untreated cattle for up to 49 d posttreatment. The percentage control remained >99% at weekly intervals between 14 and 49 d posttreatment, which is the minimum level of efficacy considered acceptable for use in the United States Cattle Fever Tick Eradication Program. The serum concentration of moxidectin in treated cattle increased to 25.6 ppb (parts per billion) within 1 d after treatment, and peaked at 47.3 ppb at 8 d posttreatment. Moxidectin sera levels remained above the estimated 100% threshold level for elimination of feeding ticks (5–8 ppb) for 44–53 d after treatment. The label claim of 50 d of prevention against reinfestation for the long-acting moxidectin formulation used in the study was supported by the efficacy and sera concentration data obtained. Based on these results, cattle could be treated at 63-d intervals with minimal risk of viable ticks detaching from treated animals. This treatment interval would be 4.5-fold longer than the presently required treatment interval of 14 d, thus leading to ≈75% reduction in gathering and handling costs of cattle incurred by producers.

Toxicity of nootkatone was determined in laboratory assays against unfed nymphs of Amblyomma americanum L., Dermacentor variabilis (Say), Ixodes scapularis Say, and Rhipicephalus sanguineus Latreille. We determined the 50% lethal concentration (LC₅₀) and 90% lethal concentration (LC₉₀) of nootkatone by recording tick mortality 24 h after exposure in treated glass vials. Nymphs were susceptible to nootkatone with LC₅₀ values of 0.352, 0.233, 0.169, and 0.197 µg/cm², and LC₉₀ values of 1.001, 0.644, 0.549, and 0.485 µg/cm² for A. americanum, D. variabilis, I. scapularis, and R. sanguineus, respectively. The LC₅₀ value for R. sanquineus was not significantly different from D. variabilis or I. scapularis. Other LC₅₀ comparisons were significantly different. The LC₉₀ for A. americanum was higher when compared with the three other tick species, which were not significantly different. Because nootkatone is volatile, we measured the amount of nootkatone recovered from duplicate-treated vials before tick exposure and from vials after tick exposure. Nootkatone recovered from vials before exposure ranged from 82 to 112% of the expected amounts. The nootkatone recovered after the 24-h exposure period ranged from 89% from vials coated with higher concentrations of nootkatone, down to 29% from vials coated with low nootkatone concentrations. Determination of the nootkatone residue after vial coating demonstrated loss of the active compound while verifying the levels of tick exposure. Toxicity of low concentrations of nootkatone to the active questing stage of ticks reported in this study provides a reference point for future formulation research to exploit nootkatone as a safe and environment-friendly tick control.

The number of tick bites received by individuals wearing either permethrin-treated or untreated summer clothing (T-shirt, shorts, socks, and sneakers) was compared during a controlled indoor study. Pathogen-free nymphal Ixodes scapularis Say were placed on the left shoe, right leg, and left arm of 15 (5/treatment group/d) human volunteers wearing untreated outfits or outfits treated with permethrin either commercially or using a do-at-home treatment kit. The number and location of ticks attached to subjects' skin were recorded 2.5 h postinfestation. Subjects wearing outfits treated with permethrin received 3.36 times fewer tick bites than subjects wearing untreated outfits. No statistically significant differences in number of tick bites were detected between commercial permethrin treatment (19.33%) and the do-at-home permethrin application method (24.67%). The success of permethrin-treated clothing in reducing tick bites varied depending on the specific article of clothing. Subjects wearing permethrin-treated sneakers and socks were 73.6 times less likely to have a tick bite than subjects wearing untreated footware. Subjects wearing permethrin-treated shorts and T-shirts were 4.74 and 2.17 times, respectively, less likely to receive a tick bite in areas related to those specific garments than subjects wearing untreated shorts and T-shirts. Ticks attached to subjects were classified as alive or dead before removal. On subjects wearing untreated outfits, 97.6% of attached nymphs were alive, whereas significantly fewer (22.6%) attached nymphs were alive on subjects wearing repellent-treated outfits. Results of this study demonstrate the potential of permethrin-treated summer clothing for significantly reducing tick bites and tick-borne pathogen transmission.

Laboratory studies were conducted to compare the response of Aedes aegypti (L.) and Aedes albopictus (Skuse) adults, uninfected and infected with four serotypes of dengue virus to a repellent containing 5% N, N-diethyl-3-methylbenzamide (deet). The results showed that mosquitoes infected with the four serotypes of dengue responded similarly to uninfected mosquitoes.

Ixodes scapularis Say, the blacklegged tick, vectors Borrelia burgdorferi Johnson et al. 1984, the bacterium that causes Lyme disease, the most important vector-borne disease in the United States. Efforts to reduce I. scapularis populations are shifting toward the development of biological control methods. Currently, only a few entomopathogenic fungal species are considered virulent to ticks. We hypothesized that these species may not represent the most abundant local taxa that would be pathogenic to ticks in situ. To identify potential entomopathogenic fungi at a study site in Westchester County, New York, we sampled soils and ticks, extracted and amplified the internal transcribed spacer region of nuclear ribosomal DNA (nrDNA), and compared sequences with those in GenBank. Over three sampling periods from June 2007 to May 2008, 70 fungal taxa were isolated and identified from soils (48 taxa) and ticks (27 taxa; 5 taxa were found both in soil and on ticks) collected in this study, encompassing species in 25 different genera. In laboratory bioassays, 15 fungal taxa were found to be significantly virulent, although none of these were previously considered common pathogens of I. scapularis. Two species, Hypocrea lixii Patouillard 1891 and Penicillium soppii K. M. Zalessky 1927, were tested in field trials by spraying suspensions on forested plots. Mean tick mortality was 71% after treatment with H. lixii, 58% after treatment with P. soppii, and 32% in the control plots. The complete diversity of entomopathogenic fungal species at this site is yet to be defined, but, in general, such fungi appear to be more common in forest habitats where I. scapularis resides than previously thought. Examination of intact fungal communities can provide information that serves as the foundation for site-specific biocontrol programs.

The potential of two bioinsecticidal formulations containing Brevibacillus laterosporus spores and azadirachtin, respectively, was assayed in laboratory and in comparative field treatments for the management of immature house flies on dairy farms. As already known for B. laterosporus, preliminary laboratory experiments with azadirachtin evidenced a concentration-dependent effect. Azadirachtin median lethal concentration (LC₅₀) value determined for second instar larvae was 24.5 µg/g diet. Applications in dairy farms were performed at dosages and concentrations predetermined in laboratory experiments, to employ the two formulations at an equal insecticidal potential. Repeated applications on the cow pen caused a significant fly development depression in areas treated with azadirachtin (63%) and B. laterosporus (46%), compared with the control. Formulations were applied at a dosage of 3 liters/m², and concentrations of 2 × 10⁸ B. laterosporus spores/ml and 25 µg azadirachtin/ml, respectively.

Bluetongue virus (BTV) is an economically important arbovirus of ruminants transmitted by Culicoides biting midges. Vector control using residual spraying or application to livestock is recommended by many authorities to reduce BTV transmission; however, the impact of these measures in terms of both inflicting mortality on Culicoides and subsequently upon BTV transmission is unclear. This study consisted of a standardized World Health Organization laboratory assay to determine the susceptibility of European Culicoides species to deltamethrin and a field trial based upon allowing individuals of a laboratory strain of Culicoides nubeculosus Meigen to feed upon sheep treated with Butox 7.5 pour-on (a deltamethrin-based topical formulation). Susceptibility in the laboratory trial was higher in colony C. nubeculosus (24-h LC₉₀ = 0.00106%), than in field populations of Culicoides obsoletus Meigen (24-h LC₉₀ = 0.00203%) or Culicoides imicola Kieffer (24-h LC₉₀ = 0.00773%). In the field, the pour-on formulation was tested with a total of 816 C. nubeculosus specimens fed upon on the thigh of treated sheep. The study revealed a maximum mortality rate of 49% at 4 d postapplication, and duration of lethal effect was predicted to be as short as 10 d, despite testing being carried out with a highly susceptible strain. The reasons for this low efficacy are discussed with reference both to the potential for lack of spread of the active ingredient on the host and feeding patterns of the major potential vector species on the sheep host. Practical implications for vector control strategies during BTV incursions are also detailed.

Various acaricide-resistant strains of Rhipicephalus (Boophilus) microplus, representative of the major resistance mechanisms found in Mexico and Brazil, were exposed to spinosad using the Food and Agriculture Organization of the United Nations larval packet test and adult immersion test (AIT). Larvae of all strains tested were found to be susceptible to spinosad. Conversely, spinosad did not show toxic activity toward engorged females used in the AIT. In vitro tests against larvae, nymphs, and adults of acaricide-susceptible Amblyomma americanum and Dermacentor variabilis revealed differences in activity that were dependent on tick species and parasitic life stage. Spinosad seemed to be a viable alternative to current acaricides available for tick control in the species tested. The larval packet test should be used for future monitoring of resistance, as the AIT did not provide useful information with this chemical. The potential benefit for the use of spinosad in integrated pest management or eradication programs is discussed.

Toxicities of (E)-cinnamaldehyde and (E)-cinnamic acid and their 41 structurally related compounds to adult Dermatophagoides farinae Hughes and Dermatophagoides pteronyssinus Trouessart (Acari: Pyroglyphidae) were examined using fabric-circle contact plus fumigant and vapor-phase mortality bioassays. Results were compared with those of two acaricides, benzyl benzoate and dibutyl phthalate. In contact plus fumigant mortality bioassays, the most toxic compounds were (E)-cinnamaldehyde, methyl (E)-cinnamate, cinnamyl acetate, and hydrocinnamaldehyde against adult D. farinae (17.5–23.3 mg/m²) and D. pteronyssinus (19.0–24.0 mg/m²), based on 24-h 50% lethal concentration (LC₅₀) values. These compounds were significantly more toxic than either benzyl benzoate (LC₅₀, 64.9 and 60.5 mg/m²) or dibutyl phthalate (218.9 and 232.3 mg/m²). The toxicity of allyl cinnamate versus benzyl benzoate was not significantly different. Structure-activity relationship indicates that structural characteristics, such as types of functional groups, carbon skeleton, and saturation, appear to play a role in determining the compound toxicities. In vapor-phase mortality bioassays, these compounds were effective against adult D. farinae in closed, but not in open containers, indicating that their mode of delivery was largely a result of vapor action. The active compounds described merit further study as potential house dust mite control fumigants with contact action in light of global efforts to reduce the level of highly toxic synthetic acaricides in indoor environments.

Triatomines are hematophagous insects and the vectors for Trypanosoma cruzi in the Americas. Although their preferred meal is blood from vertebrate hosts, unfed triatomine nymphs are often seen feeding on different arthropod species. Triatomine saliva has a wide range of activities that aid the hematophagic process. However, nothing is known about its role during hemolymphagy. In the current study, we reproduced hemolymphagy under laboratory conditions and evaluated the influence of hemolymphagy on the survival of Triatoma infestans nymphs. The effects of saliva on the activation of the prophenoloxidase cascade in the invertebrate host and the influence of the saliva on the motility of the bugs and contractions of the dorsal vessels were assessed. Hemolymphagy prolonged the survival rate of T. infestans first instars from 60 to >120 d compared with unfed nymphs. The saliva from T. infestans caused a 50% reduction in the amplitude and frequency of the dorsal vessel contractions of adult Rhodnius prolixus and induced paralysis for >10 min when the saliva was injected into second instars. T. infestans saliva was able to inhibit the activation of the prophenoloxidase cascade from a R. prolixus hemolymph, but had no effect on the phenoloxidase activity after the cascade was activated. The paralyzing molecule in the saliva was <5 kDa and probably had no proteic or lipidie characteristics. These results suggest that triatomine saliva may play an important role during hemolymphagy by inducing paralysis and suppressing immune responses in the invertebrate host. The importance of hemolymphagy for the survival of nymphs in periods when vertebrate blood is scarce is also discussed.

The European spider Tegenaria agrestis (Walckenaer) (hobo spider) has been implicated as a spider of medical importance in the Pacific Northwest since its introduction in the late 1980s. Studies have indicated that the hobo spider causes necrotic tissue lesions through hemolytic venom or through the transfer of pathogenic bacteria introduced by its bite. Bacterial infections are often diagnosed as spider bites, in particular the pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA). This study examines three aspects of the potential medical importance of hobo spiders in part of its introduced range, Washington State. First, the bacterial diversity of the spider was surveyed using a polymerase chain reaction-based assay to determine whether the spider carries any pathogenic bacteria. Second, an experiment was conducted to determine the ability of the spiders to transfer MRSA. Third, the venom was evaluated to assess the hemolytic activity. We found 10 genera of ubiquitous bacteria on the exterior surface of the spiders. In addition, none of the spiders exposed to MRSA transferred this pathogen. Finally, the hemolytic venom assay corroborates previous studies that found hobo spider venom was not deleterious to vertebrate red blood cells.

Aedes albopictus (Skuse) is a potential vector for many arboviruses, including La Crosse virus (LACV), the leading cause of pediatric encephalitis in North America. Bacteria isolated from the midgut and diverticula of field-caught female Ae. albopictus were cultured and identified using 16S ribosomal RNA gene amplification and sequencing. Members of seven and six bacterial families were identified from the midguts and diverticula, respectively, with nearly half of the isolates identified to the family Enterobacteriaceae. Many are related to bacteria identified in other invertebrates, and several may represent previously unknown species or genera. Of the 24 isolated bacteria, 12 (50%) showed a significant reduction in infectivity of LACV for Vero cells. Inhibition of infectivity ranged from 0 to 44% and was not dependent on bacterial classification. The antiviral activity of these bacteria warrants further investigation as an alternate means to interrupt the LACV transmission cycle.

The populations of Anopheles fluviatilis James (1902), a foothill vector, collected from village Tilpuri of district Udham Singh Nagar and from villages Auspur, Ismailpur, and Durgapur of district Hardwar were maintained at National Institute of Malaria Research Insectory at 28 ± 1°C and 80–85% RH. Anopheles fluviatilis sensu lato was identified for two sibling species T and U. A total of 94% of the females of both species T and U oviposited by day 4 after the blood meal. Maximum hatching, that is, 80 and 62% of the eggs of species T and U, was observed on the second and third day, respectively. For species T, mortality in second and third instars was recorded to be 144 ± 9 (N = 1,600) and 48 ± 6 (N = 1,200), whereas in species U, it was 196 ± 13 (N = 1,400) and 70 ± 8 (N = 1,000), respectively. Mortalities in second instars of species T and U were significantly higher than third instars (P = 0.05). The female and male ratio in pupal stage of species T and U was found to be 53:47 and 58:42, respectively.

Mosquitoes in the Culex pipiens complex are competent vectors of West Nile virus (WNV; family Flaviviridae, genus Flavivirus) in the laboratory, and field-collected mosquitoes have tested positive for the virus in California and elsewhere. A better understanding of Cx. pipiens complex blood-feeding patterns will help define the threat that these mosquitoes pose to human health and their role in WNV amplification in northern California. We collected blood-engorged Cx. pipiens complex mosquitoes from resting sites near and away from human habitation in Sacramento and Yolo Counties. Cytochrome c oxidase 1 gene sequences were used to identify the vertebrate species from which blood meals were taken. Of 330 engorged mosquitoes collected at 28 sites from June through August 2007 and May through August 2008, >99% fed on an avian host. Three mosquitoes contained bovine blood and none had fed on a human. American Robins (Turdus migratorius) were bitten most often, and the proportion of American Robin blood meals increased significantly over the summer. Other important avian hosts included House Finches (Carpodacus mexicanus), Barn Swallows (Hirundo rustica), Western Meadowlarks (Sturnella neglecta), and Mourning Doves (Zenaida macroura). In rural areas, Barn Swallows, Brewer's Blackbirds (Euphagus cyanocephalus), and House Sparrows (Passer domesticus) were frequent hosts. In settings near human habitation, Mourning Doves and Western Meadowlarks were common hosts. Our data indicate that in north central California mosquitoes in the Cx. pipiens complex may be more important as epiornitic than epidemic vectors of WNV.

The larvicidal activity of 11 Myrtaceae essential oils and their constituents was evaluated against Aedes aegypti L. Of the 11, Melaleuca linariifolia Sm., Melaleuca dissitiflora F. Muell., Melaleuca quinquenervia (Cav.) S. T. Blake, and Eucalyptus globulus Labill oils at 0.1 mg/ml exhibited ≥80% larval mortality. At this same concentration, the individual constituents tested, allyl isothiocyanate, α-terpinene, p-cymene, ( )-limonene, (-)-limonene, γ-terpinene, and (E)-nerolidol, resulted in ≥95% mortality. We also tested the acute toxicity of these four active oils earlier mentioned and their constituents against Daphnia magna Straus. M. linariifolia and allyl isothiocyanate was the most toxic to D. magna. Two days after treatment, residues of M. dissitiflora, M. linariifolia, M. quinquenervia, and E. globulus oils in water were 55.4, 46.6, 32.4, and 14.8%, respectively. Less than 10% of allyl isothiocyanate, α-terpinene, p-cymene, (-) -limonene, ( ) -limonene, and γ-terpinene was detected in the water at 2 d after treatment. Our results indicated that oils and their constituents could easily volatilize in water within a few days after application, thus minimizing their effect on the aqueous ecosystem. Therefore, Myrtaceae essential oils and their constituents could be developed as control agents against mosquito larvae.

Sharp declines in human and animal cases of plague, caused by the bacterium Yersinia pestis (Yersin), have been observed when outbreaks coincide with hot weather. Failure of biofilm production, or blockage, to occur in the flea, as temperatures reach 30°C has been suggested as an explanation for these declines. Recent work demonstrating efficient flea transmission during the first few days after fleas have taken an infectious blood meal, in the absence of blockage (e.g., early-phase transmission), however, has called this hypothesis into question. To explore the potential effects of temperature on early-phase transmission, we infected colony-reared Xenopsylla cheopis (Rothchild) fleas with a wild-type strain of plague bacteria using an artificial feeding system, and held groups of fleas at 10, 23, 27, and 30°C. Naive Swiss Webster mice were exposed to fleas from each of these temperatures on days 1–4 postinfection, and monitored for signs of infection for 21 d. Temperature did not significantly influence the rates of transmission observed for fleas held at 23, 27, and 30°C. Estimated per flea transmission efficiencies for these higher temperatures ranged from 2.32 to 4.96% (95% confidence interval [CI]: 0.96–8.74). In contrast, no transmission was observed in mice challenged by fleas held at 10°C (per flea transmission efficiency estimates, 0–1.68%). These results suggest that declines in human and animal cases during hot weather are not related to changes in the abilities of X. cheopis fleas to transmit Y. pestis infections during the early-phase period. By contrast, transmission may be delayed or inhibited at low temperatures, indicating that epizootic spread of Y. pestis by X. cheopis via early-phase transmission is unlikely during colder periods of the year.

Circulation of a unique genetic type of Rickettsia rickettsii in ticks of the Rhipicephalus sanguineus complex was detected in Mexicali, Baja California, Mexico. The Mexican R. rickettsii differed from all isolates previously characterized from the endemic regions of Rocky Mountain spotted fever in northern, central, and southern Americas. Rhipicephalus ticks in Mexicali are genetically different from Rh. sanguineus found in the United States.

Borrelia burgdorferi, the etiological agent of Lyme disease, is transmitted among hosts by the black-legged tick, Ixodes scapularis, a species that regularly parasitizes various vertebrate hosts, including birds, in its immature stages. Lyme disease risk in the United States is highest in the Northeast and in the upper Midwest where I. scapularis ticks are most abundant. Because birds might be important to the range expansion of I. scapularis and B. burgdorferi, we explored spatial variation in patterns of I. scapularis parasitism on songbirds, as well as B. burgdorferi infection in bird-derived I. scapularis larvae. We sampled birds at 23 sites in the eastern United States to describe seasonal patterns of I. scapularis occurrence on birds, and we screened a subset of I. scapularis larvae for presence of B. burgdorferi. Timing of immature I. scapularis occurrence on birds is consistent with regional variation in host-seeking activity with a generally earlier peak in larval parasitism on birds in the Midwest. Significantly more I. scapularis larvae occurred on birds that were contemporaneously parasitized by nymphs in the Midwest than the Northeast, and the proportion of birds that yielded B. burgdorferi-infected larvae was also higher in the Midwest. We conclude that regional variation in immature I. scapularis phenology results in different temporal patterns of parasitism on birds, potentially resulting in differential importance of birds to B. burgdorferi transmission dynamics among regions.

Density-dependent processes such as larval competition may be important regulatory factors among some mosquito species. The application of pesticides used for control may alter these density-dependent interactions with consequences for the number of survivors and associated sublethal and chronic effects on these individuals. We examined how intraspecific competition among larvae and low concentrations of malathion alter Aedes aegypti L. and Aedes albopictus Skuse adult life history traits and competence for arboviruses using Sindbis virus as a model system. Larvae were reared at densities of 150 and 300 larvae per container and in the absence or presence of 0.04 parts per million of malathion, before surviving females were exposed to an infectious blood meal containing 10⁵ plaque-forming units/ml Sindbis virus. For both species, competition and the presence of malathion reduced survival to adulthood. The presence of malathion eliminated the negative effects of competition that resulted in lengthened development time and smaller-sized adults. For Ae. aegypti, but not Ae. albopictus, high competition conditions and the presence of malathion independently and not interactively led to an increase in virus dissemination from the midgut. Our results suggest that larval competition and chemical contaminants may influence disease transmission directly by altering adult mosquito fitness and indirectly by altering vector interactions with arboviruses.

Insecticide resistance in the main malaria vectors in Africa is a major concern for malaria vector control program managers. The most common insecticides used for indoor residual spraying (IRS) and treating bed nets are becoming increasingly ineffective. The quest for safer and more effective insecticides for malaria vector control is urgent. This study sought to evaluate the efficacy of ACTELLIC 50 EC (pirimiphos methyl), an organophosphate, for IRS in Ghana, where there is high vector resistance to pyrethroids and organochlorines. Before the commencement of the study, standard World Health Organization (WHO) vector susceptibility tests against a common malaria vector, Anopheles gambiae s.l, were conducted using preparations of pyrethroids, organochlorines, carbamates, and organophosphates. The vector was found to be resistant to the pyrethroids, the organochlorines, and the carbamates, but susceptible to the organophosphates. The emulsifiable organophosphate concentrate formulation, ACTELLIC 50 EC, was then evaluated to determine the efficacy and the length of its residual effect. The wall bioassay test, using recommended cones from WHO, was conducted on sprayed surfaces with ACTELLIC 50 EC from 27 July 2009 to 16 October 2009. After 15 wk of trials on painted cement surface, it was found out that the main malaria vector, An. gambiae s.l, was susceptible to the insecticide even though the WHO Pesticide Evaluation recommends 2- to 3-mo duration of effective action. Therefore, it is recommended for use in IRS programs in this part of Ghana, where there is high vector resistance to most of the insecticides.

In summer 2008, a total of 512 Ixodes ricinus (Acari: Ixodidae) ticks was collected from vegetation in four areas at the Baltic coast of Germany and tested for the presence of Lyme disease spirochetes. Among them, 293 ticks from three areas were screened for Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), rickettsiae of the spotted fever group (Rickettsiales: Rickettsiaceae), and Babesia spp. (Piroplasmida: Babesiidae), respectively. Borrelia burgdorferi sensu lato genospecies (Spirochaetales: Spirochaetaceae) were detected in 3.1% of the tick samples. The prevalence of A. phagocytophilum was 1.0%, rickettsiae were present in 8.5%, and pathogenic Babesia spp. in 8.9% of analyzed ticks. Coinfections occurred in five ticks. With this study we report first data on the coexistence of established and emerging pathogens in questing ticks from recreational areas of northeastern Germany, indicating the need of further studies for a reliable risk assessment.

Cat fleas (Ctenocephalides felis [Bouché]) are the primary ectoparasites of dog and cat populations. In this study, we report the monthly population dynamics of Rickettsia felis and Bartonella spp. (two zoonotic pathogens that can cause human disease) in cat fleas collected from dogs and cats in Taipei, Taiwan, from December 2006 to December 2007. Natural R. felis infection in individual cat fleas was assessed by polymerase chain reaction (PCR) using pRF-, ompB-, and gltA-specific primer pairs. Samples positive by PCR were confirmed with DNA sequencing. R. felis was detected in cat fleas year round, and the average infection rate was 21.4% (90 of 420) in 2007. Cat fleas also play an important role in the transmission of Bartonella between reservoirs and other mammalian hosts. In this study, we used primer pairs specific for the Bartonella gltA and rpoB genes to detect Bartonella infections. Of the 420 cat fleas tested, 38 were positive by PCR for Bartonella. Sequence similarities to Bartonella henselae, Bartonella clarridgeiae, and Bartonella koehlerae were observed in 6.2% (26 of 420), 2.1% (9 of 420), and 0.7% (3 of 420) of the fleas, respectively. Based on the pap31 gene sequence, several amplicons of the B. henselae detected in the cat fleas could be subgrouped into three strains: Fizz/CAL-1 (n = 18), Marseille (n = 5), and Houston-1 (n = 3). These results demonstrate that cat fleas infected with R. felis are endemic to Taiwan, and highlight the role of C. felis in Bartonella transmission between reservoirs and other mammal hosts and demonstrate the genetic variability of B. henselae in Taiwan.

Ross River virus (RRV) causes the most common human arbovirus disease in Australia. Although the disease is nonfatal, the associated arthritis and postinfection fatigue can be debilitating for many months, impacting on workforce participation. We sought to create an early-warning system to notify of approaching RRV disease outbreak conditions for major townships in the Northern Territory. By applying a logistic regression model to meteorologic factors, including rainfall, a postestimation analysis of sensitivity and specificity can create rainfall cut-points. These rainfall cut-points indicate the rainfall level above which previous epidemic conditions have occurred. Furthermore, rainfall cut-points indirectly adjust for vertebrate host data from the agile wallaby (Macropus agilis) as the life cycle of the agile wallaby is intricately meshed with the wet season. Once generated, cut-points can thus be used prospectively to allow timely implementation of larval survey and control measures and public health warnings to preemptively reduce RRV disease incidence. Cut-points are location specific and have the capacity to replace previously used models, which require data management and input, and rarely provide timely notification for vector control requirements and public health warnings. These methods can be adapted for use elsewhere.

The role of lone star ticks as vectors for Rocky Mountain spotted fever (RMSF) remains poorly described. We compared the entomological inoculation rates (EIRs) for Rickettsia spp. for representative sites in Missouri and Kansas, states that frequently report RMSF each year. Hostseeking ticks were collected during 2006 and pooled tick homogenates analyzed by polymerase chain reaction to detect probable R. rickettsii, with confirmation for multiple gene targets performed on individual ticks from pools that screened positive. Of 870 adult and nymphal lone star ticks, Amblyomma americanum (L.), 0.46% contained DNA of Rickettsia rickettsii. Interestingly, two of these positive ticks were concurrently infected by R. amblyommii. More than 90% of lone star tick pools contained R. amblyommii DNA. Of 169 dog ticks that were analyzed, none were infected by R. rickettsii. The entomological inoculation rate for spotted fever group (SFG) rickettsiae within lone star ticks was an order of magnitude greater than that for dog ticks. We conclude that lone star ticks may be epidemiologically significant vectors of Rocky Mountain spotted fever and of spotted fever group rickettsiae.

Mosquito populations are sensitive to long-term variations in climate and short-term variations in weather. Mosquito abundance is a key determinant of outbreaks of mosquito-borne diseases, such as West Nile virus (WNV). In this work, the short-term impact of weather conditions (temperature and precipitation) on Culex pipiens L.-Culex restuans Theobald mosquito abundance in Peel Region, Ontario, Canada, was investigated using the 2002–2009 mosquito data collected from the WNV surveillance program managed by Ontario Ministry of Health and Long-Term Care and a gamma-generalized linear model. There was a clear association between weather conditions (temperature and precipitation) and mosquito abundance, which allowed the definition of threshold criteria for temperature and precipitation conditions for mosquito population growth. A predictive statistical model for mosquito population based on weather conditions was calibrated using real weather and mosquito surveillance data, and validated using a subset of surveillance data. Results showed that WNV vector abundance on any one day could be predicted with reasonable accuracy from relationships with mean degree-days >9°C over the 11 preceding days, and precipitation 35 d previously. This finding provides optimism for the development of weather-generated forecasting for WNV risk that could be used in decision support systems for interventions such as mosquito control.

The phlebotomine sand flies Lutzomyia (Psathyromyia) shannoni (Dyar) and Lutzomyia (Helcocyrtomyia) vexator (Coquillett) are reported for the first time from New Jersey. Both species were collected from ultraviolet CO₂-baited Centers for Disease Control light traps during a state-wide phlebotomine survey conducted in 2007 and 2008. L. shannoni has previously been reported from New Jersey; however, this observation was made in error. These collections do not represent a northern range expansion for either species.

Aedes albopictus (Skuse) and Aedes atropalpus (Coquillett) (Diptera: Culicidae) are container-dwelling mosquito species that are well established in the eastern United States. Interspecific larval competition studies have shown Ae. albopictus to be a superior competitor over many species. A laboratory experiment was conducted in artificial containers to evaluate the effects of malathion on larval interactions between Ae. albopictus and Ae. atropalpus. The survivorship of Ae. albopictus increased with increasing Ae. atropalpus densities in control but decreased with increasing Ae. atropalpus densities in the presence of malathion. Alternatively, Ae. atropalpus survivorship did not differ between control and malathion treatments. Developmental times were not affected by interspecific competition in both treatments for either species. These results show that malathion could facilitate coexistence between Ae. albopictus and Ae. atropalpus. This demonstrates how sublethal concentrations of malathion (and perhaps other pesticides with similar modes of action) can enable an inferior competitor to coexist in the same habitat with a superior competitor. This is the first report of synergistic survival of a weaker mosquito competitor in the presence of a pesticide due to condition-specific competition.

The highly sensitive nested pCS20 polymerase chain reaction assay for Ehrlichia ruminantium was negative on 506 Amblyomma variegatum from Caribbean islands where clinical heartwater has not been reported, mainly the United States Virgin Islands (18), Dominica (170), Montserrat (5), Nevis (34), St. Kitts (262), and St. Lucia (17). Positive results were obtained with positive controls (Crystal Springs strain) and A. variegatum from countries in Africa where infections are endemic, mainly Tanzania (1/37) and Burkino Faso (2/29). Positive major antigenic protein-1 enzyme-linked immunosorbent assays for E. ruminantium were obtained on convenience samples of sera from apparently healthy cattle, sheep, and goats on Dominica (0/95, 0%; 3/135, 2%; 2/57, 4%), Grenada (0/4, 0%; 1/98, 1%; 1/86, 1%), Montserrat (0/12, 0%; 0/28, 2%; 5/139, 4%), Nevis (0/45, 0%; 0/157, 0%; 0/90, 0%), Puerto Rico (0/422, 0%; 0, 0%), St. Kitts (3/86, 4%; 1/25, 0%; 0/26, 0%), and St. Lucia (0/184, 0%; 0/15, 0%; 0, 0%), respectively. The pCS20 polymerase chain reaction results indicate E. ruminantium is not present on islands where clinical heartwater does not occur. The occasional positive major antigenic protein-1B enzyme-linked immunosorbent assay results appear, then, to be false-positive reactions, and serology appears to be of limited use in testing for E. ruminantium in the Caribbean, as is the case in Africa.