The Kingdom of Saudi Arabia includes fauna from three zoogeographic regions: the Afrotropical, Oriental, and Palaearctic regions. To study the mosquito (Diptera: Culicidae) fauna of these regions in Saudi Arabia, larval collections were made at 15 sites during 2005–2006. Thirty-three species representing nine genera were found. Six species, Anopheles culicifacies Giles s.l., Anopheles subpictus Grassi s.l., Culex arbieeni Salem, Culex simpsoni Theobald, Culex univittatus Theobald, and Ochlerotatus detritus Haliday are reported for the first time for Saudi Arabia. An annotated checklist and an illustrated key to the fourth-instar larvae of the 33 species are presented, along with some remarks about problematic species. Eleven species of genus Anopheles Meigen, five species of tribe Aedini, 13 species of genus Culex L., two species of genus Culiseta Felt, one species of genus Lutzia Theobald, and one species of genus Uranotaenia Lynch Arribálzaga were recorded during the study.

Scanning electron microscopy images of the first instars of Calliphora croceipalpis Jaennicke, 1876; Chrysomya chloropyga (Wiedemann, 1818); Chrysomya marginalis (Wiedemann, 1830); and Chrysomya putoria (Wiedemann, 1830) (Diptera: Calliphoridae) are presented for the first time, and the following morphological structures are documented: pseudocephalon, antenna, maxillary palpus, facial mask, labial lobe, thoracic and abdominal spinulation, spiracular field, posterior spiracles, and anal pads. Light microscopy photographs and line illustrations are provided for the cephaloskeleton in lateral and ventral views, and the “ectostomal sclerite” and “chitinized teeth” of the cephaloskeleton are recognized as integral parts of the mouthhooks. New diagnostic features of the cephaloskeleton and the spinulation of the abdominal segments are described. These results allow refinement, clarification, and correction of earlier descriptions, which are reviewed. The relative taxonomic importance of various morphological characters of the first instars of necrophagous blow flies is discussed, and details of the cephaloskeleton and the spinulation of the abdominal segments are highlighted as the characters most useful for species identification. Finally, a key for identifying first instars of common African carrion blow flies is provided.

The result of a survey of ectoparasites infesting bats in southeastern Brazil is presented. Of 181 bats belonging to 16 species, 10 (34.1%) were infested by streblid flies (Streblidae), nine (33.5%) by spinturnicid mites (Spinturnicidae), and five (8.3%) by macronyssid mites (Macronyssidae). One species of the families Trombiculidae and Myobiidae was found. A total of 195 streblids, 178 spinturnicids, and 76 macronyssids was collected. Paratrichobius longicrus was the most abundant bat fly species (50 specimens). The spinturnicid mite Periglischrus iheringi was the most abundant ectoparasite species (159 specimens) and was recorded on three different bat species; Radfordiella desmodi was the most numerous macronyssid (69 specimens).

The adults of Periglischrus cubanus were described by Dusbábek (1968). In this study, we describe the nymphal stages of this species and make some general observations on nymphal stages in the genus Periglischrus.

Leishmaniasis is a complex of zoonotic diseases that are endemic to many Brazilian states. They are transmitted to the vertebrates by the bite of the hematophagous female sand fly (Diptera: Psychodidae) vectors. Despite the increasing occurrence of visceral and cutaneous leishmaniasis cases in large urban centers, their transmission continues to occur primarily in a wild environment and may be associated with professional activities, ecotourism activities, or both. This study investigates the ecological parameters of the sand flies present in Ibitipoca State Park, Minas Gerais, Brazil. During 2009, systematic collections of sand flies were made monthly using HP light traps installed at five sites, including three natural settings (a cave, riparian vegetation, and a rain forest), the tourist and researchers' accommodations, and a surrounding domestic livestock area. In total, 161 sand flies (seven species) were collected, the most abundant, particularly in the surrounding domestic livestock area, being Lutzomyia (Psychodopygus) lloydi (Antunes, 1937). Furthermore, a previously unidentified Lutzomyia (Sciopemyia) sp. was prevalent in the cave environment. There are no existing records of the occurrence of leishmaniasis in Ibitipoca State Park; however, the some species of the subgenus Psychodopygus are known vectors of Leishmania spp in Brazil. Hence, the presence of a species of this genus in areas surrounding the park may represent a risk to ecotourism and the local inhabitants. Our study shows the importance of regular monitoring of the various areas used by humans to determine the distribution and spread of sand fly vectors for preventive management to forestall potential risk to health and consequent effect on ecotourists.

Species identification and global positioning system coordinates of infestations of cattle fever ticks, Rhipicephalus (Boophilus) annulatus (Say) and Rhipicephalus (Boophilus) microplus (Canestrini), were determined for 790 specimens submitted to the National Veterinary Services Laboratory between 1 October 1999 and 30 September 2010. Cattle fever tick specimens obtained by personnel of the United States Department of Agriculture-Animal and Plant Health Inspection Service-Cattle Fever Tick Eradication Program from infested cattle and wildlife along the Texas/ Mexico border were submitted for identification, as required by the program. A geographic information system database was developed that incorporates location, collection, and infestation records. Submitted ticks came from 11 Texas counties and were comprised of 19.5% R. (B.) annulatus and 80.5% R. (B.) microplus. Maps produced from this study locate and define the parapatric boundary between R. (B.) annulatus and R. (B.) microplus.

Five cohorts of 100 eggs of Rhodnius neglectus Lent 1954 (Hemiptera: Reduviidae) were reared simultaneously in the laboratory under constant conditions (26 ± 1°C and 60 ± 10% RH), with mortality and fecundity data recorded weekly. We calculated stage-specific developmental times, agespecific mortality and fecundity, stage-specific and total preadult mortality, and the intrinsic rate of natural increase (r_o = 0.21), the finite population growth rate (λ = 1.23), the net reproductive rate (R_o = 314.24), and the generation time (T = 21.45 wk). Elasticity analysis showed that the dominant life-history traits determining λ were the adult female survival, and the survival of instar V nymphs (molting into adult females). Adult females dominated the stage-specific reproductive value, and the egg stage dominated the stable stage distribution (SSD). The damping ratio (p = 1.11) suggests a relatively rapid period of recovery to a disturbed SSD. Results were compared with previous values from the literature and conform relatively well, considering that environmental conditions were not always the same. Compared with two other species of the same genus, Rhodnius neivai Lent 1953 and Rhodnius prolixus Stål 1859, R. neglectus ranked higher in fecundity (total eggs/ ♀ /life) and in female longevity, intermediate in the intrinsic rate of natural increase (r_o), and lower in developmental time and mortality. By fitting a logistic model of population growth to the density field values of a spontaneous colonization of two field experimental chicken coops we estimated the intrinsic rate of natural increase and carrying capacity parameters, and compared the former with our laboratory results.

Dermanyssus gallinae is considered to be the most economically significant ectoparasite to affect egg-laying poultry in Europe. This mite can also act as a vector for a number of pathogens. The array of bacteria associated with D. gallinae mites could provide insight into the biology and population dynamics of arthropods, but at the present time little information is available. To understand the intra- and interpopulation diversity of its associated microbiota, we analyzed the whole internal bacterial community of natural populations of D. gallinae originating from two types of poultry farm habitats (standard and free-range) in two regions of France (Brittany and the Rhone-Alpes). Total DNA was extracted from individual or pooled mites, and polymerase chain reaction temporal temperature gradient gel electrophoresis of 16S rRNA was then done to separate bacterial DNA fragments associated with the host arthropod. A large diversity of bacteria was detected, but principally firmicutes and γ-Proteobacteria. Between-group analyses of temporal temperature gradient gel electrophoresis-banding patterns revealed that bacterial populations clustered into categories according to their geographic origin and the habitat specifics of the farms. Some degree of stability of bacterial populations was observed within a specific time scale. These results suggest that environmental factors either recent (e.g., poultry farming practices) or long-standing (e.g., geographic isolation) may affect the bacterial communities present in D. gallinae. Further knowledge of the microbiota associated with D. gallinae and its variation would indeed offer new perspectives for biological control methods to prevent the establishment, proliferation, and transmission of pathogenic bacteria.

The population size, age-class structure, and movement of the American cockroach, Periplaneta americana (L.) (Dictyoptera: Blattidae), were studied in three sewers in Penang, Malaysia, from September 2008 to October 2009. Eighteen to 20 glass-jar traps (two per manhole) were deployed for a 24-h period during each sampling occasion at each sewer. Adults and nymphs were active throughout the study period, with an average monthly trap catch of 57–97 adults and 79–99 nymphs. The mean proportions of adults and nymphs at the three sewers ranged from 0.47 to 0.57. Of the 2,177 male and 2,717 female cockroaches marked and released over the three sewers, recapture rates were 29.4–45.8 and 30.8–47.0%, respectively. The proportion of marked males and females did not differ significantly from the proportion of recaptured marked males and females. However, the mean number of times a marked female was recaptured was significantly greater than that of males. Of the 783 males and 1,030 females that were marked and recaptured, 19.4 and 24.7%, respectively, had moved between manholes, and significantly more females than males moved between manholes. Of the 406 recaptured marked adults that moved between manholes, 90.4% moved a distance of 2–20 m from their initial release site; one male moved 192 m, the longest distance recorded. Trap catch on each sampling occasion was positively correlated with daily mean temperature. The number of cockroach movements between manholes also was correlated with the mean daily minimum temperature.

Thirty-three Triatoma sanguisuga (LeConte) adults and nymphs were collected during June and July 2009, at five sites on Cumberland Island and two sites on Sapelo Island, Georgia, to assess genetic diversity within and between sites. All but three specimens were found in a peridomestic habitat. The entire length (699 bp) of the cytochrome oxidase II mitochondrial gene was sequenced for each specimen. Twelve haplotypes were identified, nine from Cumberland Island and three from Sapelo Island. No haplotypes were shared between the two islands, indicating there is limited or no movement of gene flow between the islands. Phylogenetic relationships among the haplotypes were determined using both neighbor-joining and maximum parsimony analyses. The phylogenetic trees from both analyses were similar, with no distinct clades on either tree devoted to haplotypes from a single island. A haplotype network structure was determined using nested clade analysis, which produced two haplotype networks, one containing only specimens found on Cumberland Island. The second network included specimens from both islands, with the ancestral haplotype from Sapelo Island. This pilot study is the first to highlight triatomine populations in the southeastern United States using the cytochrome oxidase II mitochondrial gene, and indicates strong population structuring along the Georgia Coast.

A field test of integrated vector control was conducted in a tropical urban setting with a combination of a floating, slow-release, granular formulation of Bacillus sphaericus and environmental engineering measures (renovation of roads, collective water pumps, and cesspool lids). The targets were Culex quinquefasciatus and Anopheles gambiae in the two biggest towns of Burkina Faso (West Africa). Within the intervention zone, water pumping stations were improved and the surroundings drained to prevent the accumulation of stagnant water. Roads were leveled and given either simple gutters on each side or a concrete channel on one side to drain runoff water. Garbage containers were installed to provide an alternative to the drainage channels for waste disposal. Septic tanks were modified so that they could be emptied without destroying their lid. This study showed that it is possible to implement mosquito control in a tropical urban environment with teams of young people rapidly trained to apply a biological larvicide without any tools other than an iron bar to lift cesspool lids. Environmental improvements were initially costly, but demanded little subsequent expenditure. Local inhabitants' committees were mobilized to provide people with information and monitor the efficacy of the measures. Compared with what people spent individually on mosquito prevention and malaria medicine, these measures were not expensive, but many expected the community to pay for them from existing taxes, e.g., for water treatment and disposal. The necessary funding and logistics require a municipal organization with neighborhood support, if the measures are to be effective.

Acaricidal effects of three essential oils extracted from Mexican oregano leaves (Lippia graveolens Kunth), rosemary leaves (Rosmarinus officinalis L.), and garlic bulbs (Allium sativum L.) on 10-d-old Rhipicephalus (Boophilus) microplus (Canestrini) tick larvae were evaluated by using the larval packet test bioassay. Serial dilutions of the three essential oils were tested from a starting concentration of 20 to 1.25%. Results showed that both Mexican oregano and garlic essential oils had very similar activity, producing high mortality (90–100%) in all tested concentrations on 10-d-old R. microplus tick larvae. Rosemary essential oil produced >85% larval mortality at the higher concentrations (10 and 20%), but the effect decreased noticeably to 40% at an oil concentration of 5%, and mortality was absent at 2.5 and 1.25% of the essential oil concentration. Chemical composition of the essential oils was elucidated by gas chromatographymass spectrometry analyses. Mexican oregano essential oil included thymol (24.59%), carvacrol (24.54%), p-cymene (13.6%), and γ-terpinene (7.43%) as its main compounds, whereas rosemary essential oil was rich in α-pinene (31.07%), verbenone (15.26%), and 1,8-cineol (14.2%), and garlic essential oil was rich in diallyl trisulfide (33.57%), diallyl disulfide (30.93%), and methyl allyl trisulfide (11.28%). These results suggest that Mexican oregano and garlic essential oils merit further investigation as components of alternative approaches for R. microplus tick control.

We describe the susceptibility to deltamethrin and fipronil of four sylvatic Triatoma infestans populations from the Andean valleys of Bolivia. Fifty percent lethal doses were determined from topical application of insecticide on first instars, and mortality was assessed after 24 h for deltamethrin and 48 h for fipronil. In comparison with a reference strain from Argentina, the Bolivian populations showed deltamethrin 50 percent lethal dose ratios ranging from 1.9 to 17.4. In the case of fipronil, an insecticide never used for control of T. infestans, the Bolivian populations showed even higher variation in toxic response, with relative susceptibilities ranging from 0.5 to 139.2. However, although the sylvatic T. infestans toxicological profiles differ from each other and from those of the domiciliary population studied in this work, there were no significant differences in the activities of P450 mono-oxygenases and pyrethroid esterases between the reference strain and the studied populations.

An investigation of the Indian folk remedy plant Jatropha curcas L., was performed to identify the constituents responsible for the mosquito biting deterrent activity of the oil. J. curcas seed oil is burned in oil lamps in India and parts of Africa to repel biting insects, primarily mosquitoes. The seed oil was thoroughly analyzed by ¹H NMR, ¹³C NMR, high-performance liquid chromatographyrefractive index, and gas chromatography-flame ionization detection to identify the constituents in the oil. Identified constituents, both free fatty acids and triglycerides, were evaluated for activity in Aedes aegypti (L.) (Diptera: Culicidae) biting deterrent assays. Furthermore, an oil condensation trap was used to demonstrate that free fatty acids or triglycerides are partially volatilized during the combustion process. These compounds were found to be responsible for the biting deterrency of the burned oil. Specifically, oleic, palmitic, linoleic, and stearic acids were all active at 25 nmol/cm² above that of solvent control in Ae. aegypti biting deterrent assays. Oleic, palmitic, and linoleic acids were all more active than stearic acid in the same bioassay. Evaluation of the triglycerides containing each of these fatty acids revealed that tripalmitin, tristearin, trilinolein, and triolein all demonstrated significant activity above a solvent control at 10 µg/cm², whereas tripalmitin was the most active. Due to literature reports suggesting larvicidal activity of the oil, J. curcas seed oil and its free fatty acid constituents also were evaluated against 1-d-old Ae. aegypti larvae up to 500 ppm. Oleic acid was the only fatty acid having larvicidal activity against 1-d-old Ae. aegypti larvae, with an LD₅₀ of 47.9 ppm.

Seed ethanolic extracts of 21 Brazilian plants were evaluated for ovicidal, larvicidal, and pupicidal activities against insecticide-susceptible (SS) and field-collected (FC) strains of Aedes aegypti (L.) (Diptera: Culicidae), as well as for their effects on nontarget organisms. Myracrodruon urundeuva Fr. Allemao extract was highly toxic to both mosquito strains. Schinopsis brasiliensis Engler extract showed low toxicity and was 38 – 68 times less toxic to Ae. aegypti larvae than was M. urundeuva extract. The pupicidal activity (LC₅₀) of 14 plant seed extracts ranged between 9 and 433 µg/ml, and toxicities were comparable to both mosquito strains. Piptadenia moniliformis Benth. and Luetzelburgia auriculata (Allemao) Ducke extracts showed the highest activities against pupae of FC and SS strains. None of the extracts showed 100% ovicidal activity. In addition, the active extracts did not show high acute toxicity to mice (LD₅₀ > 1.5g/ kg), except that of Enterolobium contortisiliquum (Vell.) Morong. Most of the active extracts exhibited low toxicity against brine shrimp (Artemia sp.) nauplii. The extracts of M. urundeuva, P. moniliformis, and L. auriculata are promising sources of recognized classes of insecticidal compounds with good selectivity against immature stages of Ae. aegypti.

Larval and nymphal western blacklegged tick, Ixodes pacifiais Cooley & Kohls (Acari: Ixodidae), were collected from birds, rodents, and lizards at Quail Ridge Reserve located in Napa County in northwestern California. Species from three vertebrate classes were sampled simultaneously from two transects during two consecutive spring seasons. Feeding larval and nymphal ticks were removed and preserved for counting, examination and testing for the presence of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner. Mean infestations with I. pacificus subadults on lizards were 10.0, on birds 2.9, and on rodents 1.3. I. pacificus larvae (204) collected from 10 avian species and (215) collected from two rodent species were tested for the presence of B. burgdorferi s.s. via real-time polymerase chain reaction. Three B. burgdorferi-infected larvae were taken from two Junco hyemalis and two infected larvae from one Neotoma fuscipes Baird. This is the first reported detection of B. burgdorferi in larvae feeding on a bird in western North America.

The blacklegged tick (Ixodes scapularis) is an important vector of emerging human pathogens. It has three blood-feeding stages, as follows: larva, nymph, and adult. Owing to inefficient transovarial transmission, at least for the Lyme disease agent (Borrelia burgdorferi), larval ticks rarely hatch infected, but they can acquire infection during their larval blood meal. Nymphal ticks are primarily responsible for transmitting pathogens to hosts, including humans. The transition from uninfected host-seeking larva to infectious host-seeking nymph is therefore a key aspect of human risk of infection. It can be divided into a series of steps, as follows: finding a host, taking a blood meal, becoming infected, molting, and overwintering. The chance of succeeding in each of these steps may depend on the species identity of the blood meal host. We used a Bayesian method to estimate the molting success of larval I. scapularis collected from four commonly parasitized species of birds and eight commonly parasitized small and mid-sized mammals found in the forests of Dutchess County, New York. We show that molting success varies substantially among host species; white-footed mice, veeries, and gray catbirds support particularly high molting success, whereas ticks feeding on shorttailed shrews, robins, and wood thrushes were less successful. We also show that larval molting success varies substantially between individual blood meal hosts, and that this intraspecific variability is much higher in some species than in others. The causes of both inter- and intraspecific variation in molting success remain to be determined.

The factors determining the spatial and temporal distribution of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) activity are not well understood. Here, we explore the effects of hydrological and meteorological conditions on WNV infection among Culex genus mosquitoes collected during 2001–2009 in Suffolk County, Long Island, NY. We show that WNV infection rates in assayed pools of Culex mosquitoes are associated in both space and time with hydrological and meteorological variability. Specifically, wet winter, warm and wet spring conditions, and dry summer conditions are associated with the increased local prevalence of WNV among Culex mosquitoes during summer and fall. These findings indicate that within Suffolk County, and for a given year, areas at risk for heightened WNV activity may be identified in advance by using hydrology model estimates of land surface wetness and observed meteorological conditions.

Porcine Reproductive and respiratory syndrome (PRRS) is a globally significant swine disease caused by an arterivirus. The virus replicates in alveolar macrophages of infected pigs, resulting in pneumonia in growing pigs and late-term abortions in sows. Outbreaks occur on disparate farms within an area despite biosecurity measures, suggesting mechanical transport by arthropods. We investigated the vector potential of stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), in the transmission of porcine reproductive and respiratory syndrome virus (family Arteriviridae, genus Arterivirus, PRRSV) under laboratory conditions. Stable flies were collected around PRRS-negative boar stud barns in North Carolina and tested for presence of the virus. Stable flies were collected on alsynite traps placed near the exhaust fan of the close-sided tunnel-ventilated buildings, suggesting blood seeking flies are attracted by olfactory cues. No flies were positive for PRRSV. We assessed transmission of the virus through an infective bite by feeding laboratory reared stable flies on blood containing virus and transferring them to naïve pigs for subsequent bloodmeals. Transmission of the virus to naïve pigs by infective bites failed in all attempts. The volume of blood contained within the closed mouthparts of the stable fly seems to be insufficient to deliver an infective dose of the virus. Stable flies are unlikely to transmit PRRSV from one pig to another while blood feeding. The fate of the virus after a bloodmeal remains to be determined.

The American dog tick, Dermacentor variabilis (Say) (Acari: Ixodidae), has been implicated as a potential bridging vector to humans of Francisella tularensis, the etiological agent of tularemia. Since the initial studies evaluating vector competency of D. variabilis were conducted, F. tularensis has been subdivided into subspecies and clades that differ in their geographical distribution in the United States and in the severity of infections caused in humans. Here, we demonstrate that D. variabilis nymphs efficiently acquire, transtadially maintain, and transmit each of the strains tested (clades A1b and A2, and type B). Transmission efficiency by adult females was similarly high among infection groups and ranged from 58% for type B to 89% for A2 infections. In addition, we demonstrated that transmission can occur shortly after tick attachment. These findings support the concept that D. variabilis adults may play a significant role in epizootic transmission of F. tularensis, and as a bridging vector to humans.

Tick-borne relapsing fever is caused by spirochetes within the genus Borrelia. The hallmark of this disease is recurrent febrile episodes and high spirochete densities in mammalian blood resulting from immune evasion. Between episodes of spirochetemia when bacterial densities are low, it is unknown whether ticks can acquire the spirochetes, become colonized by the bacteria, and subsequently transmit the bacteria once they feed again. We addressed these questions by feeding ticks, Ornithodoros hermsi Wheeler (Acari: Argasidae), daily on an infected mouse during low and high levels of spirochete infections. This study demonstrates that spirochete acquisition by the tick vector can occur during low levels of mammalian infection and that once a spirochetemic threshold is attained within the blood, nearly 100% of ticks become colonized by Borrelia hermsii.

Eastern equine encephalitis virus (EEEV; family Togaviridae, genus Alphavirus) epizootics are infrequent, but they can lead to high mortality in infected horses and humans. Despite the importance of EEEV to human and animal health, little is known about how the virus overwinters and reinitiates transmission each spring, particularly in temperate regions where infected adult mosquitoes are unlikely to survive through the winter. One hypothesis to explain the mechanism by which this virus persists from year to year is the spring recrudescence of latent virus in avian reservoir hosts. In this study, we tested the recrudescence hypothesis with gray catbirds (Dumatella carolinensis) captured in northern Ohio (July–August 2007). Birds were experimentally infected with EEEV on 1 October 2007. In January 2008, they were then exposed to exogenous testosterone and/ or extended photoperiod to initiate reactivation of latent EEEV infection. All birds became viremic with EEEV, with mean viremia of 6.0 log₁₀ plaque-forming units/ml serum occurring at 1 d postinoculation. One male in the testosterone, long-day treatment group had EEEV viral RNA in a cloacal swab collected on 18 January 2008. Otherwise, no other catbirds exhibited reactivated infections in cloacal swabs or blood. Antibody titers fluctuated over the course of the study, with lowest titers observed in January 2008, which corresponded with the lowest mean weight of the birds. No EEEV viral RNA was detected in the blood, kidney, spleen, brain, liver, and lower intestine upon necropsy at 19 wk postinfection.

In Japan, the prevalence of avian Plasmodium in birds and mosquitoes has been partially examined in the temperate and subtropical zones; however, mosquitoes in the Japanese subarctic zone have not been adequately investigated. In this study, mosquito collections and avian Plasmodium detections from the mosquito samples were carried out to demonstrate the avian Plasmodium transmission between vector mosquitoes and birds inhabiting in Kushiro Wetland, subarctic zone of Japan. A total of 5,657 unfed mosquitoes from 18 species and 320 blood-fed mosquitoes from eight species was collected in summer 2008, 2009, and 2010. Three Aedes esoensis that fed on Hokkaido Sika Deer and one unfed Culex pipiens group were found to be positive for avian Plasmodium by polymerase chain reaction. This is the first report of the detection of avian Plasmodium DNA from mosquitoes distributing in the subarctic zone of Japan. The blood meals were successfully identified to captive or wild animals, including seven mammalian species, four bird species, and one amphibian species. These results indicated that infected birds with avian Plasmodium inhabited and direct contacts occurred between the infected birds and mosquitoes in Kushiro Wetland, Hokkaido, Japan.

Twenty resting shelters were set on the edge of a known Culiseta breeding habitat in four groups of five to support a 4 × 4 Latin square field experiment. Collection times were 0900, 1100, 1300, and 1500 hours and systematically rotated for the order by which each group of five boxes was collected. Mosquitoes were collected from resting shelters by chloroform anesthetization. Collections were identified to species, sex, and physiological status of the females (nonblooded or blood-fed and gravid). More than 77% of the mosquitoes collected were Culiseta melanura (Coquillett). Analyses included means and SE for total collections and shelter-day (number collected per units) and means comparison by t-test and general linear model with Student—Newman—Keuls or least significant differences means tests for replicate, group, time, and interactions of time and group. There were few significant differences among or between shelter-day means but more blood-fed and gravid female Cs. melanura were collected at 1300 hours than any other time. Results confirm the effectiveness of resting shelters in a surveillance program for Cs. melanura, demonstrate the flexibility of resting shelters as a surveillance tool, and suggest that Cs. melanura will move to more acceptable resting sites during daylight.

An understanding of physiological sensitivity to temperature and its variability is important for predicting habitat suitability for disease vectors under different climatic regimes. In this study, we characterized the thermal sensitivity of larval developmental rates and survival in several Australian mainland populations of the dengue virus vector Aedes aegypti. Males developed more rapidly than females, but there were no differences among populations for development time or survival despite previously demonstrated genetic differentiation for neutral markers. Optimal development and survival temperatures were 37°C and 25°C, respectively. The values for maximal development and survival were similar to standard functions used in the container inhabiting simulation (CIMSIM) model for predicting population dynamics of Ae. aegypti populations, but CIMSIM assumed a lower optimal temperature. Heat stress experiments indicated that larvae could withstand water temperatures up to 44°C regardless of the rate at which temperature was increased. Results from development time measured under constant temperatures could predict development time under fluctuating conditions, whereas CIMSIM predicted faster rates of development. This difference acts to reduce the predicted potential number of generations of Ae. aegypti per year in Australia, although it does not influence its predicted distribution, which depends critically on the nature of the aquatic breeding sites.

For the majority of native species, human-created habitats provide a hostile environment that prevents their colonization. However, if the conditions encountered in this novel environment are part of the fundamental niche of a particular species, these low competitive environments may allow strong population expansion of even rare and stenotopic species. If these species are potentially harmful to humans, such anthropogenic habitat alterations may impose strong risks for human health. Here, we report on a recent and severe outbreak of the viciously biting and day-active mosquito Anopheles plumbeus Stephens, 1828, that is caused by a habitat shift toward human-created habitats. Although historic data indicate that the species was previously reported to be rare in Belgium and confined to natural forest habitats, more recent data indicate a strong population expansion all over Belgium and severe nuisance at a local scale. We show that these outbreaks can be explained by a recent larval habitat shift of this species from tree-holes in forests to large manure collecting pits of abandoned and uncleaned pig stables. Further surveys of the colonization and detection of other potential larval breeding places of this mosquito in this artificial environment are of particular importance for human health because the species is known as a experimental vector of West Nile virus and a potential vector of human malaria.

Control of the Chagas disease vector, Triatoma infestans, relies on the application of pyrethroid insecticides, especially deltamethrin. We performed laboratory studies to determine whether a T. infestans nymph that comes into contact with a deltamethrin-treated surface horizontally transfers the insecticide to subsequent triatomines. We found that a triatomine that walks on a deltamethrin-treated surface for a short period of time has the ability to transport the insecticide in concentrations sufficient to kill other triatomines with which it comes into contact. The effect was limited to high-density environments, and mortality as a result of secondary exposure was greater among second-instar nymphs compared with fifth-instar nymphs. Our results suggest that deltamethrin could be killing triatomines through both direct and indirect contact, although it remains unclear whether the phenomenon occurs in natural conditions.

Late instar larvae of the scuttle fly, Megaselia Scalaris Loew, were found near the right eye of a live captive-reared western hognose snake, Heterodon nasicus Baird and Girard. Dissection and removal of the snake's dorsal cranial bones revealed tissue degradation of the infected eye, the optic nerve, and the brain case; we suggest that these factors contributed to the death of this snake. This case study further demonstrates the opportunistic behavior of M. Scalaris.

Cimex lectularius L. (Hemiptera: Cimicidae) is a widespread blood feeding pest of humans around the world, including North America, and has recently undergone a resurgence. A molecular diagnostic technique applying multiplex polymerase chain reaction (PCR) was developed to distinguish bed bug eggs, leg fragments, and degraded samples from other arthropods that frequently occur in human dwellings. A 410-428-bp region of the mitochondrial DNA 16S rRNA gene was used. To design C. lectularius-specific PCR primers, DNA sequences of various bed bug samples from the United States, Canada, and Australia, along with sequences of other Cimicidae and arthropods that often occur in dwellings, were considered. Based on DNA sequence variation, one reverse PCR primer specific for C. lectularius was identified. Multiplex PCR using three primers will yield a 417- and 140-bp amplicon for C. lectularius and a single 410-428-bp amplicon for other taxa. This assay was successful in identifying C. lectularius eggs, leg fragments, and degraded samples. This technique should provide a reliable, quick, and economical technique for identifying C. lectularius, when morphological identification is not possible.

Sound plays an important role in the mating behavior of mosquitoes, including Aedes aegypti (L). Males orient to the fundamental wing beat frequency of females, and both sexes actively modulate their flight tone before mating to converge at harmonic frequencies. The majority of studies on mosquito mating acoustics have been conducted in the laboratory using tethered individuals. In this study, we present the first free-flight recording of naturally forming Ae. aegypti swarms in Thailand. We describe mating behaviors and present results on the flight tone frequency and dynamics of wild pairs in free flight. To assess the importance of these behaviors in vector control programs, especially those using genetically modified mosquitoes, it will be critical to use methods, such as those described in this work, to measure mosquito mating behaviors in the field.

Quantitative real-time polymerase chain reaction (qRT-PCR) has emerged as robust methodology for gene expression studies, but reference genes are crucial for accurate normalization. Commonly used reference genes are housekeeping genes that are thought to be nonregulated; however, their expression can be unstable across different experimental conditions. We report the identification and validation of suitable reference genes in the bed bug, Cimex lectularius, by using qRT-PCR. The expression stability of eight reference genes in different tissues (abdominal cuticle, midgut, Malpighian tubules, and ovary) and developmental stages (early instar nymphs, late instar nymphs, and adults) of pesticide-susceptible and pesticide-exposed C. lectularius were analyzed using geNorm, NormFinder, and BestKeeper. Overall expression analysis of the eight reference genes revealed significant variation among samples, indicating the necessity of validating suitable reference genes for accurate quantification of mRNA transcripts. Ribosomal protein (RPL18) exhibited the most stable gene expression across all the tissue and developmental-stage samples; α-tubulin revealed the least stability across all of the samples examined. Thus, we recommend RPL18 as a suitable reference gene for normalization in gene expression studies of C. lectularius.

One strategy suggested for the management of mosquito insecticide resistance consists of combining a pyrethroid with an insecticide that has a different mode of action. To restore the efficacy of deltamethrin (pyrethroid) against pyrethroid-resistant strain of Anopheles gambiae Giles (VKPR: homozygous Kdr), deltamethrin was combined with the neonicotinoid insecticide dinotefuran and piperonyl butoxide (PBO). Bednets impregnated with deltamethrin, dinotefuran, and PBO alone and in combination were tested in the laboratory. Knockdown (KD) and mortality were measured using WHO cone tests on susceptible and pyrethroid-resistant adult mosquitoes. The combination of deltamethrin and PBO was synergistic against resistant female An. gambiae (58.2% mortality). Both mortality and knockdown time (KDt_50/95 values) of the tricomponent mixture on the VKPR strain were similar to the insecticidal activity of deltamethrin on the pyrethroid-susceptible KIS strain (98.8 and 100% mortality, respectively). The three-compound mixture of deltamethrin PBO dinotefuran showed an insecticidal efficacy greater than the deltamethrin PBO mixture to the extent of completely restoring the efficacy of deltamethrin on pyrethroid-resistant An. gambiae.

The feeding patterns of haematophagous arthropods are of major importance in the amplification and transmission of infectious disease agents to vertebrate hosts, including humans. The establishment of new vector populations in nonnative range might alter transmission networks. The Asian tiger mosquito Aedes albopictus (Skuse) represents an example of how an invasive species can alter the risk of viral transmission to humans. Blood meal molecular identification from two sympatric mosquito species (the invasive Ae. albopictus and the native Culex pipiens) was carried out by polymerase chain reaction-based methods. Samples were collected in Barcelona metropolitan area, Spain, from June to October 2009 as part of a monitoring-control program. Blood meals were identified to the species level in 30 Ae. albopictus and 43 Cx. pipiens. Ae. albopictus acquired blood exclusively from human hosts (100%), whereas Cx. pipiens fed on a diversity of avian and mammalian hosts, including 35.7% of blood meals from humans. Based on mosquito diet, our results suggest that the Ae. albopictus invasion in Spain might increase the risk of virus transmission to humans and could support local outbreaks of imported tropical viruses such as dengue and chikungunya. However, in the studied area, the presence of this invasive species would have a negligible effect on the transmission of zoonotic agents such as West Nile virus. However, Cx. pipiens could amplify and transmit West Nile virus, but avian contribution to its diet was lower than that reported in North America. Feeding patterns of these mosquito species may help to understand the flavivirus outbreaks recently reported in southwestern Europe.