There is increasing need to apply established standards for recording data on mosquito collection events, because of the diversity of potential data providers, and the growth and interoperability of online databases designed to host these collection records. In particular, adequate taxonomic and georeference data are needed for geodatabases such as Mosquitomap ( http://www.mosquitomap.org/) that map and compare these collection points with other spatial information in a geographical information system (GIS) setting. Accurately georeferenced collection data are crucial for understanding mosquito biogeography, ecology, and the impact of environmental changes, as well as for species distribution modeling, planning mosquito surveys, and for determining disease risk. We sampled representative published reports of new mosquito species records from 1980 in North America to the present to ascertain the quality of georeference information. Our results show that authors have increased the frequency of reporting georeferences but that they vary in the precision of the georeference, and some information, such as the source, date, and datum of the georeference, are usually not given. We discuss recently established standards for recording collection events, some relevant online resources available to researchers to assist them in their georeferencing, and the data input schema developed for the Mosquitomap database. We propose that the mosquito research community adopt data standards for recording and reporting the results of mosquito collection events to increase the value of these data. In particular, we recommend authors lodge voucher specimens and use a GPS set to the WGS84 datum.

It has been two decades since the first description of Rickettsia felis, and although a nearly cosmopolitan distribution is now apparent, much of the ecology of this unique microorganism remains unresolved. The cat flea, Ctenocephalides felis, is currently the only known biological vector of R. felis; however, molecular evidence of R. felis in other species of fleas as well as in ticks and mites suggests a variety of arthropod hosts. Studies examining the transmission of R. felis using colonized cat fleas have shown stable vertical transmission but not horizontal transmission. Likewise, serological and molecular tools have been used to detect R. felis in a number of vertebrate hosts, including humans, in the absence of a clear mechanism of horizontal transmission. Considered an emerging flea-borne rickettsiosis, clinical manifestation of R. felis infection in humans, including, fever, rash, and headache is similar to other rickettsial diseases. Recent advances toward further understanding the ecology of R. felis have been facilitated by stable R. felis-infected cat flea colonies, several primary flea isolates and sustained maintenance of R. felis in cell culture systems, and highly sensitive quantitative molecular assays. Here, we provide a synopsis of R. felis including the known distribution and arthropods infected; transmission mechanisms; current understanding of vertebrate infection and human disease; and the tools available to further examine R. felis.

The etiological agent of plague, Yersinia pestis, is most commonly transmitted by the bite of infectious fleas. To date, at least 28 flea species occurring in North America have been experimentally confirmed as vectors of Y. pestis. Transmission efficiency differs among species and also between different studies of a single species. These differences may, however, in large part reflect nonstandardized experimental conditions used during the first half of the 20th century when such studies were conducted in response to the rapid spread of Y. pestis across the western United States after its introduction at the beginning of this century. The majority of these early transmission studies focused on the blocked flea mechanism of transmission, which typically does not occur until >2–3 wk after the flea becomes infected. Recent studies have challenged the paradigm that Y. pestis is usually spread by blocked fleas by demonstrating that numerous flea species, including the oriental rat flea Xenopsylla cheopis, which was the focus of the early classical studies on blocked flea transmission, are capable of “early-phase” transmission during the first few days after becoming infected and before a complete blockage can form. The aims of this review are to 1) summarize Y. pestis vector competency and efficiency studies for fleas occurring in North America, 2) discuss the implications of the results of these studies for our understanding of the dynamics of plague epizootics, 3) demonstrate why older transmission studies need to be repeated using a standardized experimental system, and 4) outline future directions for studies of fleas as vectors of Y. pestis.

Species within the subgenus Culex (Melanoconion) Theobald are the primary enzootic vectors of viruses in the Venezuelan equine encephalitis complex including Everglades virus, and probable enzootic vectors of eastern equine encephalitis and West Nile viruses. Adult females of this subgenus are often difficult or impossible to identify to species based on external morphological characters. The use of female cibarial armature allows for the identification of field-collected adult female specimens of Culex (Melanoconion). The cibarial armatures are described and illustrated for all species from the United States and a key to species using this character is presented.

Three new species of chigger mites, Neotrombicula kounickyi sp. n., Leptotrombidium angkamii sp. n., and Doloisia vlastae sp. n., are described from two species of small mammals collected in the Barun Glacier Valley, Makalu region, Nepal Himalaya. Two species, Trombiculindus mehtai Fernandes et Kulkarni, 2003 and Cheladonta ikaoensis (Sasa et al., 1951) are recorded for the first time in Nepal. Data on altitude distribution of chiggers and their host preferences are given.

Eggs of container-breeding mosquitoes are able to withstand drought conditions as an egg and hatch when submerged. Frequent rainfall can be simulated by frequent submersion, and drought conditions can be simulated by infrequent submersion. We examined the hatch response of Aedes albopictus (Skuse) eggs to simulated drought conditions. Ae. albopictus eggs from a strain originating outside Kobe, Japan, were subjected to one of three treatments; high-frequency hatch stimulation consisting of submerging the eggs in a nutrient broth mixture every 3 d, low-frequency hatch stimulation consisting of submerging the eggs every 7 d, and delayed high-frequency hatch stimulation. Eggs that were subjected to lower-frequency stimulation showed a significant decrease in hatch delay, which was the opposite of the predicted response. This decrease in hatch delay may be an example of hatch plasticity in response to drought conditions. This response could not be explained as a result of the difference in the ages of the eggs on any given stimulus. A decreased hatch delay response to potential drought conditions combined with rapid larval development may enable Ae. albopictus, whose eggs are not as desiccation resistant as some other container-breeding mosquitoes, to survive extended drought.

Aedes albopictus (Skuse) is an invasive container-dwelling mosquito and an important disease vector that co-occurs with the native mosquito, Aedes triseriatus (Say), and the predatory midge, Corethrella appendiculata (Grabham). Larval Ae. triseriatus show significantly greater antipredatory responses when compared to larval Ae. albopictus in the presence of predation cues from C. appendiculata. The potential for evolution of antipredatory behavioral responses to C. appendiculata in Ae. albopictus is unknown. We used a controlled laboratory selection experiment to test whether Ae. albopictus could evolve antipredatory behavioral responses to C. appendiculata predation. We subjected replicate Ae. albopictus populations to four generations of predation by C. appendiculata or a predator-free control treatment and compared the behavior and life history of Ae. albopictus in the two treatments in each generation. There were no differences in Ae. albopictus behavioral responses between predation and control lines in any of the four generations. There was also no evidence of differences in life histories between predation and control lines. Ae. albopictus is superior as a competitor compared with Ae. triseriatus, which it has replaced in areas where C. appendiculata are rare. Our results suggest limited potential for Ae. albopictus to evolve stronger antipredatory behavioral responses to C. appendiculata predation and imply that C. appendiculata will continue to act as an impediment to invasion by Ae. albopictus and replacement of Ae. triseriatus and to promote coexistence of these competitors.

Human victims of a massive number of stings have been steadily increasing since the invasion of Africanized honey bees (Apis mellifera) to the United States in 1990. Multiple honey bee stings may result in venom toxicity, leading to renal failure and even death. Here we tested the efficacy of methyl anthranilate as a honey bee repellent during a massive defensive response by Africanized honey bees. An aerosolized solution of 10% methyl anthranilate reduced the number of defensive bee hits to a retreating victim by 95% compared with a water control. One hundred fifty milliliters of the 10% methyl anthranilate solution sprayed onto stationary foam balls covered with black suede leather located 2 m from provoked Africanized colonies received 80% fewer stings than targets treated with water. Methyl anthranilate (100%) delivered through a UV blocking 3 mil polyethylene pouch was 100% effective in preventing Polistes colonization in wildlife observation huts and from the roof overhang of home patios. Although methyl anthranilate was not 100% effective in preventing honey bee stinging, it seemed to reduce number of stings below the average human LD₅₀, indicative of a promising tool for preventing honey bee venom toxicity and wasp colonization.

The hematophagous bug Triatoma brasiliensis (Neiva) is an important vector of Chagas disease in Brazil. We describe the mating behavior of this species in a comparative manner and analyze the effect of time elapsed since the first imaginal bloodmeal on motivation of males to mate, on female mate-rejections, and on overall mating frequency. We also evaluate whether females of this species copulate with several males in short time intervals, and we examine whether the duration of copulation may vary in the presence of other males compared with what is observed with isolated pairs. Age affected the motivation of males to perform mating attempts, whereas nutritional status affected the receptivity of females. Results showed that 70% of the females tested accepted copulating with three males over a period of 90 min, whereas the remaining 30% mated with at least two males. Mating duration in T. brasiliensis was prolonged in the presence of other males compared with observations of isolated pairs, and males of this species performed long postcopulatory associations only in the presence of other males, apparently as a mate guarding response to the presence of potential competitors.

Knowledge of the genetic variability, population structure, and evolutionary history of Triatoma infestans may be useful for developing rational vector control strategies. A 661-bp fragment of the mitochondrial gene cytochrome oxidase I (COI) was sequenced and analyzed in bugs from Argentina, Uruguay, Peru, and Bolivia, including peridomestic, domestic, Andean, and Chaco sylvatic bugs. A total of 48 polymorphic sites among 37 haplotypes were described. Nucleotide variation fluctuated among samples, with the highest nucleotide diversity observed in seven Argentinean provinces. Within this group, some populations showed patterns of variability compatible with population expansions and/or fine-scale population structure, whereas others suggested population bottlenecks and/or population admixture processes. A maximum parsimony analysis of the haplotypes showed the presence of a Bolivian/Peruvian and an Argentinean/Uruguayan clade. Bolivian sequences were further divided in Chaco sylvatic and Andean domestic and sylvatic. Two different nested clades were found within the Argentinean/Uruguayan cluster. Analysis of molecular variance (AMOVA) and K_ST^* analysis supported a strong population structure in Argentina, where genetic differentiation was correlated with geographic distance. Departures from neutrality expectations and a nested cladistic analysis suggest a recent population expansion of T. infestans in Argentina, followed by restricted gene flow and patterns of isolation by distance. This expansion could have taken place as a two-wave process, as was shown by the phylogenetic analysis and signatures of population admixture in the southernmost Argentinean populations.

Using quantitative polymerase chain reaction (QPCR), the relative transcriptional levels of cytochrome c oxidase subunit 3 (COS) were studied in Aedes aegypti in response to treatment with acetone, permethrin, and fipronil. The transcriptional levels of CO3 were significantly (P < 0.05) higher in acetone-treated Ae. aegypti compared with that in untreated samples. Using ribosomal L24, heat shock protein (HSP), and actin as reference genes, relative transcription levels of CO3 in acetone-treated Ae. aegypti were 2.88 ± 0.38-, 2.60 ± 0.60-, and 3.24 ± 0.70-fold higher, respectively, compared with that in untreated mosquitoes. Transcriptional levels of CO3 were induced significantly higher (6.54 ± 1.22-, 4.62 ± 0.74-, and 9.47 ± 3.71-fold, respectively) by permethrin at LD₁₀ compared with acetone (P < 0.05). Taken together, our results suggest that overexpression of CO3 is tightly regulated in Ae. aegypti in response to xenobiotic treatment.

Insecticide-treated vertical net barriers were used to intercept foraging sand flies. Two different nets were draped on fenced enclosures (10 by 10 m; 2 m high) in the central Jordan Valley. One enclosure was draped with a deltamethrin-impregnated net (PermaNet, 225 holes/in²). The holes of this net are sufficiently large to allow sand flies to pass through but not without coming in close contact with the mesh. The other enclosure was covered with SpiderNet (1,240 holes/in²) and sprayed with beta-cyfluthrin. Sand flies were captured inside and outside the enclosures before and after draping with the nets using CO₂-baited CDC traps or CDC light traps. Both barrier types exhibited >90% efficacy in blocking sand flies from entering the enclosures (P < 0.01). The SpiderNet exhibited high efficiency even before being sprayed with insecticide because the small mesh size physically prevented flies from passing through. In Ma'ale Adumim, a 60-m-long, 2-m-high PermaNet barrier was erected to intercept sand flies approaching houses from their natural habitats. Sand flies were monitored on all sides of the barrier using CO₂-baited CDC traps or CDC light traps. Results showed a 60% reduction in the mean number of sand flies trapped behind the net compared with the untreated areas adjacent to it (P < 0.05). Integrated vector control campaigns for reducing the burden of sand fly bites should consider vertical fine-mesh nets to reduce the numbers of sand flies arriving at inhabited areas.

The essential oil of catmint, Nepeta cataria L., contains nepetalactones, that, on hydrogenation, yield the corresponding dihydronepetalactone (DHN) diastereomers. The DHN diastereomer (4R,4aR,7S,7aS)-4,7-dimethylhexahydrocyclopenta[c]pyran-l(3H)-one, DHN 1) was evaluated as mosquito repellent, as was the mixture of diastereomers {mostly (4S,4aR,7S,7aR)-4,7-dimethylhexahydrocyclopenta[c]pyran-1(3H)-one, DHN 2} present after hydrogenation of catmint oil itself. The repellency of these materials to Aedes aegypti L. and Anopheles albimanus Wiedemann mosquitoes was tested in vitro and found to be comparable to that obtained with the well-known insect repellent active ingredient N,N-diethyl-3-methylbenzamide (DEET). DHN 1 and DHN 2 also repelled the stable fly, Stomoxys calcitrans L., in this study. DHN 1, DHN 2, and p-menthane-3,8-diol (PMD), another natural monoterpenoid repellent, gave comparable levels of repellency against An. albimanus and S. calcitrans. Laboratory testing of DHN 1 and DHN 2 using human subjects with An. albimanus mosquitoes was carried out. Both DHN 1 and DHN 2 at 10% (wt:vol) conferred complete protection from bites for significant periods of time (3.5 and 5 h, respectively), with DHN2 conferring protection statistically equivalent to DEET. The DHN 1 and DHN 2 diastereomers were also efficaceous against black-legged tick (Ixodes scapularis Say) nymphs.

The two fish species Betta splendens (Regan) and Poecilia reticulata (Peters) are known predators of Aedes aegypti (L., 1762) larvae. Both species have been used for biological control in northeastern Brazil. However, the feasibility of these fish for the control of Ae. aegypti larvae in domestic containers may be limited by their survival in chlorinated water, as supplied by the public water system. We exposed fish to three different concentrations of chlorine: 1, 1.5, and 2.0 mg/liter. All B. splendens survived at 1.0 mg/liter chlorine concentration; 72.5 and 39.3% of B. splendens survived chlorine concentrations of 1.5 and 2.0 mg/liter, respectively. In contrast, only 4.4% of P. reticulata survived at a chlorine concentration of 1.0 mg/liter. We conclude that B. splendens may be an appropriate species for biological control of Ae. aegypti in domestic water tanks.

Previous in vitro studies showed that molecules in an extract of the mite Sarcoptes scabiei variety canis De Geer could modulate the secretion of cytokines from cultured normal human epidermal keratinocytes and dermal fibroblasts in the absence of proinflammatory cytokines in the cell culture media. The purpose of this study was to investigate whether scabies extract could also modulate cytokine and chemokine secretion from epidermal keratinocytes and dermal fibroblasts in the presence of proinflammatory cytokines that are likely present in the scabietic lesion in vivo. In particular, could the downmodulating properties of this ectoparasitic mite on skin cells be maintained in the presence of proinflammatory cytokines? We found that even in the presence of the proinflammatory cytokines interleukin (IL)-1α, IL-1β, and a mixture of tumor necrosis factor (TNF) α IL-17, scabies extract still downregulated the levels of IL-8 secretion from keratinocytes and fibroblasts and of granulocyte/macrophage-colony stimulating factor (GM-CSF) secretion from fibroblasts that were induced by stimulation of the cells with proinflammatory cytokines alone. This study also showed that scabies molecules induced secretions of growth-related oncogene α. (GROα), transforming growth factor α (TGFa), and cutaneous T-cell attracting chemokine (CTACK) from keratinocytes and IL-6 and granulocyte-colony stimulating factor (G-CSF) from fibroblasts. These findings, coupled with the previous findings that molecules in scabies extract could downregulate expression of intracellular adhesion molecule-1 (ICAM-1) and E-selectin by normal dermal microvascular endothelial cells and secretion of IL-1ra from keratinocytes, suggest that multiple factors from scabies mites play a role in the characteristic delayed inflammatory response to a primary infestation with S. scabiei. These are adaptations that favor invasion of the host by the parasite.

n a previous study, the highly pathogenic avian influenza (HPAI) H5N1 viruses were isolated from blow flies collected at the Tamba Town of Kyoto prefecture during the outbreak period in March 2004. In this study, we carried out virus exposure experiments to investigate whether the H5N1 virus would survive in a blow fly, Calliphora nigribarbis. The virus exposure experiments showed that the H5N1 influenza virus was isolated from the crop and intestine of C. nigribarbis for at least 24 h, and the viruses remained viable with titers ranging from 0.5 to 4.63 TCID₅₀. This result suggests that C. nigribarbis could possibly transport the H5N1 virus over a distance of 2 km, which is the distance they can migrate within 24 h.

Several outbreaks of Rocky Mountain spotted fever have occurred in recent years in Colombian communities close to the border with Panama. However, little is known about rickettsiae and rickettsial diseases in eastern Panamanian provinces, the Darien Province and the Kuna Yala, located north of the endemic area in Colombia. In 2007, 289 ticks were collected in several towns from dogs, horses, mules, cows, and pigs. DNA was extracted from 124 Dermacentor nitens, 64 Bhipicephalus sanguineus, 43 Amblyomma ovale, 35 A. cajennense, 10 Boophilus microplus, 4 A. oblongoguttatum, and 9 A. cajennense nymphs. SYBR-Green polymerase chain reaction assays targeting a fragment of the OmpA and 16S rRNA genes were used for detection of DNA of the spotted fever group rickettsiae (SFGR) and Anaplasmataceae (Anaplasma and Ehrlichia), respectively. In total, 37.4% ticks were positive for SFGR, including 20.3% R. sanguineus, 27.9% A. ovale, 25.8% D. nitens, 50% B. microplus, 50% A. oblongoguttatum, and 100% A. cajennense. The presence of Rickettsia amblyommii DNA was confirmed by sequencing in A. cajennense, A. oblongoguttatum, A. ovale, B. microplus, and R. sanguineus. DNA of R. rickettsii was only detected in one D. nitens collected from a horse in Santa Fe, Darien Province. Prevalence of Anaplasmataceae varied from 6.3% in R. sanguineus to 26.5% in A. cajennense. DNA of Ehrlichia chaffensis was found in three D. nitens and three A. cajennense from horses. This is the first study providing molecular characterization and prevalence information on SFGR in ticks from these areas and thus will be helpful for future evaluations of the risk of rickettsial diseases for individuals living in this region.

We set out no. 17 Trinidad traps baited with hamsters at a swamp in Tennessee, where recent eastern equine encephalomyelitis (EEE) equine outbreaks had occurred, to determine which mosquito species at this site were attracted to these traps. We also set out CO₂-baited CDC light traps for comparison. Of 11 species present in CO₂-baited CDC light traps, only Culex (Melanoconion) erraticus (Dyar and Knab) blood fed on hamsters in the Trinidad traps. Significantly less Cx. erraticus mosquitoes entered Trinidad traps per trap night (mean = 17.6, median = 6.0) compared with CDC traps (mean = 38.7, median = 35.0). Advantages and disadvantages in using hamster-baited no. 17 Trinidad traps compared with CO₂-baited CDC light traps to capture Cx. erraticus mosquitoes are discussed. Further understanding of the feeding behavior and ecology of this potential vector is warranted.

Vesicular stomatitis New Jersey virus (VSNJV) is an insect-transmitted Rhabdovirus causing vesicular disease in domestic livestock including cattle, horses, and pigs. Natural transmission during epidemics remains poorly understood, particularly in cattle, one of the most affected species during outbreaks. This study reports the first successful transmission of VSNJV to cattle by insect bite resulting in clinical disease. When infected black flies (Simulium vittatum Zetterstedt) fed at sites where VS lesions are usually observed (mouth, nostrils, and foot coronary band), infection occurred, characterized by local viral replication, vesicular lesions, and high neutralizing antibody titers (>1: 256). Viral RNA was detected up to 9 d postinfection in tissues collected during necropsy from lesion sites and lymph nodes draining those sites. Interestingly, when flies were allowed to feed on flank or neck skin, viral replication was poor, lesions were not observed, and low levels of neutralizing antibodies (range, 1:8–1:32) developed. Viremia was never observed in any of the animals and infectious virus was not recovered from tissues on necropsies performed between 8 and 27 d postinfection. Demonstration that VSNJV transmission to cattle by infected black flies can result in clinical disease contributes to a better understanding of the epidemiology and potential prevention and control methods for this important disease.

The survival rate of infected vectors represents one of the fundamental components that influence the transmission dynamics of mosquito-borne diseases. Despite the occurrence of a number of studies investigating mosquito survival after infection with filarial worms, there remains conflicting evidence from both laboratory and field experiments as to the existence and mechanism for parasite-induced mortality among filarial mosquitoes. Here, we used a mixed effects meta-analytical framework to combine the data from all available vector—human host blood feeding experiments to evaluate the evidence for the impact of parasite load on the mortality rates of the three major lymphatic filariasis transmitting mosquito genera, Culex, Aedes, and Anopheles mosquitoes, over the extrinsic incubation period of parasitic infection. The results show that, despite the application of this approach, or in the case of Anopheles using a convention fixed effects logistic regression analysis supplemented with additional survival analysis of longitudinal data, no strong association between mortality rate and microfilariae (mf) uptake for either of the three mosquito genera is apparent in the combined data. Instead, a key finding is that study effects played a more crucial role in determining the levels of mortality observed in these experimental studies. This was most revealing in the case of Culex, given that the largest single study in terms of both the number of data points and range of mf intensities, in contrast to smaller studies, showed a significant positive association between mf intensity and mortality, indicating that in this genus at least, the detrimental effect of infection may be manifested only at the highest mf intakes. Although no density dependence in vector mortality was also observed for Aedes, possibly because of the use of restricted human mf intensity range in previous studies, an intriguing finding was that a significantly higher overall mortality was observed for this genus over mf intake ranges that produced much less corresponding mortality in Culex and Anopheles. The results also indicate that currently very little can be said about the survival rate of Anopheles mosquitoes infected with filarial worms because of the striking paucity of data for this genus. Further studies, using standardized methods and covering an appropriate range of mf uptake intensities and using study frameworks that allow the design and comparison of data from both experimental and field experiments, are clearly indicated if we are to reliably quantify the likely effect of filarial infection on vector survival.

Culex pipiens pipiens L. populations on Cyprus were sampled over a 6-yr period from 2002 to 2008 to evaluate the status of insecticide resistance toward the insecticides temephos, chlorpyrifos, and permethrin and to study susceptibility levels toward the recently introduced bacterial insecticide Bacillus thuringiensis subsp. israelensis De Barjac and the juvenile hormone analog, methoprene. Susceptibility to the three conventional chemical insecticides varied between different collections, with most collections showing moderate or low resistance. The 2004 Akrotiri collection had the highest temephos resistance ratio, 167-fold at the LC₉₅, although later sampling showed that the population returned to susceptibility after treatments stopped. Chlorpyrifos resistance was generally higher than temephos resistance. Four collections showed high resistance, and the resistance ratios of two collections were notably high with resistance ratios of 110- and 248-fold at the LC₉₅. Three collections showed high permethrin resistance (22.5-, 23.9-, and 86.3-fold). The frequency of elevated esterase activity in populations was estimated using a filter paper test, and frequencies varied from 0.9 to 65% among collections. The levels of temephos resistance and the frequency of elevated esterases in this survey were generally lower than in earlier reports, suggesting a decline in temephos resistance. Dose—response values for B. thuringiensis subsp. israelensis covered an approximate eight-fold range, but no resistance was detected. Methoprene values showed a 4.7-fold and 16-fold range at the LC₅₀ and LC₉₅, respectively. Two populations showed significant resistance ratios at the LC₉₅. These data are discussed in relation to the changes in larval control practices underway in Cyprus.

Theileria parva is the etiologic agent of East Coast fever, an economically important disease of cattle in sub-Saharan Africa. This protozoan parasite is biologically transmitted by Rhipicephalus appendiculatus (Neumann) (Acari: Ixodidae). An understanding of the vector—parasite interaction may aid the development of improved methods for controlling transmission. We developed quantitative polymerase chain reaction (qPCR) and nested PCR (nPCR) assays targeting the T. parva-specific p104 gene to study T. parva pathogenesis in two strains of R. appendiculatus that had previously been selected to be relatively more (Kiambu) or less (Muguga) susceptible to infection. Nymphs from both strains were fed simultaneously to repletion on acutely infected calves. Nymphs from the Kiambu strain showed significantly higher engorgement weights compared with Muguga strain nymphs. Immediately after engorgement qPCR confirmed that nymphal Kiambu ticks had significantly higher parasite loads at repletion than Muguga nymphs. By 12 d postengorgement, parasites were below quantifiable levels but could be detected by nPCR in 83–87% (Muguga and Kiambu, respectively) of nymphs. After the molt, adult feeding on naïve cattle stimulated parasite replication in the salivary glands. PCR detected significantly more infected ticks than microscopy, and there was a significant difference between the two tick strains both in the proportion of ticks that develop salivary gland infections, and in the number of parasites within infected salivary glands. These data confirm that although both tick strains were competent vectors, Kiambu is both a significantly more susceptible and a more efficient host for T. parva than Muguga. The mechanisms that contribute to the levels of susceptibility and efficiency are unknown; however, this study lays the groundwork for a comparison of the transcriptome of these tick strains, the next step toward discovering the genes involved in the tick-parasite interaction.

The aim of this study was to evaluate the presence of filamentous fungi and yeasts on the external surface of ants at hospitals. From March 2007 to February 2008, 2,899 ants were evaluated in two public hospitals in the city of Fortaleza, Ceará, in northeastern Brazil. The ants were attracted by nontoxic baits, distributed within critical and semicritical hospital areas. The fungi were identified through macro-and micromorphological analysis, biochemical profile, and growth in chromogenic medium. From this study, 5 genera and 13 species of ants were identified, from critical (8% of the collected ants) and semicritical (92%) areas, during the daytime (48%) and nighttime (52%) periods. In the mycological analysis, 75% of the ants were fungi carriers, with the species Tapinoma melanocephalum and species from the genus Pheidole having the most potential as carriers of airborne fungi (75 and 18%, respectively) and yeasts (6 and 1%, respectively). In summary, ants act as carriers of airborne fungi and yeasts, including some pathogenic species.

The response of Aedes albopictus to the BG-Sentinel, Omni-directional-Fay-Prince, and Mosquito MagnetX traps was evaluated in four suburban and four sylvatic habitats in north-central Florida to ascertain potential height preference of this species. These traps, which are primarily designed to attract diurnal mosquitoes, were set at 1 and 6 m and were evaluated during 40 trapping periods over 4 mo. We collected 45,640 mosquitoes, representing 26 species from 10 genera, the most common being Aedes albopictus, Ae. vexans, Coquilletidia perturbons, Culex nigripalpus, Aedes infirmatus, Ae. triseriatus, and Psorophora ferox. Although significantly more Ae. albopictus were captured at 1 m above ground than at 6 m, fewer were captured in sylvatic habitats than suburban habitats. Although not statistically different, the BG-Sentinel caught more Ae. albopictus compared with the other two traps regardless of locale. These results suggest that, although Ae. albopictus was captured as high as 6 m, the majority seek hosts at or below 1-m heights. This further supports prior research that, although Ae. albopictus has been shown to disseminate West Nile virus, it has not been implicated as a major vector for the virus, which is likely because of its propensity to feed on ground-dwelling hosts. The study also shows how trap type, trap heights, and environments influence sampling estimates when determining species abundance.

Aedes densonucleosis virus (family Parvoviridae, genus Brevidensovirus, AeDNV) is a mosquito pathogen that increases Aedes aegypti larval mortality and reduces adult life span. We conducted three laboratory population cage trials, each lasting 16–25 wk. We tested two broad hypotheses. First, Ae. aegypti raised in containers seeded with 10⁸ AeDNV genome equivaIents/ml (geq/ml), a concentration feasible for field application, increase AeDNV to concentrations that cause significant adult and larval mortality. Second, infected female mosquitoes disperse AeDNV to uninfected larval habitats. In hypothesis 1, we addressed the rate at which infected larvae secrete virus, how AeDNV titers change in seeded containers over time, whether AeDNV decays over time, and whether AeDNV exposed populations are reduced. In hypothesis 2, we monitored AeDNV concentrations in novel containers after oviposition by infected females. Both hypotheses were supported. Larvae increased AeDNV, secreting virus at a rate of 2.14 × 10⁶ geq/larva/d when exposed to 10⁸ geq/ml. AeDNV titers reached an asymptote of 10¹⁰ geq/ml by week 10 in seeded containers. AeDNV decayed by 1 log every 4 d as indicated by a reduction in larval mortality. Adult population size was reduced in treated populations. Infected females dispersed AeDNV to novel containers, with titers reaching 10⁸ geq/ml. The parameters were used in a Leslie-Lewis matrix model. This model predicted that AeDNV negatively affects Ae. aegypti densities and population structure and thus vectorial capacity.

Standard surveillance traps in North America for adult Aedes albopictus (Skuse) (Diptera: Culicidae), an invasive mosquito with public health implications, are currently ineffective. We compared the efficacy of the BG-Sentinel trap (BGS) with and without lures (BG-lure, octenol, and CO₂), the Centers for Disease Control and Prevention light trap (CDC) with and without lures, and the gravid trap (GT) for Ae. albopictus collection in two urban sites in New Jersey. The BGS with or without lures collected more Ae. albopictus compared with other trap configurations and was more specific for Ae. albopictus. In Camden County, the BGS with lures collected three times more Ae. albopictus than the CDC (with CO₂ only) and five times more than the GT. In Mercer County, BGS with lures collected the most mosquitoes, with 3 times more Ae. albopictus than the CDC with all lures and 50 times more than the GT. The BGS collected more male Ae. albopictus than other traps in both counties, providing further population monitoring. The GT and BGS provided a relative measure of the enzootic activity of West Nile virus in Culex spp. and the potential epidemic activity of WNV in Ae. albopictus. The BGS provides effective chemical and visual cues for host-seeking Ae. albopictus and should be used as a part of existing surveillance programs and new initiatives targeting this mosquito.

Ticks (n = 252) were collected from five wild-caught reptile species during routine trapping in the Djukbinj National Park and Fogg Dam Reserve, Northern Territory, Australia. Pooling of ticks (one to four ticks per pool), according to sex or host animal, resulted in 187 samples used for screening for the presence of Rickettsia species via molecular methods. Rickettsia DNA was detected via the amplification of the gltA, ompA, and ompB genes in 57 (34%) of the 187 tick samples, all of which contained only the tick Amblyomma fimbriatum Koch (Acari: Ixodidae). Further amplification and sequencing of nine of the positive samples (4.8%) for the gltA, ompA, ompB, 17-kDa, and 16S rRNA genes identified a Rickettsia species sharing closest identity to Rickettsia tamurae. In addition, amplification and sequencing of the 16S rRNA gene detected in the same tick samples the presence of a Francisella species closely related to other tick-borne Francisellae identified in ticks from the Northern Hemisphere.

Adult mosquitoes (Diptera: Culicidae) were collected in 2007 and tested for specific viruses, including West Nile virus, as part of the ongoing arbovirus surveillance efforts in the state of Iowa. A subset of these mosquitoes (6,061 individuals in 340 pools) was further tested by reverse transcription-polymerase chain reaction (RT-PCR) using flavivirus universal primers. Of the 211 pools of Culex pipiens (L.) tested, 50 were positive. One of 51 pools of Culex tarsalis Coquillet was also positive. The flavivirus minimum infection rates (expressed as the number of positive mosquito pools per 1,000 mosquitoes tested) for Cx. pipiens and Cx. tarsalis were 10.3 and 1.2, respectively. Flavivirus RNA was not detected in Aedes triseriatus (Say) (52 pools), Culex erraticus (Dyar & Knab) (25 pools), or Culex territans Walker (one pool). Sequence analysis of all RT-PCR products revealed that the mosquitoes had been infected with Culex flavivirus (CxFV), an insect-specific virus previously isolated in Japan, Indonesia, Texas, Mexico, Guatemala and Trinidad. The complete genome of one isolate was sequenced, as were the envelope protein genes of eight other isolates. Phylogenetic analysis revealed that CxFV isolates from the United States (Iowa and Texas) are more closely related to CxFV isolates from Asia than those from Mexico, Guatemala, and Trinidad.

Imported from Africa in the 1700s and despite frequent modern eradication efforts, Amblyomma variegatum (F.) spread through the Caribbean by cattle transport, small ruminants, and migrating birds. A. variegatum is a vector for Rickettsia africae, the causative agent of African tick bite fever, and Ehrlichia ruminantium, the causative agent of heartwater. We examined 95 A. variegatum and six Rhipicephalus (Boophilus) microplus (Canestrini) collected from cattle at an abattoir in Antigua. Engorged tick extracts adsorbed on Nobotu filter paper strips and new nested polymerase chain reaction (PCR) assays for E. ruminantium and Dermatophilus congolensis were used to evaluate these ticks for the presence of these pathogenic bacteria. Amblyomma ticks (62.4%) contained R.. africae DNA by PCR/restriction fragment length polymorphism analysis and DNA sequencing of the OmpA and 17-kDa antigen genes. Twenty Amblyomma and two Rh. microplus contained E. ruminantium DNA. No E. chaffeensis, Anaplasma phagocytophilum, Coxiella burnetii, or D. congolensis DNA was detected in these ticks. The continued presence of Am. variegatum in the Caribbean poses a significant risk of infection in cattle with E. ruminantium and in humans by R. africae. Eradication efforts are essential to prevent the further spread of Am. variegatum.

Ehodnius neglectus is frequently found in palm trees and bird nests in sylvatic environments. However, adult specimens infected by Trypanosoma cruzi have been invading houses in central Brazil. Analyzing and predicting the geographical distribution of this species may improve vector surveillance strategies for Chagas disease. Ecological niche modeling using the genetic algorithm for rule-set production (GARP) was applied to predict the geographical distribution of R. neglectus from occurrence records and a set of 23 predictor variables (e.g., temperature, precipitation, altitude, and vegetation). Additionally, the geographical distribution of R. neglectus was compared with the geographical distribution of four species of palm trees and two species of birds from the study region. The models were able to predict, with high probability, the occurrence of R. neglectus as a regular (although nonendemic) species of the Cerrado biome in central Brazil. Caatinga, Amazonian savanna, Pantanal, and the Bolivian Chaco appear as areas with lower probabilities of potential occurrence for the species. A great overlap was observed between the distribution of R. neglectus, palm trees (Acrocomia aculeata and Syagrus oleracea), and birds (Phacellodomus ruber and Pseudoseisura cristata). By including new records for R. neglectus (from both sylvatic and domestic environments), our study showed a distribution increase toward the west and northeast areas of Brazil in the “diagonal of open/dry ecoregions of South America”. These results should aid Chagas disease vector surveillance programs, given that household invasion by Rhodnius species maintains the risk of disease transmission and limits control strategies.

Mosquitoes collected during an outbreak of Rift Valley fever in Kenya from December 2006 to February 2007 were tested to isolate other mosquito-borne arboviruses circulating in the region. Twenty-seven virus isolations were made comprising seven viruses from three arbovirus families.

Two species of phlebotomine sand flies, Lutzomyia shannoni (Dyar) and Lutzomyia vexator (Coquillett), are reported for the first time from Kentucky and Ohio. L. vexator also is reported for the first time from Tennessee. These insects were found in a northeasterly band extending from southwestern Kentucky to southwestern Ohio. Both species were consistently captured from mid-July through September in 2006 and 2007 by using CO₂-baited Center for Disease Control light traps. Weekly sampling revealed that these flies are more abundant in the southern part of this band than in the northern part, but increasing densities throughout this new range indicate that the flies are currently expanding their range. Although both species have been reported further north along the Atlantic coast, and L. vexator along the Pacific coast, neither of them had been reported this far north along the Mississippi Valley. Previous reports established L. shannoni as far north as west central Tennessee and L. vexator in a similar spatial pattern in the eastern part of its range, extending as far north as northern Alabama. Whether the new records reported herein represent a northerly expansion of the geographic range of these species or are reflective of sampling changes is inconclusive. However, the former scenario could presage an increased prevalence of the diseases associated with this group of insects.