The Gulf Coast tick, Amblyomma maculatum Koch (Acari: Ixodidae), is a unique univoltine ectoparasite of seven vertebrate host classes in the Western Hemisphere that is increasingly recognized as a pest of livestock and wildlife, a vector of pathogens to humans and canines, and a putative vector of Ehrlichia ruminantium, the causal agent of heartwater, a fatal foreign animal disease of ruminants resident in the Caribbean. This review assembles current and historical literature encompassing the biology, ecology, and zoogeography of this tick and provides new assessments of changes in cyclical population distribution, habitat associations, host utilization, seasonal phenology, and life history. These assessments are pertinent to the emergence of A. maculatum as a vector of veterinary and medical importance, and its pest management on livestock and other animals.

The carrion beetle Oxelytrum discicolle (Brullé, 1840) has forensic importance in the Neotropical region. The age of their larvae can provide important information to estimate the minimum postmortem interval of the corpse. This study was designed to determine the larval stages of this species, using three morphological measurements, as follows: distance between dorsal stemmata, pronotal width, and total body length. Three instars were identified. Distance between dorsal stemmata and pronotal width were good discriminating variables to determine instars, whereas total length was widely variable. For each instar, intervals of three morphological measures are given.

Five new species of the genus Picobia are described and illustrated: 1) P. leucophaeus sp. nov. from the Laughing Gull Leucophaeus atricilla L. (Charadriiformes: Laridae) from Texas; 2) P. troglodytes sp. nov. from the House Wren Troglodytes aedon Vieillot (Passeriformes: Troglodytidae) from California; 3) P. cardinalis sp. nov. from the Northern Cardinal Cardinalis cardinalis (L.) (Passeriformes: Cardinalidae) from Texas; 4) P. carpodacus sp. nov. from the Purple Finch Carpodacus purpureus (Gmelin) (Passeriformes: Fringillidae) from California; and 5) P. psaltriparus sp. nov. from the Bushtit Psaltriparus minimus (Townsend) (Passeriformes: Aegithalidae) from Texas. Two avian species from the family Picidae (Piciformes) are recorded as new hosts for P. dryobatis (Fritsch, 1958): the Downy Woodpecker Picoides pubescens (L.) from Texas and the Ladder-backed Woodpecker Picoides scalaris (Wagler) from California. Additionally, all named species of the genus Picobia with their host associations and distributions are summarized in tabular form.

Species-specific differences encountered in the nucleotide sequences of a highly variable region of the 18S rRNA gene were used to design a multiplex polymerase chain reaction (PCR) assay for the identification of Phlebotomus papatasi Scopoli and Phlebotomus argentipes Annandale & Brunetti, vectors of Leishmania. This multiplex PCR assay uses a common forward primer and two reverse primers, which are specific for the two species. Amplification of a PCR product of size 788 bp indicates the presence of P. papatasi, whereas a product of size 677 bp indicates the presence of P. argentipes. The assay was found to be highly specific and sensitive.

Although quantitative surveillance data for immature stages of Aedes aegypti are often used to prioritize containers or specific types of containers for control, the relationship between immature and emergent adult populations under field conditions is largely unknown. We examined the relationships between abundance of III/IV instars and pupae, and emerging adult population for a series of water storage containers in southern Vietnam. A large proportion of III/IV instars failed to progress to adulthood, and the relationships between III/IV instars and adults were poor. Collected IV instars appeared to be nutritionally deprived, although their size and nutrient levels were not reliable indicators of emergence success. Conversely, pupal abundance was a good indicator of emerging adult populations, especially over the ensuing 48-h period. Although there were clear advantages of pupal surveillance over surveillance of III/IV instars for the estimation of adult mosquito productivity, there were practical limitations associated with the enumeration of pupae, and their comparatively low densities may preclude the identification of potentially productive containers.

Large-scale patterns in the seasonal abundance profiles of the arboviral mosquito vectors Culex tarsalis Coquillett and the species of the Culex pipiens complex were described based on a decade of counts from 868 New Jersey light traps located throughout the urban and agricultural areas of California. Mean seasonal abundance profiles varied markedly among hydrologic regions. For all species, abundance increased earlier and declined later in drier, warmer southern regions, but variation could not be explained fully by latitude. The observed patterns may be driven by temperature, availability of larval habitats, and for the Cx. pipiens complex, the taxonomic composition of local populations.

Reduced survival and future reproduction due to of current reproduction is a trade-off known as the cost of reproduction. Surprisingly, only a few studies have assessed the cost of reproduction in arthropod disease vectors, despite its effect on longevity, and thus on vectorial capacity. We evaluated the cost of reproduction on survival of Anopheles gambiae Giles by comparing mosquitoes that were denied exposure to the other sex, hereafter named virgins, and those that were allowed exposure to the other sex and mating, hereafter named mated. Merely 6 d of exposure to females with mating activity reduced male survival from a median of 17 d in virgins to 15 d in mated, indicating that male mating cost is substantial. The increase in mortality of mated males began several days after the exposure to females ended, indicating that mating is not associated with immediate mortality risk. Notably, body size was negatively correlated with male mortality in mated males, but not in virgins. The rate of insemination declined after 4 d of exposure to females, indicating that male mating capacity is limited and further supporting the hypothesis that mating is costly for males. Consistent with previous studies, female survival on sugar alone (median = 16 d) was shorter than on blood and sugar (median = 19 d), regardless if she was mated or virgin. Overall, survival of mated females was lower than that of virgins on a diet of blood and sugar, but no difference was found on a diet of sugar only. However, the cost of reproduction in females remains ambiguous because the difference in survival between virgin and mated females was driven by the difference between virgin (median = 19 d) and uninseminated females exposed to males (median = 17 d), rather than between virgin and inseminated females (median = 19 d). Accordingly, sperm and seminal fluid, egg development, and oviposition have negligible cost in terms of female survival. Only exposure to males without insemination decreased female survival, Nonetheless, if exposure to males under natural conditions is also associated with reduced survival, it might explain why females remain monogamous.

The effects of adult body size and nutritional regimen on the mean lethal time (LT) to death for the 50th (LT₅₀) and 90th centiles (LT₉₀) of large and small male and female Aedes albopictus (Skuse) were determined in the laboratory. The estimated LT₅₀ and the LT₉₀ for large/small females were significantly shorter for test populations receiving water (5.1/3.8 d) or water blood (6.7/5.0 d) than for those receiving sucrose (44.4/31.7 d) or sucrose blood (41.4/29.2 d). The estimated LT₅₀ and LT₉₀ for large/small males receiving water (respectively: 3.2/2.5 d and 4.8/3.8 d) were significantly shorter than for those receiving sucrose (respectively: 18.8/15.9 d and 34.9/32.6 d). The availability of sucrose to adult mosquitoes extended the maximum life span of large and small male and female Ae. albopictus by 6–8× when compared with the availability of water or water blood. Results indicate that sugar availability is more important than blood as a nutritional parameter for survival and longevity of Ae. albopictus in the laboratory.

Polymerase chain reaction analysis was performed to determine whether mosquito predators in wetland habitats feed on Anopheles gambiae sensu lato (s.l.) larvae. Aquatic mosquito predators were collected from six wetlands near Lake Victoria in Mbita, Western Kenya. This study revealed that the whole positive rate of An. gambiae s.l. from 330 predators was 54.2%. The order of positive rate was the highest in Odonata (70.2%), followed by Hemiptera (62.8%), Amphibia (41.7%), and Coleoptera (18%). This study demonstrates that the polymerase chain reaction method can determine whether aquatic mosquito predators feed on An. gambiae s.l. larvae if the predators have undigested An. gambiae s.l. in their midgut or stomach.

To assess the risk that indigenous mosquitoes in China are capable of transmitting and sustaining West Nile virus (WNV), four important Culex mosquito species, Culex tritaeniorhynchus, Culex modestus, Culex pipiens pallens, and Culex pipiens quinquefasciatus, were allowed to feed on the artificial infectious blood meal with WNV dose of 106,8 plaque-forming unit/ml and tested ≈2 wk later to determine infection and transmission rates. The results indicated that four Culex mosquitoes were competent laboratory vectors of WNV. The infection rates and transmission rates were statistical differences among different species of mosquito (χ² = 20.620, P = 0.000; χ² = 15.020, P = 0.005, respectively). The highest infection rate and transmission rate were obtained with Cx. tritaeniorhynchus (87.5 and 74.2%, respectively).

Stomoxyine flies (Stomoxys spp.) were collected in 10 localities of Thailand using the Vavoua traps. These localities represented four major ecological settings, as follows: small local dairy farms, large industrial dairy farms, a national park, and one elephant conservation area. Three species of stable flies were identified in the following proportions: Stomoxys calcitrans (91.5%), Stomoxys indicus (7.9%), and Stomoxys sitiens (0.6%). The number of flies collected differed significantly among collection sites (χ² = 360.15, df = 3, P < 0.05). The greatest number of stomoxyine flies was captured in dairy farms, Seasonal and daily activity of S. calcitrans was observed during a 1-yr period at two selected locations (Dairy Farming Promotion Organization of Thailand and Khao Kheow Open Zoo). S. calcitrans was more abundant during the rainy season (March–September), but was not associated with the total rainfall (r² = 0.0002, P > 0.05). Peak of daily flight activity of males S. calcitrans was at 1000 and 1600 h, whereas females showed an increase of activity all along the day until 1600 h. A better understanding of stomoxyine fly behavior related to patterns of daily activity will facilitate and improve the efficiency of fly control measures in private and government sectors.

The American house dust mite, Dermatophagoides farinae Hughes, and European house dust mite, Dermatophagoides pteronyssinus Trouessart, are major pests of medical importance throughout the developed world, causing atopic diseases such as asthma, rhinitis, and atopic dermatitis. Previous studies in our laboratory have shown that the behavioral responses of house dust mites toward volatiles from food sources could be assessed using a Y-tube olfactometer assay. The current study used this Y-tube assay to investigate house dust mite pheromones. A hexane extract of D. farinae, along with fractions of the extract prepared by microscale liquid chromatography over Florisil, were tested for behavioral activity. One of the chromatographic fractions was shown to be significantly attractive (P < 0.05) for D. farinae, compared with a solvent control. Coupled gas chromatography-mass spectrometry analysis of this behaviorally active fraction indicated that neryl or geranyl formate was the major component. Peak enhancement by gas chromatography, using authentic samples of the neryl and geranyl isomers prepared in high purity by chemical synthesis, confirmed the identity of the major peak as neryl formate. In Y-tube assays, male and female D. farinae and D. pteronyssinus both were significantly attracted to synthetic neryl formate at doses of 100 and 10 ng, respectively (P < 0.05). No significant differences were found for D. farinae and D. pteronyssinus when synthetic neryl formate and house dust mite extracts containing natural neryl formate were tested at the same level. Dynamic headspace collection of D. farinae and D. pteronyssinus colonies showed that neryl formate was released as a volatile organic compound by both species. Our study shows that neryl formate is an aggregation pheromone for D. farinae and D. pteronyssinus, and has the potential to be used as part of a novel lure-and-kill system for house dust mite control.

Aboveground and belowground populations of the mosquito Culex pipiens L. are traditionally classified as form pipiens and form molestus, respectively, and gene flow between forms is thought to be limited. Relatively few f. molestus populations have been found in the United States, which has hindered their study in North America. In this study, we used microsatellites to characterize a newly discovered population of f. molestus in Chicago, IL, and compared levels of genetic diversity and differentiation in aboveground and belowground populations from Chicago and New York City, NY. Levels of genetic diversity, as measured by expected heterozygosity and allelic richness, were markedly lower in both f. molestus populations. Allele frequencies were distinctly different between the two f. molestus populations, and some alleles were present in one belowground population and not the other. Pairwise F_ST values between populations indicated that f. molestus populations were highly divergent from each other, as well as from their associated aboveground populations. Cluster analysis suggested the most likely number of groups was three, with the four f. pipiens populations in one cluster, and each of the f. molestus populations in its own cluster. Admixture analysis detected a low number of hybrids, 8%, between forms. We also tested the efficacy of two assays purported to distinguish between the forms, the CQ11 assay and a restriction fragment-length polymorphism assay of the COI gene, and found neither assay reliable in this regard. Our findings support the hypothesis that f. molestus populations in Chicago and New York City arose from local aboveground populations.

Mendelian crosses were used to analyze the patterns of inheritance of Cry-toxin resistance in two colonies of Culex quinquefasciatus Say larvae resistant to bacterial toxins produced by Bacillus thuringiensis subsp. israelensis de Barjac. Resistance levels exceeded 1000-fold at 95% lethal concentration of the Cry11Aa-resistant colony (Cq11A). F₁ offspring of reciprocal crosses to a susceptible colony revealed autosomal inheritance and offspring were intermediate in resistance to the susceptible and resistant parental lines. Dose-response tests on backcross offspring were consistent with polyfactorial inheritance of resistance toward Cry11Aa and Cry4Aa Cry4Ba, whereas cross-resistance toward Cry11Ba best fit a monofactorial model. Resistance was 600-fold at 95% lethal concentration in the colony selected with Cry4A Cry4B (Cq4AB). Inheritance of resistance in F₁ offspring was autosomal and intermediate to the susceptible and resistant parents. Inheritance of Cry4Aa Cry4Ba and Cry11Ba resistance best fit a polyfactorial model in offspring of the Cq4AB backcross, whereas Cry11Aa-resistance inheritance fit a monofactorial model. Dominance values were calculated at different Cry-toxin concentrations for F₁ offspring of both resistant colonies; dominance generally decreased as treatment concentration increased. Resistance and cross-resistance remained stable in Cq11A and Cq4AB in the absence of insecticide pressure. Allelic complementation tests were complementary and suggested that Cq11A and Cq4AB evolved resistance to Cry toxins at common loci. The patterns of cross-resistance suggest cross-recognition of binding moieties by Cry11Aa, Cry4Aa Cry4Ba, and Cry11Ba in these Culex, which may be partly responsible for the toxin synergy characteristic of B. thuringiensis subsp. israelensis de Barjac.

Anopheles dirus females landing on humans inside experimental huts treated with residual applications of DDT or deltamethrin were observed during the wet season in Pu Teuy Village, Kanchanaburi Province, western Thailand. Two identical experimental huts were constructed in the fashion of typical local rural Thai homes. Pretreatment (baseline) human-landing collections (HLC) in both huts showed an early evening peak of activity between 1900 and 2000 h with no significant difference in numbers of mosquitoes captured between huts over a period of 30 collection nights. During posttreatment HLC, female mosquitoes continued to show greater landing activity inside huts fitted with insecticide-treated panels during the first half of the evening compared with the second half. A greater number (proportion) of An. dims females landed on humans in the hut treated with deltamethrin compared with DDT. Comparing pre- and posttreatment HLC, the DDT-treated hut showed a 79.4% decline in attempted blood feeding, whereas exposure to deltamethrin resulted in a 56.3% human-landing reduction. An odds ratio was performed to demonstrate the relative probability (risk) of mosquitoes entering and attempting to blood feed in the two treated huts compared with untreated control huts. Mosquitoes were ≈times less likely to land on humans inside a DDT-treated hut compared with the deltamethrin-treated hut. Although both chemicals exerted strong excitatory responses, DDT appears to have a more pronounced and significant (P = 0.002) effect on behavior than deltamethrin, resulting in greater movement away from the insecticide source and thus potential reduction of blood-feeding activity.

Chemicals can protect humans from the bites of hemophagous arthropods through three different primary actions: irritancy (excitation), repellency, or toxicity, actions that can be evaluated using a laboratory-based assay system, In this study, the deterrent and toxic actions of three synthetic pyrethroids and DDT were characterized on six field strains of Aedes aegypti from Thailand under laboratory-controlled conditions using the high throughput screening system. All six strains showed significant contact irritant responses to the three synthetic pyrethroids, but significantly weaker irritant responses to DDT, Marked repellency responses were seen in all six Ae. aegypti test strains exposed to DDT, whereas the synthetic pyrethroids resulted in greater toxicity than DDT under similar test conditions. Although significantly different in actions, irritancy and repellency may reflect and be influenced by the background insecticide susceptibility status of a particular mosquito population. Results from this study can be used to guide decision making regarding more effective Ae. aegypti adult control in Thailand.

In recent decades, the majority of human plague cases (caused by Yersinia pestis) have been reported from Africa. In an effort to reduce the risk of the disease in this area, we evaluated the efficacy of a host-targeted rodent bait containing the insecticide imidacloprid for controlling fleas on house-dwelling commensal rodents in a plague-endemic region of northwestern Uganda. Results demonstrated that the use of a palatable, rodent-targeted, wax-based bait cube was effective at reducing the prevalence of fleas on commensal rodents and flea burdens on these animals at day 7 postbait exposure, but lacked significant residual activity, allowing flea populations to rebound in the absence of additional bait applications. Our results indicate the use of a palatable host-targeted bait block containing imidacloprid was an effective technique for quickly reducing flea numbers on rodents in northwest Uganda and, thus, could be useful for lowering the potential risk of human flea bite exposures during plague outbreaks if applied continuously during the period of risk.

Field efficacy studies comparing two formulations of deet (N,N-diethyl-3-methylbenzamide) against mosquitoes were conducted on Ndendo Island, Solomon Islands. The repellent study was conducted at Pala village in November 2008, and the only mosquito species collected was Anopheles farauti Laveran. A formulation containing 35% deet in a gel provided >95% protection for 2 h, whereas a formulation containing 40% deet in ethanol in a spray applicator provided >95% for only 1 h. This field study demonstrated again that repellents containing deet provide a relatively short period of complete protection against Anopheles spp.

The horn fly, Haematobia irritans L., is an obligate blood-feeding fly and the primary insect pest parasitizing cattle in the United States. Pesticide resistance has become a substantial problem for cattle producers, and although several mechanisms of resistance are possible, target site resistance is the most important mechanism preventing control of this fly in the United States and possibly other countries. We developed a multiplex polymerase chain reaction assay to detect the known target site, pyrethroid resistance-associated mutation in the horn fly and a recently reported G262A mutation in the horn fly acetylcholinesterase, the target site for organophosphates. As expected, the pyrethroid resistance target site mutation was found in fly populations from Texas, Louisiana, Washington, Georgia, Mexico, and Brazil. This mutation was found to have a gender bias as it was more prevalent in females than males. The G262A acetylcholinesterase mutation was found in Texas, Louisiana, Washington, Georgia, and Mexico, but not Brazil. There was no gender bias in the occurrence of this mutation, and there was no correlation between the occurrence of the kdr and the G262A mutations. Unlike the case with the pyrethroid target site mutation, the presence of G262A did not appear to exclusively provide the level of resistance required to account for bioassay results. It is likely an additional mutation (s) occurs in the target site and/ or a metabolic resistance mechanism exists in organophosphate-resistant horn fly populations.

Field efficacy of an emulsifiable concentrate formulation of the entomopathogenic fungus Metarhizium anisopliae strain F52 for the control of Ixodes scapularis nymphs was evaluated at residential sites in northwestern Connecticut in 2007. Two spray applications with two rates, 3.2 × 10⁵ and 1.3 × 10⁶ spores/cm², were made: the first on 8–9 May, 2–3 wk before nymphal activity, and the second on 29 June or 2 July when ticks were active. There was no significant difference in nymphal abundance between the three treatment groups (P = 0.490) after the first application, indicating that preseason or early applications are not effective, despite a bioasaay with yellow mealworms that showed spores in the treated areas was infective for at least 1 mo postapplication. By contrast, there was a significant difference in the number of nymphs collected between the treatments and control 3 wk (F = 16.928, df = 2, P < 0.001) and 5 wk (F = 6.627, df = 2, P = 0.002) after the second application. During the 3 wk after the second application, 87.1 and 96.1% fewer ticks were collected from lower and higher rate-treated sites, respectively, and after 5 wk, tick reductions were 53.2 and 73.8%, respectively. Over one- third (36.4% of 173) of the nymphs collected from the treated sites developed mycosis from M. anisopliae. The application of M. anisopliae strain F52 could provide another tool for the integrated approach to managing ticks in the residential landscape.

A full-length cDNA encoding house dust mite allergen Der f 7 from Dermatophagoides farina. (Acari: Pyroglyphidae) from China was cloned, sequenced, and successfully expressed. A reference sequence (GenBank accession AY283292) was used to design polymerase chain reaction primers. Analysis revealed eight mismatched nucleotides in five Der f 7 cDNA clones, and the projected amino acid sequence contained six incompatible residues. These results suggest that the sequence of Der f 7 may be polymorphic. Further bioinformatic analysis revealed that the mature Der f 7 allergen had a molecular mass of ≈21.88 kDa and a theoretical isoelectric point of 4.90. Der f 7 protein secondary structure was composed of α helix (56.63%), extended strand (5.10%), and random coil (38.27%). Group 7 allergens are present in Pyroglyphidae, Acaridae, and Glycyphagidae families, and homology analysis revealed a 86% similarity between Der f 7 and Der p 7. Furthermore, a phylogenetic tree constructed of group 7 allergens from different mite species revealed that Der f 7 and Der p 7 clustered with 100% bootstrap support. Bioinformatics-driven characterization of Der f 7 allergen as conducted in this study may contribute to diagnostic and therapeutic applications for dust mite allergies.

Studies have shown that molecules in an extract made from bodies of the ectoparasitic mite, Sarcoptes scabiei De Geer, modulate cytokine secretion from cultured human keratinocytes and fibroblasts. In vivo, in the parasitized skin, these cells interact with each other by contact and cytokine mediators and with the matrix in which they reside. Therefore, these cell types may function differently together than they do separately. In this study, we used a human skin equivalent (HSE) model to investigate the influence of cellular interactions between keratinocytes and fibroblasts when the cells were exposed to active/burrowing scabies mites, mite products, and mite extracts. The HSE consisted of an epidermis of stratified stratum corneum, living keratinocytes, and basal cells above a dermis of fibroblasts in a collagen matrix. HSEs were inoculated on the surface or in the culture medium, and their cytokine secretions on the skin surface and into the culture medium were determined by enzyme-linked immunosorbent assay. Active mites on the surface of the HSE induced secretion of cutaneous T cell-attracting chemokine, thymic stromal lymphopoietin, interleukin (IL)1α, IL-1β, IL-1 receptor antagonist (IL-1ra), IL-6, IL-8, monocyte chemoattractant protein-1, granulocyte/macrophage colony-stimulating factor, and macrophage colony-stimulating factor. The main difference between HSEs and monocultured cells was that the HSEs produced the proinflammatory cytokines IL-1α and IL-1β and their competitive inhibitor IL-1ra, whereas very little of these mediators was previously found for cultured keratinocytes and fibroblasts. It is not clear how the balance between these cytokines influences the overall host response. However, IL-1ra may contribute to the depression of an early cutaneous inflammatory response to scabies in humans. These contrasting results illustrate that cell interactions are important in the host's response to burrowing scabies mites.

To determine which arthropods should be targeted for control should Rift Valley fever virus (RVFV) be detected in North America, we evaluated Culex erraticus (Dyar and Knab), Culex erythrothorax Dyar, Culex nigripalpus Theobald, Culex pipiens L., Culex quinquefasciatus Say, Culex tarsalis Coquillett, Aedes dorsalis (Wiedemann), Aedes vexans (Meigen), Anopheles quadrimaculatus Say, and Culicoides sonorensis Wirth and Jones from the western, midwestern, and southern United States for their ability to transmit RVFV. Female mosquitoes were allowed to feed on adult hamsters inoculated with RVFV, after which engorged mosquitoes were incubated for 7–21 d at 26°C, then allowed to refeed on susceptible hamsters, and tested to determine infection, dissemination, and transmission rates. Other specimens were inoculated intrathoracically, held for 7 d, and then allowed to feed on a susceptible hamster to check for a salivary gland barrier. When exposed to hamsters with viremias ≥108.8 plaque-forming units/ml blood, Cx. tarsalis transmitted RVFV efficiently (infection rate = 93%, dissemination rate = 56%, and estimated transmission rate = 52%). In contrast, when exposed to the same virus dose, none of the other species tested transmitted RVFV efficiently. Estimated transmission rates for Cx. erythrothorax, Cx. pipiens, Cx. erraticus, and Ae. dorsalis were 10, 8, 4, and 2%, respectively, and for the remaining species were ≤1%. With the exception of Cx. tarsalis and Cx. pipiens, all species tested had moderate to major salivary gland barriers. None of the C. sonorensis became infected and none of the An. quadrimaculatus tested transmitted RVFV by bite, even after intrathoracic inoculation, indicating that these species would not be competent vectors of RVFV. Although Ae. vexans from Florida and Louisiana were relatively efficient vectors of RVFV, specimens of this species captured in Colorado or California were virtually incompetent, illustrating the need to evaluate local population for their ability to transmit a pathogen. In addition to laboratory vector competence, factors such as seasonal density, host feeding preference, longevity, and foraging behavior should be considered when determining the potential role that these species could play in RVFV transmission.

Amblyomma americanum was confirmed as a competent vector in the transmission of Cytauxzoon felis to domestic cats. Infection with C. felis was produced and replicated in four domestic felines by the bite of A. americanum adults that were acquisition fed as nymphs on a domestic cat that survived cytauxzoonosis. Numerous attempts to transmit C. felis with Dermacentor variabilis at the same time were not successful. All cats upon which infected A. americanum were transmission fed exhibited disease typical of cytauxzoonosis, and the eitiologic agent's presence was confirmed. Clinical signs including fever, inappetence, depression, and lethargy were observed beginning 13 d postinfestation. Pale mucus membranes, splenomegaly, icterus, and dyspnea were also observed during the course of the disease. Rectal temperatures of the C. felis-infected principal cats fluctuated from high to subnormal before returning to the normal range. Clinical signs of cytauxzoonsis improved by 24 d postinfestation in all but one cat, with survivors remaining parasitemic and subclinically infected with C. felis. Unengorged A. americanum and D. variabilis were collected from wild habitats to determine the minimum infection rate of C. felis in ticks from an enzootic area. Infection of C. felis was found only in wild-collected A. americanum. The minimum infection rate of C. felis in A. americanum was 0.5% (one of 178) in males, 0.8% (three of 393) in nymphs, and 1.5% (three of 197) in females. We found no wild-collected D. variabilis infected with C. felis. Our results confirm that A. americanum is a primary vector of C. felis.

The cliff swallow (Petrochelidon pyrrhonota, Vieillot) could potentially play an important role in the transmission of West Nile virus (WNV) because of its breeding ecology, reservoir competence status, and potentially high natural exposure rates. These birds nest in colonies and their nests are occupied year round by swallow bugs (Oeciacus vicarius, Hovarth), hematophagus ectoparasites that feed primarily on cliff swallows. Swallow bugs are most likely exposed to WNV while feeding on infectious blood of cliff swallows and, thus, if competent vectors, could contribute to initiation and maintenance of seasonal WNV transmission. In addition, swallow bugs remain within nests year round and, therefore, if persistently infected and competent vectors, they could provide an overwintering mechanism for WNV. We tested the hypothesis that swallow bugs become infected with WNV through direct abdominal inoculation or ingestion of infectious blood meals. We observed that swallow bugs did not maintain or amplify WNV, and infectious virus titers within bugs declined over 15 d postexposure. These results suggest that swallow bugs may not be competent vectors of WNV, and therefore are unlikely to play a significant role in transmission.

Anthroponotic visceral leishmaniasis, transmitted by Phlebotomus argentipes Annandale & Brunetti (Diptera: Psychodidae) sand flies, is regarded as a major problem of public health importance in the Indian subcontinent. Understanding the feeding behavior of the vector can be used to investigate changes in human-vector contact during intervention programs. An enzyme-linked immunosorbent assay (ELISA) was modified to make it suitable to identify the origin of P. argentipes and Phlebotomus papatasi Scopoli (Diptera: Psychodidae) blood meals. The sensitivity and specificity of the precipitin ring test and ELISA were compared, as well as the stability of the tests across different stages of blood meal digestion. The ELISA was more sensitive and specific than the precipitin test for identifying the sources of blood meals. When using the ELISA method with a plate reader, it was possible to obtain 100% sensitivity and specificity. When comparing the techniques across digestion stages, it was found that there was a drop in sensitivity, 48 and 72 h postblood meal for precipitin and visually read ELISA, respectively. However, the sensitivity of the ELISA using a plate reader was not altered by the digestion time. The feeding habits of P. argentipes and P. papatasi from the Terai region of Nepal, determined by the ELISA developed, showed P. papatasi to be highly anthropophilic, and P. argentipes appeared to feed both on humans and animals, in particular bovines.

Although there is some detailed research on anaphylactic reactions to Hymenoptera venom, there continues to be little epidemiological data about the distribution, trend, and factors associated with the occurrence of Hymenoptera envenomations in humans. We describe characteristics of persons suffering Hymenoptera stings from bees, wasps, and hornets as reported to the Illinois Poison Center, and assess seasonal, climatologic, and time trends of calls for envenomations between 2002 and 2007. Mean daily temperature and mean daily atmospheric pressure were positively associated with envenomations, whereas wind speed was negatively associated with envenomations. We also observed a significant increase in calls for envenomations on summer holidays (P < 0.001). In addition, our findings showed that the number of calls for envenomations declined by nearly half after 2005 (P < 0.001) compared with previous years. Our findings indicate that the decline in bees, wasps, and hornets may be widespread, affecting both wild and commercial populations, and that the decline appears to have been rapid and sustained in recent years. Poison center data are a valuable resource for the surveillance of poisoning in humans, but our findings show that the data can be used to monitor changes in nonhuman species.

The current study investigated the occurrence of ticks and their rickettsiae in the Serra do Mar State Park, which encompasses one of the largest Atlantic rain forest reserves of Brazil. From July 2008 to June 2009, a total of 2,439 ticks (2,196 free living and 243 collected on hosts) was collected, encompassing the following 13 species: Amblyomma aureolatum (Pallas), Amblyomma brasiliense Aragão, Amblyomma dubitatum Neumann, Amblyomma fuscum Neumann, Amblyomma incisum Neumann, Amblyomma longirostre (Koch), Amblyomma naponense (Packard), Amblyomma nodosum Neumann, Amblyomma ovale Koch, Haemaphysalis juxtakochi Cooley, Ixodes aragaoi Fonseca, Ixodes loricatus Neumann, and Rhipicephalus sanguineus (Latreille). Ticks were submitted to polymerase chain reaction assays targeting portions of the rickettsial genes gltA and ompA. Polymerase chain reaction products were DNA sequenced and compared with corresponding sequences available in GenBank. Rickettsia bellii, a rickettsia of unknown pathogenicity, was detected in one A. aureolatum, one A. ovale, and three A. incisum specimens. At least 8.8% (3/34) of the free-living A. ovale ticks, 13.6% (8/59) of the A. ovale ticks collected from dogs, and 1.9% (1/54) of the R. sanguineus (Latreille) ticks were found to be infected by Rickettsia sp strain Atlantic rain forest, a novel strain that has been shown to cause an eschar-associated spotted fever in the state of São Paulo. Our results suggest that A. ovale is the vector of Rickettsia sp strain Atlantic rain forest in the state of São Paulo.

The secondary screwworm, Cochliomyia macellaria (Fabricius), was reared on either equine gluteus muscle or porcine loin muscle at 20.8°C, 24.3°C, and 28.2°C. C. macellaria needed ≈35% more time to complete development when reared at 20.8 than 28.2°C. Furthermore, larval growth and weight over time did not differ between larvae reared on equine versus porcine muscle. This study is the second in the United States to examine the development of C. macellaria and is the first to examine development of this species on muscle tissue from different vertebrate species. These data could provide significant information regarding time of colonization, including myiasis and neglect cases involving humans and animals. Furthermore, these results in comparison with the only other data set available for this species in North America indicate a fair amount of phenotypic variability as it relates to geographic location, suggesting caution should be taken when applying these data to forensic cases outside the region where this study was conducted.

A model was developed using 167 carboxamide derivatives, from the United States Department of Agriculture archival database, that were tested as arthropod repellents over the past 60 yr. An artificial neural network employing CODESSA PRO descriptors was used to construct a quantitative structure-activity relationship model for prediction of novel mosquito repellents. By correlating the structure of these carboxamides with complete protection time, a measure of repellency based on duration, 34 carboxamides were predicted as candidate mosquito repellents. There were four additional compounds selected on the basis of their structural similarity to those predicted. The compounds were synthesized either by reaction of 1-acylbenzotriazoles with secondary amines or by reaction of acid chlorides with secondary amines in the presence of sodium hydride. The biological efficacy was assessed by duration of repellency on cloth at two dosages (25 and 2.5 µmol/cm²) and by the minimum effective dosage to prevent Aedes aegypti (L.) (Diptera: Culicidae) bites. One compound, (E)-N-cyclohexyl-N-ethyl-2-hexenamide, was superior to N,N-diethyl-3-methylbenzamide (deet) at both the high dosage (22 d versus 7 d for deet) and low dosage (5 d versus 2.5 d for deet). Only one of the carboxamides, hexahydro-1-(1-oxohexyl)-1H-azepine, had a minimum effective dosage that was equivalent or slightly better than that of deet (0.033 µmol/cm² versus 0.047 µmol/cm²).

This study focused on two West Nile virus (WNV) disease outbreak years, 2003 and 2007, and included a three-county area (Larimer, Boulder, and Weld) in North Central Colorado that is hyperendemic for WNV disease. We used epidemiological data for reported WNV disease cases at the census tract scale to: 1) elucidate whether WNV disease incidence differs between census tracts classified as having high versus lower human population density (based on a threshold value of 580 persons/km²) and 2) determine associations between WNV disease incidence and habitat types suitable as development sites for the larval stage of Culex mosquito vectors. WNV disease incidence was significantly elevated in census tracts with lower human population density, compared with those with high density of human population, in both 2003 (median per census tract of 223 and 143 cases per 100,000 population, respectively) and 2007 (median per census tract of 46 and 19 cases per 100,000 population). This is most likely related, in large part, to greater percentages of coverage in less densely populated census tracts by habitats suitable as development sites for Culex larvae (open water, developed open space, pasture/hay, cultivated crops, woody wetlands, and emergent herbaceous wetlands) and, especially, for the subset of these habitats made up by irrigated agricultural land (pasture/hay and cultivated crops) that presumably serve as major producers of the locally most important vector of WNV to humans: Culex tarsalis. A series of analyses produced significant positive associations between greater coverage of or shorter distance to irrigated agricultural land and elevated WNV disease incidence. As an exercise to produce data with potential to inform spatial implementation schemes for prevention and control measures within the study area, we mapped the spatial patterns, by census tract, of WNV disease incidence in 2003 and 2007 as well as the locations of census tracts that had either low (<25th percentile) or high (>75th percentile) WNV disease incidence in both outbreak years (relative to the incidence for each year). This revealed substantial changes from 2003 to 2007 in the spatial pattern for census tracts within the study area with high WNV disease incidence and suggests a dynamic and evolving scenario of WNV transmission to humans that needs to be taken into account for prevention and control measures to stay current and represent the most effective use of available resources.

This report describes two male specimens of the sand fly species Lutzomyia shannoni (Dyar) (Diptera: Psychodidae: Phlebotominae) collected at Fort Rucker, AL, and Fort Campbell, KY, in dry ice-baited light traps during September 2005. The specimens were observed to have anomalies to the number of spines on the gonostyli. The taxonomic keys of Young and Perkins (Mosq. News 44: 263–285; 1984) use the number of spines on the gonostylus in the first couplet to differentiate two major groupings of North American sand flies. The two anomalous specimens were identified as L. shannoni based on the following criteria: 1) both specimens possess antennal ascoids with long, distinct proximal spurs (a near diagnostic character of L. shannoni in North America), 2) the sequences of the partial cytochrome c oxidase subunit 1 gene from both specimens indicated L. shannoni, and 3) the sequences of the internal transcribed spacer 2 molecular marker from both specimens indicated L. shannoni. The anomalous features are fundamentally different from each other as the Fort Rucker specimen possesses a fifth spine (basally located) on just one gonostylus, whereas the Fort Campbell specimen possesses five spines (extra spines subterminally located) on both gonostyli. Because the gonostyli are part of the external male genitalia, anomalies in the number of spines on the gonostyli may have serious biological consequences, such as reduced reproductive success, for the possessors. These anomalies are of taxonomic interest as the specimens could easily have been misidentified using available morphological keys.