One of the most significant modern day efforts to prevent and control an arthropod-borne disease during a military deployment occurred when a team of U.S. military entomologists led efforts to characterize, prevent, and control leishmaniasis at Tallil Air Base (TAB), Iraq, during Operation Iraqi Freedom. Soon after arriving at TAB on 22 March 2003, military entomologists determined that 1) high numbers of sand flies were present at TAB, 2) individual soldiers were receiving many sand fly bites in a single night, and 3) Leishmania parasites were present in 1.5% of the female sand flies as determined using a real-time (fluorogenic) Leishmania-generic polymerase chain reaction assay. The rapid determination that leishmaniasis was a specific threat in this area allowed for the establishment of a comprehensive Leishmaniasis Control Program (LCP) over 5 mo before the first case of leishmaniasis was confirmed in a U.S. soldier deployed to Iraq. The LCP had four components: 1) risk assessment, 2) enhancement of use of personal protective measures by all personnel at TAB, 3) vector and reservoir control, and 4) education of military personnel about sand flies and leishmaniasis. The establishment of the LCP at TAB before the onset of any human disease conclusively demonstrated that entomologists can play a critical role during military deployments.

The emergence of Anopheles species resistant to insecticides widely used in vector control has the potential to impact directly on the control of malaria. This may have a particularly dramatic effect in Africa, where pyrethroids impregnated onto bed-nets are the dominant insecticides used for vector control. Because the same insecticides are used for crop pests, the extensive use and misuse of insecticides for agriculture has contributed to the resistance problem in some vectors. The potential for resistance to develop in African vectors has been apparent since the 1950s, but the scale of the problem has been poorly documented. A geographical information system-based decision support system for malaria control has recently been established in Africa and used operationally in Mozambique. The system incorporates climate data and disease transmission rates, but to date it has not incorporated spatial or temporal data on vector abundance or insecticide resistance. As a first step in incorporating this information, available published data on insecticide resistance in Africa has now been collated and incorporated into this decision support system. Data also are incorporated onto the openly available Mapping Malaria Risk in Africa (MARA) Web site ( http://www.mara.org.za). New data, from a range of vector population-monitoring initiatives, can now be incorporated into this open access database to allow a spatial understanding of resistance distribution and its potential impact on disease transmission to benefit vector control programs.

We conducted experiments to investigate the importance of algal food resources for larval growth and adult emergence of Anopheles gambiae Giles s.s. in simulated larval habitats in Kenya, and in greenhouse and laboratory microcosms in the United States. In the first experiment, we used shading to reduce algal biomass, and because algal production and larval development might be a function of underlying soil nutrients, we crossed sun–shade treatments with soils of two distinct types collected near larval habitats. Shading reduced pupation rates and total adult biomass of An. gambiae by ≈50%. Soil type had no significant effect on mosquito production, but it did significantly affect concentrations of phosphorus and chlorophyll a in the surface microlayer. In a subsequent experiment conducted in the greenhouse to reduce temperature differences found between the shaded and sunlit treatments, <1% of larvae in the shaded treatments reached the pupal stage. There was a marked reduction of chlorophyll a levels as a function of shading and larval density. In a third experiment, larvae receiving material harvested from sunlit surface microlayers performed as well as those receiving liver powder, whereas those receiving surface microlayer from shaded habitats suffered >90% mortality and failed to pupate. In a fourth experiment, glucose was added to shaded microcosms to stimulate bacterial activity in the absence of algae. Bacterial growth rates were 2 to 3 times higher, and larval development was enhanced in glucose-amended treatments. However, pupation rates and adult weights in glucose-amended shaded microcosms were still poor compared with those in nonamended sunlit microcosms. Overall, these results demonstrate the importance of algal biomass in the surface microlayers of larval habitats to development and adult production of An. gambiae.

We conducted experiments in laboratory microcosms to simulate input of stemflow nutrients and flushing of metabolites in the tree hole habitats of larval Ochlerotatus triseriatus (Say). In the first experiment, we simultaneously examined the effects of nutrient additions (nitrogen, phosphorus, glucose, or combination) and flushing (removal of one-half of water volume replaced by deionized water) on mosquito production. The combination of nutrients had the greatest positive effects on mosquito production, with nitrogen (N) likely accounting for most of the increase in adult emergence and adult mass. Dilution of the nutrient pool via simulated flushing reduced mosquito growth, suggesting that the primary effect of stemflow input was nutrient addition as opposed to dilution of any latent toxic metabolites. In a second experiment, N additions were crossed with larval presence or absence to examine effects on key microbial processes. N increased leaf decay rates, soluble carbohydrate concentrations, fungal biomass and leaf-associated carbohydrase activity, but it did not stimulate bacterial productivity. Leaf decay was enhanced and bacterial production on leaves and in the water column was depressed in the presence of larvae. We conclude that the inputs of soluble N stimulated fungal growth, which made more fungal biomass available because of both its absolute increase and via the softening of the leaf particulate matter that could allow direct ingestion by larvae.

Wolbachia bacteria manipulate the reproduction of mosquito hosts via a form of sterility known as cytoplasmic incompatibility (CI), promoting the spread of infections into host populations. The rate at which an infection invades is affected by host fitness costs associated with the Wolbachia infection. Here, we examine for an effect of Wolbachia infection on the immature fitness of the Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae). In two experiments, we examine for a Wolbachia effect on immature survivorship and developmental rate, adult size, and an effect of larval nutrition on CI level. The highest survivorship can be observed in uninfected larvae, primarily because of reduced survivorship of Wolbachia-infected males. Although differences in the developmental rates are observed between the examined strains, the differences cannot be readily attributed to Wolbachia. An effect of Wolbachia on adult size is not observed. Poor male nutrition is associated with reduced fecundity and egg hatch of mates. The latter is hypothesized to explain the reduced egg hatch observed in CI crosses of malnourished males relative to well fed males. We discuss the results in relation to previously identified differences in adult fitness, naturally occurring invasions of Wolbachia, applied strategies of population replacement, and the need for additional modeling effort.

Endosymbiotic Wolbachia bacteria have been previously shown to infect laboratory colonies of the human bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), but little information exists regarding the extent of infection in natural populations. We assayed C. lectularius populations from five North American regions (California, Connecticut, Florida, New York, and Toronto, Canada) and one African region (Macha, Zambia) for Wolbachia infection by the polymerase chain reaction (PCR). Wolbachia infections were prevalent in all populations assayed (83–100%). There were no significant differences in infection frequency between geographic regions, between sexes, or between life stages (adult versus nymph). The potential utility of Wolbachia for alternative bed bug control strategies is discussed.

Onchocerciasis is a serious disease vectored by black flies in the genus Simulium that are infected with the filarial parasite Onchocerca volvulus. In the Americas, black flies of the Simulium ochraceum s.l. species complex are important vectors of this parasite. Cytological studies have suggested that this species complex consists of at least three cytotypes that inhabit distinct habitats. In this study, the NADH dehydrogenase subunit four (ND4) and internal transcribed spacer (ITS) of the ribosomal RNA gene cluster were used to explore the degree of genetic diversity among S. ochraceum s.l. populations found in the three O. volvulus foci in Mexico. Both sequence regions were found to exhibit intra- and interpopulation variation. Four different ND4 alleles were found among the populations examined. Similarly, variation was noted in the ITS domain sequences within and among populations. Variation within the ITS sequence was primarily confined to a complex microsatellite locus. Four ITS length variants were observed, two of which were only seen in flies collected from the onchocerciasis focus in northern Chiapas. These data suggest that the ND4 and ITS sequences may prove to be useful markers for exploring interactions within and among the S. ochraceum s.l. populations in Mexico.

The complete cDNA sequence encoding a Boophilus microplus (Canestrini) (Acari: Ixodidae) acetylcholinesterase (AChE3) was expressed in the baculovirus system. The recombinant AChE3 protein (rBmAChE3) was secreted as a soluble form into the cell culture medium and was identified as a functional AChE by substrate specificity and by inhibition with the AChE-specific inhibitors eserine sulfate and BW284c51. Inhibition kinetics of rBmAChE3, in the presence of the organophosphate paraoxon, revealed sensitivity comparable with that of adult, organophosphate-susceptible neural AChE. To our knowledge, this is the first report of the cloning and successful expression of a functional ixodid AChE.

Culex pipiens L. reared under diapause-inducing conditions (short daylength; 18°C) were more cold tolerant and desiccation resistant than their nondiapausing counterparts (long daylength; 18°C). Upon cold exposure (−5°C), diapausing mosquitoes reared at 18°C survived nearly twice as long as nondiapausing mosquitoes reared at 18°C and 10 times longer than nondiapausing mosquitoes reared at 25°C. Thus, rearing temperature provided partial protection against low temperature injury in nondiapausing mosquitoes, but maximum resistance to cold was attained by the diapause state. In this species, the supercooling point is not a good indicator of cold tolerance. Both diapausing and nondiapausing females had supercooling points of approximately −16°C, but diapausing as well as nondiapausing females died at temperatures well above the supercooling point, suggesting that low temperature mortality was due to indirect chilling injury. Diapause also conferred greater resistance to desiccation (1.6–2-fold increase in survival) compared with the nondiapause state. The gene encoding a 70-kDa heat shock protein, hsp70, was not up-regulated (i.e., more highly expressed) as a part of the diapause program, nor was it up-regulated by desiccation stress, but it was up-regulated during recovery from cold shock. Cx. pipiens thus differs from a number of other diapausing insect species that are known to developmentally up-regulate hsp70 during diapause.

Physical methods such as high and low temperatures were used in the past for the control of human body louse, Pediculus humanus humanus L. (Anoplura: Pediculidae). In the current study, the minimum time necessary to kill all lice after exposing them to temperatures other than those described in the literature, the mortality of lice after immersing them in water, and the survival of lice whose legs were amputated were studied. All lice died after 6 d at 6°C, after 11 d at 24°C, and after 9 d at 31°C. At −17°C, all lice were dead after 35 min, whereas at −70°C, all lice were dead after 1 min. All lice died after immersion in water within 19 h. The differences in mortality were significant but borderline between controls and lice whose two legs were amputated immediately or 24 h after feeding (3.3 versus 13.3% and 8.3 versus 21.7%). For lice whose leg was amputated 48 h after feeding, significant differences were found between controls and lice with one amputated leg (13.3 versus 48.3%), between controls and lice with two amputated legs (13.3 versus 68.3%), and between lice with one and two amputated legs (48.3 versus 68.3%).

We studied the toxicity and delayed effects of a mosquitocidal toxin (Mtx1) and a binary toxin (Bin) produced in Escherchia coli E-TH21 and Bacillus thuringiensis B-CW1, respectively, on Culex quinquefasciatus (Diptera: Culicidae). Bioassay results showed that both E-TH21 powder and B-CW1 sporulated culture were highly toxic against susceptible Cx. quinquefasciatus, with LC₅₀ values of 0.65 and 1.70 mg/liter against third and fourth instars at 48 h, respectively. After initial 48-h exposure of larvae to different concentrations of Mtx1 and Bin, significant continued mortality could be observed in larval, pupal, and emergence stages of Cx. quinquefasciatus. Importantly, the Mtx1 could induce higher cumulative larval and preadult mortalities than Bin toxin on the target mosquito. This finding is important for understanding the mode of action of Mtx1 and Bin toxins and for developing a new bioassay procedure for evaluation of toxicity of Bacillus sphaericus Neide, some strains of which produce Mtx1 and Bin, in the laboratory and field.

MyggA Natural (Bioglan, Lund, Sweden) is a commercially available repellent against blood-feeding arthropods. It contains 30% of lemon-scented eucalyptus, Corymbia citriodora (Hook.) K. D. Hill & L. A. S. Johnson (Myrtaceae), oil with a minimum of 50% p-menthane-3,8-diol. MyggA Natural also contains small amounts of the essential oils of lavender, Lavandula angustifolia Mill. (Lamiaceae), and geranium, Pelargonium graveolens L’Her. (Geraniaceae). In laboratory bioassays, MyggA Natural and C. citriodora oil exhibited 100% repellency against host-seeking nymphs of Ixodes ricinus (L.) (Acari: Ixodidae). Lavender oil and geranium oil, when diluted to 1% in 1,2-propanediol, had weak repellent activities on I. ricinus nymphs, but when diluted to 30% in 1,2-propanediol had 100% repellencies. 1,2-Propanediol (100%) had no significant repellent activity in comparison with that of the control. In field tests in tick-infested areas in central Sweden, tick repellency of MyggA Natural and C. citriodora oil was tested by the blanket-dragging technique for 4 d during a 6-d period. The repellencies (74 and 85%, respectively) on day 1 are similar (89%) to that of blankets treated in a similar manner with 19% diethyl-methyl-benzamide, based on previous work. Repellencies declined significantly from day 1 to day 6 (74 to 45% for MyggA Natural; 85 to 42% for C. citriodora oil).

To investigate the reservoir role of the lizard Psammodromus algirus for the Lyme disease spirochete, 199 lizards were trapped from April to October 2003 in El Jouza, northwestern Tunisia. In this site, the infection rate of free-living Ixodes ricinus (L.) by Borrelia was evaluated by immunofluorescence as 34.6% for adult ticks and 12.5% for nymphs. Eighty percent of P. algirus (117/146) captured during this study were infested by I. ricinus, the predominant tick species collected from lizards. The intensity of tick infestation of this host by larvae and nymphs ranged from 0.14 to 7.07 and from 1.5 to 6.58, respectively. These immature stages of I. ricinus were found on lizards in spring and the beginning of summer, with a peak of intensity during June (10.16 immature ticks by lizard). Tissue cultures from lizards and xenodiagnosis with larval I. ricinus were used to assess the infection and the ability, respectively, of infected lizards to transmit Borrelia to naive ticks. Seventeen percent of xenodiagnostic ticks (40/229) acquired B. lusitaniae while feeding on P. algirus. Therefore, we demonstrated the ability of the lizards to sustain Borrelia infection and to infect attached ticks, and we proved that P. algirus is a reservoir host competent to transmit B. lusitaniae.

Lyme borreliosis is associated with several genospecies of Borrelia burgdorferi sensu lato (s.l.) (Spirochaetales), but human disease has been associated only with Borrelia burgdorferi sensu stricto (s.s.) Johnson, Schmid, Hyde, Steigerwalt & Brenner in the western United States. Restriction fragment length polymorphism (RFLP) analysis of rrf-rrl amplicons from 124 tick and mammalian isolates from various habitats yielded 13 RFLP patterns. Of these patterns, six were patterns previously associated either with Borrelia bissettii Postic, Marti Ras, Lane, Hendson & Baranton or Borrelia burgdorferi s.s., and the remaining seven patterns belonged to diverse and previously uncharacterized Borrelia spp. Uncharacterized Borrelia spp. were cultured most frequently from Ixodes spinipalpis Hadwen & Nuttall and California kangaroo rats, Dipodomys californicus Merriam, inhabiting grasslands, and B. bissettii from I. spinipalpis and dusky-footed woodrats, Neotoma fuscipes Baird, associated with oak woodlands or chaparral. B. burgdorferi s.s. typically was isolated from host-seeking Ixodes pacificus Cooley & Kohls collected in dense oak woodlands, woodland–grass, or redwood forests. Although some isolates of B. burgdorferi s.s. were cultured from woodrats, there was no clear association of this human pathogen with any vertebrate host. These findings, along with recent evidence indicating that the western gray squirrel, Sciurus griseus Ord, may be an important reservoir of B. burgdorferi s.s. in Californian oak woodlands, suggest that our earlier hypothesis implicating an enzootic cycle involving woodrats and I. spinipalpis is insufficient to account for observed patterns of infection in nature.

The endosymbiont Wolbachia usually causes cytoplasmic incompatibility in dipteran hosts, including mosquitoes. However, some important arbovirus-transmitting mosquitoes such as Aedes aegypti (L.) are not heritably infected by Wolbachia. In Wolbachia-harboring mosquito Armigeres subalbatus Coquillett, colocalization of Wolbachia and inoculated Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV) in salivary gland (SG) cells was shown by electron microscopy. The infection rate of JEV in SGs, detected with either immunofluorescent antibody test or reverse transcription-polymerase chain reaction, did not show significant differences between Wolbachia-infected and -free colonies. It is suggested that Wolbachia did not mediate resistance of SG cells to superinfection by JEV, although both microorgamisms coexist in the same niche, i.e., the same SG cell. Therefore, a SG escape barrier may not be elevated due to Wolbachia infection, which presumably has no deleterious effects on vector competence in Wolbachia-harboring mosquitoes.

Aedes triseriatus (Say) (Diptera: Culicidae), the major vector of La Crosse (LAC) virus, efficiently transmits LAC virus both horizontally and transovarially. We compared the vector competence and transovarial transmission ability of Ae. triseriatus, Aedes albopictus Skuse, and Aedes aegypti (L.) for LAC virus. Ae. triseriatus and Ae. albopictus were significantly more susceptible to oral infection with LAC virus than Ae. aegypti. The three species also differed in oral and disseminated infection rates (DIRs). Transovarial transmission (TOT) rates and filial infection rates (FIRs) were greater for Ae. triseriatus than either Ae. albopictus or Ae. aegypti. These measures were integrated into a single numerical score, the transmission amplification potential (TAP) for each species. Differences in TAP scores were due mainly to the differences in DIRs and FIRs among these mosquitoes. Although the TAP score for Ae. albopictus was lower than that of Ae. triseriatus, it was 10-fold greater than that for Ae. aegypti.

The blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae), has a wide geographical distribution in Ontario, Canada, with a detected range extending at least as far north as the 50th parallel. Our data of 591 adult I. scapularis submissions collected from domestic animals (canines, felines, and equines) and humans during a 10-yr period (1993–2002) discloses a monthly questing activity in Ontario that peaks in May and October. The Lyme disease spirochete Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt & Brenner was detected in 12.9% of I. scapularis adults collected from domestic hosts with no history of out-of-province travel or exposure at a Lyme disease endemic area. Fifty-three isolates of B. burgdorferi were confirmed positive with polymerase chain reaction by targeting the rrf (5S)-rrl (23S) gene. Using DNA sequencing of the ribosomal species-specific rrf (5S)-rrl (23S) intergenic spacer region, all isolates belong to the pathogenic genospecies B. burgdorferi sensu stricto (s.s.). Nucleotide sequence analysis of a 218- to 220-bp amplicon fragment exhibits six cluster patterns and, collectively, these isolates branch into four phylogenetic cluster groups for both untraveled, mammalian hosts and those with travel to the northeastern United States (New Jersey and New York). Four of five geographic regions in Ontario had strain variants consisting of three different genomic cluster groups. Overall, our molecular characterization of B. burgdorferi s.s. shows genetic heterogeneity within Ontario and displays a connecting link to common strains from Lyme disease endemic areas in the northeastern United States. Moreover, our findings of B. burgdorferi in I. scapularis reveal that people and domestic animals may be exposed to Lyme disease vector ticks, which have wide-ranging distribution in eastern and central Canada.

Observations of considerable numbers of Anopheles ziemanni Grunberg resting inside human dwellings in Ahero, western Kenya, raised the question of its status in malaria transmission. To address this question, samples of this species were tested for Plasmodium falciparum infection rates and blood-feeding pattern, and data were compared with those of known vectors in the area. Although no An. ziemanni mosquitoes were infected with P. falciparum sporozoites, the proportions of this mosquito species among all anophelines captured and its human blood index compared well with those of other vectors, suggesting a possible role in the transmission of malaria. Additional studies, including both indoor- and outdoor-resting mosquito populations as well as incorporating laboratory-based parasite susceptibility tests should clarify the situation.

Mapping ordinarily increases our understanding of nontrivial spatial and temporal heterogeneities in disease rates. However, the large number of parameters required by the corresponding statistical models often complicates detailed analysis. This study investigates the feasibility of a fully Bayesian hierarchical regression approach to the problem and identifies how it outperforms two more popular methods: crude rate estimates (CRE) and empirical Bayes standardization (EBS). In particular, we apply a fully Bayesian approach to the spatiotemporal analysis of Lyme disease incidence in New York state for the period 1990–2000. These results are compared with those obtained by CRE and EBS in Chen et al. (2005). We show that the fully Bayesian regression model not only gives more reliable estimates of disease rates than the other two approaches but also allows for tractable models that can accommodate more numerous sources of variation and unknown parameters.

Six taxa of prey were identified from the gut contents of 973 Culex (Lutzia) fuscanus Wiedemann (Diptera: Culicidae) larvae collected from 19 sites in Shantou, Guangdong Province, China, from May to October 2004. Chironomidae were found in the guts of 78.6% of Cx. fuscanus larvae but mosquito larval remains in only 2.5%. The junction between prey head and thorax was the most frequent attack site in field (125/169 = 73.9%) and laboratory (187/260 = 71.9%) observations.