The benzoylurea chitin synthesis inhibitor diflubenzuron was evaluated as a rodent feed-through for the control of immature stages of Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of second instars of P. papatasi larvae that were fed feces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing 0, 8.97, 89.7, or 897 ppm diflubenzuron was evaluated. No pupation or adult emergence occurred when larvae were fed feces from hamsters that were fed diets containing diflubenzuron. The mortality of sand flies fed feces from treated hamsters was coincident with pupation of the controls, suggesting a specific effect on the larval-to-pupal molt. The results of this study suggest that a control strategy using rodent baits containing diflubenzuron for phlebotomine sand flies and zoonotic cutaneous leishmaniasis may be possible.

Infestations of the bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), are increasing around the world at an alarming rate and have become a major public health concern. The evolution of insecticide resistance could be a primary factor in explaining this resurgence. Extremely high levels of resistance to two pyrethroid insecticides, deltamethrin and λ-cyhalothrin, relative to a susceptible colony, were detected in populations collected from human dwellings in Kentucky and Ohio. Offspring of a cross between a resistant and susceptible colony had intermediate susceptibility. Evaluations of populations from across the United States indicate that resistance to pyrethroid insecticides is already widespread. Without the development of new tactics for bed bug management, further escalation of this public health problem should be expected.

We describe a new subspecies, Cameronieta torrei dusbabeki, a parasite of Pteronotus quadridens fuliginosus (Gray 1843) from Puerto Rico.

Recent experiments demonstrated that most field-collected Triatoma infestans (Klug) (Hemiptera: Reduviidae) adults from northern Argentina either never initiated flight or did so repeatedly in both sexes. This pattern could not be explained by sex, adult age, weight, weight-to-length ratio (W/L), or chance. We examined whether bugs that never initiated flight possessed developed flight muscles, and whether flight muscle mass relative to total body mass (FMR) was related to the probability of flight initiation. Approximately half of the adults that never initiated flight had no flight muscles. The absence of flight muscles was 2.4 times more frequent in males than females. Females had significantly larger flight muscle mass than males. For both sexes, the frequency of bugs with no flight muscles was spatially heterogeneous among individual collection sites. A logistic regression model of flight initiation that included both FMR and W/L provided a better fit than models including either one of these predictors. FMR is a novel predictor of flight initiation in Triatominae, with a stronger effect than W/L. The higher frequency of females initiating flight in our experiments may be explained by females having flight muscles more frequently than males, and having FMR and W/L values more suitable for flying. These findings demonstrate that individuals and natural populations of T. infestans can differ dramatically with regard to flight initiation.

In response to an identified paucity of information on the size and composition of immature Aedes aegypti (L.) (Diptera: Culicidae) populations in large field containers, we assessed net sampling and pumping/sieving methods for estimating and enumerating third (III)/fourth (IV) instar and pupal populations. Sweep net detection thresholds (number above which ≥90% chance of a positive sample) were ≤28 immatures for seven different container types (115–3000 liter jars and tanks) in the laboratory, and mean recovery percentages varied by container type (6.15–41.29 and 7.39–33.10% for III/IV instars and pupae, respectively). A pumping method or hand bailing was applied in the field for the collection of III/IV instars and pupae from 406 receptacles, of which 343 had been previously sampled via a five-sweep netting technique. Larvae were 9.30 times more prevalent than pupae, and abundance varied by container type with means of 36–537 III/IV instars and 6–53 pupae per receptacle. Sweep netting for III/IV instars effectively identified 86.2% of Ae. aegypti-positive containers, whereas sampling for pupae detected only 43.1% of positive containers. When conversion factors (inverse of laboratory recovery percentages) were applied to field net sampling data, estimates of container populations were more accurate for III/IV instars than pupae (maximum R² = 0.610 and 0.328, respectively); however, the relationship between immature abundance and emergent adult populations remains to be defined.

Aedes aegypti (L.) and Aedes albopictus (Skuse) differ in their distribution in Taiwan. The former species is distributed in the south of Taiwan, whereas the latter is found throughout the island. One possible explanation proposes that low temperatures in the winter prevent the expansion of Ae. aegypti. Hence, the impact of low temperatures on immatures of both species was studied in the laboratory and in the field. Our study showed that, under most conditions, Ae. aegypti were more sensitive to low temperatures than Ae. albopictus both in the laboratory and in the field. The survival rates at 10°C for the first and fourth instars of Ae. albopictus were significantly better than those of Ae. aegypti. At 2.5 and 5.0°C, the first instars of Ae. albopictus survived better than the same stadium of Ae. aegypti, but the fourth instars of Ae. aegypti survived better. Short exposures to low temperatures did not affect the acclimatization of Ae. aegypti immatures but longer exposures did increase the physiological adaptation to low temperatures. For Ae. albopictus, exposure to low temperatures increases the acclimatization of this species. In field experiments, Ae. aegypti larvae had a significantly higher mortality than Ae. albopictus during exposures to cold fronts in the 2004 winter. We conclude that low temperatures in northern and central Taiwan have a negative impact on the distribution of Ae. aegypti, but this factor alone is not sufficient to prevent this species from occupying the rest of Taiwan.

To determine whether the Culex (Diptera: Culicidae) mosquitoes that transmit West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in the northeastern United States seek hosts and oviposit contemporaneously, we recorded when these mosquitoes attacked caged birds and when they deposited eggs. They traversed oviposition sites most frequently ≈2 h after astronomical sunset, and eggs generally were deposited at that time. Although they most frequently approached avian hosts ≈2 h after sunset during midsummer, they are more opportunistic during mid- to late fall. Because the Culex mosquitoes that serve as the main vectors of West Nile virus in the northeastern United States quest for hosts and seek to oviposit well after sunset, insecticidal aerosols would be most effective when applied at that time.

Feeding preferences of larval container-dwelling mosquitoes are not well understood. Primary production is often absent in container systems and external inputs of animal and plant detritus supply the energy base of container food webs by supporting microorganism prey for mosquitoes. We quantified the feeding preferences of Aedes albopictus (Skuse), a non-native invasive mosquito, and Ochlerotatus triseriatus (Say), a native mosquito, when given a choice of animal and plant detritus. We tested for interpopulational (Illinois versus Florida) differences in feeding preferences, and quantified each species’ performance on these two detritus types. When given a choice, both species spent significantly more time feeding at an animal detritus patch. The Illinois populations of both species spent more time feeding at animal detritus patches than did the Florida populations, which spent more time feeding at leaf detritus than did Illinois populations. Both species reached a later instar and had higher survival when reared with animal versus leaf detritus. Ae. albopictus spent more time feeding at animal detritus and had higher survival when reared on either detritus type compared with Oc. triseriatus. Greater preference for and better performance exhibited by Ae. albopictus in high-quality food (animal detritus) may result in preemption of high quality food and may contribute to the superior competitive ability of Ae. albopictus relative to Oc. triseriatus.

The accurate specific identification of ticks is essential for the study, control and prevention of tick-borne diseases. Herein, we determined ribosomal nucleotide sequences of the second internal transcribed spacer (ITS2) of 15 Neotropical hard tick species of the genus Amblyomma Koch found in Brazil. Most of the studied ticks accidentally parasite humans and potentially act as vectors of zoonoses. Lengths of the ITS2 sequences ranged from 956 to 1,207 bp, whereas GC content varied from 62.4 to 66.9%. A matrix of ITS2 divergence was calculated with the ITS2 sequence data obtained showing divergence levels varying from 1.5 to 28.8%. The analysis indicated that this molecular marker can be useful for Amblyomma-specific identification. Phylogenetic inferences based on the ITS2 sequences were used to assess some issues in subgenus taxonomy.

The suitability of polytene chromosomes for population genetic and in situ DNA physical mapping was investigated in Johannesburg, South Africa, Culex quinquefasciatus Say and Culex pipiens L.. The most defined and consistent polytene spreads were obtained from salivary glands of early fourth instars from Cx. quinquefasciatus. A photomap is provided for a Cx. quinquefasciatus colony originating from Johannesburg. Variations in banding patterns, chromosome aberrations, and paracentric inversions within the colony and comparisons with previous published descriptions are discussed. Chromosomes of sufficient quality were obtained such that a DNA coding for an odorant binding protein could be mapped by colorimetric in situ hybridization.

Salivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva. In the current study, a collection method for midge saliva was developed. Over a 3-d period, 3- to 5-d-old male and female Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were repeatedly placed onto the collection system and allowed to deposit saliva into a filter. Salivary products were eluted from the filters and evaluated by gel electrophoresis and mass spectrometry as well as by intradermal testing and determination of clotting time. Gel electrophoresis revealed ≈55 protein spots displaying relative molecular masses from 5 to 67 kDa and isoelectric points ranging from 4.5 to 9.8. The majority of molecular species analyzed by mass spectrometry showed high convergence with salivary proteins recently obtained from a cDNA library of Culicoides sonorensis Wirth & Jones, including proteins involved in sugarmeal digestion, defense, and coagulation inhibition as well as members of the D7 family and unclassified salivary proteins. In addition, the proteome analysis revealed a number of peptides that were related to proteins from insect species other than Culicoides. Intradermal injection of the saliva in human skin produced edema, vasodilatation, and pruritus. The anticoagulant activity of the saliva was demonstrated by significantly prolonged clotting times for human platelets. The potential role of the identified salivary proteins in the transmission of pathogens and the induction of allergies is discussed.

Phytochemicals endowed with hormonal, antihormonal, or toxic activity are potential agents for insect control. Thus, we became interested in testing Brazilian plant metabolites on Chrysomya megacephala (F.) (Diptera: Calliphoridae), a public health menace that is one of the most prevalent flies in Brazilian urban areas. We tested the lignan yangambin, from the leaves of Ocotea duckei Vattimo (Lauraceae). Topical treatment of eggs and first instars with yangambin as well as feeding larvae a yangambin-treated diet resulted in inhibition of postembryonic development, morphological alteration, and oviposition reduction.

Lethal ovitraps (LOs) containing an insecticide-treated ovistrip are used as a lure-and-kill device for the container-breeding dengue vector, Aedes aegypti (L.). We aimed to affirm that the pyrethroid bifenthrin could be used effectively in LOs against Ae. aegypti in north Queensland, Australia, by quantifying oviposition in and mortality caused by LOs. Small cage experiments in which individual gravid Ae. aegypti were given a choice of LOs and untreated ovitraps revealed that although LOs were less acceptable for oviposition, they provided an average 64.6% adjusted mortality. Although 92% of mosquitoes ovipositing in LOs died, 61.8% of mosquitoes that visited but did not oviposit in an LO also died, demonstrating that lethal contact occurred without egg laying. The bifenthrin content of strips (≈0.1 mg/cm²; 7 mg/strip) did not decrease significantly after 4 wk of field exposure nor did the toxic effect of the LOs. Large cage trials with groups of 10 Ae. aegypti confirmed that bifenthrin-treated LOs provided consistent control (average adjusted mortality 79.7%). Four-week field trials in north Queensland showed that although LOs were acceptable to ovipositing Ae. aegypti (mean time to first egg 10.9 d; mean eggs 47.3), insecticide-free ovitraps were oviposited in more readily (6.8 d, 199 eggs). The number of eggs laid per mosquito in laboratory LOs allowed calculation of the number of Ae. aegypti killed in field-deployed LOs; rapid estimates can be made by simply dividing the number of eggs on the strip by 2.84. Overall, the studies demonstrated that bifenthrin-treated LOs have potential for use as a lure-and-kill device against Ae. aegypti and that they should be effective in the field for at least 4 wk. Given that untreated ovitraps were more acceptable for Ae. aegypti oviposition, the removal of alternative oviposition sites before deployment of LOs in the field should maximize their effectiveness.

Aedes aegypti (L.) (Diptera: Culicidae) is the primary vector of both dengue and yellow fever. Use of insecticides is one of the primary ways to control this medically important insect pest. However, few new insecticides have been developed for mosquito control in recent years. As a part of our collaborative effort to search for new insecticides to control mosquitoes, piperidine was used as base compound for further optimization. Herein, we report the structure–activity relationships of 33 piperidines against adult female Ae. aegypti. On the basis of 24-h LD₅₀ values after topical application, the most toxic compound was 2-ethyl-piperidine, with an LD₅₀ as low as 0.8 μg per mosquito. The toxicities of piperidine derivatives were significantly decreased when a benzyl moiety was attached to the carbon of the piperidine ring, with an LD₅₀ value as high as 29.2 μg per mosquito. The toxicity order of three moieties attached to the carbon of the piperidine ring was ethyl- > methyl- > benzyl-derivatives. When the same moiety was attached to the piperidine ring, the carbon position to which the moiety was attached conferred different toxicity and the toxicity order was second carbon > third carbon > fourth carbon. Together, these preliminary results may be useful in guiding further piperidine ring modifications in the development of potential new insecticides.

Owing to the spread of pyrethroid resistance in Anopheles gambiae s.s. (Diptera: Culicidae) and other vector mosquitoes, there is an urgent need to develop alternative insecticides to supplement the pyrethroids for malaria control. Indoxacarb is an oxadiazine insecticide initially commercialized by DuPont for control of agricultural pests. Performance against An. gambiae bearing kdr (pyrethroid and DDT resistance) or Ace-1^R insensitive acetylcholinesterase (organophosphate and carbamate resistance) mechanisms was studied using larval and adult bioassays and a simulated experimental hut system (tunnel tests) that allows fuller expression of the behavioral responses to insecticide. Larval and adult bioassays (topical application and cone tests on treated netting) showed a standard probit dosage–mortality response and no evidence of cross-resistance to the kdr and Ace-1^R resistance mechanisms. Toxic activity was slow compared with standard insecticides and additional mortality was observed. Indoxacarb induced no excitorepellency in adults. In tunnel tests, indoxacarb induced no inhibition of mosquito penetration or blood feeding through the holed netting, but it induced delayed mortality over 24–96 h. There was >90% mortality of the kdr strain on netting treated with the 500 mg/m² dosage, whereas permethin at 500 mg/m² only killed 30% of this strain. A mixture of indoxacarb and pyrethroid showed neither synergism nor antagonism. The absence of cross-resistance to current insecticides indicates that indoxacarb has malaria vector control potential as larvicide or adulticide where mosquitoes are pyrethroid resistant.

The efficacy of injectable doramectin applied at 200 μg/kg was evaluated against adult female Boophilus microplus (Canestrini) (Acari: Ixodidae) in the later stages of engorgement before detachment. Lethal levels of doramectin in the serum of treated cattle (9.7–36.6 ppb) were reached and sustained within 24 h after treatment. However, treatment at 19 or 20 d postinfestation allowed greater tick survival and reproductive capability (IF) than at 18 d postinfestation, indicating that a significant portion of the ticks were able to engorge and detach before obtaining a lethal dose of doramectin. Thus, treatment at 18 d after infestation provided significantly higher overall control (99.5%) than treatment at 19 or 20 d postinfestation (95.8 and 89.1%, respectively). Analysis of control on a daily basis demonstrated that treatment at 18 d postinfestation provided >99% on each day of the evaluation. Conversely, treatment at 19 or 20 d postinfestation produced levels of control ranging from 22.6 to 85.6% during the first 2 d of female detachment, and ≥99% control was not achieved until after the fourth day of female detachment, where it remained throughout the study. Therefore, application of injectable doramectin at ≤18 d after tick infestation was the only treatment regime considered acceptable for use in the U.S. Boophilus Eradication Program. Treatment intervals >18 d postinfestation could pose a substantial risk of dispersing viable ticks to tick-free areas outside the permanent quarantine zone that has been established along the Texas–Mexico border.

The ability of the Miller, Soberanes, and White bioassay techniques to describe amitraz susceptibility in three different strains of Boophilus microplus (Canestrini) (Acari: Ixodidae) was compared. For a susceptible strain, all techniques adequately described amitraz susceptibility by producing a full range of mortality that corresponded with increasing concentration of amitraz. However, when resistant strains were evaluated, only the Miller and the Soberanes techniques adequately estimated the dose–response relationship. Lethal concentrations were not precisely estimated when all the data were included in the analyses for every strain and technique tested. Better estimates were obtained when subsets of data around the range of interest were subjected to probit analysis. For the Soberanes technique, the slope of the probit regression was steeper for the Brazilian resistant and Texan susceptible strains compared with the heterozygous Mexican strain. The pattern was different when the same strains were tested with the Miller technique. The slopes of the regressions for the Mexican and the Texan strains did not differ significantly, but the Brazilian strain had a steeper slope than the other strains. Resistance ratios were much greater when the Soberanes technique was used than when the Miller technique was used on the same strains. However, neither technique produced enough separation between susceptible and resistant strains to develop a traditional discriminating dose (DD) test that required a concentration of 2× LC_99.9 estimate. A DD test at the LC₉₉ would be possible for both techniques. We discuss the strengths and weaknesses of the three techniques, including potential improvements to the White technique. The White technique has the greatest potential to determine the mechanisms of amitraz resistance in detailed synergist studies. Currently, only the Miller method can fulfill this task. The Miller and Soberanes techniques are well suited for the study of the epidemiology of resistance worldwide, because they use commercially available, formulated amitraz that is easy and inexpensive to obtain.

The relationship between mosquito and parasite involves a delicate balance that is influenced not only by the mosquito but also by parasite determinants. Using the biologically and morphologically similar parasites Brugia malayi and Brugia pahangi and the mosquito Armigeres subalbatus (Coquillett) (Diptera: Culicidae), it should be possible to dissect out the key elements involved in initiating or avoiding an immune response, known as melanotic encapsulation, because in this mosquito B. malayi microfilariae (mf) are melanized and destroyed, but B. pahangi mf develop normally into infective-stage larvae. Because of limitations in isolating sufficient mf from the circulation of an infected mammalian host, Brugia spp. mf that can be obtained in large numbers from the peritoneal cavity of an infected host were tested to ascertain the immune response of Ar. subalbatus to this source of mf. Results indicate that the immune response of Ar. subalbatus against intraperitoneal (i.p.) Brugia spp. mf mimics that which is observed when this mosquito is exposed to mf-infected animals, indicating that i.p. mf are similar to those mf that circulate naturally in the blood of the vertebrate host. Therefore, the i.p. mf should serve as an excellent source of material for genomic and proteomic studies designed to analyze the role of the parasite in influencing the immune response of the mosquito.

Inter- and intraspecific transfer of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) occurred infrequently when donor Culex tarsalis Coquillett fed concurrently on house finches with recipient Culex quinquefasciatus Say and Cx. tarsalis. Five of six of these house finches had WNV in blood samples collected by jugular venipuncture 30–45 min postfeeding, with titers ranging from 2.3 to 4.2 log₁₀ plaque-forming units (PFU)/ml. After 2-wk incubation at 26°C, three Cx. quinquefasciatus and one Cx. tarsalis of 230 blood-fed recipients were infected, of which one Cx. quinquefasciatus was capable of transmission. Our data indicated that infectious female mosquitoes feeding on small vertebrates create a nonpropagative viremia capable of infecting concurrently cofeeding females. The proportion of recipients infected is likely related to the amount of virus expectorated by donor females, the blood volume of the vertebrate host, and the susceptibility of the cofeeding mosquitoes.

Lizards and small rodents were live captured in Tuscany, central Italy, from May through August 2005. Prevalence of infestation by larval Ixodes ricinus L. (Acari: Ixodidae) and mean numbers of larvae per host were not significantly different for common wall lizards, Podarcis muralis Laurenti, and Apodemus spp. mice, whereas infestation levels by nymphs were significantly greater on lizards. Borrelia lusitaniae, which was previously shown to be dominant in host-seeking I. ricinus in the same study area, was detected by polymerase chain reaction (PCR) in 19.8% (95% confidence interval: 14.4, 26.0) of larval ticks and in 52.9% (27.8, 77.0) of nymphs that were collected from lizards. Moreover, 18.8% (7.2, 36.4) and 25.0 (3.2, 65.1) of lizards’ tail biopsies and blood samples, respectively, were positive for B. lusitaniae. Conversely, attached ticks and ear biopsies from Apodemus spp. mice were PCR negative. Passerine birds belonging to 10 species were live captured in March 2005, and Borrelia valaisiana was detected in 57.1% (18.4, 90.1) of I. ricinus nymphs feeding on Eurasian blackbirds, Turdus merula L. Results of this study suggest that lizards play an important role as reservoirs for B. lusitanae and may affect the dominance of this genospecies in the Mediterranean area.

Boophilus microplus (Canestrini) (Acari: Ixodidae), collected in Hidalgo County, TX, were determined to be resistant to permethrin. Discriminating dose (DD) tests at the LC₉₉ and 2× the LC₉₉ of susceptible ticks yielded lower than expected mortalities for permethrin but not for coumaphos or amitraz acaricides. Initial bioassay results confirmed the pyrethroid resistance detected in the DD assays. Two generations of selection with permethrin at a rate >60% increased the measured resistance ratios from 9.5 (7.9–11.5) to 263 (217–320). Synergist studies suggested that metabolic enzymes were involved in permethrin resistance. Native gel electrophoresis verified that the CZS9 esterase was not involved in resistance to permethrin. Polymerase chain reaction examination revealed the presence of a mutation of the sodium channel gene (Phe[arrow]Ile amino acid substitution in the S6 trans-membrane segment of domain III), in the resistant B&H population. The frequency of this mutation increased after selection with permethrin and concurrent increase in estimated resistance ratios. The B&H population was eradicated from the United States by the U.S. Department of Agriculture’s Animal and Plant Health Inspection Service Veterinary Services, Cattle Fever Tick Eradication Program through the use of the organophosphate acaricide coumaphos.

Song sparrows, Melopiza melodia, inoculated subcutaneously with either western equine encephalomyelitis virus (family Togaviridae, genus Alphavirus, WEEV) or West Nile virus (family Flaviviridae, genus Flavivirus, WNV) developed elevated viremias, and they were considered to be competent experimental hosts for both viruses. However, birds that ingested from three to 20 mosquitoes containing comparable amounts of either WEEV or WNV failed to become infected, indicating limited oral susceptibility. Comparatively few field-collected birds had antibodies against either WEEV or WNV, indicating that this species was infrequently bitten by infectious mosquitoes in nature and probably was of limited importance in viral amplification.

The hypothesis that Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) may serve as a reservoir and vector of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in California was tested by determining the ability of this tick species to become infected with the NY99 strain of WNV while feeding on viremic song sparrows, to maintain the infection transstadially, and then to transmit WNV to recipient naïve song sparrows and western fence lizards during the nymphal stage. The percentage of ticks testing positive by reverse transcription-polymerase chain reaction (RT-PCR) decreased from 77% of 35 larvae at day 6 after ticks were transferred to donor song sparrows (day of detachment) to 23% of 35 nymphs at 59 d postinfestation (≈19 d after molting to the nymphal stage). However, the percentage of ticks positive by RT-PCR from which infectious virus was recovered by Vero cell assay decreased from 59% on day 6 to 12% on day 59, even though there was no statistically significant decrease in the quantity of RNA within positive ticks. Attempts to improve the sensitivity of plaque assays by blind passage through C6/36 cell cultures were unsuccessful. These data indicated that ticks maintained viral RNA but not necessarily infectious virus over time. Nymphs from larvae that fed on song sparrows with peak viremias ranging from 7.2 to 8.5 log₁₀ plaque-forming units (PFU) per ml were used in transmission attempts. From one to seven RNA-positive nymphal ticks engorged and detached from each of four recipient song sparrows or western fence lizards. Blood samples from sparrows and lizards remained negative, indicating that transmission did not occur. An additional four lizards inoculated with 1,500 PFU of WNV developed moderate viremias, ranging from 4.2 to 5.6 log₁₀ PFU/ml. Our data and data from previous studies collectively indicated that ixodid ticks were not able to experimentally transmit WNV and therefore most likely would not be important vectors in WNV transmission cycles.

A survey of wild rodents was performed in the Morro Bay area of central coastal California to determine serological and polymerase chain reaction (PCR) prevalence of Anaplasma phagocytophilum Dumler, Barbet, Bekker, Dasch, Palmer, Ray, Rikihisa, and Rurangirwa, Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt, and Brenner, Francisella tularensis McCoy, and Yersinia pestis Yersin; to describe the ectoparasitic fauna on important vector-borne disease hosts; and to determine whether pathogen exposure was associated with infestation by ectoparasites. We trapped 411 rodents from 10 species in 2004 and 2005. Anaplasma phagocytophilum exposure was detected in 11% of all wild rodents tested, with seropositive animals in eight species. Anaplasma phagocytophilum DNA was detected by PCR amplification in Neotoma fuscipes Baird and Reithrodontomys megalotis Baird (0.6% of all rodents). Yersinia spp. exposure was identified in 3.2% of all rodents tested, with highest detected exposure in peridomestic rodents, Mus musculus L. (20%), and Rattus rattus L. (50%). No individuals tested positive for the Y. pestis pla gene by PCR. In total, 338 fleas were identified from each of 10 rodent species examined. The most abundant flea was Malareus telchinus Rothschild. Relative density of flea infestation was highest on Spermophilus beecheyi Richardson and Microtus californicus Peale. Ticks recovered from trapped animals included Ixodes angustus Neumann, Ixodes pacificus Cooley & Kohls, Ixodes spinipalpis Hadwen & Nuttall, and Dermacentor occidentalis Marx. Given the moderate climate and diversity of rodents and arthropods in the Morro Bay area, ongoing investigation of this region will be helpful in understanding disease maintenance cycles.

The succession of mosquito species and abiotic factors affecting their distribution and abundance in rice (Oryza spp.) fields was investigated over a 16-wk rice growing cycle covering the period between January and May 2006. Fifteen experimental rice plots were sampled for mosquito larvae and characterized based on rice height, number of tillers, floating vegetation cover, water depth, water temperature, turbidity, salinity, pH, dissolved oxygen, total dissolved solids, and conductivity. Microscopic identification of 3,025 larvae yielded nine mosquito species predominated by Anopheles arabiensis Patton (45.0%), Culex quinquefasciatus Say (35.8%), Anopheles pharoensis Theobald (9.0%) and Ficalbia splendens Theobald (7.1%). Other species, including Anopheles rufipes Gough, Anopheles coustani Laveran, Anonopheles maculipalpis Giles, Culex annulioris Theobald, and Culex poicilipes Theobald made up 3.1% of the total collection. Anopheles gambiae s.l., Cx. quinquefasciatus, and An. pharoensis occurred throughout the cycle, but they were more abundant up to 4 wk posttransplanting with peaks after fertilizer application. As rice plants became established, three groups of mosquitoes were recognized: the first groups included An. rufipes, Fl. splendens, and Cx. annulioris, which occurred throughout much of the second half of the rice cycle, whereas the second group included Cx. poicilipes, which was found in the middle of the rice cycle. An. coustani and An. maculipalpis formed the third group occurring toward the end of the cycle. Dissolved oxygen, number of tillers, and rice height were negatively associated with the abundance of An. arabiensis and Cx. quinquefasciatus larvae. In addition, Cx. quinquefasciatus also was associated with water depth (−ve) and turbidity ( ve). Abundance of An. pharoensis larvae was significantly associated with water temperature ( ve), the number of tillers (−ve), and rice height (−ve), whereas Fl. splendens was significantly associated with the number of tillers ( ve). The results demonstrate a complex nature of the interactions between some of the factors in the ecosystem and mosquito species abundance and calls for time-dependent and species-specific mosquito control operations.

BG-Sentinel mosquito traps were trialed as a tool for the rapid assessment (24-h collections) and routine monitoring (72-h collections) of adult Aedes aegypti L. populations in north Queensland. Analysis of Ae. aegypti collections using BG-Sentinels set in suburban Cairns for 24 h permitted the calculation of sample size for a range of precision levels. Clusters of houses with BG-Sentinels operating continuously for 15 d, with collections every 72 h, also permitted required sample size calculation. Evidence of Ae. aegypti spatial clustering at the house scale was revealed, with statistically significant effects detected for all collection days. Less variation was detected at each trap location, with only nine of 32 trap locations revealing significant clustering over time. Trap-out effects through continuous BG-Sentinel operation at a fixed location were absent. The findings support fixed position sampling at 72-h intervals for routine monitoring of Ae. aegypti populations in Cairns. Despite the relationship between collections of adult vectors and the incidence of disease remaining unknown, BG-Sentinel collections provide an alternative and less labor-intensive abundance measure for assessing risk of dengue virus transmission and success of dengue vector control programs.

The use of the postmortem interval (PMI) in practical applications of forensic entomology is based upon developmental data of blow flies (Diptera: Calliphoridae) generated under controlled environmental conditions. Careful review of the published forensic entomology data sets showed that experimental (environmental) parameters differed between studies. Despite the differences in study design, there are no empirical data on the effect of photoperiod on blow fly development; yet, photoperiod has been shown to alter some insect development and behavior among noncalliphorids. Consequently, will differences in photoperiod alter the developmental times of calliphorids, and thereby alter PMI estimates? To answer this question, we used a replicated design with precise temperature measurement to examine the effects of photoperiod on the forensically important blow fly Phormia regina (Meigen). We concluded that inaccurate temperature recordings by using set-chamber temperatures over rearing-container temperatures would have overshadowed any affect light had on development. Second, constant light increased variation in overall adult developmental time and significantly delayed development compared with cyclic light. Finally, not accounting for delayed development induced by photoperiod underestimated the initial empirical estimate of the PMI. These sources of variation need to be included in forensic estimates because this variation can compromise predictions of PMI based upon current data sets. Without pinpointing optimal photoperiods with which to test development, we must assume that potentially large sources of variability exist within current estimates of the PMI.

Nonremoval drag sampling was conducted in multiple sites in Larimer County, CO, from March to July 2006 to determine the seasonal pattern of host-seeking activity by the human-biting adult life stage of the Rocky Mountain wood tick, Dermacentor andersoni Stiles (Acari: Ixodidae). Four sites, at elevations ranging from 1,790 to 2,470 m, consistently yielded host-seeking ticks. Ticks were active from March until late June; the length of the active period ranged from 84 to 104 d. Based on site-specific linear regression models of increase and decrease in tick host-seeking activity over time, densities of D. andersoni adults were estimated to reach 50% of their peaks between 2 and 11 April, to peak between 21 and 28 April, and to fall below 50% of the peaks between 24 and 27 May. The length of the periods with tick densities exceeding 50 and 75% of the peak was 43–52 and 21–26 d, respectively. Rapid increases in tick numbers in late March to early April followed a stretch of days with daily maximum temperatures exceeding 5°C, whereas rapidly declining tick numbers in mid- to late May were associated with daily maximum temperatures consistently exceeding 20°C and daily minimum relative humidity commonly falling below 20%. Densities of D. andersoni adults exceeded 50% of the peak when daily maximum temperatures were in the 16–19°C range and daily minimum relative humidity was >20%. Finally, tick seasonality may be adapted to local climatic conditions within Larimer County; site-specific daily maximum temperatures at the time of peak tick host-seeking activity in late April were positively associated with site-specific mean daily maximum temperatures for April.

Dermatophagoides farinae Hughes (Acari: Epidermoptidae), the American house dust mite, and Tyrophagus putrescentiae (Schrank) (Acari: Acaridae), the mold mite, are medically and economically important but controlling them has proved difficult, and recolonization is commonplace. Their behavioral responses to different sources of volatile chemicals are still not fully elucidated. For the first time, the Y-tube olfactometer, which is an enclosed bioassay to resolve responses to test and control volatiles, has been successfully used with these mites. Mites were tested individually, and both T. putrescentiae and D. farinae responded to food volatiles. Y-tube olfactometers may be used to test for potential semiochemicals, thereby increasing knowledge of our behavior of astigmatic mites.

The biological processes affecting Ixodes scapularis Say survival are complex. Understanding these processes will be beneficial for predicting tick distribution and population dynamics. This research shows that the duration for which nymphal ticks are exposed to drying air is an important factor for their survival. Experimental analysis of variance results show that duration of exposure to dry air (duration) is as important as vapor pressure deficit (relative humidity) (duration, relative humidity, P < 0.0001). Ticks do not survive when exposed to dry air for long periods; however, the return of humid air within 4–8 h has as large a positive impact on tick survival, as does constant humid air. This experiment exposes nymphal ticks to conditions of suboptimal humidity for different durations and then returns them to saturated conditions that are more typical of daily relative humidity fluctuations experienced during summer in southern New England forests.

Dengue outbreaks occur regularly in parts of northern Queensland, Australia, and there is concern that these outbreaks may spread with the introduction and range expansion of the two main vectors Aedes aegypti (L.) and Aedes albopictus (Skuse). Problems encountered in separating larvae of endemic and exotic container mosquito species resulted in the development of a polymerase chain reaction diagnostic procedure that uses a restriction enzyme to cut the internal transcribed spacer region 1 of the ribosomal DNA to separate Ae. aegypti and Ae. albopictus from a number of common local container mosquito species which can be used at any stage of the life cycle, including eggs up to 8 wk of age. Identification was possible using desiccated or alcohol-preserved specimens with a response time of <24 h after receipt of the specimens.

We evaluated the effectiveness of commercially available light traps and sticky traps baited with chemical light sticks for the collection of phlebotomine sand flies. Evaluations were conducted at Tallil Air Base, Iraq, in 2003. In an initial study, a Centers for Disease Control and Prevention (CDC)-style trap with UV bulb collected significantly more sand flies than did an up-draft CDC trap, a standard down-draft CDC trap (STD-CDC), or a sticky strap with a green chemical light stick. In a subsequent study, we found that the addition of chemical light sticks to sticky traps resulted in a significant increase in the number of sand flies collected compared with sticky traps without the light sticks. These data indicate that 1) the CDC light trap with an UV bulb is an effective alternative to the standard CDC light trap for collecting phlebotomine sand flies in Iraq, and 2) that the addition of a chemical light stick to a sticky trap can result in a field-expedient tool for the collection of sand flies.

The population density of Phlebotomus papatasi Scopoli (Diptera: Psychodidae), vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the etiologic agent of zoonotic cutaneous leishmaniasis (ZCL), was assessed May–November 2005 in central Tunisia by using sticky traps. The densities of P. papatasi were found to peak in early spring and again in the autumn. The lowest densities were observed in August. Prevalence of ZCL in the governorate of Sidi Bouzid peaks in December, 3 to 3.5 mo after the fall sand fly population peak, suggesting a close temporal association with the abundance of P. papatasi.