Insecticide resistance and absence of clinical cures or vaccines for many vector-borne diseases has stimulated interest in using genetically modified arthropod vectors for disease control. Current transgenic strategies focus on vector susceptibility to pathogen infection, which is an inefficient target for pathogen transmission interference. Manipulation of vector survival is theoretically more effective, resulting in larger reductions in the expected number of human infections. A hypothetical method to manipulate vector survival is to drive mortality-inducing Wolbachia into populations. For varying patterns and degrees of induced mortality, we outline the conditions under which virulent Wolbachia introductions into vector populations are expected to succeed and quantify the resultant reduction in pathogen transmission. The most critical component to the success of this strategy is the pattern of induced mortality. For operationally feasible introductions, induced mortality must be delayed until after vector reproduction begins. If this condition is not met, introduction thresholds become exceedingly high, ranging from ≈40% to 90% of the total adult population. Delayed induced mortality patterns can reduce introduction thresholds to ≈15–45% of the total adult population. Reduction in cytoplasmic incompatibility with male age has negligible effects on introduction success regardless of the induced mortality pattern. Under proper circumstances, symbiont-induced manipulation of vector survival can theoretically result in up to 100% reduction in pathogen transmission, depending on Wolbachia parameters, magnitude and pattern of induced mortality, and duration of pathogen incubation in the vector. Our results indicate that a broadening of the current paradigm for genetic manipulation of vectors to parameters other than arthropod vector competence is justified and will reveal new research possibilities for vector-borne disease control.

The midgut ultrastucture of the third-instar of Dermatobia hominis (L., Jr.) was investigated using transmission electron microscopy (TEM). The tubular midgut bears a monolayer of epithelial cells with the plasma membrane showing multiple folding where it adjoins the basement membrane. Septate junctions bound the epithelial cells on each side. These cells have electrolucent cytoplasm containing mitochondria, vacuoles, rough and smooth endoplasmic reticula, lamellar bodies, and a prominent nucleus with dispersed chromatin. The peritrophic matrix is close to elongate microvilli, which are sometimes forked. Regenerative cells, in an undifferentiated state when closest to the basement membrane, are scattered throughout the epithelial cells. A thick basement membrane, surrounded by thick connective tissue including muscle, tracheal tubes, and extracellular matrix is linked to epithelial cells by hemidesmosome-like structures. Entero-endocrine, goblet or cuprophilic cells were not observed.

The subgenus Paraphlebotomus of Phlebotomus includes several sand fly vectors of cutaneous and visceral leishmaniasis. A new species of the subgenus Paraphlebotomus, named Phlebotomus gemetchi is described from the volcanic blisters by the shore of lake Beseka near Metahara, Upper Awash Valley, in the Rift Valley of eastern Ethiopia. The presence of one distinct character in the male genitalia (a bifurcating ‘y’-shaped spine at the middle of the style) and three characters in the female (lengths of the fourth antennal and palpal segments and labrum) easily distinguish it from the other Afrotropical species recorded in East Africa including Ethiopia. The new species is 1 of 5 species representing the subgenus in Ethiopia as well as the rest of the Afrotropical Region. The distribution and medical importance of this sand fly species requires further investigation.

Demodex injai sp. nov. is described from the hair follicles of a domestic dog in Columbus, OH in October 1996. The mites occupy follicles from the orifice down to and into the sebaceous glands. The individual host may harbor both this new species and D. canis. A comparison of these two species is provided for identification purposes.

Maxillae and mandibles of males of 44 species of 12 mosquito genera and females of three autogenous genera and two partially autogenous species were examined under light microscopy and scanning electron microscopy. The maxillae and mandibles of male mosquitoes are delicate, tape-like structures with lengths characterizing genera or higher level classification units. Five patterns are recognized: (A) long maxillae and mandibles with mandibles longer than maxillae in Anopheles; (B) long maxillae and mandibles with maxillae longer than mandibles in Toxorhynchites; (C) short or intermediate lengths of maxillae with short mandibles in Aedes, Armigeres, Culex, Ochlerotatus, Orthopodomyia, and Uranotaenia; (D) short or intermediate length of maxillae with no mandibles in Mimomyia and Tripteroides; and (E) no maxillae and mandibles in Malaya and Topomyia. Maxillary and mandibular lengths of male mosquitoes show a positive correlation. Length of maxillae and mandibles of autogenous females are reduced to the same level as conspecific males. In contrast, females of partially autogenous species have complete maxillae and mandibles as in females of anautogenous species.

The hobo spider, Tegenaria agrestis (Walckenaer), and the related Tegenaria duellica Simon are very similar European spiders that have become well established in the northwestern United States and British Columbia. The hobo spider is considered to be medically important; T. duellica is considered harmless but is often misidentified as the hobo spider. The current distribution of the hobo spider includes southern British Columbia, Washington, Oregon, Idaho, northern Utah, the western half of Montana, western Wyoming, and two small, isolated populations in Colorado. T. duellica is found mostly west of the Cascade and Coastal mountain ranges from southern British Columbia to central Oregon. In large human population centers where both species are sympatric, T. duellica is usually more common than the hobo spider. Data from a total of 1,232 hobo spiders and 395 T. duellica are included in this study.

In 1995 and 1998, outbreaks of Japanese encephalitis (JE) virus occurred for the first time in Australia. Virus isolation from pools of mosquitoes indicated Culex annulirostris Skuse was the most likely vector. It was hypothesized that wind-blown mosquitoes introduced JE from Papua New Guinea to the Torres Strait and Cape York, northern Australia. This study used levels of genetic differentiation, as indirect evidence of dispersal of mosquitoes between the two continents. The results have demonstrated that in the region incorporating Western Province in PNG, the Torres Strait and Cape York Peninsula, Cx. annulirostris is represented by a panmictic population, indicating frequent widespread dispersal throughout the region of investigation. The closely related Cx. Palpalis Taylor was also present at some locations but the population structure of this species is uncertain. This supports the hypothesis that wind-blown Cx. annulirostris and possibly Cx. Palpalis could be a possible mechanism for introduction of JE virus into Australia.

Previous studies have determined that Amblyomma cajennense (F.) produces one generation per year in Southeastern Brazil. Larvae predominate from April to July, nymphs from July to October, and adults from October to March. In the current study, we observed engorged females, their eggs, and respective larvae in the field. Engorged females were released in grass plots under natural conditions, at monthly intervals, from October 2000 to March 2001. Preoviposition periods lasted 1–2 wk, and mean egg incubation periods lasted 5–7 wk. Consequently, larvae hatched at different periods from early December 2000 to early April 2001. However, all hatched larvae stayed confined to the plot ground, under the vegetation, until late April or early May 2001, when some larvae were seen questing on top of vegetation for the first time. Thereafter, larvae were always seen on this questing position until August 2001, when all larvae had died on the plots. In addition, premolt period data of engorged larvae and nymphs held in laboratory at different temperatures were used to construct a premolt mathematical model, which was applied to reported seasonal dynamic data of A. cajennense in the field. The premolt model indicated that the field intervals between larvae and nymphs and between nymphs and adults are controlled solely by nondiapause events, such as ground temperature. We concluded that the 1-yr generation pattern of A. cajennense in Southeastern Brazil is primarily controlled by larval behavioral diapause, shown by larvae born during spring/summer months, before their activity period at next autumn.

The abundance of Ixodes scapularis Say (Ixodes dammini Spielman, Clifford, Piesman & Corwin), the vector tick of the Lyme disease spirochete and other human pathogens, is related to the presence of its primary reproductive stage host, white-tailed deer (Odocoileus virginianus Zimmerman). However, this relationship has not been quantified in terms that would guide wildlife management in areas in which the public is, or is likely to become, exposed to infected ticks. In this study, deer density and tick abundance were measured in an emergent area for Lyme disease at three spatial scales using estimation methods appropriate for each. Simple linear regression was used to relate (1) the number of ticks found on deer at tagging stations in southern Maine to harvest-derived estimates of the density of deer within the towns in which they were killed, (2) tick densities estimated from fall flagging counts to deer densities estimated from pellet group counts made within multiple transects distributed through 5.2-km² study sites, and (3) tick counts to pellet group counts within the individual transects. At the broadest scale, ticks on deer decreased with elevation and distance from the coast and increased with deer density, although deer and tick presence were only weakly related. Among the 5.2-km² study sites and within individual transects, tick abundance related more strongly to deer pellet group counts. Few ticks were collected at deer densities <7/km².

Field-collected Rocky Mountain wood ticks Dermacentor andersoni Stiles from British Columbia caused limited paralysis in hamsters at high doses, but not at lower doses, while ticks collected from Alberta failed to cause paralysis. This prompted development of a laboratory strain of ticks with a more consistent ability to cause paralysis. Progeny of ticks that paralyzed a cow near Walhachin, British Columbia were reared and assayed for paralyzing ability. A high-virulence colony was selected for four generations using female ticks that cause paralysis singly on hamsters. A low-virulence colony was concurrently selected using female ticks that failed to cause paralysis at four ticks per hamster. Logistic regression indicated that the paralyzing ability of the high-virulence colony was consistently higher than that of the low-virulence colony, but that the paralyzing ability of both colonies increased through four generations of selection. Ticks required storage at 10°C to terminate behavioral diapause. The effects of prolonged storage at 10°C on the paralyzing ability of ticks was estimated and was small compared with differences between colonies and across generations. Single ticks from the high-virulence colony caused an increasing proportion of paralysis with each generation, but only limited paralysis with no increase across generations for the low-virulence colony. Single ticks from the high-virulence colony that caused paralysis were similar in weight and percent weight gain compared with single ticks that failed to cause paralysis. Engorged weight and egg mass production after four generations of rearing in the laboratory were similar to that measured for field-collected adults. Results indicated that laboratory selection could increase paralysis, but not eliminate it. This was likely a result of an inability to select male ticks.

A sodium channel ‘kdr’-type mutation was identified in the M form of Anopheles gambiae from Burkina Faso in the tropical savannah area belt. The molecular M form of An. gambiae is found at high frequencies in the flooded rice cultivation area of Kou Valley, where the insecticide selection pressure is limited. The spread of the mutation in the M population is an ongoing process, as it increased from a frequency of 0.006 in 1999 to 0.02 in 2000. The S molecular form occurs in sympatry in our study village, with the M form at a relatively low frequency. The common ‘kdr’ mutation was previously detected in this area in the S form, and has probably invaded the M population through genetic introgression. This impacts on the question of actual levels of gene flow between the two molecular forms in tropical savannah areas. A hybrid M/S individual was identified during the course of this study, which was homozygous for the ‘kdr’ mutation. Pyrethroid resistant An. gambiae were caught in October and November, which is the time of year that the molecular S form migrates into this area as rain-fed breeding sites in the cotton fields dry out.

The sensitivity of vesicular stomatitis (VS) viruses to interferon (IFN)-mediated anti-viral effects has been well documented. Previous studies in our laboratory have shown the ability of mosquito saliva to enhance vesicular stomatitis New Jersey (VSNJ) virus infection in mice. To investigate the effect of mosquito saliva on virus replication and IFN α/β expression, virus titers were analyzed at various time points after infection in cells that were treated with mosquito salivary gland homogenate (SGH). Salivary gland treatment of mouse fibroblast cells (L929) resulted in a significant increase in virus growth kinetics compared with untreated controls. In contrast, Vero cells, which are deficient in the IFN α/β response, did not yield increased viral titers in the time points examined. Treatment of L929 cells with an IFN α/β neutralizing antibody also slightly increased virus yield. Ribonuclease protection assays revealed that induction of IFN α2 expression was reduced in L929 cells treated with SGH. Modulation of IFN α/β by mosquito saliva may be a critical determinant of the transmission and pathogenesis of VSNJ virus.

Immunosuppression of house finches was attempted by blood feeding Culex tarsalis Coquillett mosquitoes or by injecting birds with the corticosteroid dexamethasone or the immunosuppressant drug cyclophosphamide before and after inoculation with western equine encephalomyelitis or St. Louis encephalitis viruses. Mosquito bites (8–37 females blood feeding on each bird over a 3-d period) did not enhance the viremia response or increase the frequency of chronic infection. In contrast, dexamethasone and cyclophosphamide enhanced the amplitude and duration of the viremia response, but had no consistent effect on the antibody responses as measured by enzyme immunoassay or plaque reduction neutralization assay. Elevated viremias were followed by increases in the frequency of chronic infections with St. Louis encephalitis, but not western equine encephalomyelitis. Immunosuppression may provide a useful tool to study the chronic infection process of flaviviruses in vertebrates.

A polymerase chain reaction (PCR) protocol was developed to identify host bloodmeals from mosquitoes. Primers for the cytochrome b gene were designed to distinguish between mammalian and avian bloodmeals and further differentiate among four avian orders: passeriformes, falconiformes, columbiformes, and galliformes. The assay was validated by testing tissues from 18 species of passeriformes, three species of falconiformes, three species of columbiformes, and two species of galliformes. American crows were distinguished from other passeriformes by restriction enzyme digestion. Host bloodmeals from engorged mosquitoes collected in New York State were identified to avian order level. PCR was able to detect the mosquito bloodmeal for up to 3 d after feeding on a quail. Significantly, these studies use order-specific primers in a single PCR test to identify mosquito bloodmeals.

Migratory birds could introduce West Nile (WN) virus to Arkansas. The purpose of this study was to establish a cooperative arbovirus surveillance program to monitor mosquitoes and birds in Arkansas for arboviruses. Our objectives were to: 1) perform routine, multicounty collections of mosquitoes and test them for eastern equine encephalitis, St. Louis encephalitis, and WN viruses; and 2) conduct passive surveillance by testing dead wild birds for WN virus. Arbovirus surveillance was organized by the Arkansas Department of Health, University of Arkansas, and Vector Disease Control Incorporated. None of the 14,560 mosquitoes (425 pools) tested were virus positive. Two hundred forty-two dead birds from 62 counties were tested for WN virus. Four blue jays in three counties were positive. These infections are the first reported incidences of WN virus in Arkansas. Sera from five horses with suspected encephalitis all tested negative for WN, eastern equine encephalitis, and western equine encephalitis viruses.

Ticks are of considerable medical and veterinary importance because they directly harm the host through their feeding action and indirectly through vectoring many bacterial pathogens. Despite many ticks being known from Thailand, very little is known about the bacteria they may harbor. We report here the results of a survey of tick-associated bacteria in Thailand. A total of 334 individuals representing 14 species of ticks in five genera were collected from 10 locations in Thailand and were examined for the human pathogens, Borrelia, Francisella, Rickettsia, and the common arthropod endosymbionts, Wolbachia, by polymerase chain reaction (PCR) assay using specific primers. Rickettsial DNA was detected in 30% (9/30) of Amblyomma testudinarium (Koch, 1844) collected from Khao Yai National Park, Nakhon Nayok Province and 16.84% (16/95) of Hemaphysalis ornithophila (Hoogstraal and Kohls, 1959) collected from Khao Yai National Park, Nakhon Nayok Province and Khao Ang Rue Nai Wildlife Sanctuary, Chachoengsao Province. Rickettsial DNA was not detected in any of the other tick species and no DNA of Borrelia, Francisella, or Wolbachia was detected in any of 14 tick species. Phylogenetic relationships among the rickettsiae detected in this study and those of other rickettsiae were inferred from comparison of sequences of the 17-kDa antigen gene, the citrate synthase gene (gltA), and the 190-kDa outer membrane protein gene (ompA). Results indicated that the three Thai rickettsiae detected in this study represent new rickettsial genotypes and form a separate cluster among the spotted fever group rickettsiae.

Host-seeking activity of adult blacklegged ticks, Ixodes scapularis Say was monitored by flagging during winter months in Beltsville, MD. Ticks were active many days during January and February, the coldest months, with some captures made when there was 70% snow cover and temperatures as low as −2°C. Substantial numbers (70–90 ticks/h of flagging) of adult I. scapularis were captured on favorable days in January and February. The cost to treat white-tailed deer, Odocoileus virginianus (Zimmermann), using ‘4-poster’ deer self-treatment devices, was estimated on a per female tick basis. We estimated deer abundance and tick attachment rates using data from the literature, tick activity levels using weather data and drag counts of ticks, and costs to operate the devices from experience. We found that self-treatment devices need not be operated continuously from late December until the third week of February. On average, savings of half the operating costs would be realized by not operating the devices when tick activity was low.

The bont tick, Amblyomma hebraeum Koch, is the principal vector of Rickettsia africae, the agent of African tick bite fever, in southern Africa. In contrast to most other hard ticks, members of the genus Amblyomma exhibit a characteristic attack host-seeking behavior. The efficacy of topically applied 20% lotions of DEET (diethyl-3-methylbenzamide), the currently considered reference repellent substance, and KBR 3023, a recently developed piperidine compound, was evaluated against laboratory-reared A. hebraeum nymphs on treated fingers. Both substances repelled >85% of nymph attacks at 0 and 1 h postapplication. At 2, 3, and 4 h, the repellent efficacies of DEET were 84%, 68% and 71%, whereas those of KBR 3023 were 56, 55, and 54%. The observed differences between the two test substances were statistically significant only at 2 h postapplication. This first study of topical repellents against A. hebraeum suggests that 20% lotions of DEET and KBR 3023 are effective for 2 and 1 h, respectively, but that the repellent efficacies decrease thereafter.