Ixodes philipi Keirans & Kohls, 1970 is so far known only from two female specimens taken from the streaked shearwater, Puffinus leucomelas (Temminck, 1835) [presently Calonectris leucomelas (Temminck, 1835)], or the madeiran storm-petrel, Oceanodroma castro (Harcourt, 1851), in Japan. We redescribe the female and describe the male of this species. We used nine adults (four males and five females) taken from the bodies of C. leucomelas and/or soil samples of their nest burrows in Japan, indicating that C. leucomelas is the main natural host bird of I. philipi.

Midguts and salivary glands of newly hatched larvae (L₁) of Dermatobia hominis (L., Jr.) were studied using light and electron microscopy. The larval midgut has a tubular, sinusoidal form and consists of a monolayer of epithelial cells with an underlying basement membrane and a surrounding layer of connective tissue. The fine structure of the midgut shows digestive cells with short microvilli, large nuclei, and cytoplasm containing few visible organelles (mitochondria, rough endoplasmic reticulum, and free ribosomes). In the basal region, the plasma membrane of the cells is folded into a labyrinth area. Hemidesmosomes link the basal surface to the basement membrane and septet junctions are present between adjacent cells. The connective tissue circling the basement membrane contains collagen fibrils, muscle fibers, and tracheal tubes. Prominent nuclei with evident nucleoli occur in the digestive cells. The salivary gland is simple and tubular. It has a monolayer of epithelial cells surrounded by basement membrane and connective tissue. The fine structure of the salivary gland shows epithelial cells, microvilli, secretion into the lumen, septate junctions at the lateral face and a basal labyrinth region. The cell nucleus is large and the cytoplasm contains rough endoplasmic reticulum, ribosomes and mitochondria.

Three new species and one new subspecies in the subgenus Afrorhynchus of the genus Toxorhynchites are described from the Afrotropical region. With the addition of these four new taxa, 14 species and two subspecies of subgenus Afrorhynchus are presently known in the region. Keys are provided for the identification of adult males and females of the subgenus Afrorhynchus in Africa.

The morphology of all instars of Chrysomya nigripes Aubertin, a blow fly species of forensic importance, is presented with the aid of both light microscopy and scanning electron microscopy (SEM). Morphological features of the cephalopharyngeal skeleton, anterior spiracle, posterior spiracle, and dorsal spines between the prothorax and mesothorax are highlighted. No consistent features were found, even using SEM, for distinguishing the first instar of C. nigripes from that of Chrysomya megacephala (F.) or Chrysomya rufifacies (Macquart), two other commonly associated blow fly species in corpses in Thailand. Several features observed in second and third instars proved to be valuable characteristics for separating these species.

This study investigated whether Anopheles gambiae s.l. and Anopheles funestus Giles mosquito populations were distributed randomly among houses on the coast of Kenya. Sample means and variances of mosquitoes were estimated from bimonthly pyrethrum spray collections at 30 villages from July 1997 through May 1998. In total, 5,476 An. gambiae s.l. and 3,461 An. funestus were collected. The number of An. gambiae s.l. collected was highest in November/December and lowest in May. The number of An. funestus collected was highest during September/October and lowest during May. As the density of mosquitoes decreased, there was a tendency toward randomness in the distribution. The proportion of An. gambiae s.l. and An. funestus mosquitoes collected per house for each sampling period also showed patterns of clustering, with 80% of An. gambiae s.l. collected from <30% of the houses and 80% of An. funestus collected from <20% of the total houses. The total number of mosquitoes collected from any one house ranged from 0 to 121 for An. gambiae s.l. and from 0 to 152 for An. funestus. This coupled with the results of the variance to mean ratio plots suggests extensive clustering in the distribution of An. gambiae s.l. and An. funestus mosquito populations throughout the year along the coast of Kenya.

The argasid tick Ornithodoros coriaceus (Koch) is the only confirmed vector of epizootic bovine abortion (EBA) in the United States. The disease and its tick vector have historically been reported in the foothills of the Sierra Nevada Mountains and coast ranges of California. In the past two decades, the range of EBA has apparently expanded into southern Oregon and northern Nevada. Possible explanations for this expansion include 1) increased recognition and reporting of EBA in these regions; 2) widespread movement of tick-infested and EBA-infected hosts with subsequent colonization of these regions by infected ticks; and 3) widespread movement of the EBA agent, independent of tick movements, into extant tick populations in these new regions. The current study was performed to evaluate these hypotheses by examining patterns of variability in a 420-bp segment of the 16S mitochondrial rDNA gene sequence among 210 O. coriaceus individuals from 14 sites in California, Oregon, and Nevada. Sixty-three unique haplotypes were identified in the ticks tested, with 84% of the sequence variation attributable to among-population variation and 16% to within-population variation. A majority of the haplotypes were unique to their particular collection site, whereas only four collection sites shared haplotypes. Overall, very little evidence of gene flow among tick populations was detected, making it unlikely that widespread tick movement had introduced O. coriaceus and the EBA agent into new regions.

Ochlerotatus japonicus (Theobald) collected in gravid traps were held individually in modified BioQuip breeders in a darkened room under ambient conditions and readily oviposited on Styrofoam blocks. Counts were made of eggs laid on Styrofoam blocks, container sides, and water surface, and dissections allowed for counts of retained eggs to determine the fecundity of this species. In total, 32,970 eggs were oviposited by 289 females with a range of 3–249 eggs per female. The mean ± SD number of eggs oviposited per female, 114.08 ± 51.08, was significantly less than the mean number of eggs produced per female, 125.85 ± 47.89. Wing lengths ranged from 2.8 to 4.7 mm with a mean of 3.81 ± 0.38 mm. Wing length was positively correlated with total eggs formed (r² = 0.3481); wing = 3.22054 0.0047 total eggs formed. There was no monthly size difference although females collected during October produced significantly fewer eggs than females collected from June to September. A low egg viability of 40% was attributed to improper storage of the Styrofoam blocks on which eggs were oviposited.

Two stable fly species, Stomoxys calcitrans (L., 1758) and Stomoxys niger niger Macquart, 1851, co-occur in La Réunion, where they are important pests of cattle. The survival and developmental rate of the immature stages were compared at five constant temperatures from 15 to 35°C. In both species, immature survival was highest at 20–25°C and markedly decreased at 15 and 35°C. At the lower temperatures, mortality was observed mainly for S. calcitrans larvae and S. niger eggs. At the higher temperatures, mainly pupae of both species died. At all temperatures, S. calcitrans survived better than S. niger. Developmental time was highly similar in both species, decreasing from 71 d at 15°C to 13 d at 30°C in S. calcitrans and from 69 d at 15°C to 14 d at 30°C in S. niger. Developmental times increased slightly at 35°C. Surprisingly, the tropical S. niger developed slightly faster than the cosmopolitan S. calcitrans at 15–20°C; the reverse was found at higher temperatures. Temperature summation models confirmed that S. niger had a lower developmental threshold than S. calcitrans (11.3 versus 12.2°C) and higher day-degree (DD) requirements to complete development (251 versus 225 DD). Overall, the results suggest that S. calcitrans is better adapted than S. niger, in terms of adult production from eggs, in the temperature range of La Réunion.

House dust mites are found in almost all dwellings in New Zealand and are a major risk factor in the development of asthma and perennial allergic rhinitis. We studied the longevity, life stage length, and fecundity of a New Zealand strain of European house dust mite, Dermatophagoides pteronyssinus (Trouessart), at constant (23°C, 75% RH) and the fluctuating conditions typically found in dry (18–25°C, 60–38% RH) and damp (18–23°C, 70–55% RH) New Zealand dust mite microhabitats in carpets. All the adult mites placed in the “dry” conditions died within 18 d. Mites in the “damp” conditions had developmental times, oviposition, and death rates that were not significantly different from constant conditions. These mites are tolerant of fluctuating temperatures, but they are more susceptible to environments that strongly fluctuate in humidity.

The western Kenya highland has been experiencing dramatic landuse changes in the past three decades. Landuse change has been hypothesized to be one of the mechanisms for malaria epidemics in African highlands because it can alter the physical and chemical characteristics of mosquito breeding habitats. One important landuse change in western Kenya highland is deforestation. The current study examined the effects of forestation or deforestation on the survivorship of Anopheles gambiae larvae and colonization of other aquatic insects in larval habitats in Kakamega forest (elevation 1,500–1,700 m above sea level), western Kenya. We found that the survivorship of An. gambiae larvae was reduced from 55 to 57% in habitats fully exposed to sunlight (open habitats) to 1–2% in habitats with full forest canopy coverage (forest habitats) and partial canopy coverage (forest edge habitats) in two out of three trials. The average daily water temperature of the open habitats was ≈3–3.4°C higher than the forest habitats. Insect species in the orders of Diptera, Coleoptera, and Odonata colonized the larval habitats, but the three habitat types differed greatly in the animal assemblage. Canonical correspondence analysis found that water temperature and amount of leaf litter were the significant variables associated with animal assemblages. Redundancy analysis revealed that openness and the presence of predatory animals were significantly related to An. gambiae survivorship. This result suggests that deforestation facilitates the survival of the immature stage of An. gambiae in the highland.

We studied the aquatic mosquito habitats in and around enzootic foci of Venezuelan Equine Encephalitis virus (VEE) in western Venezuela. Specimens were sampled for 5 mo in three types of vegetation: tall lowland tropical forests, short inundated/secondary growth forests, and pastures/herbaceous vegetation around forests. Ground pools, flooded pastures, swamps, ponds, and canals predominated. We used a multivariate statistical approach to quantitatively assess the relationships of mosquito species with broad categories of the landscape, and with environmental variables within each aquatic habitat. Twenty-four mosquito species in the genera Aedes, Psorophora, Culex, Mansonia, and Uranotaenia were collected. Species richness was higher in the tall forests than in other types of vegetation. Discriminant Function Analysis showed a strong association between landscape category and mosquito species assemblage and identified Culex erraticus Dyar & Knab and Mansonia titillans Walker as indicator species of open areas, and Aedes serratus Theobald as an indicator of tall forests. M. titillans, Uranotaenia geometrica Theobald, Cx. erraticus, and Culex dunni Dyar were associated with unshaded, warm, vegetated waters in flooded pastures and swamps, whereas Ae. serratus, Aedes fulvus (Wiedemann), Psorophora albipes Theobald, Psorophora ferox (Humboldt), Culex caudelli Dyar & Knab, and Culex pedroi Sirivanakarn & Belkin were associated with small, shaded ground pools within the tall forests. Culex coronator Dyar & Knab was associated with partially exposed sites within short forests. These results allowed us to interpret better our previous studies on mosquito adult spread in the study area and their possible role as VEEV disseminators.

We compared the distribution of sympatric Ixodes scapularis Say and Amblyomma americanum (L.) within several suitable forested habitats at different spatial scales and characterized differences in microhabitat features accounting for the observed distribution of questing ticks. We used automatic data loggers placed in the shrub and litter layers to contrast mesoclimate and microclimate conditions experienced by questing ticks. Larger numbers of I. scapularis were collected at sites where the forest canopy was more fragmented and where the canopy contained more hardwood species than pitch pine, Pinus rigida Mill. Dominance of pine in the canopy affected the character of the shrub layer vegetation and composition of the litter layer, which concomitantly affected the microclimate conditions experienced by questing ticks. Pitch pine-dominated habitats were drier and hotter than those under a broad-leaved canopy, and questing ticks experienced increased saturation deficit in the later spring and summer in pine forest sites. The shrub layer vegetation seemed to have a moderating effect on the microclimate experienced by questing ticks and subtle differences in vegetation structure resulted in substantially different conditions as encountered by questing ticks over the space of a few meters. In contrast to questing I. scapularis, all three stages of questing A. americanum exhibited poor relationships with microclimate variables recorded in the litter and shrub layers. Further research is required to determine which environmental conditions and which habitats are most likely to support this species.

One potentially important target of malaria vector control is the immature stages of anopheline mosquitoes. To design efficient larval control methods, mechanisms regulating mosquito productivity in natural habitats must be understood. We examined the relationships between pupal occurrence of Anopheles gambiae s.l. and size and stability of larval habitats for a period of 1 yr in western Kenya. We also examined relationships between abundance of indoor resting anophelines and habitat availability. Habitat size was measured by the total water volume (cubic meters). Habitat stability was defined as the number of occurrences when water was continuously present in a habitat for 6 d. Pupal occurrence was indicated by the number of days that pupae were observed in a habitat during the study period. We found that habitat stability and pupal occurrence were positively correlated with habitat size. When habitat size fell below ≈1 m³, habitat stability and pupal occurrence decreased rapidly. Habitat availability was significantly correlated with the density of indoor resting mosquitoes in houses near to larval sites. These results suggest that habitat size is an important determinant of habitat stability, pupal occurrence, and adult mosquito abundance.

In an effort to determine whether female Culex pipiens L. and Culex restuans Theobald mosquitoes (Diptera: Culicidae) are attracted to crow, Corvus brachyrhynchus, uropygial gland secretions, CDC miniature light traps (baited with CO₂ but with the lights removed) were placed at ≈1.5- and 5-m elevations, in 10 trees in a woodlot near Niagara Falls, Canada. These traps were assigned either a bird odor or a blank control. Bird odors were created by attaching cotton swabs coated with crow uropygial gland secretions to the trap intake. A significantly greater number of Cx. pipiens/restuans were found in the 5-m traps compared with 1.5-m traps, with a significant number attracted to the bird odor over the no odor traps at the 5-m elevation, but not at 1.5 m. We also found more Aedes vexans (Meigen) in the 1.5-m traps than the 5-m traps; however, presence or absence of bird odor did not influence the distribution of Ae. vexans.

Nepetalactone, the primary component of catnip oil, was compared with the repellent N,N-diethyl-3-methylbenzamide (deet) for its ability to affect the host-seeking ability of Aedes aegypti (L.). A triple cage olfactometer was used to bioassay each substance and to assess its attraction inhibition (spatial repellent) attributes when combined with the following attractants: carbon dioxide, acetone, a blend of l-lactic acid and acetone, and human odors. Repellent tests were conducted with each substance against female Ae. aegypti, Anopheles albimanus Weidemann, and Anopheles quadrimaculatus Say. Catnip oil and deet were both weakly attractive to Ae. aegypti, catnip oil was the better spatial repellent, whereas deet was a more effective contact repellent in tests with all three species of mosquitoes.

Nocturnal behavior of nonblood-fed females of Aedes aegypti (L.) and Aedes albopictus (Skuse) was studied using an automatic recording device equipped with a photoelectric sensor. Carbon dioxide, heating, and the contrast of the black and white colors were used as attractive cues for mosquitoes. The nocturnal host-seeking activity positively correlated with the increasing light intensity in both species. Ae. aegypti was found to be more sensitive to light than Ae. albopictus. The threshold of light intensity for the activation of the nocturnal host-seeking activity was <0.1 lx (≈0.01 foot candle) in Ae. aegypti and >10 lx (≈1 foot candle) in Ae. albopictus. Complete darkness during the daytime deactivated the host-seeking activity of both species, irrespective of their increasing flight activity controlled by their intrinsic circadian rhythms. This finding suggested that visual cues are indispensable for host-seeking behavior. The eye parameter value, the product of the ommatidial diameter, and the interommatidial angle were significantly larger in Ae. aegypti than those in Ae. albopictus, indicating that the eye of Ae. aegypti is more adapted to a darker environment.

Isozyme electrophoresis studies were conduced on Anopheles oswaldoi sensu lato from three states in Amazonian (Brazil), including Acre, where this taxon has been incriminated as a potential human malaria vector. The 13 enzymes analyzed yielded a total of 20 loci. Of these, 10 were monomorphic in the three samples. Diagnostic loci were not found. The measures of genetic variability showed mean number of alleles per locus between 1.5 and 2.0, percentage of polymorphic loci of 40%, and mean heterozygosities from 0.031 to 0.062. The inbreeding coefficient F_IS had a moderate mean value (0.109), which resulted from heterozygote genotype deficiencies and the presence of rare alleles in the homozygote state. Mean F_ST value (0.0502) and genetic distance values (Nei’s genetic similarity values of 0.994–0.997; Roger’s genetic similarity values of 0.963–0.968) were very low among the three samples of An. oswaldoi s. l. Preliminary results of identifications of male genitalia indicated that An. oswaldoi s. s. Peryassú and Anopheles konderi Galvão & Damasceno of the An. oswaldoi complex coexist sympatrically in Sena Madureira, Acre, and São Miguel, Rondônia, whereas in the sample from Coari, Amazonas, only An. konderi was identified. The isozyme results do not suggest differences between these species, particularly in areas of sympatry.

This survey was carried out to determine the seroprevalence of nasal infestation by sheep bot fly, Oestrus ovis L., and to identify the risk factors associated with the disease in flocks in southwestern Spain. In total, 5,878 sera samples of adult sheep were collected at random in 551 farms from four provinces in the southwestern Spain: Badajoz, Cáceres, Córdoba, and Sevilla. Sera were tested by enzyme-linked immunosorbent assay for O. ovis antibodies, by using a crude L2 larval as antigen. The seropositive mean prevalence was 69.30%, and mean percentage of optical densities was 61.83%. There were significant differences between the provinces studied; Córdoba and Sevilla were the provinces with more infested animals and higher seroprevalences. The correlation between seroprevalence and percentage of antibodies by farms was significant. There were only 18 farms free of seropositive animals, and 115 of the total 551 farms had all sampled animals seropositive, an indication of the high importance of this parasitosis in the investigated areas. Altitude, latitude, flock size, and ovine population density were the potential risk factors associated with the detection of O. ovis antibodies. Those animals breeding in regions located at low altitudes (<500 m), meridian latitudes (<39.5° N), and on farms with medium-to-large flock size (>250 sheep) and high ovine population density (>100 sheep per km²) were more likely to be seropositive. These findings confirm that these studied factors should be considered as potential risk factors to the presence of O. ovis in ovines from southwestern Spain.

Nineteen strains and species of entomopathogenic fungi were examined for their potential as biological control agents of the house fly, Musca domestica L. Bioassays against larvae, pupae, and adult males and females in small containers indicated five strains of Beauveria bassiana (Bals.) Vuill. were relatively more virulent. Three of the virulent strains, chosen for efficiency of production in a rice-based, diphasic fungal production system, were further evaluated by mixing spore-contaminated rice residue with sugar and tested as a food bait for adult flies in a 33-m³ walk-in chamber. Up to 90% mortality was obtained within 15 d of exposure. The results indicate bait formulations of the most virulent and aggressive strain should be further developed and studied.

A fundamental principal of resistance management is that the more complex and potent a toxin mixture, the slower resistance will develop to the mixture in an insect population. Thus, to develop more complex and potent mosquitocidal bacteria, we genetically engineered Bacillus thuringiensis subsp. morrisoni PG-14 and Bacillus thuringiensis subsp. jegathesan, to synthesize, respectively, the binary (Bin) toxin of Bacillus sphaericus or a combination of Bin and the Cyt1A protein of Bacillus thuringiensis subsp. israelensis. Engineering these two larvicidal bacteria in general significantly improved their efficacy against fourth instars in comparison with their wild-type parental strains. For B. thuringiensis subsp. morrisoni PG-14, which naturally synthesizes Cyt1A, synthesis of Bin improved efficacy nine-fold (LC₅₀ from 4.5 to 0.5 ng/ml) against Culex quinquefasciatus Say, although no improvement was observed (LC₅₀ of 2 ng/ml for both strains) against Aedes aegypti L. For B. thuringiensis subsp. jegathesan, cosynthesis of Bin plus Cyt1A in combination with its normal complement of endotoxins improved efficacy 17-fold (LC₅₀ from 34 to 2 ng/ml) against Cx. quinquefasciatus and 3.2-fold (LC₅₀ from 68 to 21 ng/ml) against Ae. aegypti. Addition of Bin alone to B. thuringiensis subsp. jegathesan did not improve toxicity (LC₅₀ from 68 to 65 ng/ml) against Ae. aegypti, indicating that Cyt1A synergized the activity of the endotoxins in this strain against Ae. aegypti. These results demonstrate that mosquitocidal efficacy of these strains and likely their resistance management properties can be improved significantly by increasing their toxin complexity and the amount of toxin they synthesize.

We studied the profile of permethrin resistance in populations of head lice infesting children 6–12 yr old in schools and their homes in and around Buenos Aires, Argentina. Five permethrin-resistant populations with different levels of resistance were collected: Hogar Loyola (HL), Republica de Turquia (RT), Hogar Mitre (HM), Guardia de Honor (GH), and Ricardo Guiraldes (RG). One susceptible population, Bandera Argentina (BA), also was collected. Their level of resistance was evaluated, and results showed resistance ratios of 13 for HL, 16 for RT, 22 for HM, 61 for GH, and 69 for RG. To elucidate the possible involvement of the cytochrome P450 monooxygenase system in conferring permethrin resistance, ethoxycoumarin-O-deethylase (ECOD) activity was measured in abdomens of individual third instars and adults by using a fluorometric assay. The ECOD activity was lower in the susceptible BA population (4.7 ng per louse) than in the resistant ones (13.7 ng per louse for RG, 12.3 ng per louse for GH, 8.6 ng per louse for RT, and 8.2 ng per louse for HL). ECOD activity was significantly correlated with the level of resistance in the field populations (r = 0.97, P = 0.0009), suggesting a role for cytochrome monooxygenase P450 system in permethrin resistance by head louse, Pediculus humanus capitis De Geer.

Cell-free culture supernatants of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin were able to induce mortality in several tick species, including adult Ambylomma americanum (L.), Ambylomma maculatum Koch, and Ixodes scapularis Say. Four lines of experimental evidence support the hypothesis that oxalic acid secretion by B. bassiana, coupled to a reduction in the pH of the medium, act as potent acaricidal factors during pathogenesis. 1) Acaracidal activity of culture supernatants was retained after treatments including boiling and protease digestion, but was lost after dialysis. 2) Metabolite analyses indicated oxalate to be the major secreted organic compound present in the active culture supernatants. 3) Treatment of ticks with the pure compound oxalate at pH 4.0 resulted in almost 80% mortality in adult A. americanum ticks within 14 d, whereas treatment of ticks with oxalate at pH 7.0, or with formate, citrate, or phosphate at both pH 4 and 7 resulted in <10% mortality, even after 28 d. 4) Cell-free culture supernatants from B. bassiana mutants with decreased oxalate production displayed lower acaricide activity than wild type.

Laboratory bioassays were conducted to determine the activity of 15 natural products isolated from essential oil components extracted from the heartwood of Alaska yellow cedar, Chamaecyparis nootkatensis (D. Don) Spach., against Ixodes scapularis Say nymphs, Xenopsylla cheopis (Rothchild), and Aedes aegypti (L.) adults. Four of the compounds from the essential oil have been identified as monoterpenes, five as eremophilane sesquiterpenes, five as eremophilane sesquiterpene derivatives from valencene and nootkatone, and one as a sesquiterpene outside the eremophilane parent group. Carvacrol was the only monoterpene that demonstrated biocidal activity against ticks, fleas, and mosquitoes with LC₅₀ values after 24 h of 0.0068, 0.0059, and 0.0051% (wt:vol), respectively. Nootkatone from Alaska yellow cedar was the most effective of the eremophilane sesquiterpenes against ticks (LC₅₀ = 0.0029%), whereas the nootkatone grapefruit extract exhibited the greatest biocidal activity against fleas (LC₅₀ = 0.0029%). Mosquitoes were most susceptible to one of the derivatives of valencene, valencene-13-aldehyde (LC₅₀ = 0.0024%), after 24 h. Bioassays to determine residual activity of the most effective products were conducted at 1, 2, 4, and 6 wk after initial treatment. Residual LC₅₀ values for nootkatone did not differ significantly at 4 wk posttreatment from the observations made at the initial 24-h treatment. The ability of these natural products to kill arthropods at relatively low concentrations represents an alternative to the use of synthetic pesticides for control of disease vectors.

The phenomenon of immunoglobulin uptake from the midgut into the hemolymph has been observed in both argasid and ixodid ticks, but the mechanism of the uptake is not known. Using capillary feeding of adult Amblyomma americanum (L.) female ticks, we investigated the role of immunoglobulin G Fc and Fab fragments in the uptake process. Intact IgG and fragments labeled with ¹²⁵I were introduced alone or as a mixture with a 10-fold molar excess of unlabeled compounds into ticks, and then the concentration of the compounds after 3 and 6 h of continuous feeding was assessed in the hemolymph. Our results showed that the uptake of Fc fragments was ≈4 times higher than Fab fragments at 6 h of the feeding and that Fc but not Fab fragments competed with uptake of intact IgG. Glycosylation of the immunoglobulins did not seem to be required for uptake. There was not a statistically significant difference between the uptake of untreated IgG and IgG treated either with periodate or N-glycanase, and untreated Fc fragments or deglycosylated Fc fragments had equal activity in competition experiments with IgG. The uptake into hemolymph of IgG and Fc fragments, but not Fab fragments, was dependent on the pH of the feeding solution and showed a maximum at ≈pH 7.0. In addition, tick midgut cells bound Fc fragments with high affinity, estimated at 21 nM; the interaction with Fab fragments seemed nonspecific. Our results suggest that Fc fragments are the major determinant for the specific uptake of immunoglobulin G into tick hemolymph.

The ability of the invading NY99 strain of West Nile virus (WNV) to elicit an elevated viremia response in California passerine birds was critical for the effective infection of Culex mosquitoes. Of the bird species tested, Western scrub jays, Aphelocoma coerulescens, produced the highest viremia response, followed by house finches, Carpodacus mexicanus, and house sparrows, Passer domesticus. Most likely, few mourning, Zenaidura macroura, or common ground, Columbina passerine, doves and no California quail, Callipepla californica, or chickens would infect blood-feeding Culex mosquitoes. All Western scrub jays and most house finches succumbed to infection. All avian hosts produced a lower viremia response and survived after infection with an endemic strain of St. Louis encephalitis virus. Culex species varied in their susceptibility to infection with both viruses, with Culex stigmatosoma Dyar generally most susceptible, followed by Culex tarsalis Coquillett, and then Culex p. quinquefasciatus Say. Populations within Culex species varied markedly in their susceptibility, perhaps contributing to the focality of WNV amplification. Transmitting female Cx. tarsalis expectorated from six to 3,777 plaque-forming units (PFU) of WNV during transmission trials, thereby exposing avian hosts to a wide range of infectious doses. Highly susceptible house finches and moderately susceptible mourning doves were infected by subcutaneous inoculation with decreasing concentrations of WNV ranging from 15,800 to <0.3 PFU. All birds became infected and produced comparable peak viremias on days 2–3 postinoculation; however, the rise in viremia titer and onset of the acute phase of infection occurred earliest in birds inoculated with the highest doses. WNV virulence in birds seemed critical in establishing elevated viremias necessary to efficiently infect blood feeding Culex mosquitoes.

The effect of prefeeding Dermacentor andersoni Stiles on naïve and previously challenged cattle before conducting virulence assays by using hamsters was examined. In an initial observational trial, ticks prefed on a previously challenged heifer did not cause paralysis after transfer to hamsters, whereas ticks from the same lot that were prefed on a naïve heifer did cause paralysis. Subsequent paired trails were conducted to simultaneously compare paralysis after feeding on naïve and previously challenged heifers. In the first trial, prefeeding ticks on a previously challenged heifer did not result in loss of virulence. In a second trial, ticks fed on a previously challenged heifer had greatly diminished virulence after transfer to hamsters. This also occurred in a third trial after the previously challenged animal had two prior exposures to virulent D. andersoni. Weight gains of ticks that did not cause paralysis after transfer to hamsters were higher after feeding on previously challenged cattle, indicating that loss of virulence was not due to failure of ticks to reattach and feed. Ticks gained little weight on paralyzed hamsters due to the reduced time spent feeding on hamsters when paralysis occurred. Results indicate that assaying ticks for virulence requires prefeeding on naïve hosts. If these are not available, rabbits, a resistant host, can be used for prefeeding ticks.

The capacity of ticks to transmit a fungus was examined by analyzing tick saliva, host tissue from feeding sites, and host blood for presence of Scopulariopsis brevicaulis (Sacc.) Bainier, an internal mycosymbiont of the American dog tick, Dermacentor variabilis (Say), and lone star tick, Amblyomma americanum (L.). Although >85% of ticks were infected with S. brevicaulis, conidia presence was low (0–5% of samples) in microscopic observations and mycological culturing of saliva expressed from larvae, nymphs, and adults. Additionally, the recovery of S. brevicaulis from blood and tissue feeding sites from a rabbit where S. brevicaulis-positive adult ticks had attached and fed was not increased compared with control tissue where no feeding occurred, indicating that transmission does not occur by the blood-feeding route. Tick mouthparts were found to contain S. brevicaulis in addition to Penicillium glabrum (Wehmer) Westling, but these agents were sparse in isolations from the feeding sites, which makes it unlikely that ticks act frequently as a mechanical fungal vector.

In California, dense woodlands have been recognized as important biotopes where humans are exposed to the nymphal stage of the western blacklegged tick, Ixodes pacificus Cooley & Kohls, the primary vector of the Lyme disease spirochete Borrelia burgdorferi sensu stricto (s.s.), in the far-western United States. To identify the principal reservoir host(s) of this spirochete, and of closely related spirochetes in the B. burgdorferi sensu lato (s.l.) complex, in dense woodlands in Mendocino County, California, ≈50 species of birds and mammals, including wood rats and kangaroo rats, were evaluated as potential hosts for vector ticks and borreliae in 2002 and 2003. Although polymerase chain reaction (PCR) and sequencing analyses revealed that many vertebrate species had been exposed to one or more members of the B. burgdorferi s.l. spirochetal complex, only the western gray squirrel, Sciurus griseus, fulfilled the major criteria for a reservoir host of B. burgdorferi s.s. Ear-punch biopsies from eight of 10 squirrels collected from five separate woodlands were PCR-positive for B. burgdorferi s.s., 47% of I. pacificus larvae (n = 64) and 31% of nymphs (n = 49) removed from squirrels contained B. burgdorferi s.l., and the engorgement status of I. pacificus larvae was associated positively with acquisition of spirochetes. Overall, 83 and 100% of the amplicons sequenced from PCR-positive I. pacificus larvae and nymphs, respectively, were identified as B. burgdorferi s.s. Among the five remaining positive I. pacificus larvae, three contained B. bissettii and two had uncharacterized B. burgdorferi s.l. Borrelia burgdorferi s.s. was detected in one of five larvae and zero of two nymphs of the Pacific Coast tick, Dermacentor occidentalis Marx, that likewise had been removed from squirrels. The rickettsial agent of human anaplasmosis, Anaplasma phagocytophilum, was detected in the blood or ear biopsies of two squirrels and in one (1.6%) of 64 I. pacificus larvae and two (4.1%) of 49 nymphs obtained from squirrels. The one rickettsial-positive larva was coinfected with B. burgdorferi s.s. The apparently high reservoir potential of S. griseus for B. burgdorferi s.s., plus the fact that the geographic distribution of this squirrel coincides well with that of most reported human cases of Lyme disease in this region, indicated that it may be essential for maintaining foci of B. burgdorferi s.s. in certain types of woodlands. The findings with respect to A. phagocytophilum, although of less certain significance, suggest that S. griseus could serve as a secondary host of this rickettsia.

Populations of Escherichia coli obtained by feeding larval house flies, Musca domestica L. and stable flies, Stomoxys calcitrans (L.), persisted through the pupal stage. The abundance of E. coli in house fly pupae increased initially then declined before adult emergence. Abundance of E. coli in stable fly pupae increased through pupal development and remained high. Infected stable fly pupal cases typically contained more E. coli than house fly pupal cases. A greater proportion of emerging adult house flies were infected with E. coli compared with stable flies; however, the abundance of E. coli on infected flies was similar between species. Adult flies contained 0.04–0.19% of the E. coli in the pupal cases. The proportion of infected house fly adults and the amount of E. coli on the infected flies were related to the levels of E. coli in the pupal cases; however, these relationships did not occur with the stable fly. Results suggest that retention of E. coli from larval to adult house flies could play a role in the transmission and spread of E. coli, whereas stable fly adults probably play a minor role in E. coli spread. However, pupae of both species have potential to act as reservoirs for E. coli.

Venezuelan equine encephalitis (VEE) complex alphaviruses are serious health threats in the Americas and regularly infect humans living in or near Amazonian rain forests. As part of a larger surveillance program, we placed six hamster-baited mosquito traps in a disturbed white sand forest of northeastern Peru for 3d. Virus isolations from hamster serum and trapped mosquito pools demonstrated that a VEE subtype IIICalphavirus was transmitted to a hamster by the mosquito Culex (Melanoconion) gnomatos Sallum, Hutchings & Ferreira. This species, like the other seven proven VEE complex alphavirus vectors, is a member of the Spissipes section of this subgenus. The composition of mosquitoes collected at the site over the sampling period was typical for the region.

To determine the vector competence of Culicoides sonorensis Wirth & Jones midges for vesicular stomatitis virus (VSV)-New Jersey, insects were experimentally infected per os and sampled over time. Viral replication, as determined by in situ hybridization, was seen in epithelial, neural, and hemolymph cell types throughout the insect. Spatial and temporal distribution of virus was determined by immunohistochemical examination of sequentially sampled insects. Tissues of the alimentary canal were infected in a temporal pattern that paralleled the route of digestion/absorption: foregut and midgut by day 1, surrounding hemolymph and Malpighian tubules by day 3, and finally the midgut/hindgut junction, hindgut, and rectal region by day 5. The circulation of virus in the hemolymph by day 3 coincided with infection of the dermis and fat bodies, the salivary glands, eyes, cerebral and subthoracic ganglia, and the ovaries. Oviduct epithelium and ovarial sheaths were infected by day 3, followed by infection of the developing oocytes by day 5. Interestingly, neural infections were seen in the subabdominal ganglia innervating the midgut in 33% of insects by 1 d postfeeding in the absence of positive staining in the hemolymph or surrounding tissues. A retrograde axonal transport infection route for these ganglia is discussed. The disseminated, productive, noncytolytic infection in Culicoides is consistent with that of an efficient biological vector for VSV. Virus readily replicated throughout the insect, passing both midgut and salivary gland infection barriers and reaching transmission-related organs in 3 d. Establishing the competence of this insect vector for VSV provides the foundation for animal transmission studies in the future. The possibility of horizontal, transovarial, and mechanical transmission is discussed.

Until 2001, the Chinese Territory of Macao had not registered any autochthonous dengue cases, despite the abundance of Aedes albopictus (Skuse), a known vector. This work describes a bioecological characterization of the local Ae. albopictus adult population, with the purpose of estimating the receptivity of Macao to dengue introduction. In the wet seasons of 1997 and 1998 and the dry season of 1998, Ae. albopictus was the most abundant human-biting mosquito. Daily biting rates of 314 mosquitoes per person were recorded in the wet season with a reduction to 94 in the dry season. Ae. albopictus was mainly exophagic and exophilic and had a human blood index of 44%. The parity rate of field-collected mosquitoes was 57%. Daily survival rate ranged from 91 to 97%. Estimates of vectorial capacity ranged from 144 to 880, depending on what parameter values were used. These estimates indicated a great receptivity for the introduction of dengue viruses, as the 2001 outbreak came to prove.

The ultrastructural features of West Nile virus (WNV) replication and dissemination in orally infected Culex pipiens quinquefasciatus Say were analyzed over a 25-d infection period. To investigate the effects of virus replication on membrane induction, cellular organization, and cell viability in midgut and salivary gland tissues, midguts were dissected on days 3, 7, 14, and 21, and salivary glands were collected on days 7, 14, 21, and 25 postinfection (d.p.i.) for examination by transmission electron microscopy (TEM). Whole mosquito heads were embedded for TEM analysis 14 d.p.i. to localize WNV particles and to investigate the effects of replication on nervous tissues of the brain. Membrane proliferation was induced by WNV in the midgut epithelium, midgut muscles, and salivary glands, although extensive endoplasmic reticulum swelling was a unique feature of salivary gland infection. TEM revealed WNV-induced pathology in salivary glands at 14, 21, and 25 d.p.i., and we hypothesize that long-term virus infection of this tissue results in severe cellular degeneration and apoptotic-like cell death. This finding indicates that the efficiency of WNV transmission may decrease with mosquito age postinfection.

Reservoir competence for the Lyme disease spirochete, Borrelia burgdorferi, was tested for six species of native North American birds: American robin, gray catbird, brown thrasher, eastern towhee, song sparrow, and northern cardinal. Wild birds collected by mist netting on Fire Island, NY, were held in a field laboratory in cages over water and locally collected larval ticks were placed on the birds, harvested from the water after engorgement, and tested for infection by direct fluorescent-antibody staining after molting to the nymphal stage. American robins were competent reservoirs, infecting 16.1% of larvae applied to wild-caught birds, compared with 0% of control ticks placed on uninfected laboratory mice. Robins that were previously infected in the laboratory by nymphal feeding infected 81.8% of applied larvae. Wild-caught song sparrows infected 4.8% of applied larvae and 21.1% when infected by nymphal feeding. Results suggest moderate levels of reservoir competence for northern cardinals, lower levels for gray catbirds, and little evidence of reservoir competence for eastern towhees or brown thrashers. Lower infection rates in larvae applied to wild-caught birds compared with birds infected in the laboratory suggest that infected birds display temporal variability in infectiousness to larval ticks. Engorged larvae drop from birds abundantly during daylight, so the abundance of these bird species in the peridomestic environment suggests that they might contribute infected ticks to lawns and gardens.

To evaluate their relative importance in tick-borne disease transmission in New Jersey, host-seeking Amblyomma americanum (L.) and Ixodes scapularis Say adults and nymphs were collected during spring activity periods in 2003 and 2004 to determine relative frequencies at which these ticks were encountered from an area known to be hyperendemic for Lyme disease. Although similar numbers of the two species were encountered during early spring of both years, A. americanum were encountered more often later in the season and exhibited a longer activity period than I. scapularis. A. americanum nymphs were collected at frequencies between 2.6 and 7.3 times higher than I. scapularis nymphs. Polymerase chain reaction (PCR) analysis of 121 A. americanum adults yielded infection prevalences of 9.1% for Borrelia lonestari, 12.3% for Ehrlichia chaffeenensis, and 8.2% for E. ewingii, and coinfection prevalences of 4.1% for E. chaffeensis/E. ewingii and 0.8% for E. chaffeensis/B. lonestari. Infection prevalences in 147 I. scapularis adults were 50.3% for B. burgdorferi, 6.1% for Anaplasma (Ehrlichia) phagocytophilum, and 1.4% for a recently described novel Borrelia species, whereas the coinfection prevalences were 2.7% for B. burgdorferi/A. phagocytophilum, 0.7% for B. burgdorferi/novel Borrelia, and 0.7% for A. phagocytophilum/novel Borrelia. The B. burgdorferi infection prevalence in I. scapularis was considerably higher than that in A. americanum. However, the higher A. americanum encounter frequencies compared with I. scapularis may result in increased risk of acquiring exposure to A. americanum-transmitted pathogens. The potential public health implications of these results are discussed.

Lyme disease, caused by Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt & Brenner; babesiosis, caused by Babesia microti Franca; and human granulocytic ehrlichiosis, caused by Anaplasma phagocytophilum Bakken & Dumler have been reported in Wisconsin, mainly in the endemic areas of the northwestern part of the state. People exposed to blacklegged ticks, Ixodes scapularis Say, from this region can potentially contract one or all of these diseases concurrently. Within the past several years, there have been cases of Lyme disease reported from southeastern Wisconsin, an area that contains deer, mice, and similar vegetation found at regions with high endemicity. However, past white-tailed deer surveys suggested no existence of I. scapularis in southeastern Wisconsin. However, in 1996, we identified questing adult I. scapularis in the southernmost part of the Southern Kettle Moraine State Forest, Walworth County. To determine whether an established population of I. scapularis exists in this region, we performed a detailed survey of the abundance of host-seeking I. scapularis adults and the presence and abundance of subadults feeding on white-footed mice, Peromyscus leucopus Rafinesque. We also tested for possible infections of B. burgdorferi, Ba. microti, and A. phagocytophilum in ticks and B. burgdorferi harbored by mice. In 1997 and 1998, a total of 249 P. leucopus mice and 118 questing adult I. scapularis ticks, in addition to 157 larvae and seven nymphs feeding on mice, were collected and their locations were recorded from the Nordic trails of the Southern Kettle Moraine State Forest. Only one P. leucopus and its attached engorged I. scapularis nymph were infected with B. burgdorferi, whereas none of the engorged larvae attached to mice were infected. However, 4.2% of questing adult I. scapularis were infected with B. burgdorferi. The abundance of questing adult I. scapularis was 1.6 ticks per hour. The prevalence of subadult ticks on mice was 27%, with a mean intensity on infested mice of 2.0. I. scapularis adults were not infected with either Ba. microti or A. phagocytophilum. A unique strain of B. burgdorferi s.l. (W97F51) was discovered, showing 33 nucleotide substitutions and one codon insertion in a 567-bp fragment of the OspB gene, compared with Borrelia bissettii (strain Ca389). The sequences of ospA, ospB, ospC, fla, and rrs genes and the rrf-rrl intergenic spacer region were compared between W97F51 and other B. burgdorferi s.l. species. Although W97F51 was most genetically related to B. bissettii, the genetic identity of W97F51 was less than that of B. bissettii conspecifics. This study documents the existence of an established population of I. scapularis and the presence of B. burgdorferi with a novel strain in southeastern Wisconsin.

Carios kelleyi (Colley & Kohls 1941), a tick associated with bats and bat habitats, has been reported to feed on humans, but there is little published data regarding the presence of vector-borne pathogens in these ticks. C. kelleyi nymphs and adults were collected from residential and community buildings in Jackson County, Iowa, and tested by polymerase chain reaction for Rickettsia, Borrelia, Bartonella, Coxiella, and Anaplasma. Rickettsia DNA was detected in 28 of 31 live ticks. Sequences of the 17-kDa and rOmpA genes suggest that this agent is a novel spotted fever group Rickettsia. Transstadial and transovarial transmission of this Rickettsia were demonstrated. The flagellin gene of a Borrelia, closely related to B. turicatae, was detected in one of 31 live ticks. The 16S-23S intergenic spacer region of Bartonella henselae also was detected in one of 31 live ticks. Coxiella or A. phagocytophilum DNA were not detected in these ticks.

The effects of antemortem ingestion of ethanol by domestic pigs, Sus scrofa L., on postmortem insect successional patterns and the development of Phormia regina (Meigen) were studied during summer 2003 in Blacksburg, VA. Insect samples were collected from the carcasses of ethanol-treated and untreated pigs for 10 d postmortem during two successional studies. In total, 32 insect taxa were collected during the two studies, with 29 and 27 taxa observed on the carcasses of ethanol-treated and untreated pigs, respectively. The earliest arrivers to both carcass types were dipterans. This group was represented by six families, with P. regina and Phaenicia coeruleiviridis (Macquart) being the most common calliphorids. Beetles in six families were collected on the carcasses of ethanol-treated pigs, but only three of the families were collected on carcasses of the untreated pigs. Permutation analyses to test the null hypothesis of no similarity between successional patterns of insect taxa from carcasses of ethanol-treated and untreated pigs showed that the successional patterns were similar between carcass types in the first (P = 0.003) and the second (P = 0.01) studies. The results of the development study of P. regina maggots in the field show that there was a significant difference between the distributions of length for maggots reared on loin tissue from ethanol-treated and untreated pigs. Maggots that fed on tissue from ethanol-treated pigs took ≈11.9 h longer to reach the pupal stage than maggots that fed on tissue from untreated pigs. The longer developmental time for maggots on tissue from ethanol-treated pigs was due mainly to the longer postfeeding period of the third instar.

In total, 1,324 Culex pipiens pipiens L. female mosquitoes were collected at Ft. Hancock, Monmouth County, New Jersey, from January to March 2001–2003. Mosquitoes were held in an insectary at 27°C and a photoperiod of 16:8 (L:D) h for 6 to 21 d after which they were tested in 34 pools. West Nile viral RNA was detected in one pool by a TaqMan reverse transcription-polymerase chain reaction assay; however, infectious virus could not be isolated using either Vero cell plaque assay or C6/36 mosquito cells. Twenty females dissected in January and March 2003 confirmed ovarian diapause status. We suggest that the mode of infection in this pool of overwintering females may have been due to vertical (transgenerational) transmission.

A total of 191 adult sand flies belonging to the genus Sergentomyia were collected from seven villages in Karimnagar and Warangal districts of Andhra Pradesh State, India, after an outbreak of encephalitis due to Chandipura virus (CHPV). Fifteen pools, each containing two specimens, were tested by reverse transcriptase-polymerase chain reaction and sequencing. One pool of Sergentomyia from Kolanur village in Karimnagar District was positive for CHPV.

I found Carios kelleyi Cooley & Kohls (formerly Ornithodoros kelleyi) (Acari: Ixodida: Argasidae) on five species of bats (Eptesicus fuscus, Myotis lucifugus, Myotis ciliolabrum, Myotis evotis, and Myotis volans) in the prairies of southeastern Alberta and north central Montana. This is the first occurrence of ticks on bats in Canada. This is also the first record of C. kelleyi on M. evotis and M. volans. Dates of parasitization ranged from 22 May to 17 October, and the number of ticks per host ranged from 1 to 44 (median two ticks per host). Number of parasitized individuals was significantly higher in rock-roosting E. fuscus compared with building-roosting E. fuscus, rock- and building-roosting M. lucifugus, and rock-roosting M. ciliolabrum and M. evotis. A significant sex bias was noted in rock-roosting E. fuscus in 2000 with significantly more females parasitized. The number of ticks per bat host was highest in 2001, corresponding to the driest and hottest summer of the survey.

While estimating the prevalence of the Dermacentor variabilis (Say) symbiont (DVS) in dog ticks on Martha‘s Vineyard, MA, we identified DNA that may represent a heretofore unrecognized Francisella sp. Polymerase chain reaction targeting a portion of the 16S rDNA specific for DVS yielded an amplicon that was only 96.6% similar to that of DVS accessioned in GenBank. Phylogenetic analysis of the 16S and 23S rDNA genes suggests the presence of a distinct bacterium closely related to the other endosymbionts of Dermacentor spp. Fifty-five percent of dog ticks tested from three sites in Massachusetts showed evidence of infection with this new agent, called Dermacentor variabilis francisella (DVF), whereas 100% tested positive for DVS. All larval progeny of dog ticks known to contain DVF also showed evidence of colonization, demonstrating that this agent may be maintained by transovarial transmission. Coinfection of ticks with both Francisella species did not seem to interfere with transmission.

Blood fed nymphal Ixodes scapularis Say infected with Borrelia burgdorferi were dissected to obtain salivary gland and midgut extracts. Extracts were inoculated into C3H/HeJ mice, and ear, heart, and bladder were cultured to determine comparative infectivity. Aliquots of extracts were then analyzed by quantitative polymerase chain reaction to determine the number of spirochetes inoculated into mice. A comparative median infectious dose (ID₅₀) was determined for both salivary gland and midgut extract inoculations. Our data demonstrated a statistically significant difference (P < 0.002) in the ID₅₀ derived from salivary gland (average = 18) versus midgut (average = 251) extracts needed to infect susceptible mice. A rationale for the differential infectivity of salivary and midgut derived spirochetes is discussed.