This study compared the reliability of population estimates of adult horn fly, Haematobia irritans (L.), obtained using different sample units. Mean-variance relationships were similar for abundance estimates obtained by counting flies on the sunny sides of cattle, on the upper body, and on the whole animal. Precision varied among the sample units, and was lowest for estimates obtained using the sunny side. Abundance estimates obtained using the sunny side and upper body sample units were related to estimates obtained using the whole body sample unit. However, the proportion of flies in the upper body and sunny side sample units declined with increasing fly density. Seasonal movement toward the belly accounted for this decline. This movement resulted in bias in estimating rates of change based on counting flies on the sunny side and upper body sample units. Rates of change based on sampling the sunny side were more biased than estimates based on the upper body sampling unit. Bias in estimating rates of change was examined using an analytical model compared with field data, and resulted from changes in the proportion of flies occupying the sample unit. Bias also increased with increasing actual rates of change. The implication of these findings for studying horn fly populations are discussed.

A triple cage olfactometer provided with insect traps was used for evaluating behavioral responses of Stomoxys calcitrans (L.) females to human skin and breath, CO₂, and L-lactic acid analogs. After demonstrating there were no significant differences caused by cage location or time of day, 3 sets of 3 olfactometer tests were performed in a day, every 2 h beginning at 0900 hours. When a human hand was used as attractant, the attraction (expressed as percentage of trapped flies) increased as a function of the time; an inverted U-shaped relationship between attractancy and air speed was observed; and variation in fly density in the range 25–75 per cage did not affect the attraction response. When human breath was used as attractant the attraction increased linearly as a function of time and it was exhalation frequency dependent; when air flow was absent the highest response was observed; and 24- to 38-h-old flies were more attracted than younger and older. When CO₂ was tested, activation and orientation and probing behavior were concentration dependent with flows ranging between 0.0001 and 0.038 liter s⁻¹, but attraction was not. No attraction was observed with 10, 100, or 1,000 μg of compounds related to L-lactic acid and several synthetic human odors and related compounds, although orientation was often observed.

Fecundity, oogenesis, oviposition, and percentage egg hatch were quantified for the blood-feeding midge Forcipomyia (Lasiohelea) townsvillensis (Taylor). Data are similar to that reported for other species of blood-feeding Forcipomyia. Eggs rarely developed from a partial blood meal but invariably developed after a single, complete blood meal. Results suggest that this species is anautogenous. Oviposition media were investigated and a successful medium and holding chamber type identified. Longevity of adults in the laboratory was studied and indicates the possibility for >1 gonotrophic cycle to occur. Adult survival at different relative humidities showed midges can survive 35–98% RH. Rearing of larvae in the laboratory and culture media are discussed. The data supplied in this paper provide the basis for the laboratory culture of F. (L.) townsvillensis.

Phlebotomus papatasi (Scopoli), the vector of Leishmania major (Yakimoff & Schokhor), feeds on plants in desert habitats in the Jordan Valley. At the end of the dry summer, the life span of sand flies is short and the amount of sugars in their guts is small. In this season the plants are under the stress of heat and dehydration. This stress arrests the photosynthesis and decreases the amounts of the main end products, sucrose and starch. We presumed that the paucity of sugars in the sand fly plant tissue diet resulted from the arrest of photosynthesis. To test this assumption, we compared the feeding of sand flies on branches of Capparis spinosa (L.) that had been kept for 24 h in darkness and on branches cut after a normal day of photosynthesis. In darkness, the branches had lost more than half of their sugar content. Afterward they were fed upon overnight by 45.2% of female and 14.3% of male sand flies. A higher proportion of 81.0% females and 38.7% males fed on branches from natural conditions and these fed flies were significantly heavier. Laboratory experiments also showed that plant tissue meals of P. papatasi often include starch grains. Such grains were found also in 50% of field-caught males and females. The nutritive potential of plant tissues was demonstrated by the 33-d median survival of P. papatasi series that had been maintained on fresh C. spinosa branches and water.

Lutzomyia longipalpis (Lutz & Neiva) is the primary vector of visceral leishmaniasis in Venezuela. An analysis of alleles at seven enzyme-encoding loci among four populations from different geographic and epidemiological regions revealed strong genetic substructuring. Isozyme analysis indicated that L. longipalpis in Venezuela is a complex of at least two subspecies. Possible differences in population size during their evolutionary histories, varying colonization histories and geological events may explain discrepancies in the patterns of variation observed at genetic markers between these four populations.

The susceptibility to two coindigenous Plasmodium vivax Grassi & Feletti phenotypes VK210 and VK247 of three colonized Anopheles albimanus Wiedemann strains (white-striped, green and brown) from southern Mexico was investigated. Mosquitoes of the three strains were simultaneously fed with P. vivax-infected patient blood and examined 1 wk later for the presence of oocysts. The circumsporozoite protein phenotype type (VK210 and VK247) was determined by immunoflorescence of salivary gland sporozoites using monoclonal antibodies. The proportions of specimens infected and the number of oocyst per mosquito indicated that all mosquito strains were more susceptible to the phenotype VK210 than to VK247, but the white-striped strain was more susceptible to both parasite phenotypes than the other two strains.

To assess the Lyme borreliosis vector population density we set up a methodology for sampling the Ixodes ricinus L. population host questing on the vegetation. We focused on the collection of the nymphal stage, which is the principal stage of disease transmission to humans. This study was carried out in Rambouillet forest (Yvelines, France) where seven study areas were demarcated. These areas are maximally homogeneous for plant species using a finer scale than the phytosociological classification as defined by the method of landscape diagnostics. Out of 23 collections performed from March 1997 to May 1998, 2,906 I. ricinus nymphs were collected. The sampling technique chosen was the cloth lure technique. The technical parameters were studied and fixed (cloth type, cloth size, sample size, researcher position). It appeared that toweling was the best cloth type to optimize the number of ticks collected; the position of the researcher had no effect on tick samples. To satisfy the criteria for correct sampling, we studied representativity, randomness, and nonselectivity of our methodology. The spatial distribution of nymphs in a homogeneous area was close to random and thus very few subsamples were needed to obtain a relative density which was representative. No significant differences were found between random samples and following transect samples; and nonselectivity was totally satisfied because we only worked on questing nymphs. We grouped the samples that presented no significant differences to attribute a density index, which varied from 0 to 5. This methodology, applied with the same parameters, offers potential for producing comparable results from studies in different geographical areas and at different times of the years.

Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are associated with a variety of reproductive alterations in their hosts, the best known being cytoplasmic incompatability. The Wolbachia pipientis assemblage has been divided into two major groups (A and B) and 12 subgroups. We report herein the first systematic survey of Wolbachia in mosquitoes, and the first survey classifying Wolbachia infections by subgroup. Wolbachia were detected in 28.1% of 89 wild-caught mosquito species, based on a polymerase chain reaction assay using ftsZ and wsp gene primers. Infections were found in all major disease vector genera except Anopheles. Nine of the 12 Wolbachia subgroups were represented. Group B Wolbachia strains showed more phylogenetic concordance with their host taxa than group A strains. Of the 25 positive mosquito species, five were superinfected with group A bacteria strains (AA), eight were superinfected with A and B strains (AB), and one was superinfected with group B strains (BB). The widespread distribution of Wolbachia among mosquito species further supports their potential importance in the genetic control of disease vectors.

Colonization of Phlebotomus neglectus Tonnoir, the major vector of visceral leishmaniasis, in Greece is reported for the first time. Starting with wild-caught specimens, a small closed colony was established that was maintained for 17 mo or 10 generations. Gonotrophic discordance, stenogamic mating behavior, low fecundity, and dormancy because of low temperature were the most important findings that characterized the colony.

Human monocytic ehrlichiosis (HME) is a sometimes fatal, emerging tick-borne disease caused by the bacterium Ehrlichia chaffeensis. It is frequently misdiagnosed because its symptoms mimic those of the flu. Current evidence indicates that Amblyomma americanum (L.), the lone star tick, is the major vector of HME. To determine if E. chaffeensis is present in ticks at Aberdeen Proving Ground, MD, questing A. americanum ticks were collected from 33 sites. Nucleic acid was extracted from 34 adult and 81 nymphal pools. Sequences diagnostic for E. chaffeensis from three different loci (16S rRNA, 120-kDa protein, and a variable-length polymerase chain reaction [PCR] target, or VLPT) were targeted for amplification by the PCR. Fifty-two percent of the collection sites yielded pools infected with E. chaffeensis, confirming the presence and widespread distribution of E. chaffeensis at Aberdeen Proving Ground. Analysis with the both the 120-kDa protein primers and the VLPT primers showed that genetic variance exists. A novel combination of variance for the two loci was detected in two tick pools. The pathogenic implications of genetic variation in E. chaffeensis are as yet unknown.

Estimates of absolute density were determined over a 5-yr period (1990–1994) for a population of Ixodes scapularis Say located in Westchester County, NY, by mark-release-recapture (nymphs and adults) and removal (larvae) methods. Density estimates for larvae ranged from 5.2 to 16.5/m² and averaged 11.5/m². Values for nymphs varied as much as fourfold among successive years, ranging from 0.5 to 2.3/m² and averaging 1.2/m², whereas adult density ranged from 0.3 to 0.4/m², averaging 0.33/m². Natural mortality of nymphs and adults was measured in experimental cages during population estimation periods, and indicated that survival declined linearly over the short-term and did not significantly influence estimates. Drag sampling efficiency, the proportion of the estimated population obtained in a single sample, averaged 6.3% among all stages. Efficiency was not significantly different among stages and was independent of tick density within a given life stage. The population estimation techniques employed in this study are well suited for use with I. scapularis and can provide data that offer insights into mortality patterns in individual populations.

Terrestrial invertebrate carcasses are an important resource for insects developing in pitcher plants. However, little is known of the role of these carcasses in other containers, which also receive leaf fall and stemflow inputs. This experiment investigated effects of accumulated invertebrate carcasses as a resource for two competing mosquitoes, Aedes albopictus (Skuse) and Aedes aegypti (L.), whether either species differentially benefited from accumulated carcasses, and if such a benefit affected interspecific competition. First, we measured accumulation of invertebrate carcasses in standard containers at a field site. We then used a replacement series with five different species ratios at the same total density, and varied the input of invertebrate carcasses [dead Drosophila melanogaster (Meigen)] in three levels: none, the average input from our field site, or the maximum input recorded at our field site. Survivorship, development time, and mass were measured for each mosquito species as correlates of population growth, and were used to calculate a population performance index, λ′. There were strong positive effects of invertebrate carcass additions on all growth correlates and λ′. Differences in performance between species were pronounced in small or no carcass additions and absent in large inputs of invertebrate carcasses, but there was little evidence that inputs of invertebrate carcasses altered the competitive advantage in this system. These results suggest that terrestrial invertebrate carcasses may be an important resource for many types of container communities, and large accumulations of dead invertebrates may reduce resource competition between these mosquitoes, thus favoring coexistence. We propose that the total amount of resource, including accumulated invertebrate carcasses, may explain observed patterns of replacement involving these mosquitoes.

The objective of this study was to determine the influence of the blood meal source on the life cycle and reproductive development of female Triatoma infestans (Klug, 1834), Triatoma brasiliensis Neiva, 1911, Triatoma sordida (Stal, 1859), and Triatoma pseudomaculata Corrêa & Espínola, 1964. In all triatomine species studied the life cycle was shorter for the groups fed on mice than for those fed on pigeons, the range of differences being between 1.5 times (T. pseudomaculata and T. infestans) and 2.4 times (T. brasiliensis). The mortality rate of nymphs during the life cycle tended to be greater in insects fed on pigeons than in those fed on mice, the differences for T. brasiliensis being statistically significant. Females of T. sordida and T. pseudomaculata had a greater fecundity than those of T. infestans and T. brasiliensis independently of the blood meal source. The differences of fecundity observed probably reflect differences in the availability of blood in the silvatic ecotopes of these species, meals being more frequent for T. infestans and T. brasiliensis, which live at high densities in association with rodents in highly stable ecotopes. Because T. sordida and T. pseudomaculata live in more unstable ecotopes with fewer sources of blood they form small sparse colonies and invest more energy in reproduction than maintenance.

The survival and dispersal of adult Aedes notoscriptus (Skuse) were estimated in Brisbane, Queensland, Australia, where this species has been identified as a vector of Ross River and Barmah Forest viruses and shown to be orally susceptible to dengue virus types 1–4. Standard mark-release-recapture methods were used. Before the field study, laboratory trials showed that marking Ae. notoscriptus with fluorescent powder had no effect on survivorship. Female recapture rates of 10.6 and 2.3% over an 8-d period were achieved for two cohorts of marked Ae. notoscriptus released at the same field site. No males were recaptured over the 8-d period. The probability of daily survival was calculated using the Saul model as 0.77 and 0.79 for blue- and pink-marked females, respectively. For blue- and pink-marked females, the mean distance traveled was 105.2 and 179.9 m, and the maximum distance traveled was 195 and 238 m, respectively. The results indicate that protected harborage sites and shade influenced the distribution of Ae. notoscriptus within the study site.

Stage-specific survivorship curves were constructed for the immature stages of a native, predatory mosquito, Toxorhynchites rutilus (Coquillet), by regular censuses in the summer and fall of water-holding treeholes and tires. Survival from egg to adult ranged from 1.8 to 5.6%, and survivorship patterns were significantly heterogeneous between seasons and container types. The probability of death was highest in the first and fourth larval instars. Rainfall and drought were relatively unimportant risk factors during this study, but the probabilities of disappearance of eggs and first and fourth instars were significantly higher in the presence of large conspecifics, suggesting that cannibalism is a major source of mortality.

Permethrin-impregnated fabric has been shown to be an effective repellent against various tick species. However, some tick species are not repelled by this chemical. In Hyalomma dromedarii (Koch), permethrin exposure is reported to actually enhance the tick’s attachment behavior. This study evaluated the histological effects of permethrin exposure on the salivary glands and neuroendocrine organs of unfed, virgin H. dromedarii ticks of uniform age. Three fabric treatments consisting of unwashed-untreated (control), washed after treatment (0.125 mg [AI]/cm²) and unwashed-treated were used after 5- and 10-min exposure times for unfed, unmated females. For all of the organs examined, the cellular structure of treated ticks differed from controls as evidenced by increases in cellular activity, as well as significant increases in the size of the cells of the organs under study (P < 0.05). These data conclusively demonstrate that an unexpected enhanced attachment response observed in this tick species after permethrin exposure is the direct result of increased neurosecretory and salivary gland activity induced by that exposure.

The 2362 strain of Bacillus sphaericus, which produces a binary toxin highly active against Culex mosquitoes, has been developed recently as a commercial larvicide. It is being used currently in operational mosquito control programs in several countries including Brazil, France, India, and the United States. Laboratory studies have shown that mosquitoes can develop resistance to B. sphaericus, and low levels of resistance have already been reported in field populations in Brazil, France, and India. To develop tools for resistance management, the Cyt1A protein of Bacillus thuringiensis subsp. israelensis De Barjac was evaluated for its ability to suppress resistance to B. sphaericus in a highly resistant population of Culex quinquefasciatus Say. A combination of B. sphaericus 2362 in a 10:1 ratio with a strain of B. thuringiensis subsp. israelensis that only produces Cyt1A reduced resistance by >30,000-fold. Resistance was suppressed completely when B. sphaericus was combined with purified Cyt1A crystals in a 10:1 ratio. Synergism was observed between the Cyt1A toxin and B. sphaericus against the resistant mosquito population and accounted for the marked reduction in resistance. However, no synergism was observed between the toxins against a nonresistant mosquito population. These results indicate that Cyt1A could be useful for managing resistance to B. sphaericus 2362 in Culex populations, and also provide additional evidence that Cyt1A may synergize toxicity by enhancing the binding to and insertion of toxins into the mosquito microvillar membrane.

From 1990 through 1997, Ixodes scapularis Say larvae and nymphs were sampled between May and October along a 400-m segment of a nature trail in a Lyme disease endemic site in northern Illinois. Ticks were removed from Peromyscus leucopus mice and collected via tick drags at ≈3-wk intervals. Mouse population estimates along the trail varied from 2, in the spring of 1996 following a year of drought, to >200 in 1993, the wettest year on record. During the 8-yr period, there were major droughts during the summers of 1991 and 1995. Cumulative degree-days were positively correlated with the number of ticks collected on drags in the same year and negatively correlated with larval tick populations for the following year (P < 0.05). Cumulative rainfall was positively correlated with larval tick abundance for the following year. This was most readily apparent by examination of the larval density on captured mice. In the year following each of two drought years, larval densities were significantly depressed compared with the 8-yr average at the site.

The horn fly, Hematobia irritans (L.), is an important pest of livestock because the adult stage of both sexes are aggressive blood-feeders. Remarkably, even though horn fly adults feed recurrently on their hosts as ectoparasites, these flies lack the ADP-responsive antiplatelet aggregation and vasodilatory antihemostatic systems described for other blood-feeding Diptera. Horn fly salivary gland extracts do interfere with the normal coagulation process as demonstrated by the recalcification time assay. Using this as a baseline, the effects of saliva on recalcification time, activated partial thromboplastin time, prothrombin time, and thrombin time were measured to determine which arm(s) of the coagulation cascade might be impacted. Factor-deficient plasma assays also were used to measure possible perturbations in clotting. Gland-free saliva delayed the recalcification time as well as the activated partial thromboplastin time, prothrombin time, and thrombin time. Saliva also further delayed clotting times of plasmas deficient in factor V, factor VIII, and factor XIII, indicating that other factors in the coagulation cascade were inhibited. Although horn fly saliva did not alter the ability of deficient plasma reconstituted with factor X to clot, it did inhibit deficient plasma reconstituted with factor II (thrombin). Antithrombin activity in saliva was confirmed by its ability to interfere with thrombin hydrolysis of fibrinogen, its normal substrate, and by its inhibition of thrombin action on a chromagenic substrate that mimics the hydrolytic site of fibrinogen. Thus, horn fly saliva contains a factor that specifically targets thrombin, a key component in the coagulation cascade. While the biochemical mechanisms of inhibition may vary, this antihemostatic characteristic is shared with other zoophilic Diptera such as black flies, Simulium spp., and tsetse, Glossina morsitans morsitans Westwood, that feed on ungulates.

We used a quantitative competitive polymerase chain reaction (PCR) (QC-PCR) to determine bacterial loads in 669 fleas collected in areas of confirmed and suspected plague epizootics. Fleas were collected out of rodent burrows (67.9%) and off of captured animals (24.1%) and rodent carcasses (8.1%). An initial PCR screening assay indicated that 12.1% (81/669) of all fleas were positive for Yersinia pestis. Fleas collected from burrows had significantly lower (χ² = 264.9, P < 0.0001) infection rates (6.8%) but significantly higher (Student t-test, P < 0.0001) bacterial loads (mean = 10^5.6 Y. pestis per flea) than fleas collected off of rodent carcasses (infection rate = 92.6%; mean bacterial load = 10^4.8 Y. pestis per flea). None of the fleas collected off of captured animals were positive for Y. pestis by PCR, although seven of the 176 captured animals were serologically positive for Y. pestis.

“Push-pull” is a behavior manipulation strategy in which behavior-modifying stimuli are integrated with a pest control agent. We evaluated the efficacy of an insecticide bait in combination with attractants (“pull”), repellents (“push”), or both (“push-pull”) using a hydramethylnon-based bait, feces-contaminated surfaces as an attractant, and methyl neodecanamide-treated surfaces to repel cockroaches. Both adult males and first-instar German cockroaches, Blattella germanica (L.), chose shelters nearest the attractant-treated surfaces and farthest from the repellent-treated surfaces. Food consumption was highest from food nearest the preferred shelters, and mortality was highest when the insecticide bait was near the preferred shelter. These patterns were more apparent in first instars than in adults. Our results from large arena studies in the laboratory show that the push-pull strategy can be used to displace pests from resources or commodities that are to be protected, and simultaneously lure the pest to an attractant source coupled with a pest control agent. Concentrating cockroaches into a limited area should facilitate the precision-targeting of the pest population and promises to reduce insecticide use.

We examined the potential for Phlebotomus papatasi (Scopoli), Phlebotomus duboscqi (Neveu-Lemarie), Phlebotomus sergenti (Parrot) , and Sergentomyia schwetzi (Adler, Theodor, & Parrot) to transmit Rift Valley fever (RVF) virus. After feeding on hamsters that had been inoculated with RVF virus, P. papatasi, P. sergenti, and S. schwetzi became infected and developed disseminated infections. All P. papatasi and P. duboscqi inoculated with RVF virus developed high-titer infections. In contrast, only 41% of the inoculated S. schwetzi contained detectable virus, and infected individuals contained significantly less virus than the two Phlebotomus species. Although 50% of the inoculated P. duboscqi transmitted RVF virus to hamsters, only 14% of P. papatasi and none of the S. schwetzi transmitted this virus. Additional studies are needed to determine the role of sand flies as vectors of RVF virus.

A 2-yr study was conducted in a northwestern Florida state park and recreation area to determine tick species composition, seasonal abundance, and spatial distribution. Risk of tick attachment to park visitors was also assessed relative to tick abundance from several habitats. Tick collections consisted of weekly cloth drag samples obtained from ground and emergent vegetation along a deer trail, a walking trail located in a 3-mo-old control burned area of forest, a nonburned walking trail, picnic area, and campsites. Sampling was conducted from February 1994 through February 1996. Ticks collected during the study were as follows in descending order of abundance: Ixodes scapularis Say, Amblyomma americanum (L.), A. maculatum Koch, and Dermacentor variabilis (Say). Significantly more (<0.05) adult ticks were recovered from vegetation compared with ground samples only for I. scapularis. Adult questing I. scapularis were collected from October through May (peak December). A. americanum adults were collected from March through August (peak May). Adult A. maculatum were collected during August and September and adult D. variabilis were collected July and August. Larval and nymphal stages of questing A. americanum were collected from June through November (peak July) and February through October (peak September), respectively. A. maculatum nymphs were collected from February through April, June, and September (peak March). No nymphs of I. scapularis were collected. Only four larvae of D. variabilis were collected during the study, all during February 1995. No I. scapularis or A. maculatum larvae were collected from vegetation or ground drags. The greatest risk for tick attachment was in campsites where ≈60% of all adult and nymphal host-seeking ticks were collected. The next greatest risk area was the walking trail located in the control burned area where an additional 28% of ticks were obtained.

A total of 145 ectoparasites was found on five pups of the Indiana bat, Myotis sodalis, from a roost in Warren County, in southwestern Ohio. Most were Macronyssus crosbyi, the common mite of this host, but three males, two nymphs and a nonengorged female of Paraspinturnix globosa were found. Males and nymphs of this species have not previously been reported. In addition, two female adults of Paraspinturnix globosa were found in the anus of one adult female. The male, female deutonymph and nongravid engorged female of P. globosa are described for the first time.

In 1997, Aedes albopictus (Skuse) was discovered in Peoria, IL, a known focus of La Crosse (LAC) virus transmission. This accidental introduction provided an opportunity to determine whether Ae. albopictus would reemerge in the spring or summer and, if successful overwintering occurred, to follow changes in the geographic range of Ae. albopictus, and to compare its distribution to that of the local treehole mosquito and LAC vector Aedes triseriatus (Say). In 1998, 25 oviposition traps were placed in and around the area of the initial finding of Ae. albopictus, with adult collections by aspirators and larval collections from water-holding containers used to identify areas of additional activity. Ae. albopictus successfully survived the mild 1997–1998 El Niño winter, and expanded its range during 1998. By September 1998, Ae. albopictus oviposited in all 25 traps, including traps near and in wooded sites. Intensity of oviposition activity (number of eggs per positive trap) ranged from 20–40, lower than the range for Ae. triseriatus, which was as high as 175 eggs per infested trap in mid-August. Prevalence of Ae. albopictus increased through September, where as the prevalence of Ae. triseriatus declined starting in mid-July. Although direct competition between the two mosquito species cannot be inferred based on this surveillance effort, a gradual range expansion was observed, and Ae. albopictus eggs were collected in traps where initially only Ae. triseriatus oviposition activity was detected.

In the field, the female horn fly obtained blood meals from the bovine host before oviposition. The female flies moved to the lower portion of the hind legs as the legs were spread, and the tail was raised before excretion. Females deposited their eggs on the manure after excretion. The fly flew onto the manure pat for oviposition as the cow walked forward after excretion. It appeared that flies located on the belly received some stimuli for oviposition from the cow just before excretion. The horn fly deposited most of its eggs during the day with occasional deposition at night. Egg deposition increased at 10°C. Fewer flies were observed on manure pats that contained >90% or <84% water. The location of horn flies on the cow and environmental factors associated with oviposition in the field are discussed.

A reverse transcription-polymerase chain reaction (RT-PCR) was assessed in laboratory tests to detect the presence of singleAedes aegypti (L.) orEretmapodites quinquevittatus Theobald mosquitoes infected with Rift Valley fever virus in pools of mosquitoes, 50–600 in size, from laboratory colonies or mixed field collections. The viral RNA was detected in all pools containing infected mosquitoes and was shown to be as sensitive as infant mice but more sensitive than Vero cell cultures for virus detection. Pools diluted down to the equivalent of 1:16 000 mosquitoes were also positive by RT-PCR. RNAs from 4 other phleboviruses were negative, there were no false positives and the procedure followed, with the 2 particular primers chosen, gave consistently clear bands of the PCR products on agarose gels without nested PCR being necessary.

The responses of adult female blacklegged ticks,Ixodes scapularis Say, to urine from white-tailed deer,Odocoileus virginianus (Zimmermann), belonging to 4 reproductive categories (doe in estrous, doe out of season, reproductive [dominant] buck, young buck) and to a mixture of urine from nondominant bucks in rut, young bucks out of rut, and nonestrous does were studied in laboratory behavioral bioassays. In high humidity (≈95% RH) in a glove box there were no statistically significant arrestment responses to any of the 5 types of urine, but an avoidance response was observed to urine from dominant reproductive bucks. When ticks were tested at ≈50% RH, with samples of all 5 types of urine in the glove box, significant arrestant responses by the ticks were elicited by urine from does in estrous and by dominant reproductive bucks. When tested without other types of urine in the glove box, the urine mixture elicited an arrestant response at 50% RH. In some circumstances, adultI. scapularis may possibly use deer urine as a chemical cue in selecting host-ambush sites.

Anopheles flavirostris (Ludlow) from Lombok Island, Indonesia, was crossed withAn. minimus species A (CM strain) from Thailand and species E (ISG strain), a new sibling species, from Japan, to determine genetic compatibility. We also compare the scanning micrographs of female cibarial armature of these three species. BothAn. minimus CM and ISG females crossed withAn. flavirostris males produced eggs without embryos. OneAn. flavirostris female crossed with anAn. minimus CM male deposited unhatchable eggs with an 18% embryonation rate. The scanning micrographs of the cibarial armature clearly show significant differences amongAn. flavirostris and the two sibling species of theAn. minimus complex. These results support the specific status ofAn. flavirostris. Previous records ofAn. minimus Theobald in this country are also discussed.

We examined the role of lesser mealworm,Alphitobius diaperinus (Panzer), in the transmission of an enteric disease of turkeys caused by a coronavirus. Turkey coronavirus (TCV) from two sources was studied, one isolate (NC95) was embryo propagated, the second was TCV infected material from turkeys diagnosed with poult enteritis mortality syndrome (PEMS). Beetles were fed virus-infected feces mixed with chicken feed. Transmission of virus was effectively halted by surface sterilization of the beetles. Turkey poults administered beetle homogenates infected with TCV PEMS that had not been surface sterilized had reduced weight gains and 50% mortality. Mortality and weight gains were not effected in the NC95 group. Virus isolation procedures were performed to determine NC95 viability at varying time intervals. Beetles were dissected and the guts removed 1, 12, and 24 h after the initial viral feeding. Whole beetles were also examined for comparison. Whole beetles and beetle guts were homogenized and injected into turkey eggs for embryo propagation. Direct immunofluorescence was used to determine the presence of TCV.A. diaperinus were capable of mechanical transmission of TCV. However, only turkey embryos receiving whole beetle and beetle gut homogenates within 1 h of feeding on the virus were positive for TCV. Laboratory studies demonstrating PEMS transmission byA. diaperinus are continuing.

Drag sampling is a commonly used method to obtain relative estimates of the density of questing nymphalIxodes scapularis Say andI. pacificus Cooley & Kohls ticks, which are primary vectors of Lyme disease spirochetes to humans in North America. However, the efficiency of drag sampling in determining absolute population densities of questing nymphs has not been evaluated previously. Therefore, we assessed the efficiency of a single drag-sampling occasion to estimate the total population size of questingI. pacificus nymphs in a leaf-litter habitat in California. Repeated daily removal sampling was carried out in four areas, each covering 300 m², on 17 occasions over a 23-d period in the spring of 1999. In total, 573I. pacificus nymphs were collected, of which 55 (9.6%) were collected on the initial sampling occasion and 20 (3.5%) on the last occasion. The total population size of questing nymphs, i.e., the intersection with the horizontal axis of a linear regression of daily nymphal catch rates on the number of nymphs caught previously, was estimated to be 936. Thus, the efficiency of the initial sampling occasion to estimate the total population size was 5.9% (4.8, 5.0, 5.8, and 9.1%, respectively, for the four individual sampling areas). Further, the overall mean efficiencies of the two, five, and 10 first removal sampling occasions to estimate the absolute nymphal density was 5.2, 4.7, and 4.3%, respectively, and 13 sampling occasions were required to collect 50% of the estimated total nymphal population.