Both sexes of two new Brazilian phlebotomine sand flies of the genus Nyssomyia Barretto 1962 [ = Lutzomyia (Nyssomyia) sensu Barretto], Nyssomyia delsionatali n. sp. and Nyssomyia urbinattii n. sp., presenting close affinity with Nyssomyia antunesi (Coutinho), are described and illustrated. N. delsionatali n. sp was captured on the edge of a riparian Amazonian forest on the Juruena river in the northwest of Mato Grosso state and N. urbinattii n. sp in a riparian Amazonian forest on the Teles Pires river between Mato Grosso and Pará states. Some measurements of both sexes of N. antunesi and illustration of the male genitalia and the female spermathecae as well as an identification key for males and females of the genus Nyssomyia are provided.

A fixed precision sampling plan was developed for off-host populations of adult Rocky Mountain wood tick, Dermacentor andersoni (Stiles) based on data collected by dragging at 13 locations in Alberta, Canada; Washington; and Oregon. In total, 222 site-date combinations were sampled. Each site-date combination was considered a sample, and each sample ranged in size from 86 to 250 10 m² quadrats. Analysis of simulated quadrats ranging in size from 10 to 50 m² indicated that the most precise sample unit was the 10 m² quadrat. Samples taken when abundance <0.04 ticks per 10 m² were more likely to not depart significantly from statistical randomness than samples taken when abundance was greater. Data were grouped into ten abundance classes and assessed for fit to the Poisson and negative binomial distributions. The Poisson distribution fit only data in abundance classes <0.02 ticks per 10 m², while the negative binomial distribution fit data from all abundance classes. A negative binomial distribution with common k = 0.3742 fit data in eight of the 10 abundance classes. Both the Taylor and Iwao mean-variance relationships were fit and used to predict sample sizes for a fixed level of precision. Sample sizes predicted using the Taylor model tended to underestimate actual sample sizes, while sample sizes estimated using the Iwao model tended to overestimate actual sample sizes. Using a negative binomial with common k provided estimates of required sample sizes closest to empirically calculated sample sizes.

In September 2008, Aedes (Stegomyia) albopictus (Skuse 1894), a mosquito of great medical importance, was reported for the first time in a district of Athens, Greece. The following year, its distribution and population abundance over time around its first detection site was investigated. A network of 50 ovitraps was established in a surrounding area of 25 km² and monitored for 17 mo from 17 August 2009 through 31 December 2010. Weekly servicing of the ovitraps and subsequent laboratory treatment of the collected mosquito eggs, revealed that Ae. albopictus is the dominant container breeding Aedes species and is widespread in the entire study area. The seasonal monitoring showed that this mosquito is continuously active from mid spring until the end of December with a considerably high oviposition activity recorded during summer and fall. Moreover, comparison of its activity during late August and end of December, between the two consecutive surveillance years, indicated a significant increase of population abundance the second year. The current study suggests the necessity of setting up in the city of Athens an integrated surveillance program in a greater scale for Asian tiger mosquito's effective control.

Culex pipiens L. (Diptera: Culicidae) and Culex restuans Theobald are the primary enzootic and bridge vectors of West Nile virus in the eastern United States north of 36° latitude. Recent studies of the natural history of these species have implicated catch basins and underground storm drain systems as important larval development sites in urban and suburban locales. Although the presence of larvae in these habitats is well-documented, the influence of abiotic factors on the ecology of Culex larvae developing in them remains poorly understood. Therefore, we examined the effects of multiple abiotic factors and their interactions on abundance of Culex larvae in catch basins in the Chicago, IL, metropolitan area. Low precipitation and high mean daily temperature were associated with high larval abundance, whereas there was no correlation between catch basin depth or water depth and larval abundance. Rainfall was an especially strong predictor of presence or absence of larvae in the summer of 2010, a season with an unusually high precipitation. Regression tree methods were used to build a schematic decision tree model of the interactions among these factors. This practical, visual representation of key predictors of high larval production may be used by local mosquito abatement districts to target limited resources to treat catch basins when they are particularly likely to produce West Nile virus vectors.

The spread of exotic mosquito species into new environments can introduce shifts in mosquito populations and potentially alter public health risks to mosquito-borne diseases. The successful establishment of exotic species may occur due to their competitive advantage over other cohabitating species. We hypothesized that the recently introduced exotic mosquito Aedes japonicus japonicus (Theobald) would be a more effective competitor than Aedes atropalpus (Coquillett) and Aedes triseriatus (Say), and an equal competitor to Culex pipiens (L.) based on larval abundance data within tire habitats. Impacts of competition were measured using the larval developmental rate and survival of larvae, adult mortality, wing length, and sex ratio. We found that intraspecific competition acted strongest against Ae. japonicus versus the other three resident mosquito species by delaying larval development and increasing adult mortality. Interspecific competition was generally weak and significant main effects were only detected for species and density. Overall, our results show that larval competition between Ae. japonicus and the three resident species was weak when present, indicating that other ecological or behavioral factors may be influencing the invasion success for Ae. japonicus in North America.

Species diversity and seasonal abundance of muscoid flies (Diptera: Muscidae) developing in biosolid cake (dewatered biosolids) stored at a wastewater treatment facility in northeastern Kansas were evaluated. Emergence traps were deployed 19 May through 20 October 2009 (22 wk) and 27 May through 18 November 2010 (25 wk). In total, 11,349 muscoid flies were collected emerging from the biosolid cake. Stable flies (Stomoxys calcitrans (L.)) and house flies (Musca domestica (L.)), represented 80 and 18% of the muscoid flies, respectively. An estimated 550 stable flies and 220 house flies per square-meter of surface area developed in the biosolid cake annually producing 450,000 stable flies and 175,000 house flies. Stable fly emergence was seasonally bimodal with a primary peak in mid-July and a secondary peak in late August. House fly emergence peaked with the first stable fly emergence peak and then declined gradually for the remainder of the year. House flies tended to emerge from the biosolid cake sooner after its deposition than did stable flies. In addition, house fly emergence was concentrated around midsummer whereas stable fly emergence began earlier in the spring and continued later into the fall. Biosolid age and temperature were the most important parameters affecting emergence for house flies and stable flies, whereas precipitation was not important for either species. This study highlights the importance of biosolid cake as a larval developmental habitat for stable flies and house flies.

The waste artificial larval rearing media of New World screwworms, Cochliomyia hominivorax (Coquerel) were evaluated to determine their effectiveness as oviposition attractants. Various concentrations of waste larval media resulting from rearing screwworm larvae in gel and cellulose fiber-based artificial diets tested over a 4-wk period attracted varying number of gravid screwworm flies to oviposit. Three-day-old waste medium with concentrations of 10 and 25% were most attractive to gravid female flies for oviposition and resulted in the most oviposition. One and 7-d-old wastes at all concentrations were less attractive for oviposition than the 3d-old media. The fresh (0-d-old), 14-d- and 28-d-old waste media were the least attractive substrates for oviposition. The waste from the cellulose fiber-based diet resulted in significantly more oviposition compared with waste from the gel-based diet. Microorganisms growing in the waste media probably produce metabolites that attract gravid screwworm flies to oviposit. Use of the waste products of appropriate age and dilution as oviposition substrates would enhance oviposition in mass production colony cages.

Resolution of systematic relationships among members of the Culex pipiens (L.) complex has important implications for public health as well as for studies on the evolution of sibling species. Currently held views contend that in California considerable genetic introgression occurs between Cx. pipiens and Cx. quinquefasciatus Say, and as such, these taxa behave as if they are a single species. Development of high throughput SNP genotyping tools for the analysis of Cx. pipiens complex population structure is therefore desirable. As a first step toward this goal, we sequenced 12 gene fragments from specimens collected in Marin and Fresno counties. On average, we found a higher single nucleotide polymorphism (SNP) density than any other mosquito species reported thus far. Coding regions contained significantly higher GC content (median 54.7%) than noncoding regions (42.4%; Wilcoxon rank sum test, P = 5.29 × 10^-5). Differences in SNP allele frequencies observed between mosquitoes from Marin and Fresno counties indicated significant genetic divergence and suggest that SNP markers will be useful for future detailed population genetic studies of this group. The high density of SNPs highlights the difficulty in identifying species within the complex and may be associated with the large degree of phenotypic variation observed in this group of mosquitoes.

Anopheles arabiensis Patton is one of the principal vectors of malaria in sub-Saharan Africa, occupying a wide variety of ecological zones. This species is increasingly responsible for malaria transmission in Africa and is becoming the dominant vector species in some localities. Despite its growing importance, little is known about genetic polymorphisms in this species. Multiple sequences of various gene fragments from An. arabiensis isolates from Cameroon were obtained from GenBank. In total, 20 gene fragments containing single-nucleotide polymorphisms (SNPs) at moderate density were selected for direct sequencing from field collected specimens from Tanzania and Zambia. We obtained 301 SNPs in total from the 20 gene fragments, 60 of which were suitable for Illumina GoldenGate SNP genotyping. A greater number of SNPs (n = 185) was suitable for analysis using Sequenom iPLEX, an alternative high-throughput genotyping technology using mass spectrometry. An SNP was present every 59 (± 44.5) bases on average. Overall, An. arabiensis from Tanzania and Zambia are genetically closer (mean F_ST = 0.075) than either is to populations in Cameroon (F_{ST TZ-CM} = 0.250, F_ST,ZA-CM = 0.372). A fixed polymorphism between East/southern and Central Africa was identified on AGAP000574, a gene on the X chromosome. We have identified SNPs in natural populations of An. arabiensis. SNP densities in An. arabiensis were higher than Anopheles gambiae s.s., suggesting a greater challenge in the development of high-throughput SNP analysis for this species. The SNP markers provided in this study are suitable for a high-throughput genotyping analysis and can be used for population genetic studies and association mapping efforts.

Complex biological events occur during the developmental process of the mosquito Anopheles gambiae (Giles). Using cDNA expression microarrays, the expression patterns of 13,440 clones representing 8,664 unique transcripts were revealed from six different developmental stages: early larvae (late third instar/early fourth instar), late larvae (late fourth instar), early pupae (<30 min after pupation), late pupae (after tanning), and adult female and male mosquitoes (24 h postemergence). After microarray analysis, 560 unique transcripts were identified to show at least a fourfold up- or down-regulation in at least one developmental stage. Based on the expression patterns, these gene products were clustered into 13 groups. In total, eight genes were analyzed by quantitative real-time polymerase chain reaction to validate microarray results. Among 560 unique transcripts, 446 contigs were assigned to respective genes from the An. gambiae genome. The expression patterns and annotations of the genes in the 13 groups are discussed in the context of development including metabolism, transport, protein synthesis and degradation, cellular processes, cellular communication, intra- or extra-cellular architecture maintenance, response to stress or immune-related defense, and spermatogenesis.

Four cattle parasiticides of the avermectin/milbemycin class were examined for lethal and sublethal effects on the zoophilic, African malaria vector Anopheles arabiensis. Ivermectin, moxidectin, doramectin, and eprinomectin were mixed with bovine blood and provided to laboratoryreared An. arabiensis in a membrane feeder. Ivermectin and eprinomectin were lethal to An. arabiensis at low concentrations (LC₅₀s of 7.9 ppb and 8.5 ppb, respectively). While the lethality of doramectin (LC₅₀ of 23.9 ppb), was less than that of ivermectin and eprinomectin, it markedly reduced egg development. The concentration of moxidectin required to reduce survivorship and egg production in An. arabiensis was >100 fold greater than for ivermectin or eprinomectin. Moxidectin was weak in its actions against An. arabiensis relative to the other three chemicals. These results suggest that cattle treated with ivermectin or eprinomectin in the prescribed range of low dosages as parasiticides have blood toxic to zoophilic malaria vectors. Regionally coordinated, seasonal treatment of cattle could suppress An. arabiensis populations, thereby reducing malaria transmission. Doramectin (although less toxic) would have population level effects on egg production if used in this manner.

The major components of geranium (Geranium maculatum L.) oil and their mixtures were tested against female Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae). Chemical analysis by gas chromatography coupled to mass spectrometry revealed four major constituents: citronellol (38%), geraniol (16%), citronellyl formate (10.4%), and linalool (6.45%) (concentration expressed as percentage of total). Topical application demonstrated that the most potent component was citronellol and geraniol, with LD₅₀ values 9.7 and 12.7 µg/insect, respectively. Linalool and Citronellyl formate were less toxic with LD₅₀ values 24.7 and 38.5 µg/insect, respectively. Toxicity of these four major constituents in the same proportion as the natural oil, was greater than whole oil and each individual component. Removal of any four constituents produced a decreased in effectiveness. The absence of citronellol caused the greatest decrease in toxicity (DL₅₀ from 2.2 to 10.9 µg/insect), leading us to conclude that this constituent is the major contributor to oil toxicity. The knowledge of the role of each constituent in the toxicity of the whole oil gives the possibility to create artificial blends of different constituents for the development of more effective control agents.

Using the in vitro rearing system in conjunction with the hair tuft bioassay, NYDA and NYDA without fragrances formulations (92% wt:wt dimeticones) were 100% ovicidal (0% of treated eggs hatched) after an 8-h exposure of the eggs of the human head louse (Pediculus humanus capitis De Geer) following the manufacturer's instructions. Comparatively, 78 and 66% of eggs similarly exposed hatched after distilled deionized water or Nix (1% permethrin) treatments, respectively. NYDA and NYDA without fragrances formulations were also statistically and substantially more ovicidal than either distilled deionized water or Nix treatments after 10, 30 min, and 1 h exposures. Only the 10 min exposure of eggs to NYDA and NYDA without fragrances formulations resulted in hatched lice that survived to adulthood (5–8% survival). Of the lice that hatched from eggs exposed to NYDA formulations for 10 min, there were no significant differences in the time it took them to become adults, female fecundity or the viability of eggs laid by surviving females. The longevity of adults, however, was reduced after the 10 min treatments of eggs with NYDA and NYDA without fragrances formulations compared with either the distilled deionized water or Nix treatments.

Culex quinquefasciatus Say mosquitoes flourish in belowground stormwater systems in the southern United States. Recent evidence suggests that oviposition-site-seeking females may have difficulties locating, entering, and ovipositing inside permanent water chambers when surface entry through pickholes in manhole covers are sealed. It remains unknown, however, if newly emerged adults are able to detect cues necessary to exit these partly sealed systems via lateral conveyance pipes or if they perish belowground. Fourth instar Cx. quinquefasciatus were placed within proprietary belowground stormwater treatment systems to determine the percentage of newly emerged adults able to escape treatment chambers via a single lateral conveyance pipe. Overall, 56% of deployed mosquitoes were captured in adult exit traps with an ≈1:1 male:female ratio. The percentage of adults captured varied significantly among chambers, but was not associated with structural site characteristics such as the chamber depth or the length and course of conveyance pipe to the exit trap. Empirical observations suggested that longbodied cellar spiders, Pholcus phalangioides (Fuesslin), ubiquitous in these structures, may have reduced adult trap capture. Findings demonstrate that newly emerged Cx. quinquefasciatus can exit subterranean chambers under potentially difficult structural conditions but suggest that a portion may perish in the absence of surface exit points in manhole shafts.

Because of resistance to current insecticides and to environmental, health, and regulatory concerns, naturally occurring compounds and their derivatives are of increasing interest for the development of new insecticidal compounds against vectors of disease-causing pathogens. Fifty-eight compounds, either extracted and purified from plants native to China or synthetic analogs of curcumin, were evaluated for both their larvicidal activity against Aedes aegypti (L.) and their ability to inhibit binding of cholesterol to Ae. aegypti sterol carrier protein-2 in vitro. Of the compounds tested, curcumin analogs seem especially promising in that of 24 compounds tested five were inhibitors of Ae. aegypti sterol carrier protein-2 with EC₅₀ values ranging from 0.65 to 62.87 µM, and three curcumin analogs exhibited larvicidal activity against fourth instar Ae. aegypti larvae with LC₅₀ values ranging from 17.29 to 27.90 µM. Adding to the attractiveness of synthetic curcumin analogs is the relative ease of synthesizing a large diversity of compounds; only a small fraction of such diversity has been sampled in this study.

Seven different strains of Aedes aegypti (L.), including a genetically diverse laboratory strain, three laboratory-selected permethrin-resistant strains, a standard reference strain, and two recently colonized strains were fed on human blood containing various concentrations of ivermectin. Ivermectin reduced adult survival, fecundity, and hatch rate of eggs laid by ivermectin-treated adults in all seven strains. The LC₅₀ of ivermectin for adults and the concentration that prevented 50% of eggs from hatching was calculated for all strains. Considerable variation in adult survival after an ivermectin-bloodmeal occurred among strains, and all three permethrin-resistant strains were significantly less susceptible to ivermectin than the standard reference strain. The hatch rate after an ivermectin bloodmeal was less variable among strains, and only one of the permethrin-resistant strains differed significantly from the standard reference strain. Our studies suggest that ivermectin induces adult mortality and decreases the hatch rate of eggs through different mechanisms. A correlation analysis of log-transformed LC₅₀ among strains suggests that permethrin and ivermectin cross-resistance may occur.

Fleas are vectors for a number of pathogens including Yersinia pestis, yet factors that govern interactions between fleas and Y. pestis are not well understood. Examining gene expression changes in infected fleas could reveal pathways that affect Y. pestis survival in fleas and subsequent transmission. We used suppression subtractive hybridization to identify genes that are induced in Xenopsylla cheopis (Rothschild) (Siphonaptera: Pulicidae) in response to oral or hemocoel infection with Y. pestis. Overall, the transcriptional changes we detected were very limited. We identified several genes that are likely involved in the production or removal of reactive oxygen species (ROS). Midgut ROS levels were higher in infected fleas and antioxidant treatment before infection reduced ROS levels and resulted in higher bacterial loads. An ROS-sensitive mutant strain of Y. pestis lacking the OxyR transcriptional regulator showed reduced growth early after infection. Our results indicate that ROS may limit Y. pestis early colonization of fleas and that bacterial strategies to overcome ROS may enhance transmission.

Japanese encephalitis virus (JEV) transmission in northern Australia has, in the past, been facilitated by Culex annulirostris Skuse feeding on domestic pigs, the primary amplifying hosts of the virus. To further characterize mosquito feeding behavior in northern Australia, 1,128 bloodmeals from Cx. annulirostris were analyzed using a double-antibody enzyme-linked immunosorbent assay. Overall, Cx. annulirostris obtained >94% of blood meals from mammals, comprising marsupials (37%), pigs (20%), dogs (16%), and cows (11%), although the proportion feeding on each of these host types varied between study locations. Where JEV activity was detected, feeding rates on pigs were relatively high. At the location that yielded the first Australian mainland isolate of JEV from mosquitoes, feral pigs (in the absence of domestic pigs) accounted for 82% of bloodmeals identified, representing the first occasion that feeding on feral pigs has been associated with JEV transmission in Australia. Interestingly, <3% of Cx. annulirostris had fed on pigs at locations on Badu Island where JEV was detected in multiple pools of mosquitoes in a concurrent study. This suggests that either alternative hosts, such as birds, which comprised 21% of blood meals identified, or infected mosquitoes immigrating from areas where domestic pigs are housed, may have contributed to transmission at this location. Because Cx. annulirostris is both an opportunistic feeder and the primary JEV vector in the region, environmental characteristics and host presence can determine JEV transmission dynamics in northern Australia.

Studies of mosquito preferences for avian hosts have found that some bird species are at greater risk than others of being fed upon by mosquitoes. The ecological factors that determine this interspecific variation in avian host use by mosquitoes have been little studied, despite the possibility that such variation may influence spatial and temporal patterns of the occurrence of mosquito-borne pathogens. Our objective was to identify ecological variables associated with the avian host forage ratios estimated from a previous study of mosquito feeding patterns in Tuskegee National Forest, AL. We used species' characteristics derived from the literature to develop multiple linear regression models for the forage ratios of Culiseta melanura (Coquillett) and Culex erraticus (Dyar & Knab) for avian hosts. We found that habitat-edge association and body mass of avian host species were the best predictors of forage ratios of Cx. erraticus for avian hosts. Although no avian host traits were inferred to be strong predictors of forage ratios of Cs. melanura, body mass had the greatest importance weight among those considered. Our results suggest that characteristics of avian hosts may predict their levels of use by some mosquito species.

Our study compared different estimates of adult mosquito abundance (Pupal Demographic Survey [PDS], Human Landing Collection [HLC], Number of Bites declared by Citizens during interviews [NBC]) to the mean number of eggs laid in ovitraps. We then calculated a disease risk threshold in terms of number of eggs per ovitrap above which an arbovirus epidemic may occur. The study was conducted during the summers of 2007 and 2008 in the Emilia-Romagna region of Italy where a Chikungunya epidemic occurred in 2007. Ovitrap monitoring lasted from May to September, while adult sampling by means of PDS, HLC, and NBC was repeated three times each summer. Based on calculated rate of increase of the disease (R₀) and the number of bites per human per day measured during the outbreak, we estimated that only 10.1% of the females transmitted the Chikungunya virus in the principal focus. Under our conditions, we demonstrated that a positive correlation can be found between the females' density estimated by means of PDS, HLC, and NBC and the mean number of eggs in the ovitraps. We tested our hypothesis during the 2007 secondary outbreak of CHIKV in Cervia, and found that R₀ calculated based on the number of biting females estimated from the egg density was comparable to the R₀ calculated based on the progression of the human cases. The identification of an epidemic threshold based on the mean egg density may define the high risk areas and focus control programs.

Lyme disease (LD) is emerging in Canada because of the northward expansion of the geographic range of the tick vector Ixodes scapularis (Say). Early detection of emerging areas of LD risk is critical to public health responses, but the methods to do so on a local scale are lacking. Passive tick surveillance has operated in Canada since 1990 but this method lacks specificity for identifying areas where tick populations are established because of dispersion of ticks from established LD risk areas by migratory birds. Using data from 70 field sites in Quebec visited previously, we developed a logistic regression model for estimating the risk of I. scapularis population establishment based on the number of ticks submitted in passive surveillance and a model-derived environmental suitability index. Sensitivity-specificity plots were used to select an optimal threshold value of the linear predictor from the model as the signal for tick population establishment. This value was used to produce an “Alert Map” identifying areas where the passive surveillance data suggested ticks were establishing in Quebec. Alert Map predictions were validated by field surveillance at 76 sites: the prevalence of established I. scapularis populations was significantly greater in areas predicted as high-risk by the Alert map (29 out of 48) than in areas predicted as moderate-risk (4 out of 30) (P < 0.001). This study suggests that Alert Maps created using this approach can provide a usefully rapid and accurate tool for early identification of emerging areas of LD risk at a geographic scale appropriate for local disease control and prevention activities.

The cattle tick, Rhipicephalus (Boophilus) spp. is a key vector of protozoa that cause bovine babesiosis. Largely eradicated from most of the United States, the cattle tick continues to infest south Texas, and recent outbreaks in this area may signal a resurgence of cattle tick populations despite current management efforts. An improved understanding of the dynamic ecology of cattle fever ticks along the U.S.-Mexico border is required to devise strategies for sustainable eradication efforts. Management areas of the cattle tick overlap considerably with dense, wide infestations of the non-native, invasive grass known as giant reed (Arundo donax L.). Here we show that stands of giant reed are associated with abiotic and biotic conditions that are favorable to tick survival, especially when compared with other nearby habitats (open pastures of buffelgrass (Pennisetum ciliare) and closed canopy native forests). Overhead canopies in giant reed stands and native riparian forests reduce daily high temperature, which was the best abiotic predictor of oviposition by engorged females. In sites where temperatures were extreme, specifically open grasslands, fewer females laid eggs and the resulting egg masses were smaller. Pitfall trap collections of ground dwelling arthropods suggest a low potential for natural suppression of tick populations in giant reed stands. The finding that A. donax infestations present environmental conditions that facilitate the survival and persistence of cattle ticks, as well or better than native riparian habitats and open grasslands, represents an alarming complication for cattle fever tick management in the United States.

Aedes aegypti (L.) (Diptera: Culicidae) female mosquitoes infected systemically with Sindbis virus (SINV) took longer than uninfected mosquitoes to locate and fully engorge on blood. On days 7 and 14 postexposure, blood feeding took 1.3 and 1.5 times longer in mosquitoes with a disseminated SINV infection, respectively. SINV dissemination did not affect the average weight of unfed Ae. aegypti, but did result in a 10 and 12% increase in blood imbibed compared with mosquitoes without a positive SINV dissemination and non-SINV-exposed mosquitoes, respectively. Ae. aegypti mosquitoes with a disseminated SINV infection fed an average of 4 h sooner than uninfected mosquitoes when offered a bloodmeal contained inside a DEET (N,N-diethyl-3-methylbenzamide) saturated (30%) bovine sausage casing. Together, these results indicate that behavioral changes in mosquito host-seeking, blood feeding and sensitivity to DEET occurred in mosquitoes after SINV infection and dissemination.

Necrophagous blow fly species (Diptera: Calliphoridae) are the most important agents for estimating the postmortem interval (PMI) in forensic entomology. Nevertheless, the oviposition preferences of blow flies may cause a bias of PMI estimations because of a delay or acceleration of egg laying. Chrysomya megacephala (F.) and C. rufifacies (Macquart) are two predominant necrophagous blow fly species in Taiwan. Their larvae undergo rather intense competition, and the latter one can prey on the former under natural conditions. To understand the oviposition preferences of these two species, a dual-choice device was used to test the choice of oviposition sites by females. Results showed when pork liver with and without larvae of C. rufifacies was provided, C. megacephala preferred to lay eggs on the liver without larvae. However, C. megacephala showed no preference when pork liver with and without conspecific larvae or larvae of Hemipyrellia ligurriens (Wiedemann) was provided. These results indicate that females of C. megacephala try to avoid laying eggs around larvae of facultatively predaceous species of C. rufifacies. However, C. rufifacies showed significant oviposition preference for pork liver with larvae of C. megacephala or conspecific ones when compared with pork liver with no larvae. These results probably imply that conspecific larvae or larvae of C. megacephala may potentially be alternative food resources for C. rufifacies, so that its females prefer to lay eggs in their presence. When considering the size of the oviposition media, pork livers of a relatively small size were obviously unfavorable to both species. This may be because females need to find sufficient resources to meet the food demands of their larvae. In another experiment, neither blow fly species showed an oviposition preference for pork livers of different stages of decay. In addition, the oviposition preferences of both species to those media with larvae were greatly disturbed in a dark environment. If we removed the larvae that had previously fed on the pork liver and let the females choose, no oviposition preference was observed; but both species still showed a preference for the larger media in the dark. This suggests that female blow flies can use visual cues to recognize larvae on the media and decide on their oviposition site. Our studies point out the effects of some biotic and abiotic factors which were previously overlooked, and remind us to reevaluate these effects on oviposition, especially when using insect developmental data to estimate PMIs.

In a previous study, the presence of tick-borne encephalitis virus (TBEV) in questing Ixodes ricinus L. ticks and in field derived ticks that engorged on small mammals (n = 9,986) was investigated at four sites located in a TBE area in Switzerland. Two of these sites were already recognized as TBE foci (Thun and Belp) and the screening of ticks revealed the presence of TBEV in ticks at a third site, Kiesen, but not at the fourth one, Trimstein. The aim here was to test another approach to detect TBE endemic areas. Sera from 333 small mammals (Apodemus flavicollis, A. sylvaticus, Myodes glareolus) captured in 2006 and 2007 at the four sites were examined for the presence of antibodies against TBEV using immunofluorescence and avidity tests. Overall the prevalence of antibodies against TBEV in rodents reached 3.6% (12/333). At two sites known as TBE foci, Thun and Belp, anti-TBEV antibodies were detected in 9.9% (9/91) and 1.6% (1/63) of rodent sera, respectively. At the third site, Kiesen, recently identified as a TBE focus by the detection of TBEV in ticks, anti-TBEV antibodies were detected in 1.8% (2/113) of rodent sera. Finally, at Trimstein, none of the examined rodent sera had antibodies against TBEV (0/66). This study shows another approach to detect TBE foci by testing antibodies in small mammal sera that is less time-consuming and less expensive than molecular tools.