The mosquito Aedes (Stegomyia) aegypti (L.), which occurs widely in the subtropics and tropics, is the primary urban vector of dengue and yellow fever viruses, and an important vector of chikungunya virus. There is substantial interest in how climate change may impact the bionomics and pathogen transmission potential of this mosquito. This Forum article focuses specifically on the effects of temperature on the bionomics of Ae. aegypti, with special emphasis on the cool geographic range margins where future rising temperatures could facilitate population growth. Key aims are to: 1) broadly define intra-annual (seasonal) patterns of occurrence and abundance of Ae. aegypti, and their relation to climate conditions; 2) synthesize the existing quantitative knowledge of how temperature impacts the bionomics of different life stages of Ae. aegypti; 3) better define the temperature ranges for which existing population dynamics models for Ae. aegypti are likely to produce robust predictions; 4) explore potential impacts of climate warming on human risk for exposure to Ae. aegypti at its cool range margins; and 5) identify knowledge or data gaps that hinder our ability to predict risk of human exposure to Ae. aegypti at the cool margins of its geographic range now and in the future. We first outline basic scenarios for intra-annual occurrence and abundance patterns for Ae. aegypti, and then show that these scenarios segregate with regard to climate conditions in selected cities where they occur. We then review how near-constant and intentionally fluctuating temperatures impact development times and survival of eggs and immatures. A subset of data, generated in controlled experimental studies, from the published literature is used to plot development rates and survival of eggs, larvae, and pupae in relation to water temperature. The general shape of the relationship between water temperature and development rate is similar for eggs, larvae, and pupae. Once the lower developmental zero temperature (10–14°C) is exceeded, there is a near-linear relationship up to 30°C. Above this temperature, the development rate is relatively stable or even decreases slightly before falling dramatically near the upper developmental zero temperature, which occurs at ∼38–42°C. Based on life stage-specific linear relationships between water temperature and development rate in the 15–28°C range, the lower developmental zero temperature is estimated to be 14.0°C for eggs, 11.8°C for larvae, and 10.3°C for pupae. We further conclude that available population dynamics models for Ae. aegypti, such as CIMSiM and Skeeter Buster, likely produce robust predictions based on water temperatures in the 16–35°C range, which includes the geographic areas where Ae. aegypti and its associated pathogens present the greatest threat to human health, but that they may be less reliable in cool range margins where water temperatures regularly fall below 15°C. Finally, we identify knowledge or data gaps that hinder our ability to predict risk of human exposure to Ae. aegypti at the cool margins of its range, now and in the future, based on impacts on mosquito population dynamics of temperature and other important factors, such as water nutrient content, larval density, presence of biological competitors, and human behavior.

Simulium (Gomphostilbia) leparense sp. nov. is described from females, males, and pupae collected from Peninsular Malaysia. This new species is assigned to the ceylonicum species-group of the subgenus Gomphostilbia, and is characterized by the female and male scuta covered with dark-brown short hairs, smaller number of male upper-eye facets, presence of shiny paired spots on the male abdominal segments 2–8, and absence of grapnel-shaped hooklets on the pupal abdominal segment 9. The male and pupa of S. capillatum Takaoka, which was originally described from larvae collected from Sarawak and Sabah, are described for the first time. Keys to identify all 32 species of the Simulium ceylonicum species-group including 27 species from other countries are provided for females, males, pupae, and mature larvae.

Four new species of the feather mite family Proctophyllodidae are described from two species of the family Mimidae (Passeriformes): Proctophyllodes apanaskevichi sp. n. (Proctophyllodinae) and Amerodectes dumetellae sp. n. (Pterodectinae) from the Gray Catbird, Dumetella carolinensis (L., 1766), and Proctophyllodes gallowayi sp. n. (Proctophyllodinae) and Metapterodectes toxostomae sp. n. (Pterodectinae) from the Brown Thrasher, Toxostoma rufum (L., 1758). A. dumetellae and M. toxostomae represent the first records of mites of the subfamily Pterodectinae on mockingbirds.

The medically important spiders Loxosceles intermedia Mello-Leitão and Loxosceles laeta (Nicolet) are segregated in Curitiba, southern Brazil, where L. intermedia is more abundant and widespread than L. laeta. Because they share similar microhabitat preferences and wander in search of web sites, agonistic encounters are likely to occur. The purposes of this study were to describe agonistic interactions and interpret their consequences for the relative abundances and spatial segregation of L. intermedia and L. laeta. Experimental contests were performed between residents and intruders. Asymmetries between contestants included sex, age, species, weight, and residence status. Nine behavioral categories were defined. Through discriminant analyses, it was possible to differentiate spider sex, species, and residence based on their agonistic behaviors. Intruders, juveniles, and L. intermedia individuals were better characterized by exploratory behaviors, whereas L. laeta females were differentiated by aggressiveness. By performing a multiple logistic regression, with winning or defeat as a dependent variable of sex, age, species, size, weight, and residence, it was possible to say that residents and L. intermedia individuals had the highest winning odds in contests,whereas juveniles had lower winning odds than adults. Advantages of the prior residence may help to explain the predominance of L. laeta in old colonization sites, whereas the higher winning odds of L. intermedia and less aggressive behavior toward conspecifics may lead to a successful establishment of dense populations in new sites. A better understanding of agonistic interactions as a mechanism of spacing, segregation, and species replacement among spiders may be helpful for control purposes.

The distribution of Culicoides sonorensis Wirth and Jones was examined in Alberta, Canada. Sampling was conducted weekly using blacklight traps at eight locations in 2009, and 10 locations during 2010–2012. Nine supplemental sites were sampled twice annually during both 2011 and 2012. Abundance of C. sonorensis was consistently greatest at a site near the U.S. border, and declined in a northerly direction. Mean annual abundance at this site ranged from 6.4- to >1,000-fold greater across positive sites. Data from a less extensive survey conducted during 2002–2006 were included in the remaining analyses. C. sonorensis was distributed below a diagonal spanning 49° 30′ N, 113° 0′ W to 51° 21′ N, 110° 40′ W. The relationship between the proportion of weekly samples positive and mean annual abundance at a site was determined and indicated that the proportion of positive samples could be used as a surrogate measure of abundance to overcome issues associated with the extreme variation in abundance. A series of logistic regression models were developed and evaluated to determine the effects of spatial (latitude and longitude), climatic (historic temperature and precipitation during the warmest quarter), and weather (temperature during the sample interval and spring precipitation) on abundance as measured by the proportion of positive samples. Spatial and climatic variables set the overall level of abundance, while weather variables added seasonal fluctuations within years, and also fluctuations between years. These data will be useful for long-term monitoring of C. sonorensis and as a baseline for detecting shifts in abundance that might occur because of climate change.

The impact of climate change on the distribution of Aedes albopictus (Skuse) was analyzed in northern Japan, where chronological distribution records are incomplete. We analyzed local climate data using linear regression of the thermal suitability index (TSI) for the mosquito and mean annual temperature as functions of time. In northern Japan, thermal conditions since the early 20th century have become increasingly suitable for Ae. albopictus, more as a result of decreasing coldness in the overwintering season than increasing warmth in the reproductive season. Based on recent discovery records of Ae. albopictus in the northern border range, we determined thermal criteria for estimating when its persistent establishment became thermally possible. Retrospective analyses indicated that those criteria were reached in most coastal lowlands of northern Honshu before the accelerated temperature increase after the mid-1980s and the first records of this species after 1990; at some sites, temperature criteria were reached during or before the early 20th century. Expansion of the thermally suitable range after 1990 was supported only for inland areas and the northernmost Pacific coast. The estimated expansion rate was ≈26 km per decade. Our analyses also demonstrated the importance of local climate heterogeneity (apart from north—south or altitudinal temperature gradients) in determining the expansion pattern.

Mosquito eggs laid on water surfaces typically hatch spontaneously soon after the embryos within them become fully formed first-instar larvae. However, we have found that Anopheles gambiae Giles, an important vector of malaria in Africa, exhibits delayed hatching until the water surface is agitated, a feature overlooked in most laboratory colonies. Agitation within 24 h postoviposition, before embryonation was complete, failed to stimulate delayed postembryonic hatching of isolated eggs on the following day (day 2), when <1% had hatched spontaneously. However, 5 min of water agitation of these dormant pharate first-instar larvae on day 2 resulted in an almost immediate hatch of 63.3 versus 0% of nonagitated controls, plus another 3.9 versus 0.3%, respectively, during the following 24 h. With daily agitation, installment hatching occurred mainly during 2–6 d postoviposition. The mean cumulative hatch after 7 d of daily agitation was 83.1 versus 1.1% of nonagitated eggs. Experiments with eggs in groups demonstrated that egg density and activity of already-hatched larvae had no stimulatory effect. Eggs stored 1–4 wk at 25.5 or at 15.5°C, and then agitated daily for 6 d at 25.5°C, showed a gradual decline in viability. Viability was sustained longer at the lower temperature. Implications of agitation-induced egg hatching for rainy-season and dry-season ecology of An. gambiae are discussed. Suspended hatching and cool storage already are proving convenient for efficient mass rearing and accurate modeling of weather-based population dynamics.

Gravid screwworm flies, Cochliomyia hominivorax (Coquerel), are attracted to the volatiles from waste larval rearing media to deposit eggs. Studies were conducted to identify volatile chemicals from the waste larval media and determine their effectiveness to attract gravid flies to oviposit. Volatiles were collected using solid-phase microextraction method, and five active chemicals, namely, dimethyl disulfide, dimethyl trisulfide, phenol, p-cresol, and indole, were identified using gas chromatography—mass spectrometry. In electroantennography studies, antennae of gravid screwworm flies, Cochliomyia macellaria (F.), responded positively to each of the identified compounds. A synthetic blend of these five compounds in the ratio of 335:200:57:1:12 was prepared and tested for its effectiveness to attract both C. hominivorax and C. macellaria using laboratory bioassay methods. Significantly more gravid C. macellaria were attracted to and landed on substrates treated with 10-fold diluted blends compared with those landed on substrates treated with ethanol only (as control). Only a few young females and young and old males were attracted to the substrates treated with the synthetic blend. The C. hominivorax females laid significantly more eggs on substrates treated with waste media, 10-fold diluted blend, and 100-fold diluted blend than on substrates with undiluted blend or ethanol. Similarly, C. macellaria deposited significantly more eggs on substrates treated with waste media, 10-fold diluted blend, and 100-fold diluted blend compared with substrates with undiluted blend or ethanol. C. macellaria females deposited significantly less amount of eggs than did C. hominivorax females. These results indicate that the synthetic blend of five compounds identified may serve as an oviposition attractant for C. hominivorax as well as for C. macellaria.

Yellow fever virus is a reemerging infection responsible for widespread, sporadic outbreaks across Africa. Although Aedes aegypti (L.) is the most important vector globally, in East Africa, epidemics may be vectored by Aedes bromeliae (Theobald), a member of the Aedes simpsoni (Theobald) species complex. The Ae. simpsoni complex contains 10 subspecies, of which Ae. bromeliae alone has been incriminated as a vector of yellow fever virus. However, morphological markers cannot distinguish Ae. bromeliae from conspecifics, including the sympatric and non-anthropophilic Aedes lilii (Theobald). Here, we used three sequenced nuclear markers to examine the population structure of Ae. simpsoni complex mosquitoes collected from diverse habitats in Rabai, Kenya. Gene trees consistently show strong support for the existence of two clades in Rabai, with segregation by habitat. Domestic mosquitoes segregate separately from forest-collected mosquitoes, providing evidence of habitat partitioning on a small spatial scale (<5 km). Although speculative, these likely represent what have been described as Ae. bromeliae and Ae. lilii, respectively. The observation of high levels of diversity within Rabai indicates that this species complex may exhibit significant genetic differentiation across East Africa. The genetic structure, ecology, and range of this important disease vector are surprisingly understudied and need to be further characterized.

A field strain of Aedes aegypti (L.) was collected from Puerto Rico in October 2008. Based on LD₅₀ values by topical application, the Puerto Rico strain was 73-fold resistant to permethrin compared with a susceptible Orlando strain. In the presence of piperonyl butoxide, the resistance of Puerto Rico strain of Ae. aegypti was reduced to 15-fold, suggesting that cytochrome P450-mediated detoxification is involved in the resistance of the Puerto Rico strain to permethrin. To determine the cytochrome P450s that might play a role in the resistance to permethrin, the transcriptional levels of 164 cytochrome P450 genes in the Puerto Rico strain were compared with that in the Orlando strain. Of the 164 cytochrome P450s, 33 were significantly (P < 0.05) up-regulated, including cytochrome P450s in families four, six, and nine. Multiple studies have investigated the functionality of family six and nine cytochrome P450s, therefore, we focused on the up-regulated family 4 cytochrome P450s. To determine whether up-regulation of the four cytochrome P450s had any functional role in permethrin resistance, transgenic Drosophila melanogaster Meigen lines overexpressing the four family 4 P450 genes were generated, and their ability to survive exposure to permethrin was evaluated. When exposed to 5 μg per vial permethrin, transgenic D. melanogaster expressing CYP4D24, CYP4H29, CYP4J15v1, and CYP4H33 had a survival rate of 60.0 ± 6.7, 29.0 ± 4.4, 64.4 ± 9.7, and 11.0 ± 4.4%, respectively. However, none of the control flies survived the permethrin exposure at the same concentration. Similarly, none of the transgenic D. melanogaster expressing CYP4J15v1 or CYP4H33 ?5 survived when they were exposed to permethrin at 10 μg per vial. However, transgenic D. melanogaster expressing CYP4D24 and CYP4H29 had a survival rate of 37.8 ± 4.4 and 2.2 ± 2.2%, respectively. Taken together, our results suggest that CYP4D24 might play an important role in cytochrome P450-mediated resistance to permethrin.

Studies were made on the ribosomal DNA intergenic region, comprising complete internal transcribed spacer (ITS)-1, 5.8S, and ITS-2 sequences, of populations of the triatomine Panstrongylus megistus, the most important vector of Chagas' disease in Brazil since Triatoma infestans eradication. Specimens were from 26 localities of Rio Grande do Sul, Santa Catarina, Paraná, São Paulo, Minas Gerais, Bahia, and Sergipe states. In total, 21 ITS-1 and 12 ITS-2 haplotypes were found. Nucleotide differences were higher in ITS-1 (3.00%) than in ITS-2 (1.33%). The intergenic region was 1,513–1,522-bp-long (mean 1,516.9 bp), providing 26 combined haplotypes. The combination of microsatellites found in both ITSs may be of applied usefulness, to assess interpopulation specimen exchange and potential recolonizations after vector elimination by control implementation. Network results suggest that São Paulo maybe considered one of the spreading centers of this species. Molecular clock datation suggests that P. megistus populations are diversifying at least since 4.54 million years ago, with diversification still ongoing today by geographical isolation of populations. Evidence is provided about the relationship of genetic diversity with geographical spread that characterizes a major vector and explains its ability to colonize distant areas and different ecotopes, including human habitats, and consequently its importance in Chagas' disease epidemiology.

Behavioral bioassays remain a standard tool in the discovery, development, and registration of arthropod repellents. Tick repellent bioassays are generally uncomplicated, but their results can be affected by basic variables (e.g., dimensions of testing materials, substrate, timing, temperature) of the assay. Using lone star tick, Amblyomma americanum (L.), nymphs in climbing bioassays, we tested for the effects of substrate, solvent, and drying time on tick responses. In dose-response tests, the widely used repellents N,N-diethyl-3-methyl benzamide (deet) and 1-methylpropyl-2-(hydroxyethyl)-l-piperidinecarboxylate (picaridin) were applied to filter paper strips and challenged by ticks at 10, 20, 30, 40, and 120 min after application. At 10-min drying time, repellency at the intermediate concentration 500 nmol repellent/cm² filter paper was significantly lower for ethanol solutions of deet and picaridin (0 and 10% ticks repelled, respectively) than for solutions of deet and picaridin in acetone (96.7 and 76.7% ticks repelled, respectively). Repellency was greatest for both the acetone and ethanol solutions of deet and picaridin when challenged 120 min after application, and at shorter drying times at the highest concentration tested (2,000 nmol compound/cm²). The repellency of picaridin relative to deet differed at some combinations of solvent and drying time but not others. In dose-response tests using different paper substrates and a drying time of 10 min, both ethanol and acetone solutions of deet differed in repellency, depending on both the paper substrate and the solvent. However, there were no differences in repellency between ethanol and acetone solutions of deet applied to nylon organdy in an in vitro and in an in vivo (fingertip) bioassay. When deet in solution with various proportions of ethanol:water was applied at 2,000 nmol deet/cm² filter paper, the proportion of ticks repelled decreased as the proportion of water in the test solutions increased. Somewhat similar results were seen for solutions of deet in an acetone solvent. Water absorbed from the atmosphere may affect the efficacy of repellents in solution with anhydrous ethanol. Overall, results obtained from bioassays that differ in seemingly minor ways can be surprisingly different, diminishing the value of comparing studies that used similar, but not identical, methods. Nylon organdy or another similar thin cloth may be preferable to filter papers and copier paper for minimizing solvent-related differences. When a paper substrate is used, acetone may be the more suitable solvent if the solubility of the test compound and other factors allow.

The monitoring of the susceptibility of fleas to insecticides has typically been conducted by exposing adults on treated surfaces. Other methods such as topical applications of insecticides to adults and larval bioassays on treated rearing media have been developed. Unfortunately, baseline responses of susceptible strains of cat flea, Cteno cephalides felis (Bouchè), except for imidacloprid, have not been determined for all on-animal therapies and new classes of chemistry now being used. However, the relationship between adult and larval bioassays of fleas has not been previously investigated. The adult and larval bioassays of fipronil and imidacloprid were compared for both field-collected isolates and laboratory strains. Adult topical bioassays of fipronil and imidacloprid to laboratory strains and field-collected isolates demonstrated that LD₅₀s of fipronil and imidacloprid ranged from 0.11 to 0.40 nanograms per flea and 0.02 to 0.18 nanograms per flea, respectively. Resistance ratios for fipronil and imidacloprid ranged from 0.11 to 2.21. Based on the larval bioassay published for imidacloprid, a larval bioassay was established for fipronil and reported in this article. The ranges of the LC₅₀s of fipronil and imidacloprid in the larval rearing media were 0.07–0.16 and 0.11–0.21 ppm, respectively. Resistance ratios for adult and larval bioassays ranged from 0.11 to 2.2 and 0.58 to 1.75, respectively. Both adult and larval bioassays provided similar patterns for fipronil and imidacloprid. Although the adult bioassays permitted a more precise dosage applied, the larval bioassays allowed for testing isolates without the need to maintain on synthetic or natural hosts.

Resistance to the organophosphate insecticide chlorpyrifos was evaluated in females from six strains of Aedes aegypti (L.) that expressed high levels of cross-resistance to eight pyrethroid insecticides. Relative to LC₅₀ and LC₉₀ at 24 h of a susceptible New Orleans (NO) strain, three strains were highly resistant to chlorpyrifos (Coatzacoalcos, resistance ratio [RRLC₉₀] = 11.97; Pozarica, RRLC₉₀ = 12.98; and Cosoleacaque, RRLC₅₀ = 13.94 and RRLC₉₀ = 17.57), one strain was moderately resistant (Veracruz, RRLC₉₀ = 5.92), and two strains were susceptible (Tantoyuca and Martinez de la Torre, RRLC₅₀ and RRLC₉₀ < 5) in bottle bioassays according to Centers for Disease Control and Prevention. Furthermore, high levels of α- or β-esterase activity in the sample populations were correlated with resistance, suggesting that esterase activity may be a mechanism causing the development of organophosphate resistance in these populations. Overall, the populations in this study were less resistant to chlorpyrifos than to pyrethroids. Rotation of insecticides used in control activities is recommended to delay or minimize the occurrence of high levels of resistance to chlorpyrifos among local populations of Ae. aegypti. The diagnostic dose and diagnostic time for chlorpyrifos resistance monitoring was determined to be 85 µg per bottle and 30 min, respectively, using the susceptible NO strain.

Pyroglyphid house dust mites are the most common cause of allergic symptoms in humans. An assessment was made of the toxicity of basil, Ocimum basilicum L, essential oil, 11 basil oil constituents, seven structurally related compounds, and another 22 previously known basil oil constituents to adult American house dust mites, Dermatophagoides farinae Hughes. The efficacy of four experimental spray formulations containing basil oil (1, 2, 3, and 4% sprays) was also assessed. Results were compared with those of two conventional acaricides benzyl benzoate and N,N-diethyl-3-methylbenzamide. The active principles of basil oil were determined to be citral, a-terpineol, and linalool. Citral (24 h LC₅₀,1.13 µg/cm²) and menthol (1.69 µg/cm²) were the most toxic compounds, followed by methyl eugenol (5.78 µg/cm²). These compounds exhibited toxicity greater than benzyl benzoate (LC₅₀, 8.41 µg/cm²) and N,N-diethyl-3-methylbenzamide (37.67 µg/cm²). Potent toxicity was also observed with eugenol, menthone, spathulenol, α-terpineol, nerolidol, zerumbone, and nerol (LC₅₀,12.52–21.44 µg/cm²). Interestingly, the sesquiterpenoid α-humulene, lacking only the carbonyl group present in zerumbone, was significantly less effective than zerumbone, indicating that the α,β-unsaturated carbonyl group of zerumbone is a prerequisite component for toxicity. These compounds were consistently more toxic in closed versus open containers, indicating that their mode of delivery was largely a result of vapor action. Basil oil applied as 3 and 4% sprays provided 97 and 100% mortality against the mites, respectively, whereas permethrin (cis:trans, 25:75) 2.5 g/liter spray treatment resulted in 17% mortality. Our results indicate that practical dust mite control in indoor environments can be achieved by basil oil spray formulations (3 and 4% sprays) as potential contact-action fumigants.

Orientia tsutsugamushi is a pathogen transmitted by Leptotrombidium that causes scrub typhus. To develop an infection mouse model, a mite-derived isolate of O. tsutsugamushi was established from a laboratory-maintained colony of Leptotrombidium chiangraiensis (O. tsutsugamushi Lc-1). This Lc-1 isolate was initially presented to ICR (CD-1) mice by feeding an infected Lc chigger on the ear of a mouse. Once the Lc-1 was adapted to the ICR mice, quantitative real-time polymerase chain reaction was used to investigate O. tsutsugamushi genomic equivalent copies in tissues and sera. Furthermore, times to onset of the signs of infection are reported in this study. This study provides information useful for future research on this host-pathogen interaction and the associated vaccine efficacy trials.

In 2005, established populations of Aedes albopictus (Skuse) were discovered in the Torres Strait, the region that separates Papua New Guinea from northern Australia. This increased the potential for this species to be introduced to mainland Australia. Because it is an arbovirus vector elsewhere, we undertook laboratory-based infection and transmission experiments to determine the potential for Ae. albopictus from the Torres Strait to become infected with and transmit the four major Australian endemic arboviruses—Murray Valley encephalitis virus, West Nile virus Kunjin strain (WNV_KUN), Ross River virus (RRV), and Barmah Forest virus—as well as the exotic Japanese encephalitis virus. Ae. albopictus is susceptible to infection with all viruses, with infection rates ranging between 8% for WNV_KUN and 71% for RRV. Transmission rates of ≈25% were observed for RRV and Barmah Forest virus, but these were <17% for Murray Valley encephalitis virus, WNV_KUN, and Japanese encephalitis virus. Given its relative vector competence for alphaviruses, we also examined the replication kinetics and extrinsic incubation periods required for transmission of RRV and Chikungunya virus. Despite lower body titers, more mosquitoes reared and maintained at 28°C became infected with and transmitted the virus than those reared and maintained at 22°C. The minimum time between Ae. albopictus consuming an infected bloodmeal and transmitting Chikungunya virus was 2 d at 28°C and 4 d at 22°C, and for RRV, it was 4 d, irrespective of the temperature. Given its opportunistic feeding habits and aggressive biting behavior, the establishment of Ae. albopictus on the Australian mainland could have a considerable impact on alphavirus transmission.

The house fly (Musca domestica L.) is a well-established vector of human pathogens, including Campylobacter spp., which can cause infection of broiler chicken flocks, and through contaminated broiler meat can cause outbreaks of campylobacteriosis in humans. We investigated whether Campylobacter jejuni (Jones) could be transferred between life stages of M. domestica (larvae-pupae-adults) and determined bacterial counts of C. jejuni at different time points after bacterial exposure. C. jejuni was transmitted from infected larvae to pupae, but not to the adult stage. Infected larvae maintained at 25°C had mean bacterial numbers of 6.5 ± 0.2 SE log₁₀ (colony forming units [CFU]/g) that subsequently dropped to 3.6 ± 0.3 SE log₁₀ (CFU/g) 8 h after infection. Pupae originating from infected larvae contained mean bacterial numbers of 5.3 ± 0.1 SE log₁₀ (CFU/g), and these numbers dropped to 4.8 ± 0.1 SE log₁₀ (CFU/g) 24 h after pupation. The decline in C. jejuni numbers during pupal development coincided with increased expression of antimicrobial peptides, including cecropin, diptericin, attacin, and defensin, in the larva-pupa transition stage and a later second peak in older pupae (4 or 48 h). Conversely, there was a reduced expression of the digestive enzyme, lysozyme, in pupae and adults compared with larvae.

In Israel, sand flies are the vectors of Leishmania Ross and mosquitoes are the vectors of West Nile Virus. In the Judean Desert and Tiberias, the sand fly Phlebotomus sergenti Parrot is the vector of Leishmania tropica (Wright) and the rock hyrax (Procavia capensis Pallas) is considered the main reservoir animal. The main vectors of West Nile Virus are Culex pipiens L. and Culex perexiguus Theobald. Bloodmeals of engorged field-caught female sand flies and mosquitoes are an important source for defining host preferences. Recent progress in DNA molecular techniques has enabled the accurate identification of blood sources within the arthropod gut. In this study, we applied molecular approach for species-specific identification based on polymerase chain reaction and nucleotide sequence analysis of polymorphic regions along two mitochondrial genes, 12S and 16S rRNA. The research was carried out on 261 engorged female sand flies collected in the Judean Desert and Tiberias and 50 engorged female mosquitoes collected in Tel-Aviv and Arava. Species identification of bloodmeals was successful in 92% of the samples. Rock hyrax was the most abundant host in bloodmeals of P. sergenti, while human blood was found in only seven (3%) females. L. tropica DNA was detected in three P. sergenti females from Tiberias that contained rock hyrax blood. Avian sequences were detected in 67% (10 of 15) of the identified bloodmeals from Cox. perexiguus and in 10% (3 of 29) of the identified meals from Cx. pipiens. Human sequences were found in 14% of the identified bloodmeals from Cx. pipiens. The successful analysis of the majority of the bloodmeals performed on wild sand flies and mosquitoes suggests that bloodmeal identification can be applied as one of the routine procedures in vector surveillance programs.

Trypanosoma and Leishmania infections affect wild and domestic animals and human populations. The growing process of deforestation and urbanization of Atlantic Rainforest areas has given rise to introduction of humans and domestic animals to the sylvatic cycles of Trypanosoma and Leishmania species. Serological, parasitological, and molecular surveys among wild and domestic animals in the Corrego do Veado Biological Reserve, which is an Atlantic Rainforest fragment in the state of Espírito Santo, southeastern Brazil, were evaluated. In total, 154 wild animals of 25 species and 67 domestic animals (47 dogs and 20 horses) were sampled. All the domestic animals were serologically negative for anti-Leishmania infantum chagasi antibodies and negative in parasitological approaches. Only the Order Chiroptera presented positive blood cultures and cryopreserved isolates. The phylogenetic trees based on SSU rDNA and gGAPDH genes confirmed the occurrence of Trypanosoma dionisii and provided the first record of Trypanosoma cruzi marinkellei in southeastern Brazil. The studies conducted in Atlantic Rainforest remaining trees provide the knowledge of parasite diversity or detect parasites that can accelerate the loss of hosts diversity.

Changes in the structure of managed red pine forests in Wisconsin caused by interacting root- and stem-colonizing insects are associated with increased abundance of the blacklegged tick, Ixodes scapularis Say, in comparison with nonimpacted stands. However, the frequency and variability of the occurrence of tick-borne pathogens in this coniferous forest type across Wisconsin is unknown. Red pine forests were surveyed from 2009 to 2013 to determine the prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum in questing I. scapularis nymphs. Polymerase chain reaction analysis revealed geographical differences in the nymphal infection prevalence (NIP) of these pathogens in red pine forests. In the Kettle Moraine State Forest (KMSF) in southeastern Wisconsin, NIP of B. burgdorferi across all years was 35% (range of 14.5–53.0%). At the Black River State Forest (BRSF) in western Wisconsin, NIP of B. burgdorferi across all years was 26% (range of 10.9–35.5%). Differences in NIP of B. burgdorferi between KMSF and BRSF were statistically significant for 2010 and 2011 and for all years combined (P < 0.05). NIP of A. phagocytophilum (human agent) averaged 9% (range of 4.6–15.8%) at KMSF and 3% (range of 0–6.4%) at BRSF, and was significantly different between the sites for all years combined (P < 0.05). Differences in coinfection of B. burgdorferi and A. phagocytophilum were not statistically significant between KMSF and BRSF, with an average of 3.4% (range of 1.7–10.5%) and 2.5% (range of 0–5.5%), respectively. In 2013, the density of infected nymphs in KMSF and BRSF was 14 and 30 per 1000m², respectively, among the highest ever recorded for the state. Differences in the density of nymphs and NIP among sites were neither correlated with environmental factors nor time since tick colonization. These results document significant unexplained variation in tick-borne pathogens between coniferous forests in Wisconsin that warrants further study.

As the most common primary colonizer of carrion, adult blow flies (Diptera: Calliphoridae) play an important role in initiating arthropod-mediated breakdown of soft tissue; however, their timing is highly variable. This variability complicates the estimation of precolonization intervals or periods of insect activity by forensic entomologists. In this study, the size of the adult blow fly on swine carcasses was compared with various environmental conditions including time of day, temperature, wind speed, and light levels. Four trials were conducted: two in August and September 2008, one in January 2009, and one in February-March 2010. Of the measured variables, time of day was the only consistent factor explaining the population size of blow fly on a carcass, although precipitation and high winds affected winter-active Calliphora vicina Robineau-Desvoidy. Male flies were also collected, suggesting that carcasses may play additional roles in adult blow fly ecology beyond that of a simple oviposition site. For both sexes of flies, a strong diel pattern of behavior emerged, which could be useful in estimating precolonization intervals by considering the environmental conditions at a scene, and thus forensic entomologists may be better able to estimate the likelihood of adult activity at a carcass.

Tick-borne diseases pose significant risks to U.S. military personnel who conduct operations, both domestic and abroad. To determine the feasibility of protecting personnel from tick vectors during short-term field deployments, acaricides Cypermethrin (Demon WP, Syngenta, Greensboro, NC) and lambda-cyhalothrin (Surrender Pestabs, CSI, Pasadena, TX) were applied to plots within two separate field sites on Camp Blanding Joint Training Center in Starke, FL, from May to June 2011. We analyzed their effectiveness in reducing tick counts for 6 wk after application. In total, 8,193 ticks were identified and counted, of which >99% were a mix of nymphs and adult-stage Amblyomma americanum (L.). Our results indicate that both Cypermethrin and lambda-cyhalothrin were effective in significantly reducing tick numbers and preventing entry into treated plots for 6 wk after application. Thus, these two acaracides can be used to effectively suppress tick populations and provide residual protection in small geographic areas of recreation or public health significance.

Forensically important calliphorids can be reared on a mixture of beef liver and agar. Small pieces of meat, especially fresh or frozen beef liver, will desiccate in 2–6 h, but this simple-to-make feeding substrate remains moist for at least 12 h at 25 and 30°C without desiccation, even in small (5 g) amounts. We determined the survivorship of small numbers of Chrysomya megacephala (F.) (first-instar larvae to adult eclosion) raised on 5 g of liver agar and fresh beef liver. We found that all larvae raised on 5 g of liver died due to desiccation, but survivorship on 5 g of liver agar was equivalent to that on larger (50 g) pieces of either liver agar or beef liver.

The infestation of human or animal tissues by fly larvae has been given distinctive terminology depending on the timing and location of colonization. Wounds and orifices colonized by Diptera in a living human or animal are typically referred to as myiasis. When the colonization occurs after death, it is referred to as postmortem colonization and can be used to estimate the minimum postmortem interval. What happens when the human, as in the case presented here, has a necrotic limb while the human remains alive, at least for a short period of time? The case presented here documents perimortem wound colonization by Lucilia eximia (Wiedemann) and Chrysomya rufifacies (Macquart) and the considerations for approximating development temperatures and estimating the time of colonization (TOC). This represents the first record of L. eximia in human myiasis in the United States and the first record of the co-occurrence of L. eximia and C. rufifacies in human myiasis in the United States. The TOC was estimated using both ambient and body temperature. Insect colonization before death complicates the estimation of TOC and minimum postmortem interval and illustrates the problem of temperature approximation in forensic entomology casework.

We assessed the nontarget effects of ultra-low-volume (ULV) aerial adulticiding with two new water-based, unsynergized pyrethroid formulations, Aqua-K-Othrine (FFAST antievaporant technology, 2% deltamethrin) and Pesguard S102 (10% d-phenothrin). A helicopter with GPS navigation technology was used. One application rate was tested per formulation that corresponded to 1.00 g (AI)/ha of deltamethrin and 7.50 g (AI)/ha of d-phenothrin. Three beneficial nontarget organisms were used: honey bees (domesticated hives), family Apidae (Apis mellifera L.); mealybug destroyers, family Coccinellidae (Cryptolaemus montrouzieri Mulsant); and green lacewings, family Chrysopidae (Chrysoperla carnea (Stephens)). No significant nontarget mortalities were observed. No bees exhibited signs of sublethal exposure to insecticides. Beehives exposed to the insecticidal applications remained healthy and productive, performed as well as the control hives and increased in weight (25–30%), in adult bee population (14–18%), and in brood population (15–19%).