Dr. Marcel Leclercq was a pioneer in the field of forensic entomology. He has provided his knowledge of insect biology to many forensic cases, and most of them have found the way to publication. Most of the articles he has written were focused on individual cases, and despite the abundance of entomoforensic investigations he conducted, no synthesis has been published. This article summarizes 36 yr of forensic entomological investigations in temperate Europe, mainly in Belgium. Dr. Leclercq's work includes 132 entomological cases involving 141 human corpses found in various death scenes. Under certain conditions, insect specimens found at death scene can provide information on when (postmortem interval estimation), where and how a person died. More or less 100 insect species associated with a dead body have been identified by Dr. Leclercq.

The spread of leishmaniasis to areas where it was previously considered nonendemic has been recently found in the New and Old Worlds, and climate changes are suspected as a crucial factor responsible for this spread. Ambient temperature is known to significantly affect the metabolism of sand flies and their developmental times, but little is known about the effect of temperature on the Leishmania life cycle in vectors. This study assesses the effect of temperature on the development of two closely related New World Viannia species, Leishmania braziliensis and Leishmania peruviana, in the permissive vector Lutzomyia longipalpis, and on the development of New and Old World Leishmania infantum in its natural vectors Lu. longipalpis and Phlebotomus perniciosus, respectively. The mountain species L. peruviana developed well in sand fly females kept at 20°C, whereas at 26°C, most infections were lost during the defecation of bloodmeal remains; this suggests an adaptation to the slower metabolism of sand flies living at lower ambient temperature. On the contrary, L. infantum and L. braziliensis developed well at both temperatures tested; heavy late-stage infections were observed in a majority of sand fly females maintained at 20°C as well 26°C Frequent fully developed infections of L. infantum and L. braziliensis at 20°C suggest a certain risk of the spread of these two Leishmania species to higher latitudes and altitudes.

Dermacentor raskemensis Pomerantzev, 1946 is one of the rare Asian species in this genus. The immature stages of this species have never been described. Reexamination of D. raskemensis holdings stored in the United States National Tick Collection revealed a collection lot containing reared nymphs and larvae of this species. This collection made it possible for us to find numerous nymphs and larvae of D. raskemensis among previously unidentified material collected in the field. Both immature stages of D. raskemensis are described here for the first time. Nymphs of D. raskemensis can be distinguished from those of other Dermacentor species in the region by small spiracular plate, relatively short and obtuse lateral projections of basis capituli dorsally, relatively short spurs on coxa I and the internal spur is characteristically very broadly rounded at its apex, and very small spur on coxa IV, whereas larvae of D. raskemensis can be distinguished from other Dermacentor by relatively short and obtuse lateral projections of basis capituli, ≈6 denticles in the median files on hypostome, and relatively short, broad, and rounded spur on coxa I. The nymphs and larvae of D. raskemensis studied originate from Afghanistan, India, Iran, and Pakistan, where they were collected from pikas and other small mammals.

A mark-release-recapture study was conducted to estimate the adult population size, migration, and dispersal patterns of male and female Aedes (Stegomyia) polynesiensis (Marks) in a valley of Moorea, a volcanic island, and a motu (islet) on the atoll of Tetiaroa, two settings typical of the Society Islands. Aedes polynesiensis recapture rate was high for females and low for males. The distribution of Aedes species in the valley was heterogeneous. Marked individuals dispersed to most parts of the motu and over great distances in the valley for some females. The study provides insights into the field dynamics of Ae. polynesiensis populations and confirms that more efficient sampling methods are warranted. There was no evidence of active migration between motus on the atoll, suggesting that Tetiaroa is a suitable site for small-scale initial open releases of Wolbachia incompatible insect technique and other sterile insect technique-like suppression or replacement strategies.

Movement of male Dermacentor andersoni (Stiles) was examined among 54 pairs of artificially infested donor and recipient cattle during a 3-yr period. The number of males declined at a rate independent of the initial level of infestation, while the rate of decline of females on the donor animals tended to increase with initial infestation level. Male tick movement to recipient cattle was observed in 26 of 54 (48%) of the animal pairs, but varied among years and trials. Movement tended tobe greater during April compared with May and June. The daily probability of movement averaged (SD) 0.067 (0.082), and the number of males moving per day averaged (SD) 0.083 (0.228). Logistic and Poisson regression models were developed and indicated that movement was determined by interactions between the number of males on the donor animals, differences in the number of females on the donor and recipient cattle, temperature, and female age. These models can be used to incorporate movement into tick population models.

The dispersal of and the existence of memory in sand flies were measured in a transmission area of cutaneous leishmaniasis, in the municipality of Bandeirantes, Paraná, Brazil. Sand flies were caught in a rural area, with Shannon trap installed in the forest and three Falcão traps installed in a human-inhabited environment (HIE) and three others in an impacted environment presently uninhabited by humans (EUH), from 1800 to 0600 hours. The captured sand flies were marked with yellow, blue, or red fluorescent powder, according to the environments where they were captured. All marked sand flies were released at 0700 hours at a point between the three environments. The recaptures were made with 28 Falcâo traps, distributed in the environments from for 10 consecutive days. The sand flies recaptured were examined under a stereomicroscope and later identified. It was concluded that sand flies are able to disperse over an average distance of 73 m, reaching 130 m in 24 h, showing that: 1) the sand flies were attracted with different intensities to each environment, and the ability to move among different environments allows the existence of enzootic cycle of Leishmania; 2) the sand flies possess a spatial memory, olfactory memory, or both, that enable them to return to the environment where they were captured initially, although the distances were different.

Otobius megnini (Dugès), often referred to as the spinose ear tick, is a one-host tick infesting the ear canal of a variety of ungulate species. The objective of this study was to develop sampling methods for collecting free-living stages of O. megnini and to collect preliminary data on habitat distribution. Sampling occurred during 2010 and 2011 at Fossil Rim Wildlife Center (FRWC) near Glen Rose, TX, where tick presence has been observed in the ears of a variety of ungulates. To sample for nonfeeding adult stage presence, a debris-filtering method was developed using screens of varying gauges. To sample host-seeking larval-stage tick presence, a carbon dioxide (CO₂ ) trap was developed using compressed CO₂ released through tubing along with cotton fabric used for tick attachment. Both methods proved successful with adults and larvae collected from animal shelters, with larvae also collected from oak leaf litter away from any structure. Locating both life stages off the host is the first crucial step toward the management of this tick species at facilities such as FRWC.

Effects of laterite cover soil with different characteristics on survival of buried eggs, third instar larvae, and pupae of Musca domestica (L.) were studied experimentally. Soil treatments were loose dry soil, loose wet soil, compacted dry soil, and compacted wet soil (CWS). Eggs, third instar larvae, and pupae were buried under 30 cm of the different soil treatments and placed under field conditions until adults emerged. Rearing medium was provided for eggs and larvae, and control treatments of all stages were unburied immatures placed on soil surface. Egg and pupal survival to adult were significantly affected by the cover soil treatments, but third instars were more resilient. Wet soil treatments (loose wet soil and CWS) resulted in significantly reduced pupal survival, but increased survival of eggs. However, CWS significantly reduced adult emergence from buried eggs. Though emergence of house flies buried as eggs was significantly reduced, some were able to hatch and emerging first instar larvae developed to pupation. Although cover soil does not completely prevent fly emergence, it did limit development and emergence of buried house flies.

The effects of four temperatures (15, 23.5, 28, and 35°C) on the biological characteristics of the rove beetle Paederus fuscipes Curtis were studied, and its cuticular permeability also was measured. Specimens successfully developed to adulthood at each temperature tested, but development time of each preadult stage significantly decreased with increasing temperature. Both egg and L1 stages required at least 80 degree days above a threshold of ≈10°C to develop to the subsequent stage. The lengthy development time and high survival rate of preadults at 15°C suggests that P. fuscipes can survive in a harsh environment during cold weather by hibernating, and this ability could allow preadults to succeed ecologically in temperate countries. However, adult longevity was short, and no fecundity was recorded at 15°C. At 28°C, P. fuscipes exhibited a high survival rate of adults, which had a longer life span and high fecundity; thus, the population had the highest intrinsic rate of increase (0.0788 ± 0.0051 d^-1) and the shortest mean generation time (48.57 ± 1.43 d) at 28°C. At this temperature, the population might reach a size that could facilitate invasion into residential areas. However, in the absence of a hygric environment, P. fuscipes was unable to survive despite favorable temperature. Unlike in adults and pupae, high cuticular permeability values were found in the larval stages. This indicates that larvae are highly susceptible to desiccation, and it explains why the distribution of P. fuscipes is restricted to moist habitats.

The presence of predators can have dramatic consequences on prey communities, not only by the direct effects of consumption but also through sublethal effects. We investigated the survival rate and subsequent life history of the mosquito Culex pervigilans Bergroth under the influence of its major predator, the backswimmer Anisops wakefieldi White. We established a field experiment with various treatments: 1) control without predators, 2) free-roaming A. wakefieldi (with one, three, or nine A. wakefieldi per container), 3) caged A. wakefieldi (empty cage without predators, with one, three, or nine A. wakefieldi in each cage, and 4) A. wakefieldi cues (with cues concentrations of one, three, or nine A. wakefieldi). Cx. pervigilans eggs were then taken from these four experimental treatments and reared in two different laboratory conditions: 1) in clean water without any traces of predators, or 2) in water with the same treatments as in field. The survival rate of Cx. pervigilans was significantly reduced by the presence of predators or their cues. Even after a brief exposure to waters containing predators or residual cues, the subsequent progeny and the ontogeny of the remaining survivors were still affected. The percentage of eggs that hatched, and the resulting mosquito population, was influenced by the presence of predators or their cues. Our results suggest that sublethal effects may be carried by surviving individuals primarily through the effects of stress, perhaps by epigenetic mechanisms. We may expect to observe similar plasticity in species or populations with high temporal or spatial variability in predation.

The stable fly, Stomoxys calcitrans (L.), is a cosmopolitan pest of livestock and humans. The pestiferous nature and painful bite cause stress to cattle and other animals. The stress and resulting avoidance behaviors manifest as reductions in weight gain or milk production in cattle; estimated annual economic loss in the United States exceeds US$2 billion. Understanding the population genetics of stable flies could provide information on their population dynamics, origins of outbreaks, and geographical patterns of insecticide resistance, resulting in a tactical advantage for developing management strategies. Previous studies, mostly on a local scale, reported a high level of gene flow between locations. Here, we report results wherein amplified fragment length polymorphism was used to determine genetic diversity of stable fly samples consisting of 11–40 individuals from 12 locations representing the United States, Canada, and Panama. The Analysis of Molecular Variance showed that the majority of genetic diversity was within groups; very little was among groups. The F_ST and G_ST values were low (<0.4), Nm values high (>1.0). The tests of neutrality suggested population expansion, and no genetic differentiation was found between locations. These results show that stable flies have a high level of gene flow on a continental scale, with limited isolation owing to distance or geographical barriers.

Resistance to the insecticides deltamethrin and malathion and the enzymes associated with metabolic resistance mechanisms were determined in four field populations of Aedes aegypti (L.) from western Venezuela during 2008 and 2010 using the bottle assay and the microplate biochemical techniques. For deltamethrin, mortality rates after 1 h exposure and after a 24-h recovery period were determined to calculate the 50% knock-downconcentration (KC₅₀) and the lethal concentration (LC_50,) respectively. For malathion, mortality was recorded at 24 h to determine the LC_50. For deltamethrin, resistance ratios of knock-down resistance and postrecovery were determined by calculating the RRKC₅₀ and RRLC₅₀, comparing the KC₅₀ and LC₅₀ values of the field populations and those of the susceptible New Orleans strain. Knock-down resistance to deltamethrin was moderate in the majority of the populations in 2008 (RRKC₅₀ values were between 5- and 10-fold), and only one population showed high resistance in 2010 (RRKC₅₀ >10-fold). Moderate and high postrecovery resistance to deltamethrin was observed in the majority of the populations for 2008 and 2010, respectively. There was significantly increased expression of glutathione-S-tranferases and mixedfunction oxidases. All populations showed low resistance to malathion in 2008 and 2010 with significantly higher levels of α-esterases for 2008 and 2010 and β-esterases for 2008.

The lesser mealworm, Alphitobius diaperinus (Panzer), is an insect that lives in poultry houses, and high infestations may cause economic losses to producers. The control of this insect is usually done with insecticides; however, many of these chemicals have no effect on lesser mealworm. Therefore, control alternatives are needed. The aim of this study was to evaluate the influence of Cunila angustifolia (Benth) oil on larvae and adults of A. diaperinus. In vitro tests used larvae and adults of A. diaperinus distributed in petri dishes with 0, 1, 5, and 10% of oil in a single dose. In vivo tests were performed in poultry houses with five treatments: 0, 5, and 10% and chemical insecticide (cypermethrin) in a single application, and a group with 5% of oil applied twice 15d apart. In vitro, oil bioactivity showed an efficacy of 100% both for larvae and adults, when tested at concentrations of 5 and 10%. A reduced number of larvae were observed using 1% of oil; however, it was not effective against adults as compared with the control group. In vivo, the oil effectiveness against lesser mealworm was verified by larva and adult reduction in all concentrations compared with control (0%) throughout the experiments, with better efficacy when used at 5% with two applications. Therefore, we concluded that the oil of C. angustifolia has larvicidal and insecticidal effect against A. diaperinus larvae and adults, in vitro and in vivo.

The effect on locomotor activity, the repellency, and the knock-down produced by 10 monoterpene alcohols were evaluated on first-instar nymphs of Rhodnius prolixus and Triatoma infestans, vectors of Chagas disease. A video tracking technique was used to evaluate locomotor activity and repellency by exposure to papers impregnated with monoterpenes. Eugenol on R. prolixus and (S)-cis-verbenol on T. infestans did not modify the locomotor activity. The remaining monoterpenes produced hyperactivity on both species, although the concentration required was at least a 1,000 times higher than that of deltamethrin (positive control). Carvacrol, eugenol, and geraniol resulted as repellent as N,N-diethyl-m-toluamide (positive control) for both species. A similar result was observed for almost every monoterpene on T. infestans. Knock-down effect was evaluated by exposing the nymphs in closed recipients. The order of increasing toxicity on R. prolixus was (KT₅₀ values in min): geraniol (213.7) < α-terpineol (164.5) < linalool (124.2) < carvacrol (111.6) < eugenol (89.8) < thymol (78.9), and on T. infestans: α-terpineol (289.8) < eugenol (221.3) < carvacrol (164.2) < linalool (154.9) < thymol (96.7). All monoterpenes were less toxic than the positive control, dichlorvos (3.6 min for R. prolixus and 3.9 min for T. infestans). After 7 h of exposure, (-)-carveol, citronellol, and menthol (on both species) and geraniol (on T. infestans) produced <50% of knock-down. After these results, it is worthwhile to explore more deeply the potential of these compounds as tools for controlling Chagas disease vectors.

Culex pipiens pallens (L.) is the most common mosquito in houses of central and northern China. It is the primary vector of lymphatic filariasis and Japanese encephalitis. The flight range of mosquitoes is an important factor predicting the risk area of transmission of mosquito-borne pathogens to vertebrate hosts. The flight performance of Cx. pipiens pallens was measured with a 26-channel computer-monitored flight-mill system. We found that females had longer flight capability than males for total flight distance (TFD) and total flight duration (TFDr), and females flew faster than males based on mean flight velocity. No significant difference in flight capability was found between different age-groups in males. However, certain age-groups of females showed significant differences in TFDr and TFD. Specifically, TFD and TFDr tended to be shortest for 5- and 6-d-old females. These significant differences in flight capability between ages and genders provide insights to determine the size of operational area to achieve effective control of Cx. pipiens pallens and minimize the risk of the related mosquito-borne epidemic diseases of lymphatic filariasis and Japanese encephalitis.

Ultralow volume droplets of DUET, prallethrin, and sumithrin at a sublethal dose were applied to unfed (nonbloodfed) and bloodfed female Aedes aegypti L. and Aedes albopictus (Skuse) in a wind tunnel. Control spray droplets only contained inert ingredients. Individual mosquitoes were videotaped before, during, and after spraying and various behaviors analyzed. During the spray periods of all three pesticide treatments, mosquitoes spent a greater percentage of time moving, and the distance moved was greater than for mosquitoes in the control treatments. In the postspray period, the percent of time moving increased for mosquitoes exposed to all pesticide treatments compared with the controls. After treatment, all females spent more time walking compared with controls, with unfed Ae. aegypti females walking more after exposure to DUET and sumithrin than after exposure to prallethrin and the control. Pesticide exposure increased flying in both species. Sumithrin exposure increased activity and velocity of unfed mosquitoes more than bloodfed mosquitoes. DUET and sumithrin treatments enhanced activity of Ae. aegypti females more than Ae. albopictus females.

The bloodmeal hosts used by Culex tarsalis Coquillett collected along the Salton Sea in Coachella Valley, CA, during 1998–2002 were identified using sequences of the cytochrome c oxidase I gene identified from Barcode of Life database. Overall, 265 (83.3%) of 318 bloodmeals were identified, of which 76.6% fed on birds, 18.1% on mammals, and 5.3% on reptiles. Forty-seven different hosts were identified, none of which comprised >12.5% of the total. Although Cx. tarsalis exhibits specific host-seeking flight patterns, bloodmeals seemed to be acquired opportunistically, thereby limiting potential arbovirus transmission efficiency in species-rich environments.

Alphaviruses are arthropod-borne pathogens that infect a range of hosts. In humans and other mammals, alphavirus infection can cause severe disease. In mosquito hosts, however, there are generally few symptoms. Little is known about the cellular responses of mosquitoes that allow them to cope with infection. In this investigation, a six-plex tandem mass tagging proteomic approach was used to study protein accumulation changes in the midgut of Anopheles gambiae (Giles) (Diptera: Culicidae) mosquitoes infected with o'nyong-nyong virus (Togaviridae, Alphavirus). Five hundred thirty-six nonredundant proteins were identified. Twenty-two were found in significantly different quantities in infected midguts compared with controls. Of interest, analysis revealed molecular pathways possibly targeted by virus proteins, such as those involving TAF4 and DNA polymerase phi proteins. Also identified was an FK506-binding protein. FK506-binding protein orthologs have been described as conserved host resistance factors, which suppress dengue and West Nile virus infection in human HeLa cells. This investigation constitutes the first study of the midgut-specific proteome of An. gambiae in relation to alphavirus infection. Our findings offer insight into mosquito immunity, including factors that possibly contribute to the different pathological outcomes observed in vertebrate and insect hosts.

Ticks serve as both vectors and the reservoir hosts capable of transmitting spotted fever group Rickettsia by horizontal and vertical transmission. Persistent maintenance of Rickettsia species in tick populations is dependent on the specificity of the tick and Rickettsia relationship that limits vertical transmission of particular Rickettsia species, suggesting host-derived mechanisms of control. Tick-derived molecules are differentially expressed in a tissue-specific manner in response to rickettsial infection; however, little is known about tick response to specific rickettsial species. To test the hypothesis that tissue-specific tick-derived molecules are uniquely responsive to rickettsial infection, a bioassay to characterize the tick tissue-specific response to different rickettsial species was used. Whole organs of Dermacentor variabilis (Say) were exposed to either Rickettsia montanensis or Rickettsia amblyommii, two Rickettsia species common, or absent, in field-collected D. variabilis, respectively, for 1 and 12 h and harvested for quantitative real time-polymerase chain reaction assays of putative immune-like tick-derived factors. The results indicated that tick genes are differently expressed in a temporal and tissue-specific manner. Genes encoding glutathione S-transferase 1 (dvgst1) and Kunitz protease inhibitor (dvkpi) were highly expressed in midgut, and rickettsial exposure downregulated the expression of both genes. Two other genes encoding glutathione S-transferase 2 (dvgst2) and β-thymosin (dvβ-thy) were highly expressed in ovary, with dvβ-thy expression significantly downregulated in ovaries exposed to R. montanensis, but not R. amblyommii, at 12-h postexposure, suggesting a selective response. Deciphering the tissue-specific molecular interactions between tick and Rickettsia will enhance our understanding of the key mechanisms that mediate rickettsial infection in ticks.

Mosquito feeding behavior determines the degree of vector- host contact and may have a serious impact on the risk of pathogen transmission, including that of the West Nile virus (WNV). To measure the role of Culex mosquitoes as WNV vectors, host-seeking females were collected using animal-baited traps containing live birds (quail) or mammals (rabbits) and CO₂-baited Center for Disease Control and Prevention traps placed in several wetland areas in the Czech Republic. Culex pipiens (L.) and Culex modestus (F.) were the most frequently collected species. Although Cx. modestus did not distinguish between baits, Cx. pipiens was collected significantly more frequently in bird-baited traps. Based on mitochondrial DNA analysis of bloodmeals from engorged females collected by CO₂-baited traps situated within reed beds, a diverse group of birds were the predominant hosts (93.7%), followed by mammals (4.2%) including humans, and amphibians (2.1%). Among birds, Anseriformes were fed upon most frequently by Cx. modestus, whereas Cx. pipiens fed most frequently on Passeriformes. To measure the infection risk and confirm the distribution of mosquito species in various biotopes, transects of CO₂-baited CDC traps were operated from wetland reed beds into upland vegetated areas. Even though both Culex species occurred in all biotopes sampled and frequently dispersed hundreds of meters away from fishpond shore vegetation, the spatial distribution of Cx. modestus was significantly associated with reed beds at wetlands. The first detection of WNV (subtype RabV) in Cx. modestus in Bohemia and confirmation of WNV presence in Cx. pipiens in Moravia together with observed feeding behavior supports the presumed role of both Culex species in the avian-to-avian enzootic WNV cycle and in avian-to-mammal transmission in the Czech Republic.

Although Culicoides biting midges act as a vector of important human and domestic animal diseases, their ecology is poorly understood. The lack of proper identification systems of Culicoides larvae is one of the main obstacles to progress in research. Based on mitochondrial sequences of 19 Japanese Culicoides species, we designed a universal primer set to amplify the partial sequence of the mitochondrial cytochrome c oxidase I (cox 1). The polymerase chain reaction product amplified from extracted DNA of Culicoides larvae using the primer set was directly sequenced, and species identification based on the variation at cox1 was conducted. Using the molecular identification system, we sorted 243 specimens of field-collected larvae from the southern part of Japan into 10 species including Culicoides arakawae (Arakawa), Culicoides oxystoma Kieffer, and Culicoides brevitarsis Kieffer, which are regarded as vectors of important livestock animal diseases. Eight species of Culicoides larvae, including C. arakawae and C. oxystoma, were recovered from active paddy fields and an abandoned paddy field. The result suggests that paddy fields contribute to breeding a variety of Culicoides species and maintenance and spread of Culicoides-borne pathogens. In contrast, larvae of C. brevitarsis were collected from cattle dung in pastures. The molecular identification system described herein using nucleotide sequences successfully achieved larval identification and will be useful for a better understanding of larval habitats of Culicoides biting midges.

We evaluated Aedes atlanticus Dyar and Knab, Aedes infirmatus Dyar and Knab, Aedes vexans (Meigen), Anopheles crucians Wiedemann, Coquillettidia perturbans (Walker), Culex nigripalpus Theobald, Mansonia dyari Belkin, Heinemann, and Page, and Psorophora ferox (Von Humboldt) from Florida to determine which of these species should be targeted for control should Rift Valley fever virus (RVFV) be detected in North America. Female mosquitoes that had fed on adult hamsters inoculated with RVFV were incubated for 7–21 d at 26°C, then allowed to refeed on susceptible hamsters, and tested to determine infection, dissemination, and transmission rates. We also inoculated mosquitoes intrathoracically, held them for 7 d, and then allowed them to feed on a susceptible hamster to check for a salivary gland barrier. When exposed to hamsters with viremias ≥10^7.6 plaque-forming units per milliliter of blood, at least some individuals in each of the species tested became infected; however, Cx. nigripalpus, An. crucians, and Ae. infirmatus were essentially incompetent vectors in the laboratory because of either a midgut escape or salivary gland barrier. Each of the other species should be considered as potential vectors and would need to be controlled if RVFV were introduced into an area where they were found. Additional studies need to be conducted with other geographic populations of these species and to determine how environmental factors affect transmission.

The Gulf Coast tick, Amblyomma maculatum Koch, has become increasingly important in public health for its role as a vector of the recently recognized human pathogen, Rickettsia parkeri. More recently, these ticks were also found to harbor a novel spotted fever group rickettsia, “Candidatus Rickettsia andeanae.” First identified in Peru, and subsequently reported in ticks collected in the United States, Chile, and Argentina, “Ca. R. andeanae” remains largely uncharacterized, in part because of the lack of a stable isolate. Although the isolation of “Ca, R. andeanae” was recently described in DH82, Vero, and Drosophila S2 cells, its stability in these cell lines was not shown. To evaluate “Ca. R. andeanae” transmission and pathogenicity in vertebrates, as well as further describe biological characteristics of this candidate species to fulfill criteria for its establishment as a new species, availability of a stable isolate is essential. Here we describe the propagation of “Ca. R. andeanae” by using a primary culture derived from naturally infected A. maculatum embryos. Subsequent passage of the “Ca. R. andeanae” isolate to ISE6 (Ixodes scapularis embryonic) and Vero (African green monkey kidney epithelial) cell lines demonstrated limited propagation of the rickettsiae. Treatment of the infected primary cells with tetracycline resulted in cultures negative for “Ca. R. andeanae” by polymerase chain reaction and microscopy. Establishment of an isolate of “Ca. R. andeanae” will promote further investigation into the significance of this tick-associated rickettsia, including its role in spotted fever and interactions with the sympatric species, R. parkeri in A. maculatum.

The host-vector-parasite interactions in Chagas disease peridomestic transmission cycles in the United States are not yet well understood. Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) infection prevalence and bloodmeal sources were determined for adult and immature triatomine (Hemiptera: Reduviidae) specimens collected from residential settings in central Texas. Sequenced cytochrome b DNA segments obtained from triatomine digestive tract identified nine vertebrate hosts and one invertebrate host in four triatomine species (Triatoma gerstaeckeri, Triatoma indictiva, Triatoma protracta, and Triatoma sanguisuga). The broad range of wild and domestic host species detected in triatomine specimens collected from residential sites indicates high host diversity and potential movement between the sylvatic and peridomestic settings. Domestic dogs appear to be key in the maintenance of the peridomestic transmission cycle as both a blood host for the triatomine vectors and a potential reservoir for the parasite. The high rate of T. cruzi infection among triatomine specimens that were collected from inside houses, outside houses, and dog kennels (69, 81, and 82%, respectively) suggests a current risk for Chagas disease vector-borne transmission for humans and domestic animals in residential settings in Texas because of overlap with the sylvatic cycle.

Long-term success of ongoing malaria control efforts based on mosquito bed nets (long-lasting insecticidal net) and indoor residual spraying is dependent on continuous monitoring of mosquito vectors, and thus on effective mosquito sampling tools. The objective of our study was to identify the most efficient mosquito sampling tool(s) for routine vector surveillance for malaria and lymphatic filariasis transmission in coastal Kenya. We evaluated relative efficacy of five collection methods—light traps associated with a person sleeping under a net, pyrethrum spray catches, Prokopack aspirator, clay pots, and urine-baited traps—in four villages representing three ecological settings along the south coast of Kenya. Of the five methods, light traps were the most efficient for collecting female Anopheles gambiae s.l. (Giles) (Diptera: Culicidae) and Anopheles funestus (Giles) (Diptera: Culicidae) mosquitoes, whereas the Prokopack aspirator was most efficient in collecting Culex quinquefasciatus (Say) (Diptera: Culicidae) and other culicines. With the low vector densities here, and across much of sub-Saharan Africa, wherever malaria interventions, long-lasting insecticidal nets, and/or indoor residual spraying are in place, the use of a single mosquito collection method will not be sufficient to achieve a representative sample of mosquito population structure. Light traps will remain a relevant tool for host-seeking mosquitoes, especially in the absence of human landing catches. For a fair representation of the indoor mosquito population, light traps will have to be supplemented with aspirator use, which has potential for routine monitoring of indoor resting mosquitoes, and can substitute the more labor-intensive and intrusive pyrethrum spray catches. There are still no sufficiently efficient mosquito collection methods for sampling outdoor mosquitoes, particularly those that are bloodfed.

Recent releases of mosquitoes infected with different Wolbachia types into Australia have led to conditions that have not been observed naturally and that have not been considered theoretically. In an ongoing public health campaign, two Wolbachia infection types have been introduced, and the infections differ in their fitness cost imposed on the host mosquito. The dynamics of each infection type as it interacts with the indigenous uninfected host population have been previously examined via models and empirical tests. Here we conduct a modeling analysis to predict the dynamics when the two infection types do not remain geographically isolated. The results demonstrate the potential replacement and loss of the more costly infection type, which may not be the desired public health outcome.

For organisms lacking parental care and where larval dispersal is limited, oviposition site selection decisions are critical fitness-enhancing choices. However, studies usually do not consider the interdependence of the two. In this study, we evaluated the effect of food level on the oviposition behavior of Aedes albopictus (Skuse) in the presence or the absence of a nonlethal predator (caged dragonfly nymph). We also attempted to quantify the perceived cost of predation to ovipositioning mosquitoes. Mosquitoes were presented with oviposition cups containing four levels of larval food (fermented leaf infusion) with or without a caged libellulid nymph. By titrating larval food, we estimated the amount of food needed to attract the female mosquito to oviposit in the riskier habitat. As expected, oviposition rate increased with food level and decreased in the presence of a predator. However, the effect of food level did not differ between predator treatments. By calculating the difference in the amount of food for points of equal oviposition rate in the predator-present and predator-absent regression lines, we estimated the cost of predation risk to be 1950 colony-formingunits per milliliter. Our study demonstrated the importance of considering the possible interdependence of predation risk and food abundance for oviposition-site-seeking insects. This study also quantified the perceived cost of predation and found it to be relatively low, a fact with positive implications for biological control.

Human head and body lice have been classified into three phylogenetic clades (Clades A, B, and C) based on mitochondrial DNA. Based on nuclear markers (the 18S rRNA gene and the PM2 spacer), two genotypes of Clade A head and body lice, including one that is specifically African (Clade A2), have been described. In this study, we sequenced the PM2 spacer of Clade C head lice from Ethiopia and compared these sequences with sequences from previous works. Trees were drawn, and an analysis of genetic diversity based on the cytochrome b gene and the PM2 spacer was performed for African and non-African lice. In the tree drawn based on the PM2 spacer, the African and non-African lice formed separate clusters. However, Clade C lice from Ethiopia were placed within the African Clade A subcluster (Clade A2). This result suggests that recombination events have occurred between Clade A2 lice and Clade C lice, reflecting the sympatric nature of African lice. Finally, the PM2 spacer and cytochrome b gene sequences of human lice revealed a higher level of genetic diversity in Africa than in other regions.

The residual efficacy of pyriproxyfen against Aedes aegypti (L.) was examined by treating 2-liter buckets with a range of rates of Sumilarv 0.5G (1 00, 10, 1, and 04 mg product/liter or nominal dose of 500, 50, 5, and 0.5 ppb active ingredient) under semifield conditions. Approximately every 2 wk, pupal emergence inhibition (EI) was measured by using Cairns colony Ae. aegypti. Pooled water samples from the five replicate buckets were analyzed for active pyriproxyfen by using ultra-high-pressure liquid chromatography with tandem mass spectrometry detection. A strong dose—response in EI was exhibited, with the 0.1 mg/liter giving ≈50% EI for only the initial week, whereas the 10 and 100 mg/liter doses produced EI > 90% for 8 and 40 wk, respectively. Measurable levels of active ingredient were detected in the 100, 10, and 1 mg/liter treatments, with measured starting concentrations of just 1-2-1.4% of the delivered (active ingredient) dose. Pyriproxyfen was detected in the 100 mg/liter treatment through the entire course of the trial (60 wk).

Lipoptena deer keds (Diptera: Hippoboscidae) are blood-sucking ectoparasites of mammals, and only one species, Lipoptena cervi (L.), has previously been reported from the Republic of Korea. A study of Siberian roe deer Capreolus pygargus Pallas on Jeju Island, south of the Korean mainland, led to the discovery of a second species of deer ked, Lipoptena fortisetosa Maa, for Korea. In total, 518 deer keds were collected from 19 adult and juvenile deer examined from May to October 2012. The infestation of all of the deer examined and the occurrence on newly born deer suggest a high prevalence and abundance of L. fortisetosa parasitizing Siberian roe deer throughout Jeju Island. This deer represents a new host species for L. fortisetosa.