The yellow fever mosquito, Aedes aegypti (L.) (Diptera: Culicidae), is well recognized for its extensive adaptation to diverse ecological conditions and for genetic variation. Recognizing the importance of strain variation of this mosquito, researchers have established a large number of laboratory strains. Some of the popular strains have been used for research for years in many laboratories around the world. However, the exact origins of many of these strains are unknown. In this review, publications and archival records were examined to report the early laboratory mosquito rearing practices around the world and to identify the origins of selected strains. The records showed that inter-laboratory sharing of strains was already underway in the early part of the 20th century because of the ease of breeding Ae. aegypti and of sending eggs by mail. It also was found that the four strains established in major U.S. institutions by the mid-1930s, including the “ROCK” (short for Rockefeller) strain, had been derived from Cuba, Nigeria, Philippines, or Puerto Rico, all known for a long history of transmission of yellow fever virus or dengue virus rather than from North America. The strains used for research in Europe were primarily derived from West Africa, but strains of Asian, Caribbean, and South American origins also were used for comparative experiments among geographic strains. Neglected issues related to strain designation and original source identification in scientific publications were found and their relevance to current research is discussed.

Phlebotomine sand flies are distributed across nearly all faunal regions of the world, represented by over 800 species, of which many are important vectors of human pathogens. Brazil is currently faced with the expansion and urbanization of leishmaniases, with an increase in the numbers of human cases and seropositive dogs in various medium-sized to large cities. The objective of the current study was to survey the phlebotomine sand fly species in an area endemic for American cutaneous leishmaniasis (ACL) and American visceral leishmaniasis (AVL), i.e., the municipal district of Santa Luzia, lying within the metropolitan region of Belo Horizonte in the Brazilian State of Minas Gerais. Sand flies were collected monthly in 2004–2005 using modified Falcão light traps hung in the peridomiciles of houses and surrounding wooded areas in the district of Baronesa. A total of 1,552 sand flies belonging to seven species was collected, and an interesting pattern of the distribution of the most abundant species relative to the sampling locality was revealed. In the wooded areas Lutzomyia whitmani (Antunes & Coutinho) predominated, whereas in the urban area Lutzomyia longipalpis (Lutz & Neiva) was the most abundant species. These results indicate two possible epidemiological patterns of Leishmania transmission in Santa Luzia: one for American cutaneous leishmaniasis associated predominantly with wooded areas, and another for AVL, with transmission principally occurring around human habitations.

Patterns of mosquito dispersal are important for predicting the risk of transmission of mosquito-borne pathogens to vertebrate hosts. We studied dispersal behavior of Culex erraticus (Dyar & Knab), a potentially significant vector of eastern equine encephalitis virus (EEEV) that is often associated with foci of this pathogen in the southeastern United States. Using data on the relative density of resting adult female Cx. erraticus around known emergence sites in Tuskegee National Forest, Alabama, we developed a model for the exponential decay of the relative density of adult mosquitoes with distance from larval habitats through parameterization of dispersal kernels. The mean and 99th percentile of dispersal distance for Cx. erraticus estimated from this model were 0.97 and 3.21 km per gonotrophic cycle, respectively. Parameterized dispersal kernels and estimates of the upper percentiles of dispersal distance of this species can potentially be used to predict EEEV infection risk in areas surrounding the Tuskegee National Forest focus in the event of an EEEV outbreak. The model that we develop for estimating the dispersal distance of Cx. erraticus from collections of adult mosquitoes could be applicable to other mosquito species that emerge from discrete larval sites.

The effect of temperature and humidity on the survival and water loss of the tropical bed bug, Cimex hemipterus (F.), was studied using two field-collected strains. Insects were exposed to temperatures ranging from 20 to 45°C and relative humidities (RHs) of 33, 75, and 100%. C. hemipterus survived longest under the interaction of low temperature (20°C) and high RH (75–100%). Survival and water loss were significantly affected (P < 0.01) by temperature and RH (either singly, or in interaction). Strain and sex significantly (P < 0.01) influenced bed bug survival, but not on water loss. Eggs, first instars, and adults reached their upper thermal lethal limit within 1 h at 39°C, 44°C, and 46°C, respectively. The survival and water loss profiles showed that starved C. hemipterus started to die after losing 35–45% of their body weights.

The exposure of sublethal doses (25% emergence inhibition) of diflubenzuron (DFB) (0.0002 mg/liter) and azadirachtin (AZD; 0.0079 mg/liter) insect growth regulators was provided to late third—early fourth instars of Culex quinquefasciatus till emergence of adults to find out the effects on various biological and life table attributes of adults, and comparison was made with control (CTL) as well. Life expectancy of both males and females of DFB survivors was significantly less in comparison with AZD and CTL ones; however, it was nonsignificantly different between CTL and AZD. In all treatments, males were shorter lived than females of their respective treatments. DFB survivors showed significant reduction in number of blood-fed females, resulting in minimum of oviposition days, total number of egg rafts, total number of eggs, eggs per egg raft, eggs per female life span, and eggs per female per day than survivors of both AZD and CTL significantly, but these parameters did not differ significantly between CTL and AZD, except eggs per egg raft. The net reproductive rate, intrinsic rate of increase, and finite rate of increase were significantly less in DFB than rest of the survivors, whereas doubling time for generation was significantly high; however, difference in mean generation was not significant among them. This study reveals that DFB has the potency to affect severely the various life table attributes at sublethal dose, but AZD does not have such effectiveness at similar sublethal dose against Cx. quinquefasciatus.

Disordered urbanization and deforestation are the main activities proposed as causal factors of re-emergence of American cutaneous leishmaniasis caused by Leishmania braziliensis. The purpose of this work was to investigate, in the hyperendemic area of Argentina, the distribution of Phlebotomine sand flies at the modified primary vegetation-crop interface, as one of the potential sites where the effects of changing landscape on sand fly populations may be manifested. Twenty samplings were made between June 2004 and August 2005. The traps to catch sand flies were set on two consecutive nights every month (except in 5 mo, where it became every 15 d). The relationship between sand fly abundance and meteorological and landscape variables was analyzed using non-metric multidimensional scaling and Kendall's correlation coefficients. Lutzomyia neivai (Pinto) was the most abundant species, followed by Lutzomyia migonei (França), Lutzomyia cortelezzii (Brèthes), Lutzomyia shannoni (Dyar), and Lutzomyia punctigeniculata (Floch and Abonnenc). Traps located close to modified areas collected the greatest numbers of sand flies, whereas traps located in the least modified area (adjacent to the primary vegetation) collected the fewest. There was a strong negative correlation between the abundance of sand flies and precipitation. This study shows that even small modifications in the landscape led to an increase in sand fly abundance, mainly Lu. neivai, a Leishmania braziliensis vector. This underscores the need for recommendations about the risk of American cutaneous leishmaniasis before any environmental intervention is done in an endemic area, as well as for the monitoring of sand fly population dynamics at the site of intervention, before, during, and after the process.

Competitive interactions may facilitate or repel invaders into new communities, and these interactions may depend on other environmental conditions such as the presence of pesticides. Malathion is widely used in controlling agricultural pests and mosquitoes worldwide. Small amounts of malathion, previously considered inconsequential, may in fact increase in lethality when combined with biotic stressors in aquatic systems. We tested whether low concentrations of malathion (0.11 ppm) that are often detected in aquatic systems, affect competition between two invasive mosquito species Aedes albopictus (Skuse) and Aedes japonicus Theobald. There were no survivors of Ae. japonicus larvae in malathion. There was a significant negative effect of Ae. japonicus density on Ae. albopictus survival, but this effect was absent in the presence of malathion. There was also a moderate negative effect of Ae. japonicus density on Ae. albopictus female size, but this effect was absent in the presence of malathion. These findings indicate that pesticide-mediated alterations in competition and species-specific differences in susceptibility to pesticides could play a role in enhancing invasive potential of Ae. albopictus.

Acquisition of ticks by bird hosts is a central process in the transmission cycles of many tick-borne zoonoses, but tick recruitment by birds has received little direct study. We documented acquisition of Ixodes scapularis Say on birds at Fire Island, NY, by removing ticks from mist-netted birds, and recording the number of ticks on birds recaptured within 4 d of release. Eight bird species acquired at least 0.8 ticks bird^-1 day^-1 during the seasonal peak for at least one age class of I. scapularis. Gray Catbirds, Eastern Towhees, Common Yellowthroats, and Northern Waterthrushes collectively accounted for 83% of all tick acquisitions; and six individuals apportioned among Black-billed Cuckoo, Gray Catbird, Eastern Towhee, and Common Yellowthroat were simultaneously infested with both larvae and nymphs. Bird species with the highest acquisition rates were generally ground foragers, whereas birds that did not acquire ticks in our samples generally foraged above the ground. Tick acquisition by birds did not differ between deciduous and coniferous forests. Among the 15 bird species with the highest recruitment rates, acquisition of nymphs was not correlated with acquisition of larvae. Tick acquisition rates by individual bird species were not correlated with the reservoir competence of those species for Lyme borreliae. However, birds with high tick acquisition rates can contribute large numbers of infected ticks, and thus help maintain the enzootic cycle, even if their levels of reservoir competence are relatively low.

Cat fleas, Ctenocephalides felis (Bouché) (Siphonaptera: Pulicidae), are common ectoparasites of companion animals that negatively impact their hosts directly by causing dermatitis and blood loss during feeding and indirectly through the potential transmission of disease causing agents. We isolated and characterized seven novel microsatellite loci from a partial genomic library of the cat flea enriched for di-, tri-, and tetranucleotide repeats. We screened these loci in cat fleas from two laboratory colonies and one wild-caught population collected at a temporary animal shelter (Parker coliseum) in Baton Rouge, LA. Six loci were polymorphic, with two to 15 alleles per locus and an average observed heterozygosity of 0.21 across populations. Although the two laboratory cat flea colonies were isolated from each other for many years, they did not significantly differ in their genotypic composition. The cat flea population from Parker coliseum was genetically different from the laboratory colonies, but also showed high degrees of inbreeding. Multilocus genotypes of the polymorphic loci were sufficient to assign over 85% of cat fleas to their population of origin. Genetic markers for flea population identity will allow further studies to examine the origins and movement of cat fleas with important genetic traits such as insecticide resistance or pathogen susceptibility. The use of microsatellites also could determine if there are host-specific strains of cat fleas and add insight into the development of the different subspecies of C. felis.

Rhodnius ecuadoriensis is an important vector of Chagas disease in Ecuador. Whereas only sylvatic and peridomestic populations are common in Manabí province, this species occupies domestic, peridomestic, and sylvatic habitats in Loja province where high reinfestation of houses was observed. To explore the existence of phenetic changes linked to the domiciliation of the species, this study set out to analyze the wing and antennal phenotypes of R. ecuadoriensis in these two provinces where the vector presents different affinity for domestic habitats. The antennal phenotype and the wing size and shape distinguish the two geographical populations of R. ecuadoriensis. In Manabí, sylvatic and peridomestic specimens were very similar. In Loja, sylvatic and nonsylvatic (domestic and peridomestic) populations showed distinctive characteristics. Remarkable sexual dimorphism of wing and antenna, exclusive of domestic specimens, and high metric disparity in the wing shape of the domestic females point out the existence of a particular situation in this habitat. The results of this phenotypic analysis and previous evidence of behavioral differences support the hypothesis of disruptive selection acting upon R. ecuadoriensis populations.

We examined the effects of environmental regulation of gene transcription on the accuracy of a transcriptional profiling method for determining insect age. In combined temperature/nutrition treatments, Aedes aegypti (L.) mosquitoes were maintained in the laboratory at three different temperatures (20, 26, and 32°C), and larvae were fed on low, medium, and high diet regimens. Adult mosquitoes of distinct size classes were produced. Transcription of three age-responsive genes (Ae-15848, Ae-8505, and Ae-4274) was measured from 1-, 10-, and 19-d-old specimens using a quantitative reverse-transcription polymerase chain reaction method incorporating dual-labeled TaqMan probes. Temperature had a significant effect on transcript abundance for two of the model genes (Ae-15848 and Ae-8505), and transcription of model genes was unaffected by the main effect of larval diet level; however, significant temperature by diet level interactions were observed. Total RNA yield from individual mosquitoes varied according to adult age and temperature, and when combined with wing length, provided a useful predictor variable in age prediction models. More accurate age predictions were achieved from models generated at the same temperature as test mosquitoes; however, whereas significant differences in mean predicted ages were observed between 1- and 10-d-old mosquitoes, differences between 10 and 19 d were nonsignificant. This study highlights the effect of environmental regulation on gene transcription age grading and the need to identify additional gene biomarkers of age to improve the classification of older mosquitoes.

Magnesium is crucial for baculovirus transmission in Culex nigripalpus (Theobald) and Culex quinquefasciatus (Say) larvae. However, the mechanistic role of magnesium in baculovirus transmission is unknown. To investigate the possible role of host response factors in baculovirus transmission, suppression subtractive hybridization was used to identify genes differentially transcribed after magnesium exposure in Cx. quinquefasciatus larvae. Suppression subtractive hybridization was performed in both directions enriching for cDNAs differentially transcribed between a nonmagnesium larval control and magnesium (15 mM MgSO₄) treatment of Cx. quinquefasciatus larvae held for 1 h at 27°C. Clones from differentially transcribed genes were evaluated by sequencing, and relative gene transcription levels were analyzed using quantitative real-time polymerase chain reaction quantitative real-time polymerase chain reaction. Target transcripts up/downregulated by magnesium included Cx. quinquefasciatus troponin C, isocitrate dehydrogenase, allergen, cytochrome b5, chymotrypsinogen, apolipophorins, tryptase gamma, carboxylesterase, prolylcarboxypeptidase, imaginai disc growth factor, aldehyde dehydrogenase, tropomyosin-1, chitotriosidase, heat shock protein 70 B2, inorganic phosphate cotransporter, and many other hypothetical protein genes. Magnesium can alter gene transcription in a vector mosquito population, and understanding this process can provide insight into the mechanistic role of magnesium in baculovirus transmission.

The aim of the study was to establish a model of the environmental fate of German cockroach (Blattella germanica L.) allergens Bla g 1 and Bla g 2 in feces under controlled and field conditions. Temporal decline (3, 6, and 9 mo) of allergens Bla g 1 and Bla g 2 in the feces protected from cleaning was measured under laboratory and experimental household conditions. The influence of environmental temperature (15, 20, 25, 30, and 35°C) and moisture (53, 75, 85, and 100% RH) on allergen degradation was estimated for 3, 6, and 9 mo. Bla g 1 was more stable than Bla g 2 and the proteins. The proteins and Bla g 2 contents were correlated negatively with the decomposition time; Bla g 1 was not. However, when the content of Bla g 1 in control and exposed tubes was compared, the decrease after exposure was significant at exposure in 35°C, 53 and 100% RH. In laboratory, the shortest half-life (16–38 d) of Bla g 2 was at high temperature and humidity (100% RH at 35°C), whereas the longest half-life (340 d) was at 25°C and 85% RH. In the apartment, the half-life was 406 d. The results indicate that Bla g 1 and Bla g 2 allergens can persist in feces for several months under usual household humidity and temperature.

A major expenditure in vector biology laboratories is the rearing of mosquitoes. Most mosquito colonies require substantial effort to maintain, including frequent bloodmeals for optimal performance. Successful cryopreservation of mosquitoes continues to be elusive. Although using diapause as a storage mechanism is an option for mosquito preservation, several obstacles include the lack of a well-characterized diapause or the inability of some species to enter diapause. Thus, other options for preservation are needed. To address this issue, we investigated the use of long-term low-temperature storage in the absence of diapause for adults of the northern house mosquito, Culex pipiens L. Our results indicate that although male longevity is not substantially increased by cold storage, female longevity is dramatically increased by storage at lower temperatures. When mated before storage, females remain reproductively viable after at least 10 wk of storage, although at reduced levels. These results indicate that cold storage without diapause induction is a viable option for colony maintenance in vector biology laboratories.

Chagas disease is a major health concern in Latin America, and Triatoma infestans (Klug, 1834) is responsible for the majority of cases of Chagas disease in the continent. After the discovery of deltamethrin highly resistant populations in the neighboring areas to Yacuiba (Bolivia) and Salvador Mazza (Argentina), we studied T. infestans populations north and southward, with the aim of describing the range of the resistant area. In addition, tests were conducted to describe the susceptibility to fipronil in deltamethrin-resistant populations. Tarija populations were highly resistant to deltamethrin, showing that the resistant area is greater than previously reported. Argentinean and Paraguayan populations were susceptible or presented moderate to low levels of deltamethrin resistance. Resistance to fipronil was found in Bolivian populations, although this insecticide was effective against Argentinean populations. This study shows that the Argentinean and Paraguayan populations are currently under successful control with deltamethrin. However, continuous resistance monitoring is necessary. We found that fipronil is a viable option for Argentina, whereas the resistance found in Bolivia implies that new formulations are needed to control T. infestans in Bolivia. Further research is required to find new alternatives of control in those areas that are currently suffering from high infestation rates.

The dispersal and survival of laboratory-reared Aedes albopictus Skuse males were investigated during the summer of 2007 in three Northern Italy urban localities by mark-release-recapture techniques. Two marking methods were compared: one group of males was dusted with fluorescent pigments on the body (FP), and the other group was obtained from a strain whose natural infection of Wolbachia had been removed (WB0). FP- and WB0-marked males were released as adults and pupae, respectively, in one fixed station at each locality. Recaptures were performed by skilled technicians, within a radius of 350 m from the release site, on days 4, 5, and 7 after the release, and the males were collected while flying around the technician's body or in swarms. Recapture rates ranged from 0.63 to 4.72% for FP males and from 2.39 to 11.05% for WB0 males. The mean distance traveled for WBO males was significantly higher than for FP males; no difference was observed between the dispersal distance measured for the males recaptured on human host versus males recaptured while swarming. No further increase of the dispersal occurred during the postrelease period investigated (from day 4 to day 7 after release). The mean survival rate at the release was 0.51 for FP-marked males and 0.81 for WBO males. The data obtained are discussed for their significance in planning sterile insect technique programs against Ae. albopictus.

Behavioral responses of Aedes aegypti male populations developed for Release of Insects Carrying a Dominant Lethal (RIDL) technology and a Malaysian wild-type population of two age groups (4–5 and 8–10 d old) were tested under laboratory conditions against chemical irritants and repellents using the high-throughput screening system device. Results indicate that all male Ae. aegypti test populations showed significant (P < 0.01) behavioral escape responses when exposed to alphacypermethrin, DDT, and deltamethrin at the test dose of 25 nmol/cm². In addition, all populations showed significant (P < 0.05) spatial repellent responses to DDT, whereas alphacypermethrin and deltamethrin elicited no directional movement in the assay. These data suggest that genetic modification has not suppressed expected irritancy and repellency behavior. Age effects were minimal in both contact irritant and spatial repellent assays. The magnitude of irritant response, based on percentage responding, was stronger in the RIDL test cohorts as compared with the wild-type Malaysian population, but the impact, if any, that this increased behavioral sensitivity might have on the success of a RIDL strategy has yet to be defined. Information of the type reported in the current study is vital in defining the effects of genetic modification on vector behavior and understanding how these behaviors may influence the success of RIDL technology as they relate to other vector control interventions implemented in the same disease-endemic locale.

Female Culex quinquefasciatus Say (Diptera: Culicidae) mosquitoes were exposed to sublethal amounts of prallethrin, sumithrin, and piperonyl butoxide applied as ultralow volume (ULV) droplets in a wind tunnel. Mosquitoes were video recorded before, during, and after treatment, and the number and size of droplets on their bodies were later determined using a compound microscope. A positive correlation was found between mosquitoes that spent more time flying during the time of spraying and number of droplets on their bodies. Excitation, in the form of increased speed and duration of flight, was immediate in mosquitoes exposed to prallethrin, whereas exposure to sumithrin did not increase their exposure to the ULV droplets. The location of droplets on mosquitoes, the effects of droplet volume, and subsequent mortality are discussed.

Two commercial formulations of Beauveria bassiana (Balsamo-Crivelli) Vuillemin were applied to residential sites in Old Lyme, CT, for the control of nymphs of the blacklegged tick, Ixodes scapularis, in 1999 and 2000. The pyrethroid bifenthrin was applied to other premises for comparison with B. bassiana. A wood chip barrier was installed and maintained at six of the treated properties. In 1999, control of I. scapularis nymphs ranged from 74.5 to 83.0% on lawns without wood chips and from 88.9 to 90% on lawns with wood chip barriers. As a control check, no ticks (n = 23) collected at pretreatment or control sites died from B. bassiana, although 15 of 18 nymphs from treated lawns developed mycoses. Control of I. scapularis on the lawns in 2000 with the two B. bassiana products was lower, as follows: 38.0 and 58.7% without the barrier and 56.9 and 55.1% with the wood chip barrier. Posttreatment differences in nymphal numbers between treatments and control were significant (P = 0.005 and P = 0.039, 1999 and 2000, respectively). The bifenthrin provided 86 and 87% control each year, respectively. The application of Metarhizium anisopliae (Metschnikoff) Sorokin strain F52 to 9 residential sites in Westport and Weston, CT, in 2002 provided significant (P = 0.034; P = 0.039) reductions in nymphal tick abundance with 55.6 and 84.6% fewer ticks on lawn and woodland plots, respectively. These results suggest the application of entomopathogenic fungi could provide another approach for the control of J. scapularis nymphs in residential or similar landscapes.

We have developed a novel test cage and improved method for the evaluation of mosquito repellents. The method is compatible with the United States Environmental Protection Agency, 2000 draft OPPTS 810.3700 Product Performance Test Guidelines for Testing of Insect Repellents. The Biogents cages (BG-cages) require fewer test mosquitoes than conventional cages and are more comfortable for the human volunteers. The novel cage allows a section of treated forearm from a volunteer to be exposed to mosquito probing through a window. This design minimizes residual contamination of cage surfaces with repellent. In addition, an air ventilation system supplies conditioned air to the cages after each single test, to flush out and prevent any accumulation of test substances. During biting activity tests, the untreated skin surface does not receive bites because of a screen placed 150 mm above the skin. Compared with the OPPTS 810.3700 method, the BG-cage is smaller (27 liters, compared with 56 liters) and contains 30 rather than hundreds of blood-hungry female mosquitoes. We compared the performance of a proprietary repellent formulation containing 20% KBR3023 with four volunteers on Aedes aegypti (L.) (Diptera: Culicidae) in BG-and conventional cages. Repellent protection time was shorter in tests conducted with conventional cages. The average 95% protection time was 4.5 ± 0.4 h in conventional cages and 7.5 ± 0.6 h in the novel BG-cages. The protection times measured in BG-cages were more similar to the protection times determined with these repellents in field tests.

Insecticidal properties of natural products may present alternatives to the use of synthetic molecule pesticides that are of diminishing effectiveness due to resistance. Three compounds, thymoquinone, nootkatone, and carvacrol, components of Alaska yellow cedar, Chamaecyyparis nootkatensis (D. Don) Spach, and incense cedar, Calocedrus decurrens (Torr.), essential oils, have been shown to have biological activity against a variety of mosquito and tick species. Although these components act as both repellents and insecticides, how they function in either capacity is unknown. Their use as mosquito control insecticides would be greatly increased if their mode of action is not the same as that of currently used commercial products. This study compared the lethal dosages for nootkatone, carvacrol, and thymoquinone by using colony strains of Anopheles gambiae Giles with known mutations at three different target sites. The altered target sites evaluated were the sodium channel para-locus mutation (L1014 F KDR) that confers permethrin resistance, the ACE-1 gene that confers organophosphate and carbamate resistance, and a γ-aminobutyric acid receptor mutation of the Rdl locus conferring dieldrin resistance. Significant increases in lethal dose were not observed in any of the mosquito strains for any of the compounds tested compared with the doses required of chemicals with known modes of action at the mutated sites. Although the mode of action was not determined, this screening study indicates that none of these compounds interact at the target sites represented in the test mosquito strains. These compounds represent a different mode of action than existing chemicals currently used in mosquito control.

The toxicity of permethrin to different life stages of Culex quinquefasciatus (Say) was examined in three field insecticide resistant strains, HAmCq^G0, MAmCq^G0, and BAmCq^G0; two permethrin selected strains, HAmCq^G8 and MAmCq^G6; and a laboratory susceptible strain, S-Lab. The levels of tolerance to permethrin differed among the developmental stages of Culex mosquito larvae with an ascending order from first to fourth instar. A developmentally regulated pattern of permethrin resistance in Culex mosquito larvae was identified, once again increasing from first to fourth instar. Adult mosquitoes had relatively lower levels of resistance in general compared with their larval stages, suggesting that different mechanisms may be involved in the response to insecticide selection pressure between larvae and adults. Inheritance of resistance to permethrin was examined using reciprocal crosses between resistant HAmCq^G8 and susceptible S-Lab strains; the resulting logarithm concentration-probit response curves for reciprocal F₁ lines were similar to those for the susceptible strain, with degree of dominance values of -0.2 to -0.3, suggesting that the inheritance of permethrin resistance in Cx. quinquefasciatus is incompletely recessive. No significant difference was found in the values of LC₅₀ and the slope of the log concentration—probit lines between reciprocal F₁ lines, indicating that the resistance to permethrin was inherited autosomally. Both χ² goodness-of-fit test and log concentration—probit line analysis on responses of backcross and self-bred progeny to permethrin indicated that permethrin resistance in the HAmCq^G8 strain did not follow a monogenic gene inheritance model, revealing that permethrin resistance in Cx. quinquefasciatus is controlled by more than one gene.

The effect of exposing Triatoma infestans to chickens treated with Cypermethrin pour-on combined with piperonyl butoxide (PBO) was studied. Four groups of treated chickens and one control group were used. Each treatment received 1 or 2 ml of the Cypermethrin formulation with and without PBO. Independent groups of nymphs were fed 1, 7, 15, 30, and 45 d after the treatment application. Blood intake was estimated after each feeding occasion. Up to 15 d after the pour-on application, high mortality was observed in all nymphs fed on treated chickens (>93%±12), and lower than the nymphs of the control group (<33%±15). After 30 d of the pour-on application, there was significantly different mortality between the treatment with 1 ml (80%±9) and 2 ml (>96%±5); no difference was observed between groups with or without PBO addition. After 45 d of the pour-on application, the treatments did not show significant differences (77%±7), although all treatments showed higher mortality than the control group (10%±9). Up to 45 d after the pour-on application, blood intake by nymphs exposed to treated chickens (0.85±0.96 mg/nymph) was lower than blood intake by nymphs exposed to control chickens (6.7±5 mg/nymph). This study shows that Cypermethrin pour-on produces high mortality and reduces the blood intake of third-instar nymphs of T. infestans up to 45 d after the insecticide application to chickens. PBO did not produce a detectable effect.

The house fly, Musca domestica L. (Diptera: Muscidae), is involved in phoretic movement of pathogenic agents, but it has a very efficient defense mechanism against infection. It is believed that antimicrobial peptides play a significant role in the defense system of the house fly. Here, we isolated a peptide from the immunized house fly pupae, measured its molecular mass (3987.6 Da) by matrix-assisted laser desorption ionization/ time of flight-mass spectrometry, and determined its amino acid sequence by using the Procise Protein Sequencing System (Applied Biosystems, Foster City, CA). The peptide was confirmed as a member of the insect defensin family. It displayed high activity against gram-positive bacteria but lower activity against gram-negative bacteria and fungi. Reverse transcription-polymerase chain reaction showed that the house fly defensin gene was constitutively expressed in naive pupae and strongly up-regulated after immunization. House fly defensin is an amphiphilic peptide with a structure similar to that of the CSαβ scaffold of insect defensin A from the flesh fly, Phormia terraenovae Robineau-Desvoidy. To our knowledge, this is the first isolated and characterized house fly antimicrobial peptide, and our work may provide useful information for developing pharmacologically active antimicrobial agents.

Saliva from blood-sucking arthropods modulates host homostasis and immunity, making salivary components potential candidates to be used against pathogens transmitted by these biting insects. Functional characterization of salivary molecules is fundamental to gain a better understanding into their roles during blood feeding and to determine under which conditions such molecules are expressed in the insect saliva. In the current study, we investigated the expression profile of 10 salivary genes from the sand fly Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae), a principal vector of Leishmania major. Our analyses using quantitative polymerase chain reaction were aimed at defining whether diet or senescence influences the expression of P. papatasi salivary gland-expressed genes in laboratory-reared female sand flies. Our results demonstrate that at least one of the most abundant salivary transcripts, SP44, is consistently modulated by either senescence or diet. In contrast, another abundant transcript, SP32, was expressed without any influence from the diet received or the age of the sand fly. Differential expression of the other eight transcripts was not consistently regulated by either diet or age, suggesting that other factors may have a greater influence on differential expression of these salivary gland proteins.

Antibodies against arthropod saliva have shown to be a good marker of bite exposure. Because Anopheles albimanus Wiedemann (Diptera: Culicidae) is the principal malaria vector in Haiti, we evaluated the immune response against salivary gland extract (SGE) of this species in malaria-positive and malaria-negative subjects from this country. The results showed that the level of anti-SGE immunoglobulin (Ig)G antibodies was higher in patients with clinical malaria than those in malaria uninfected people living in the same region. In addition, a significant positive correlation between the level of anti-An. albimanus IgG and IgM antibody levels was observed. These results suggest that antibodies against An. albimanus saliva, especially IgG, are useful markers of mosquito bite exposure in Haiti.

Ornithodoros hermsi Wheeler (Acari: Argasidae) is the vector of Borrelia hermsii, the primary cause of tick-borne relapsing fever in North America. This tick is one of the smallest Ornithodoros species involved with the biological transmission of spirochetes; yet, the amount of blood ingested while feeding is unknown. Therefore, we determined the amount of blood O. hermsi ingested during a bloodmeal to establish its potential for spirochete acquisition while feeding on an infected host. Ticks at different developmental stages were weighed before and after feeding and the volume of blood ingested was calculated. Females ingested the most blood, averaging ≈15 µl per meal, but late-stage nymphs took in the most blood in proportion to unfed body weight. A cohort of nymphs was weighed three more times during the 48 h after feeding, which demonstrated that O. hermsi may have excreted coxal fluid ranging from 24–36% of the bloodmeal weight. We also developed a quantitative polymerase chain reaction method to determine the number of spirochetes ingested and maintained within the ticks after feeding. The density of spirochetes in ticks having just engorged was slightly less than in the host's blood. In the first 5 d after feeding, the number of spirochetes within the ticks declined from the number initially ingested but then remained constant through 15 d. These observations establish a basis for future studies to determine the minimum number of spirochetes required in the host's blood to allow O. hermsi to become persistently infected and transmit during subsequent bloodmeals.

Ectoparasites were sampled from small mammals collected in West Java, West Sumatra, North Sulawesi, and East Kalimantan, Indonesia, in 2007–2008 and were screened for evidence of infection from bacteria in the Rickettsaceae family. During eight trap nights at eight sites, 208 fleas were collected from 96 of 507 small mammals trapped from four orders (379 Rodentia; 123 Soricomorpha; two Carnivora; three Scandentia). Two species of fleas were collected: Xenopsylla cheopis (n = 204) and Nosopsyllus spp. (n = 4). Among the 208 fleas collected, 171 X. cheopis were removed from rats (Rattus spp.) and 33 X. cheopis from shrews (Suncus murinus). X. cheopis were pooled and tested for DNA from rickettsial agents Rickettsia typhi, Rickettsia felis, and spotted fever group rickettsiae. R. typhi, the agent of murine typhus, was detected in X. cheopis collected from small mammals in West Java and East Kalimantan. R. felis was detected in X. cheopis collected from small mammals in Manado, North Sulawesi. R. felis and spotted fever group rickettsiae were detected in a pool of X. cheopis collected from an animal in East Kalimantan. Sixteen percent of the X. cheopis pools were found positive for Rickettsia spp.; four (10.8%) R. typhi, one (2.7%) R. felis, and one (2.7%) codetection of R. felis and a spotted fever group rickettsia. These data suggest that rickettsial infections remain a threat to human health across Indonesia.

Four types of commercial mosquito control traps, the Mosquito Magnet Pro (MMP), the Sentinel 360 (S360), the BG-Sentinel (BGS), and the Mega-Catch Ultra (MCU), were compared with a standard Centers for Disease Control and Prevention (CDC) light trap for efficacy in collecting phlebotomine sand flies (Diptera: Psychodidae) in a small farming village in the Nile River Valley 10 km north of Aswan, Egypt. Each trap was baited with either carbon dioxide (CO₂) from combustion of butane gas (MMP), dry ice (CDC and BGS traps), light (MCU and S360), or dry ice and light (CDC). Traps were rotated through five sites in a 5 × 5 Latin square design, repeated four times during the height of the sand fly season (June, August, and September 2007) at a site where 94% of sand flies in past collections were Phlebotomus papatasi (Scopoli). A total of 6,440 sand flies was collected, of which 6,037 (93.7%) were P. papatasi. Of the CO₂-baited traps, the BGS trap collected twice as many P. papatasi as the MMP and CDC light traps, and at least three times more P. papatasi than the light-only MCU and S360 traps (P < 0.05). Mean numbers (±SE) of P. papatasi captured per trap night were as follows: BGS 142.1 (±45.8) > MMP 56.8 (±9.0) > CDC 52.3 (±6.1) > MCU 38.2 (±6.4) > S360 12.6 (±1.8). Results indicate that several types of commercial traps are suitable substitutes for the CDC light trap in sand fly surveillance programs.

The purpose of this investigation was to identify the mosquito (Diptera: Culicidae) vectors of West Nile virus (WNV; family Flaviviridae, genus Flavivirus) during an epizootic WNV outbreak in eastern Puerto Rico in 2007. In June 2006, 12 sentinel chicken pens with five chickens per pen were deployed in six types of habitats: herbaceous wetlands, mangrove forests, deciduous forests, evergreen forests, rural areas, and urban areas. Once WNV seroconversion in chickens was detected in June 2007, we began trapping mosquitoes using Centers for Disease Control and Prevention (CDC) miniature (light/CO₂-baited) traps, CMT-20 collapsible mosquito (CO₂- and ISCA SkinLure-baited) traps, and CDC gravid (hay infusion-baited) traps. We placed the CDC miniature traps both 2–4 m and >30 m from the chicken pens, the collapsible traps 2–4 m from the pens, and the gravid traps in backyards of houses with sentinel chicken pens and in a wetland adjacent to an urban area. We found numerous blood-engorged mosquitoes in the traps nearest to the sentinel chickens and reasoned that any such mosquitoes with a disseminated WNV infection likely served as vectors for the transmission of WNV to the sentinels. We used reverse transcriptase-polymerase chain reaction and isolation (C636) on pools of heads, thoraxes/abdomens, and legs of collected blood-engorged mosquitoes to determine whether the mosquitoes carried WNV. We detected WNV-disseminated infections in and obtained WNV isolates from Culex nigripalpus Theo (minimum infection rate [MIR] 1.1–9.7/1,000), Culex bahamensis Dyar and Knab (MIR 1.8–6.0/1,000), and Aedes taeniorhynchus (Wied.) (MIR 0.34–0.36/1,000). WNV was also identified in and isolated from the pool of thoraxes and abdomens of Culex quinquefasciatus Say (4.17/1,000) and identified in one pool of thoraxes and abdomens of Culex habilitator Dyar and Knab (13.39/1,000). Accumulated evidence since 2002 suggests that WNV has not become endemic in Puerto Rico.

To determine whether Borrelia burgdorferi sensu lato (s.l.) influences tick survival under thermohygrometric stress, Ixodes ricinus (L.) (Acari: Ixodidae) questing ticks were tested under various relative humidities (13, 32, 51.5, 61, and 89% RH) at two different temperatures (12.5 and 25°C) and investigated for Borrelia infection. Survival rate of females was highest (77.6%), followed by males (51.6%), and nymphs (43.2%). The thermohygrometric factor that most importantly determined survival was saturation deficit (SD). As SD increased, tick survival rate decreased in all stages. Among the 1,500 ticks tested for B. burgdorferi s.l., 34.8% (n = 522) were infected. Adult infection rate (39.6%) was higher than that of nymphs (25.5%). Infection load in real-time polymerase chain reaction ranged from 1 to 1.2 million spirochetes per tick. B. afzelii (39.7%), B. burgdorferi sensu stricto (12.1%), B. garinii (37.9%), B. myamotoi (3.6%), and B. valaisiana (23.8%) were recorded. B. garinii infected significantly less nymphs than adults whereas B. afzelii displayed the opposite trend. Survival rate of nymphal and adult I. ricinus was significantly enhanced by infection by B. burgdorferi s.l. (χ²: nymph, P = 0.008; adult, P = 0.021). In adults, a negative effect of infection on tick survival was observed when spirochete load overcame a threshold estimated at 160,000 spirochetes per tick but not in nymphs. Moreover, ticks infected by B. afzelii survived better than other ticks (infected by other genospecies or not). The results here indicate that infection by B. burgdorferi s.l., and more specifically infection by B. afzelii, confers survival advantages to l. ricinus under challenging thermohygrometric conditions.

Stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), an economically important pest of livestock and humans, were observed parasitizing prefledged American white pelicans, Pelecanus erythrorhynchos (Pelecaniformes: Pelecanidae), in a pelican breeding colony in northeastern Montana where die-offs attributed to West Nile virus (family Flavividae, genus Flavivirus, WNV) have occurred since 2002. Engorged and unengorged flies were collected off nine moribund chicks. Of 29 blood-engorged flies testing positive for vertebrate DNA, all 29 contained pelican DNA. Virus isolation was performed on 60 pools (1,176 flies) of unengorged flies using Vero cell plaque assay. Eighteen pools were positive for WNV for an estimated infection rate of 18.0 per 1,000 flies. Fifty-four percent (36/67) of abdomens from blood-engorged flies tested positive for WNV. Pelican viremia levels from the blood-engorged fly abdomens revealed that at least one of the ill pelicans circulated a viremia capable of infecting Culex mosquito vectors. Stable flies may be involved in WNV transmission within the pelican breeding colony by serving as either a mechanical vector or as a source for oral infection if ingested by predators.

The potential of Gafanhoto Park as an American cutaneous leishmaniasis (ACL) focus was evaluated by examination of sand fly vectors of the Leishmania parasite. This forest remnant is located in a periurban area of Divinópolis, Brazil, where autochthonous cases of ACL have been reported. Sand fly populations were monitored over a 2-yr period (2006–2008) by using light traps (HP and Shannon). During systematic collections with HP traps, 824 specimens in total (342 males and 482 females) of 21 species were captured. Most prevalent species were as follows: Brumptomyia brumpti (Larrouse), Lutzomyia aragaoi (Costa Lima), Lutzomyia lutziana (Costa Lima), Lutzomyia sordellii (Shannon & Del Ponte), and Lutzomyia whitmani (Antunes & Coutinho). Using Shannon traps, 257 specimens representing 15 species were collected (159 females and 98 males), with a high prevalence of L. whitmani and Lutzomyia neivai (Pinto), both vectors of Leishmania braziliensis (Vianna). To ascertain the level of natural infection, a sample of females captured in Shannon traps was assayed for the presence of Leishmania by using polymerase chain reaction-restriction fragment length polymorphism, where 39% of insects were positive. The most infected species was L. whitmani (29 sand flies; 18.2%), followed by L. neivai (21; 13.2%), Lutzomyia christenseni (Young & Duncan) (five; 3.1%), Lutzomyia pessoai (Coutinho & Barreto) (three; 1.9%), L. aragaoi (one; 0.6%), Lutzomyia fischeri (Pinto) (one; 0.6%), Lutzomyia lenti (Mangabeira) (one; 0.6%), L. lutziana (one; 0.6%), and Lutzomyia monticula (Costa Lima) (one; 0.6%). The finding of potential and incriminated vectors naturally infected with Leishmania reinforces the need of epidemiologic surveillance in the area.

Anopheles culicifacies is the main vector for transmission of Plasmodium vivax malaria in the Indian subcontinent. A strain of An. culicifacies isolated from its natural niche displayed complete refractoriness to P. vivax by melanotic encapsulation of ookinetes. Prophenoloxidases are key components of the phenoloxidase cascade that leads to recognition and melanization of invading organisms. We isolated and cloned prophenoloxidase-encoding acppo6 gene of An. culicifacies and analyzed its expression profile under various regimens of immune challenge. The acppo6 was differentially expressed during various stages of larval development. The acppo6 transcription was also up-regulated in response to bacteria and Plasmodium vinckei petteri challenge. The transcript levels of the acppo6 gene were higher in naive adult refractory female mosquitoes as compared with female susceptible mosquitoes. Furthermore, the induction of acppo6 in the susceptible strain upon Plasmodium infection was negligible as compared with that of the refractory strain. The observation is suggestive of the role of acppo6 in effectuating a melanotic response in Plasmodium-incompetent naturally occurring refractory An. culicifacies strain.

This is the first confirmed record of the genus Lipoptena Nitzsch and its species, Lipoptena cervi (Linnaeus), from the Republic of Korea. A total of five females and 10 males was collected from eight of 29 Korean water deer, Hydropotes inermis argyropus Swinhoe, from Gangwon and Gyeongsangbuk Provinces and Ulsan Metropolitan Area from May through October 2008. An updated checklist of Korean hippoboscids contains nine species in six genera (Hippobosca Linnaeus, Icosta Speiser, Lipoptena, Ornithoctona. Speiser, Ornithomya Lattreille, and Ornithophila Rondani). Hosts, collection records, and repositories are also noted.

Funnel traps are often used to sample for the presence of Aedes aegypti (L.) (Diptera: Culicidae) larvae in subterranean aquatic habitats. These traps are generally ≥15 cm in diameter, making them impractical for use in subterranean sites that have narrow (10-cm) access ports, such as those in standard-sized septic tanks. Recent research indicates septic tanks may be important habitats for Ae. aegypti in Puerto Rico and the Caribbean. To sample mosquito larval populations in these sites, a miniaturized funnel trap was necessary. This project describes the use of a smaller funnel trap for sampling larval populations. The effects of larval instar (third and fourth) and population density on trap efficacy also are examined. The trap detected larval presence 83% of the time at a larval density of 0.011 larvae per cm² and 100% of the time at densities ≥0.022 larvae per cm². There was a significant trend of increasing percentage of recaptured larvae with higher larval population densities. Although the miniaturized funnel trap is less sensitive at detecting larval presence in low population densities, it may be useful for sampling aquatic environments with restricted access or shallow water, particularly in domestic septic tanks.

Despite knowledge of parasite biology being essential for host-parasite system functioning to be understood, the life histories of many parasites remain little studied. One example being the hippoboscid Crataerina pallida (Olivier in Latreille, 1812) (Diptera: Hippoboscidae), a nest ectoparasite of the common swift, Apus apus L. (Aves: Apodidae). The factors influencing adult parasite emergence remain unclear. Whether temperature affects emergence was studied by exposing overwintering pupae to differing temperature regimes. At higher temperatures, greater numbers of adults emerged. This indicates that adult hatching is temperature mediated and may be enhanced by host presence. The relationship between C. pallida and their hosts is thus close.

During spring and fall 2009, 60 wild turkeys (Meleagris gallopavo) harvested by Tennessee hunters were surveyed for Borrelia spp. by sampling their blood, tissue, and attached ticks. In both seasons, 70% of turkeys were infested with juvenile Amblyomma americanum; one spring turkey hosted an adult female Ixodes brunneus. Polymerase chain reaction assays followed by DNA sequencing indicated that 58% of the turkeys were positive for the spirochete Borrelia miyamotoi, with tissue testing positive more frequently than blood (P = 0.015). Sequencing of the 16S–23S rRNA intergenic spacer indicated ≥99% similarity to previously published sequences of the North American strain of this spirochete. Positive turkeys were present in both seasons and from all seven middle Tennessee counties sampled. No ticks from the turkeys tested positive for any Borrelia spp. This is the first report of B. miyamotoi in birds; the transmission pathways and epidemiological significance of this high-prevalence spirochetal infection remain uncertain.

From September 2008 to March 2010, 397 ticks (315 larvae, 33 nymphs, 23 females, and 26 males) were collected from captive and free-living wildlife species in northeastern Brazil. Six tick species were identified, including Amblyomma auricularium (Conil) on Tamandua tetradactyla (L), Amblyomma dubitatum Neumann on Hydrochaeris hydrochaeris (L.), Nectomys rattus (Pelzen) and T. tetradactyla, Amblyomma parvum Aragão on T. tetradactyla, Amblyomma rotundatum Koch on Boa constrictor L., Chelonoidis carbonaria (Spix), Kinosternon scorpioides (L.) and Rhinella jimi (Stevaux), Amblyomma varium Koch on Bradypus variegatus Schinz, and Rhipicephalus sanguineus (Latreille) on Lycalopex vetulus (Lund). Nectomys rattus and T. tetradactyla are new hosts for A. dubitatum. This study extends the known distribution of A. dubitatum in South America and provides evidence that its geographical range has been underestimated because of the lack of research. Four (A. dubitatum, A. parvum, A. rotundatum, and R. sanguineus) of six tick species identified in this study have previously been found on humans in South America, some of them being potentially involved in the transmission of pathogens of zoonotic concern.