Many mosquito species are important vectors of human and animal diseases, and others are important nuisance species. To facilitate communication and information exchange among professional groups interested in vector-borne diseases, it is essential that a stable nomenclature be maintained. For the Culicidae, easily identifiable genera based on morphology are an asset. Major changes in generic concept, the elevation of 32 subgenera within Aedes to generic status, and changes in hundreds of species names proposed in a recent article demand consideration by all parties interested in mosquito-borne diseases. The entire approach to Aedini systematics of these authors was flawed by an inordinate fear of paraphyletic taxa or Paraphylyphobia, and their inability to distinguish between classification and cladistic analysis. Taxonomists should refrain from making taxonomic changes based on preliminary data, and they should be very selective in assigning generic names to only the most important and well-defined groups of species.

Sandflies are vectors of several pathogens, constituting serious health problems. Lutzomyia longipalpis (Lutz & Neiva, 1912) is the main vector of Leishmania chagasi, agent of visceral leishmaniasis. They synthesize a thick bag-like structure that surrounds the bloodmeal, named peritrophic matrix (PM). One of the major roles of PM in blood-fed insects includes protection against ingested pathogens by providing a defensive barrier to their development. We used traditional and modern morphological methods as well as biochemical and immunolabeling tools to define details of the PM structure of the Lu. longipalpis sandfly, including composition, synthesis, and degradation. The kinetics of PM formation and degradation was found to be related to the ingestion and time of digestion of the bloodmeal. The midgut changes its size and morphology after the blood ingestion and during the course of digestion. A striking morphological modification takes place in the midgut epithelium after the stretching caused by the bloodmeal, revealing a population of cells that was not observed in the unfed midgut. The transmission and scanning electron microscopies were used to reveal several morphological aspects of PM formation. The PM looks thicker and well formed 24 h after the bloodmeal. Presence of chitin in the PM was demonstrated by immunolabeling with an α-chitin monoclonal antibody. SDS-polyacrylamide gel electrophoresis showed at least five protein bands with molecular masses of 38.7-135 kDa, induced by the protein-free diet. Mouse polyclonal antiserum was produced against PMs induced by protein-free meal and used in Western blotting, which revealed at least three associated proteins.

The noncellular peritrophic matrix (PM) that forms around the food bolus in the midgut of many arthropod species may influence the fate of ingested microbes. In mosquitoes, PMs have been identified in the pupal as well as larval and adult stages. In pupae, the PMs surround the meconium, the sloughed larval midgut epithelium. Meconial PM1 (MPM1) forms early in the pupal stadium, and a second meconial PM (MPM2) sometimes forms around the time of adult emergence. A recent study suggests that MPMs contribute to the sterilization of the adult midgut by sequestering microorganisms ingested during the larval stage, which, along with remaining meconial material, are egested after adult emergence. We have compared MPM1 formation and patterns of meconial degeneration in representative species in five mosquito genera and identified a temporal association between MPM1 formation, meconial degeneration, and apolysis. Ultrastructural study of MPM1 and MPM2 in Aedes aegypti (L.) revealed that MPM1 seems to be structurally different from either the larval or adult PMs, whereas MPM2 more closely resembles PM formed around a bloodmeal in adult females. Our results are consistent with the microbial sequestration role.

An adult male of the tick Amblyomma rotundatum Koch was collected on a naturally infested lizard, Tropidurus sp. (Squamata: Tropiduridae), at Monte Negro, State of Rondônia, Brazil. The tick’s identity was confirmed morphologically and by analysis of the second internal transcribed spacer of the ribosomal DNA. This is the third known male specimen of A. rotundatum, a species that has been shown to reproduce exclusively by parthenogenesis. The two previously reported male specimens seemed to be partially teratological or gynandromorphic, whereas the present specimen shows no visible genetic or developmental anomaly.

Species of Demodex live in hair follicles and sebaceous glands of many species of mammals. Demodex macaci n .sp. is described (all life stages) from a captive rhesus monkey, Macaca mulatta (Zimmermann, 1780). Mites were found at all levels of the hair follicles and occasionally in sebaceous glands of the skin of the perineal region. This is the first description of a demodectic hair follicle mite from the primate family Cercopithecidae.

I define two new species groups of Afrorhynchus from the Afrotropical Region and examine the taxonomic structure of the genus in the region. Keys are provided for the identification of adult males and females of all known Afrotropical Toxorhynchites.

Adult survival and reproduction were compared between two Stomoxys species that co-occur in La Réunion, the cosmopolitan Stomoxys calcitrans (L.), and the tropical Stomoxys niger niger Macquart. In a first experiment, mean longevity and fecundity were determined at five constant temperatures from 15 to 35°C, after development at the same temperatures. Adult longevity was greatest at 20°C in S. calcitrans and at 15°C in S. niger. Adult S. niger survived longer than adult S. calcitrans, especially at 15°C. At 35°C, all flies died within 3 d. Reproduction occurred only within the 20–30°C range, and S. niger laid more eggs than S. calcitrans. In both species, lifetime fecundity tended to decrease when temperature increased, because of the shortening of the oviposition period. In a second experiment, adults were maintained at 15°C after development at 25°C. The higher temperature during development significantly increased adult longevity in S. calcitrans but not in S. niger. Reproduction occurred at 15°C, with notable fecundity in S. calcitrans (22 eggs per female) but not in S. niger (<1 egg per female). Using previous results on immature survival and developmental time in the two species, several life history parameters were compared at each temperature. Generation time decreased with increasing temperature and was highly similar in both species. Concurrently, the intrinsic rate of increase (r) increased with temperature from 15 to 30°C. At 15°C, r was higher in S. calcitrans, but within the 20–30°C range, r was higher in S. niger. The results suggest 1) S. niger has evolved a strategy of survival without any reproduction during the tropical winter, in contrast with S. calcitrans that breeds more continuously; and 2) S. niger may outnumber S. calcitrans in warm areas, at least when development occurs in media of poor quality.

We live-trapped small mammals and flagged vegetation within wooded natural and residential landscapes to examine how any observed differences in small mammal species composition may influence Ixodes scapularis Say burdens and the abundance of host-seeking ticks. Two years of live trapping showed that Eastern chipmunks, Tamias striatus, were captured with significantly greater frequency in some residential areas than white-footed mice, Peromyscus leucopus, whereas the proportion of white-footed mouse captures was higher or similar to chipmunk captures in the undeveloped natural areas. Both mice and chipmunks seemed to adapt well to managed residential landscapes, with residential sites yielding similar or significantly greater numbers of captures compared with undeveloped sites. In areas where chipmunk captures outnumbered mice, larval tick burdens on mice were either higher or no different than in areas where few or no chipmunks were captured, in contrast to previous studies suggesting that alternate hosts should reduce larval burdens on mice. Chipmunks apparently play an important role in the Lyme disease transmission cycle in these residential settings.

Studies were carried out at a malaria epidemic-prone highland site in western Kenya to determine the effects of deforestation-caused microclimate change on the duration of the gonotrophic cycle of the malaria vector Anopheles gambiae Giles. Gonotrophic cycle duration was measured using field-collected F₁ A. gambiae females. Average ambient temperature in the deforested area of Kakamega (elevation 1,430–1,580 m above sea level), western Kenya, was 0.5°C higher than that of the forested area over a 10-mo period. During the dry season, deforested areas showed an increased mean indoor temperature of 1.8°C, and a shortened duration of the mosquitoes’ first and second gonotrophic cycles, by 1.7 d (59%) and 0.9 d (43%). During the rainy season, the average indoor temperature of houses located in the deforested area was 1.2°C higher than in houses in the forested area. The duration of the first and second gonotrophic cycles was shortened by 1.5 d (17%) and 1.4 d (27%), respectively, in the deforested highland site. A shorter mosquito gonotrophic cycle implies increased daily biting frequency and thus increased vectorial capacity. Together with evidence that deforestation reduced A. gambiae larva-to-adult developmental time and increased larval and adult survivorship, this study suggests that deforestation can further enhance malaria transmission potential in the highlands through increased indoor temperature and shortened gonotrophic cycle durations of A. gambiae mosquitoes.

New data on the feeding patterns of Haemagogus (Haemagogus) janthinomys Dyar from different geographical regions of Brazil, by using the precipitin test as the bloodmeal-identifying tool, are presented. The following antisera were used: bird, dog, human, rodent, cattle, horse, and opossum. The origins of 287 bloodmeals were identified, whereas 33 specimens were negative to the antiserums tested. Among the reactive specimens, 174 (60.6%) fed on only one food source, of which 35.1% originated from birds, 19.5% from rodents, 12.6% from humans, 10.3% from cattle, 10.3% from opossums, 7.5% from dogs, and 4.6% from horses. One hundred six (37.0%) mosquitoes fed on two sources, of which the most common combinations were bird rodent (16.0%), bird human (10.4%), and horse human (9.4%). Seven (2.4%) mosquitoes fed on three different hosts. Our results suggest that Hg. janthinomys is more eclectic and opportunist than previously known in relation to its hosts and that such patterns are probably highly adaptive to a temporally and spatially variable environment.

The phlebotomine sand flies Lutzomyia (Nyssomyia) intermedia (Lutz & Neiva) and Lutzomyia (Nyssomyia) whitmani (Coutinho & Antunes) are important vectors of Leishmania (Vianna) braziliensis, the etiological agent of American cutaneous leishmaniasis. In some areas, both species occur in sympatry, and their relative roles as vectors in these areas are not clear. We studied the nocturnal activity and biting rhythms of both species in Posse, a locality in Rio de Janeiro State, Brazil. Our results show differences between the activity patterns of Lu. intermedia and Lu. whitmani that might be epidemiologically important. Although the activity profiles vary between seasons and microhabitats (peridomestic versus forest), the two species show marked differences in their tendencies to bite humans in the early morning (0400–0600 hours), with Lu. whitmani showing higher feeding rates than Lu. intermedia.

We studied the oviposition site preference and egg hatchability of Anopheles gambiae Giles with water collected from farmlands, forests, and natural wetlands. Water types significantly affected oviposition preference. Mosquitoes deposited significantly more eggs in rainwater in both the dry and wet seasons than waters from forests and wetlands, suggesting that An. gambiae prefers water with few impurities for oviposition. In the dry season, An. gambiae females also deposited significantly more eggs in waters from farmlands than those from forests and natural wetlands, but these differences were not statistically significant during the wet season. In both indoor and natural conditions, egg mortality in natural wetland habitats was significantly higher than in farmland habitats. The average water temperature in natural wetland habitats was significantly lower than farmland habitats in the natural conditions, but it remained the same under indoor experimental conditions, suggesting that factors other than water temperature play an important role in egg hatchability. Together with the findings from previous studies on the effects of land cover on larval survivorship, our results support the hypothesis that variations in habitat conditions induced by different land cover types contribute to the heterogeneous spatial distribution of An. gambiae larvae in the western Kenya highland.

We studied the geographical distribution of species, chromosomal, and molecular forms of the Anopheles gambiae Giles (Diptera: Culicidae) complex in 23 sites in Cameroon, Central Africa. Almost all the specimens collected in the four northern-most arid sites were Anopheles arabiensis. Anopheles melas was found in a rural locality surrounded by mangrove swamps, on the Atlantic Coast. In total, 1,525 An. gambiae s.s. females were identified down to their molecular form, and inversion polymorphisms on polytene chromosomes were scored from 186 half-gravid females. The Forest chromosomal form, with standard arrangements almost fixed on both arms of chromosome-2, was the only one observed in the southern, more humid localities. Karyotypes typical of Savanna and Mopti were recorded northwards, in the humid savannas of the Adamawa Province. The molecular forms M and S were widespread throughout Cameroon, and assort independently from the chromosomal forms. S-form populations were characterized by karyotypes typical of Forest and Savanna chromosomal forms, and M-form populations were characterized by karyotypes typical of Forest, Savanna, and Mopti. No M/S hybrid patterns were detected, although M and S mosquitoes were sympatric in 15 sites, providing further evidence for positive assortative mating within molecular forms. The observed ecogeographical distribution of M and S was peculiar: the ecological parameters involved in this distribution still need to be clarified as well as the possible role of competitive exclusion between chromosomally homosequential molecular forms. No difference was observed in host preference or in Plasmodium falciparum infection rates between sympatric M and S populations.

Since 1996, tsetse (Glossina spp.) control operations, using odor-baited traps, have been carried out in the Luke area of Gurage zone, southwestern Ethiopia. Glossina morsitans submorsitans Newstead was identified as the dominant species in the area, but the presence of Glossina fuscipes Newstead and Glossina pallidipes Austen also was recorded. Here, we refer to the combined number of these three species and report the work undertaken from October 2002 to October 2004 to render the control system more efficient by reducing the number of traps used and maintaining the previously reached levels of tsetse occurrence and trypanosomiasis prevalence. This was done by the design and implementation of an adaptive tsetse population management system. It consists first of an efficient community-participatory monitoring scheme that allowed us to reduce the number of traps used from 216 to 127 (107 monitoring traps and 20 control traps). Geostatistical methods, including kriging and mapping, furthermore allowed identification and monitoring of the spatiotemporal dynamics of patches with increased fly densities, referred to as hot spots. To respond to hot spots, the Luke community was advised and assisted in control trap deployment. Adaptive management was shown to be more efficient than the previously used mass trapping system. In that context, trap numbers could be reduced substantially, at the same time maintaining previously achieved levels of tsetse occurrences and disease prevalence.

Pyrethroid resistance in Boophilus microplus (Canestrini) (Acari: Ixodidae) was studied by correlating discriminating-dose (DD) bioassay results and esterase activity or the frequency of a sodium channel mutation known to be involved in pyrethroid resistance in nine field strains of B. microplus from Yucatan, Mexico. Two tick strains (P67 and B74) were identified as susceptible to cypermethrin, deltamethrin, and flumethrin by DD, one strain (P65) was susceptible to cypermethrin and resistant to deltamethrin and flumethrin, and six strains were resistant to cypermethrin, deltamethrin, and flumethrin (T11, M10, C54, R49, B71, and T66). By using polymerase chain reaction, only 6.0 and 6.2% of resistance allele (R) was found in the susceptible strains (P67 and B74, respectively). In the T66 strain, with 100% of larval survival to the DD of pyrethroids as measured by the larval packet test (LPT), 98.0% of the gene pool contained the R allele. Positive correlations between the larval survival and the percentage of the R allele were found (deltamethrin r² = 0.8875, P < 0.01; cypermethrin r² = 0.8563, P < 0.01; and flumethrin r² = 0.8491, P < 0.01). There were no significant correlations between the level of larval survival and esterase-based hydrolytic activity. It was concluded that within the B. microplus populations studied, resistance to flumethrin, deltamethrin, or cypermethrin was because of the novel sodium channel mutation (Phe→Ile amino acid substitution in the S6 transmembrane segment of domain III), and there was a correlation between tick mortality by pyrethroid exposure (larval survival) and the presence of R allele. It was not determined whether enhanced esterase-based hydrolytic activity was involved in pyrethroid resistance in the populations tested.

Culicoides obsoletus (Meigen) and Culicoides scoticus Downes & Kettle (Diptera: Ceratopogonidae) are sibling species of the Obsoletus group. This group comprises species of biting midges that are suspect vectors of bluetongue virus (family Reoviridae, genus Orbivirus, BTV) and African horse sickness virus (family Reoviridae, genus Orbivirus, AHSV). BTV and AHSV have been isolated several times from females of this group, although it has not been possible to determine the particular species harboring the virus, because of the inability to clearly identify the females of each species based on morphology. Both sexes of C. obsoletus and C. scoticus midges from Catalonia and the Balearics (Spain) were sequenced for the mitochondrial gene cytochrome oxidase subunit I (COI), and these sequences were analyzed to determine intra- and interspecific genetic variability. Species-specific primers for C. obsoletus and C. scoticus were designed and a polymerase chain reaction (PCR)-based diagnostic assay based on the COI gene and using a hemi-nested PCR technique was developed for reliably distinguishing the females of both species. The species-specific PCR diagnostic was compared with morphological discrimination of C. obsoletus and C. scoticus females. The morphologic characters were not fully reliable.

The epidemiology of human and animal trypanosomiasis is determined to a large extent by the number of infected tsetse flies in a specific area. In the field, a substantial proportion of infected flies carry mixed trypanosome infections. The way in which these tsetse flies acquire a mixed infection is not fully understood. In particular, the susceptibility of tsetse flies to sequential infection with trypanosomes is not well understood. Accordingly, laboratory studies were made of the effects of age and prior infection on the probability of Glossina morsitans morsitans (Westwood) developing an infection of Trypanosoma congolense and Trypanosoma brucei brucei after feeding on infected mice. Results of these experiments clearly showed that 20–30-d-old G. m. morsitans can still pick up and develop a mature infection in the mouthparts/hypopharynx for T. congolense or in the salivary glands for T. b. brucei. However, their ability to acquire infection was significantly lower compared with teneral flies. Furthermore, 20–30-d-old flies that already carry a mature T. congolense or T. b. brucei infection remained at least as susceptible to a secondary trypanosome infection compared with noninfected flies of the same age. The immunological and epidemiological repercussions of those findings are discussed.

Sentinel pigeons, Columba livia, were installed in lard-can traps at heights of 1.5 m and 7.6–9.1 m within differing canopy cover classes in New York City. Adult mosquitoes were collected weekly from July to October 2002, as were serum samples from each pigeon. Culex pipiens L. and Culex restuans Theobald comprised 97% of mosquitoes collected and were most numerous in canopy-level, forested traps. The West Nile virus (family Flaviviridae, genus Flavivirus, WNV) seroconversion rate was significantly greater for pigeons in canopy-level traps, although seroconversions occurred concurrently with human cases in the city and were of little prognostic value to public health agencies. Our results indicate that sentinel pigeons were most effective for monitoring enzootic transmission of WNV when placed in single-sentinel caging 7.6–9.1 m above ground level.

Populations of Aedes aegypti (L.) and Aedes polynesiensis (Marks) on Moorea Island, French Polynesia, the local vectors of dengue and filariasis, respectively, were sampled by landing/biting collection at nine localities on the east, north, and west coasts, during the late dry season, early and late wet season (September–May) 2003 and 2004, to investigate epidemiologically important features of the populations and compare them between regions and months. Biting rates of both species tended to increase (but not always significantly) for each species in each region each month from the late dry season to the late wet season, and the north coast region had significantly higher biting rates of Ae. aegypti. Insemination rates of Ae. polynesiensis females (94.5–98.8%) were consistently greater than those of Ae. aegypti (87.5–93.5%) throughout the study, but there was no significant difference in the insemination rates of either species between months and regions. Parity rates were generally highest in north coast samples and lowest in west coast samples for both species and generally increased (albeit not always significantly) by month for each species, with a range of 52.9–88.8% for Ae. polynesiensis and 28.6–53.6% for Ae. aegypti, although the high gravid rate (15.8–45.9%) of Ae. aegypti samples (reflecting its feeding more than once in a gonotrophic cycle) confounded both intraspecific and interspecific comparisons. Filarial infection was rare in Ae. aegypti, although both W. bancrofti and D. immitis were recorded, and infection rates in Ae. polynesiensis increased through the study period in each region for both filarias, with up to 4.6% infected and 1.4% infective for W. bancrofti and up to 6.3% infected and 2.5% infective for D. immitis. For W. bancrofti, infection rates were significantly lower on the west coast and also in the dry season, whereas rates for infective stages were significantly greater in the late wet season. For D. immitis there was no significant difference in infection rates between regions, but rates were significantly greater in the late wet season. Rainfall in all months sustains populations of both vectors and explains the relatively few significant differences between seasons; however, the wet season may provide for increased vector abundance and longevity, and present a potentially increased risk for transmission. Although the differences shown between regions also were limited in a statistical sense, there were increased risks for the northern and eastern regions, where both locals and tourists are concentrated and where the seaports and airport are located, and these areas should be priority targets for disease surveillance and vector control.

Two hundred fifty New Jersey field-collected Ixodes scapularis Say ticks and 17 Colorado Ixodes spinipalpis Hadwen & Nuttall ticks were tested using three separate multiplex real-time polymerase chain reaction (PCR) assays. One assay targets the rrs-rrlA IGS region of Borrelia spp. to detect Borrelia burgdorferi sensu lato (s.l.) and Borrelia miyamotoi s.l. The second assay targets the ospA region of B. burgdorferi s.l. to detect B. burgdorferi sensu stricto (s.s.), Borrelia bissettii, and Borrelia andersonii. The final assay targets the glpQ region of B. miyamotoi s.l. to differentiate B. miyamotoi LB-2001 and Borrelia lonestari. A testing scheme combining these tests yielded 18% of tested I. scapularis ticks surveyed from New Jersey positive for B. burgdorferi s.s., 3.2% I. scapularis ticks positive for B. miyamotoi LB-2001, and 41.2% I. spinipalpis ticks positive for B. bissettii surveyed from Colorado.

To meet the need for high-throughput sample testing, DNA extraction kits based on the 96-well plate format have been developed for use with blood and tissue samples. These methods have not been applied to DNA extractions from ticks. To meet this need, we developed a high-throughput method for DNA extraction and polymerase chain reaction (PCR) testing of tick samples. A liquid-handling robot was used to extract DNA in a 96-well binding column plate with vacuum manifold. The quantity, purity, and quality of DNA recovered from Ixodes scapularis Say, 1821 nymphs with this method were reproducible and comparable with existing manual DNA extraction techniques. The DNA yield from pools of five nymphal ticks averaged 0.432 ± 0.04 μg (95% CI). The robot also prepared real-time PCR reactions in 96-well plates, directly from the extracted DNA. A modification of the existing P20 tool resulted in accurate pipetting of 1- to 2-μl volumes with a reproducibility of ±0.038 μl when dispensing 1.0 μl. By using this process, 96 samples can be extracted and tested while reducing human labor to ≈30 min.

One of the most daunting challenges for Chagas disease surveillance and control in Mexico is the lack of community level data on vector distributions. Although many states now have assembled representative domestic triatomine collections, only two triatomine specimens had been collected and reported previously from the state of Guanajuato. Field personnel from the state’s Secretaría de Salud conducted health promotion activities in 43 of the 46 counties in the state and received donations of a total of 2,522 triatomine specimens between 1998 and 2002. All specimens were identified, and live insects examined for Trypanosoma cruzi. In an effort to develop fine-scale distributional data for Guanajuato, collection localities were georeferenced and ecological niches were modeled for each species by using evolutionary-computing approaches. Five species were collected: Triatoma mexicana (Herrich-Schaeffer), Triatoma longipennis (Usinger), Triatoma pallidipennis (Stål), Triatoma barberi (Usinger), and Triatoma dimidiata (Latreille) from 201 communities located at elevations of 870–2,200 m. Based on collection success, T. mexicana had the broadest dispersion, although niche mapping indicates that T. barberi represents the greatest risk for transmission of Chagas disease in the state. T. dimidiata was represented in collections by a single adult collected from one village outside the predicted area for all species. For humans, an estimated 3,755,380 individuals are at risk for vector transmission in the state, with an incidence of 3,500 new cases per year; overall seroprevalences of 2.6% indicate that 97,640 individuals are infected with T. cruzi at present, including 29,300 chronic cases.

The first known males of spider Loxosceles mulege Gertsch & Ennik from the xerophilous shrub vegetation zone of Baja California Sur, Mexico, are described.

Japanese encephalitis virus (JEV) infection in mosquitoes was monitored at Guandu Nature Park in Taipei City from September 2002 to December 2004. In total, 30,386 female mosquitoes consisting of six genera and 14 species were processed for virus in 1,229 pools by using Flavivirus NS5 gene sequences detected by reverse transcription-polymerase chain reaction (PCR) and nested PCR assay. Overall, 101 pools were positive, including 95, 1, 4, and 1 for Culex tritaeniorhynchus Giles, Culex sitiens Wiedemann, Culex rubithoracis (Leicester), and Aedes vexans nocturnus (Theobald), respectively.