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[Frequency and spatial distribution of Aedes aegypti and Aedes albopictus (Diptera, Culicidae) in Rio de Janeiro, Brazil].

We evaluated the intra- and peri-domiciliary distribution of Aedes aegypti and Ae. albopictus in urban, suburban, and rural districts in the municipalities of Nova Iguaçu and Rio de Janeiro, Brazil, from August 2002 to July 2004. Mostly Ae. aegypti females and males were captured in urban areas (56%) and indoors (78%), suggesting a preference by this species to rest inside houses and in areas with high human density, a behavior that favors vector-human contact. Meanwhile, Ae. albopictus females and males were much more common in rural areas (93%) and outdoors (90%), demonstrating their preference to rest in areas with more vegetation, reducing both the probability of vector-human contact and thus their potential role in dengue transmission in the area.

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Convergent habitat segregation of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in southeastern Brazil and Florida.

During the rainy season of 2001, the incidence of the dengue vectors Aedes aegypti and Ae. albopictus was examined in different habitats of two cities (Rio de Janeiro and Nova Iguaçu) in Rio de Janeiro State, Brazil, and in two cities (Palm Beach and Boca Raton) in Florida. Oviposition trap collections were performed in urban, suburban, and rural habitats in both areas. Our hypothesis that the abundances and frequencies of occurrence of Ae. aegypti and Ae albopictus are affected in opposite ways by increasing urbanization was only partially supported. City, habitat, and their interaction significantly affected the abundance of both species. Cities with high abundance of Ae. aegypti also had a high abundance of Ae. albopictus. The two species were most abundant in the cities of Rio de Janeiro state and the lowest in Boca Raton. Habitat had a significant but opposite effect on the abundances of Ae. aegypti and Ae. albopictus. In general, Ae. aegypti was most prevalent in highly urbanized areas and Ae. albopictus in rural, suburban, and vegetated urban areas in Rio de Janeiro state and Florida. However, abundances of the two species were similar in most suburban areas. Analyses of frequencies of occurrence showed an unexpected high level of co-occurrence of both species in the same oviposition trap. Despite the different geographical origins of Ae. albopictus in Brazil and the United States, the habitats used by this recent invader are remarkably similar in the two countries.

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Differential identification of Ascogregarina species (Apicomplexa: Lecudinidae) in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) by polymerase chain reaction.

We report 2 polymerase chain reaction (PCR)-based methods for distinguishing morphologically similar gregarine species based on amplification of variable regions of the internal transcribed spacer region of ribosomal DNA. The gregarines we investigated were Ascogregarina barretti (Vavra), A. culicis (Ross), and A. taiwanensis (Lien and Levine), parasites of the mosquitoes Ochlerotatus triseriatus (Say), Aedes aegypti (Linnaeus), and Ae. albopictus (Skuse), respectively. These 3 important vector mosquitoes often utilize the same container habitats, where larval development and infection by the parasite occurs, leaving ample opportunity for cross-species gregarine infection. Because previous studies have shown that the parasites A. culicis and A. taiwanensis variably affect fitness in both normal and abnormal mosquito hosts, distinguishing parasite infection and species is important. The task is complicated by the fact that these 2 parasite species are virtually identical in morphology, whereas A. barretti is morphologically distinct. Of the 2 PCR-based assays reported here, the first provides a rapid, sensitive, and straight-forward means of general ascogregarine detection based on a single PCR amplification. The second method provides a means of differentiation between A. culicis and A. taiwanensis based on a species-specific PCR assay. Together, these assays allow whole mosquitoes to be tested for the presence of Ascogregarina species as well as identification of both A. culicis and A. taiwanensis singly or in dual infections.

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The significance of western equine encephalomyelitis viral infections in Aedes trivittatus (Diptera: Culicidae) in Iowa. I. Variation in susceptibility of Aedes trivittatus to experimental infection with three strains of western equine encephalomyelitis virus.

F1 pregnancy obtained from field-collected Aedes trivittatus were evaluated for susceptibility to infection with western equine encephalomyelitis (WEE) virus by intrathoracic inoculation and by oral imbibition of virus-blood suspensions through a membrane. Mosquitoes were uniformly susceptible to infection by intrathoracic inoculation of three strains of WEE virus, but minimum infective doses varied as much as 2,000 to 12,000-fold between strains by membrane feeding. Dose-response data obtained by membrane feeding also indicated that field strains of A. trivittatus were quite heterogeneous in their susceptibility to WEE virus since some individual mosquitoes could be infected by ingestion of low virus concentrations while others could not be infected by a 20,000-fold increase in virus concentration. Moreover, A. trivittatus showed a greater affinity for a WEE viral strain isolated from this species than for a WEE viral strain isolated from Culex tarsalis, even though the site, date of collection, and passage history of these isolates were identical. Field strains of A. trivittatus were relatively refractory to oral infection with WEE virus.

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Differences in host choice between the sibling species of treehole mosquitoes Aedes triseriatus and Aedes hendersoni.

Adult treehole mosquitoes were collected by vacuum-sweeping of vegetation in urban, suburban, and rural woodlots in northern Indiana. The sibling species Aedes triseriatus and Ae. hendersoni were identified by polyacrylamide gel electrophoresis. Blood meals were identified by the modified precipitin method. Ae. triseriatus fed predominantly on chipmunks and deer, and Ae. hendersoni fed mainly on tree squirrels and racoon. The relative rates of feeding on the major hosts were variable depending on the location of collection, and probably reflected differences in host density. No blood-feeding on humans was detected.

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Dengue 3 virus infection of Aedes albopictus and Aedes aegypti: comparison of parent and progeny candidate vaccine viruses.

DEN-3 parent (strain CH53489) and progeny candidate vaccine (clone 24/28) viruses were compared in their abilities to interact with Aedes aegypti and Ae. albopictus. The parent and progeny virus were equivalent in their ability to infect, to replicate in, and to be transmitted by both species of mosquitoes. The candidate vaccine DEN-3 clone was temperature sensitive and had small plaque morphology. These phenotypic markers remained stable during mosquito passage. Thus, temperature sensitivity and small plaque morphology are not reliable biological markers for attenuation.

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Malathion resistance in Aedes aegypti and Culex quinquefasciatus after its use in Aedes aegypti control programs.

The continued widespread use of malathion in Aedes aegypti control programs in Latin America has generated insecticide resistance to this chemical in Culex quinquefasciatus but not in Ae. aegypti. To determine the extent of this resistance, the susceptibility of Cx. quinquefasciatus and Ae. aegypti from several countries to malathion was evaluated. Bioassay results indicated that all Ae. aegypti strains evaluated from Cuba, Venezuela, Costa Rica, and Jamaica were susceptible to malathion in spite of the historical use of this insecticide in Ae. aegypti control programs in these countries. In contrast, a high level of resistance to this insecticide was found in Cx. quinquefasciatus from Venezuela, Colombia, Brazil, and Cuba. Synergist assays indicated that neither esterases nor mixed-function oxidases (MFOs) were involved as the resistance mechanism to malathion in any of the Ae. aegypti strains tested. In Cx. quinquefasciatus, synergist assays confirmed that esterases played an important role in malathion resistance but MFOs were not involved in causing malathion resistance in this species. Biochemical assays showed that both resistance mechanisms were present in the Ae. aegypti and Cx. quinquefasciatus populations. Acrylamide electrophoresis gels revealed that all Ae. aegypti strains had a strongly staining, clear band, named A4, and had a relative mobility (Rm) value of 0.7. Analysis if the results of this study suggested that malathion could continue to be used for the emergency control of Ae. aegypti, the mosquito vector for dengue and dengue hemorrhagic fever in the Americas, but that malathion is probably not effective for the control of adult Cx. quinquefasciatus in urban areas. Therefore, control operations should integrate nonorganophosphate insecticides such as pyrethroids for control of these 2 species found in the urban environment.

Acetylcholinesterase↗

The energetic costs of diving in Aedes aegypti and Aedes albopictus pupae.

Undisturbed mosquito pupae rest at the water surface and respond to passing shadows or vibrations by diving. Pupae do not feed and rely solely on energy stored from the larval stage. The ability of a newly emerged adult mosquito to survive, and therefore to transmit disease, depends on these energy reserves. Earlier studies of diving behavior in Aedes aegypti, Ae. albopictus, and Ae. triseriatus pupae provided evidence that pupae sense their state of buoyancy and modify their diving behavior accordingly. With strong stimulation pupae tend to dive to a depth where they become neutrally or negatively buoyant and commonly rest on the bottom. This behavior, as well as the tendency to rest when not disturbed, may logically be viewed as energy-conserving. The results of these studies also generated the hypothesis that the diving behavior displayed by these container-breeding mosquitoes helps them avoid being washed from their container by overflowing water during rainfall. Rainfall stimulates diving and logically, prolonged, heavy rainfall stimulates excessive diving, a likely drain on energy reserves. Our objectives were to determine, in Ae. aegypti and Ae. albopictus, the energetic costs associated with resting behavior, with frequent diving, and with buoyancy reduction. Using survival rate, mean survival after adult emergence, and measurement of total calories, we found a clear energetic cost associated with frequent diving. In contrast, relative to diving, essentially no energy cost was associated with buoyancy reduction, that is, pupae behave in response to variations in buoyancy in a way that does not impact significantly on energy reserves.

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Susceptibility of adult field strains of Aedes aegypti and Aedes albopictus in Singapore to pirimiphos-methyl and permethrin.

The susceptibilities of field strains (Fl) of Aedes aegypti and Ae. albopictus adult females to pirimiphos-methyl and permethrin were investigated and compared with a susceptible laboratory strain (S) by using a filter paper (12 x 15 cm) impregnation method. The resistance ratio between the 50% lethal concentration values (RR50) of the Fl and the S strains shows that the RR50 of Fl Ae. aegypti for pirimiphos-methyl was 1.5 and the RR50 of Fl Ae. aegypti for permethrin was 12.9. The RR50 of Fl Ae. albopictus for pirimiphos-methyl was 1.4, and that for permethrin was 1.8. This indicates that the field strain Ae. aegypti, but not Ae. albopictus, has developed resistance to permethrin. However, both species are still susceptible to pirimiphos-methyl. Therefore, control with pirimiphos-methyl will still be effective.

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Polygamy: the possibly significant behavior of Aedes aegypti and Aedes albopictus in relation to the efficient transmission of dengue virus.

The polygamous behavior of male Aedes aegypti (L.) and Ae. albopictus (Skuse) was investigated by co-habiting a newly-emerged male and females in a 30 cm3 cage (1 male: 20 females) for up to 5 consecutive days. As determined by insemination rates, the results indicated that one Ae. aegypti and Ae. albopictus male could successfully mate with 1.10 (0-4), 4.10 (1-8), 5.40 (4-8), 5.10 (2-8), 5.15 (3-9) and 0.20 (0-3), 1.70 (0-3), 2.35 (1-4), 2.30 (0-4), 2.35 (1-4) Ae. aegypti and Ae. albopictus females, respectively on day 1,2,3,4 and 5 consecutively. The possibly significant role of their polygamy in relation to dengue virus transmission is discussed.

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Distribution of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) in small towns and villages of Penang Island, Malaysia--an ovitrap survey.

An ovitrap survey was carried out in April and May of 1975 at twenty-one locations scattered throughout Penang Island excluding the City of Georgetown. The results showed the ubiquitous presence of Ae. albopictus. Ae. aegypti was recorded only in Tanjung Tokong Lama, an area adjacent to the City of Georgetown. In the survey, Ae. albopictus females were found to prefer the outdoor ovitraps than indoor ones. The effectiveness of ovitrap in Aedes survey was discussed.

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Laboratory and semi-field evaluation of Mosquito Dunks against Aedes aegypti and Aedes albopictus larvae (Diptera: Culicidae).

Laboratory bioassays and semi-field studies were conducted on the efficacy and longevity of Mosquito Dunks (7,000 ITU/mg Bti) in order to determine the concentration-response relationship and the effectiveness on the potency of the Bti product against Aedes mosquito species based on the WHO protocol standard methods and to determine the longevity of release for this product against Ae. aegypti mosquito larvae in water storage containers. This bio-potency study with the late 3rd instar larvae of Ae. aegypti and Ae. albopictus was carried out according to WHO standard protocols. The six concentrations of the Bti product used in each test were replicated 4 times with 25 mosquito larvae. Probit analysis was then used to determine the LC50 and LC95 which was equated with dosages of 1.02 and 1.86 ppm for Ae. aegypti; and 0.39 and 0.84 ppm for Ae. albopictus, which reveals a potency of 382.95 and 303.74 ITU/mg, respectively. The semi-field evaluation of this product in 200-liter earthen jars against 3rd instar larvae of Ae. aegypti showed satisfactory control of greater than 80% at 11 weeks post-treatment.

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Growth characteristics of ChimeriVax-Den vaccine viruses in Aedes aegypti and Aedes albopictus from Thailand.

Four chimeric yellow fever (YF) 17D-dengue (DEN) candidate vaccine viruses (ChimeriVax-DEN; Acambis, Cambridge, MA) were characterized in Aedes aegypti and Ae. albopictus mosquitoes collected from Thailand. The four vaccine viruses contained the relevant prM and E genes of wild-type dengue viruses (DENV; serotypes 1-4) substituted for the equivalent genes in the YF vaccine virus (17D) backbone. Each chimera conferred protection against the homologous DENV serotype; a tetravalent mix of all four chimeras stimulates an immune response against all serotypes. Field-collected mosquitoes from Thailand were fed on blood containing each of the viruses under study and held 21 days after infection. Infection and dissemination rates were based on antigen detection in the body or head tissues, respectively. All four wild-type DENV serotypes infected and disseminated, but the candidate vaccine viruses were highly attenuated in mosquitoes with respect to infection and especially with respect to dissemination. Considering the low level viremias anticipated in humans vaccinated with these viruses, it is predicted that the risks of infection and transmission by mosquitoes in nature is minimal.

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Differences in the larval alarm reaction in populations of Aedes aegypti and Aedes albopictus.

The time spent submerged during the larval alarm reaction was measured for several strains of Aedes aegypti and Ae. albopictus from different locations. Differences between the 2 species were not significant. Differences in strains within the species were highly significant. Greater differences were seen among strains of Ae. albopictus than among strains of Ae. aegypti. These differences represent an additional indication of extensive local differentiation in Ae. albopictus.

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In-vivo staining of Aedes vigilax, Aedes aegypti and Culex annulirostris larvae with Giemsa and other vital dyes.

Of 11 stains tested in the laboratory with larval Aedes aegypti, Giemsa at dosages of 6 mg/liter for 24 hr and 21 mg/liter for 3 hr proved most satisfactory for in-vivo staining. At these rates, Giemsa caused some mortality especially in first instars of Ae. vigilax, Ae. aegypti and Culex annulirostris but effectively was retained in all adults examined up to 3 days posteclosion. The method was deemed satisfactory for investigations of larval movement of Ae. vigilax, Cx. sitiens and Ae. alternans in salt marsh in southeast Queensland.

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Interspecific mating between Louisiana strains of Aedes albopictus and Aedes aegypti in the field and laboratory.

Interspecific mating between Aedes albopictus males and Ae. aegypti females was detected in the field using mark-release-recapture techniques. By 3 days after the release of virgin Ae. aegypti females into a field site containing only Ae. albopictus, 100% of the captured females were inseminated. Laboratory investigations indicated that male Ae. albopictus were very proficient at inseminating Ae. aegypti females and that Ae. aegypti males rarely inseminated Ae. albopictus females, especially if Ae. aegypti females were available. Most of the Ae. aegypti females inseminated by Ae. albopictus males contained only small amounts of dead sperm in their spermathecae, while inseminated females from the converse interspecific mating and from intraspecific matings contained only large amounts of live sperm. The results are discussed in relation to the decline in Ae. aegypti densities observed since the introduction of Ae. albopictus into the southern USA.

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Aedes bahamensis: its invasion of south Florida and association with Aedes aegypti.

The exotic mosquito, Aedes bahamensis, is now well-established in south Florida, where it is widely distributed throughout Dade and southern Broward Counties in both urban and rural areas east of the Everglades. When discarded automobile tires were sampled in areas near human habitation, larvae and pupae of Ae. bahamensis were frequently found in association with immature Ae. aegypti. Elsewhere, however, Ae. bahamensis generally occurred in the absence of Ae. aegypti. The persistence of Ae. bahamensis populations at specific sites was documented in egg collections from ovitraps and in larval samples from the water retained in discarded tires.

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