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Dispersal of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in an urban endemic dengue area in the State of Rio de Janeiro, Brazil.

Experimental releases of female Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus were performed in August and September 1999, in an urban area of Nova Igua u, State of Rio de Janeiro, Brazil, to estimate their flight range in a circular area of 1,600 m where 1,472 ovitraps were set. Releases of 3,055 Ae. aegypti and 2,225 Ae. albopictus females, fed with rubidium (Rb)-marked blood and surgically prevented from subsequent blood-feeding, were separated by 11 days. Rb was detected in ovitrap-collected eggs by atomic emission spectrophotometry. Rb-marked eggs of both species were detected up to 800 m from the release point. Eggs of Ae. albopictus were more numerous and more heterogeneously distributed in the area than those of Ae. aegypti. Eggs positively marked for Rb were found at all borders of the study area, suggesting that egg laying also occurred beyond these limits. Results from this study suggest that females can fly at least 800 m in 6 days and, if infected, potentially spread virus rapidly.

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Preliminary data on the performance of Aedes aegypti and Aedes albopictus immatures developing in water-filled tires in Rio de Janeiro.

A monthly survey of Aedes aegypti and Aedes albopictus immatures in discarded tires at a site in metropolitan Rio de Janeiro showed that Ae. albopictus was much more abundant in the rainy season, but Ae. aegypti abundance showed a less clear seasonal pattern. Pupal masses for Ae. albopictus showed a seasonal trend. In contrast, Ae. aegypti pupae did not show any clear trend in weight. Large Ae. albopictus pupae were found in the warmer months, when water volume was higher, pH lower and larval abundance lower. Further studies should be carried out to assess how seasonal variations in body size may impact vector competence of these species in Brazil.

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Insecticide susceptibility of Aedes aegypti and Aedes albopictus across Thailand.

Aedes aegypti (L.) and Aedes albopictus (Skuse), two important vectors of dengue fever and dengue hemorrhagic fever, were collected from Mae Sot, Nakhon Sawan, Nakhon Ratchasima, Surat Thani, and Phatthalung, Thailand, from July 2003 to April 2004. The patterns of insecticide susceptibility to temephos, malathion, and permethrin of both Ae. aegypti and Ae. albopictus larvae were determined. Ae. aegypti from all study sites were resistant to permethrin, they but were susceptible to malathion. Resistance to temephos was detected in all strains of Ae. aegypti, except those from Nakhon Ratchasima. Ae. albopictus larvae had low levels of resistance to all three insecticides, except Mae Sot and Phatthalung strains, which were resistant to permethrin.

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Development and survival of immature Aedes albopictus and Aedes triseriatus (Diptera: Culicidae) in the laboratory: effects of density, food, and competition on response to temperature.

Effects of food, density, and heterospecific interactions on temperature-dependent development of Aedes albopictus (Skuse) and Aedes triseriatus (Say) larvae and pupae were described using a degree-day model. Under all conditions, the predicted number of degree-days (DD(T0)) to complete larval development was less, and the threshold temperature (T0) for initiation of larval development was higher for Ae. albopictus than for Ae. triseriatus. The DD(T0) for both species was food and density dependent. However, the per capita food ration appeared to exert a greater influence on the developmental times of Ae. triseriatus immatures, whereas for Ae. albopictus effects of density were not completely eliminated by an increase in the per capita food ration. The presence of heterospecific larvae did not prolong the DD(T0) of either species. At the low food ration, DD(T0) for both species were significantly greater under conspecific conditions. The DD(T0) for Ae. triseriatus increased directly as the proportion of conspecific larvae increased. However, when a higher per capita food ration was provided, conspecific effects on DD(T0) were mitigated. For Ae. albopictus, estimated DD(T0) values for larvae reared in pure culture were significantly higher than when heterospecific larvae were present, regardless of the per capita amount of food provided. Survivorship of immatures was density and food-dependent for both species. Ae. albopictus exhibited higher immature survivorship under all conditions relative to Ae. triseriatus. For both species, survival probabilities were lowest under conditions of high density and low food. Addition of food improved survival for both species. The presence of heterospecific larvae exerted a differential effect on the survivorship response of Ae. albopictus immatures to temperature. Lowest probability of survival for Ae. albopictus was occurred at low temperature when the proportion of heterospecific larvae in containers was the highest. In contrast, at high temperatures, survivorship of immatures improved, but was lowest for pure species cultures. For Ae. triseriatus, survivorship of immatures was consistently lower for pure species cultures regardless of the temperature. Standing crop production of adults of both species was primarily food rather that density-dependent. Under field conditions, Ae. triseriatus were predicted to initiate development sooner and exhibit faster population growth early in the season than Ae. albopictus. However, because of the shorter DD(T0), Ae. albopictus population growth was predicted to surpass that of Ae. triseriatus populations later in the season. Based on optimal DD(T0) values from laboratory experiments, spring emergence dates of Ae. triseriatus females in western North Carolina for 1989 and 1990 were predicted from accumulated degree-days calculated from local air temperature records. Predicted emergence dates were congruent with results of a previous survey for the same locality, indicating that accumulated degree-days can be used to accurately predict the seasonal occurrence of Ae. triseriatus. The utility of the degree-day approach in predicting the phenology of Ae. albopictus remains to be established.

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Density and distribution of Dirofilaria immitis (Nematoda: Filarioidea) third-stage larvae in Aedes sollicitans and Aedes taeniorhynchus (Diptera: Culicidae).

Numbers and the distribution of third-stage larvae (L3) were investigated in Aedes sollicitans (Walker) and Aedes taeniorhynchus (Wiedemann) female mosquitoes fed Dirofilaria immitis (Leidy) infectious-blood with densities of microfilariae (mf) ranging from approximately 7,100-43,400 mf/ml. At each microfilarial density, a maximum of 63-66 infective larvae were recovered from an Ae. sollicitans alive on day 15 after infection. In comparison with Ae. taeniorhynchus, Ae. sollicitans averaged greater numbers of L3 and from 1.4 to 2.4 times more L3 in the head and labium per infected female. The trend was for greater numbers of L3 to be found in the labium than in the head of Ae. sollicitans, but there were no significant differences between numbers of L3 recovered from these sites in differentially infected females. However, numbers of L3 recovered from the head versus the labium of differentially infected Ae. taeniorhynchus varied significantly with the infectious blood microfilarial density. At the two lowest and highest microfilarial densities, greater numbers of L3 were recovered from the head and labium, respectively. Variations among species in the female body size, blood meal size, and retention of L3 may be factors responsible for differences observed between the total numbers and percentage distribution of D. immitis L3 recovered from the simultaneous blood-fed Ae. sollicitans and Ae. taeniorhynchus.

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Pupal mass and wing length as indicators of fecundity in Aedes albopictus and Aedes geniculatus (Diptera: Culicidae).

Many studies investigating the ecology and evolution of mosquitoes rely on morphometric measurements related to body size to estimate fecundity. Determining the most reliable estimators of fecundity is therefore an important methodological consideration. We compared the relative accuracy of pupal mass and wing length in predicting the fecundity (number of mature stage IV follicles of the first gonotrophic cycle) for (1) Aedes albopictus (Skuse) reared in the laboratory, (2) Aedes geniculatus (Oliver) reared in the laboratory, and (3) A. geniculatus that completed their larval development in water-filled tree holes (their natural habitat) in the field. In all cases, pupal weight and wing length provided highly significant and equally accurate indicators of fecundity. Furthermore, the measurement error (1 - repeatability) for both traits was < or = 1%. Our results indicate that either pupal mass or wing length can be used with confidence to predict fecundity in these two aedine mosquitoes.

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New findings on the developmental process of Ascogregarina taiwanensis and Ascogregarina culicis in Aedes albopictus and Aedes aegypti.

Infection in different stages of larvae of Aedes aegypti and Ae. albopictus with Ascogregarina taiwanensis and A. culicis, respectively, revealed that the oocysts of Ascogregarina spp. are able to infect any instar and can complete their life cycle within 9.5 +/- 1 days. When early instars ingested oocysts, parasite development was synchronized to larval-pupal ecdysis and oocyst dissemination occurred at the time of adult emergence, oviposition, or both. The parasites also developed normally when infecting 2nd, 3rd, and early 4th instars and oocysts were released only during oviposition. The parasitic development stopped at the gamont stage when oocysts were ingested by late 4th instars (6 days old). The release of sporozoites in the midgut of any larval stage started within 45 min of oocyst ingestion. About 98% of oocysts of both A. taiwanensis and A. culicis were emptied within 2-3 h of their ingestion in their respective hosts. The oocysts of both species remained viable on desiccated filter paper stored at 27 degrees C and 65 +/- 5% relative humidity, indicating that the oocysts were resistant to dryness. The oocysts of A. culicis could survive up to 6 months, whereas those of A. taiwanensis survived up to 4 months. These biological characteristics relating to parasite development might enhance the distribution of Ascogregarina spp. widely in nature and facilitate the species to be considered for biological control of Aedes mosquitoes in the future.

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Susceptibility of laboratory and field-collected Aedes aegypti and Aedes albopictus to Bacillus thuringiensis israelensis H-14.

Susceptibility levels of a few laboratory-cultured and dengue-endemic area field-collected strains of Aedes aegypti and Aedes albopictus to Bacillus thuringiensis israelensis (Bti) at different storage ages were studied. The susceptibility of laboratory-cultured World Health Organization (WHO) Bora Bora reference, Vector Control Research Unit (VCRU), and Fumakilla Malaysia Berhad (FMB) strains of Ae. aegypti to Bti was examined. The sensitivity to Bti decreased with storage age. The median lethal concentration (LC50) for Bti increased by 2-3 times after 2 years compared to a fresh sample (3-6 months of storage). However, after the 2-year storage period, Bti still provided very good efficacy against all laboratory-cultured susceptible strains of Ae. aegypti and Ae. albopictus. The observed 95% lethal concentration values were about 20 times lower than the recommended concentration (6,000 international toxic units (ITU)/liter). Results obtained from the study against the dengue-endemic area field-collected strains of Ae. aegypti and Ae. albopictus confirmed the effectiveness of the Bti after storage for 2 years (18-24 months). For Ae. aegypti, the Ujung Batu strain was the most susceptible to Bti, whereas the Sungai Nibong strain showed the most tolerance. Susceptibility of laboratory-cultured strains varied; the Air Itam strain of Ae. albopictus was the most susceptible to Bti, whereas the Kampung Serani strain was the most tolerant among the field strains. However, the laboratory strain of Ae. albopictus was more susceptible than all the field strains.

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A field trial of competitive displacement of Aedes polynesiensis by Aedes albopictus on a Pacific atoll.

Prior laboratory studies and field observations suggested that it might be possible to reduce the size of the population of, or eliminate, Aedes polynesiensis by the introduction of Aedes albopictus. The former mosquito is the principal vector of nonperiodic filariasis caused by Wuchereria bancrofti and the latter is a closely related species refractory to the development of human filariae. The practicability of such competitive displacement was studied by a field trial on a remote coral atoll where there was an established population of A. polynesiensis. Three strains of Al albopictus were liberated at separate localities on the atoll and their fate was followed for 4 years. One strain disappeared within 12 months after release and the other two disappeared within 48 months. It was not clear whether A. albopicuts failed to become established because the strains were unsuitable, the general environment was inappropriate, or A. polynesiensis was present in such numbers that A. albopictus rarely succeded in mating with its own species.

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Transmission of Ross River virus by Aedes polynesiensis and Aedes aegypti.

Laboratory studies were carried out with two geographic strains of Aedes polynesiensis and one strain of Aedes aegypti to determine whether they could transmit Ross River virus (RRV). Both species were shown to be good vectors of RRV, but Ae. polynesiensis was the most susceptible. Ae. polynesiensis represents a new vector for this virus and the epidemiologic implications of RRV spread by both mosquito species are discussed.

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Transovarial and trans-stadial transmission of California encephalitis virus in Aedes dorsalis and Aedes melanimon.

California encephalitis (CE) virus was transmitted transovarially by its natural vectors, Aedes dorsalis and Aedes melanimon following infection by intrathoracic inoculation. Virus was recovered from adult Ae. melanimon reared from eggs that were stored for up to 19 months and exposed to repeated freezing and thawing. Neither time since oviposition nor storage conditions affected infection rates in surviving embryos. Survival rates were highest in eggs stored at 4 degrees C. Transovarial infection with CE virus did not affect survival of embryos, larvae or adults. However, transovarially infected larvae took longer to develop than did their uninfected siblings.

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Evaluation of the efficiency of transovarial transmission of California encephalitis viral strains in Aedes dorsalis and Aedes melanimon.

California encephalitis (CE) virus was transmitted transovarially by its natural vector, Aedes melanimon. Vertical transmission rates ranged from 13-26% in geographical populations of Ae. melanimon infected with CE virus by intrathoracic inoculation. No consistent pattern of transmission rates was detected for location or time of year mosquito collection. Vertical transmission rates ranged from 9-29% in Aedes dorsalis inoculated with CE viral strains isolated from Ae. melanimon collected in California. The month or year of viral isolation had no effect on the efficiency of vertical transmission. However, a viral strain isolated from the Owens Valley was less efficiently transmitted than strains from the Sacramento Valley, and strains from the San Joaquin Valley were intermediate in efficiency. Filial infection rates were highest in first ovarian cycle progeny and declined with increasing ovarian cycles in both Ae. dorsalis and Ae. melanimon.

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Transovarial transmission of dengue viruses by mosquitoes: Aedes albopictus and Aedes aegypti.

Transovarial transmission of all four dengue serotypes was demonstrated in Aedes albopictus mosquitoes. The rates of such transmission varied with the serotype and strain of virus. In general, the highest rates were observed with strains of dengue type 1 and the lowest with dengue type 3. Surprisingly, despite the use of viral strains of the four dengue serotypes which gave the highest rates with Ae. albopictus, transovarial transmission was observed in Aedes aegypti only with dengue type 1, and then only at a relatively low rate. Five different strains of Ae. aegypti were employed, including one that was known to be relatively susceptible to oral infection with dengue viruses. The findings support the view that Ae. aegypti, while of major importance from the point of view of transmission of dengue to man, may be relatively unimportant in the overall natural history of dengue viruses.

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Ingestion of immune bloodmeals and infection of Aedes fowleri, Aedes mcintoshi, and Culex pipiens with Rift Valley fever virus.

Rift Valley fever (RVF) virus infection, dissemination, and transmission rates were determined for Aedes fowleri, Aedes mcintoshi and Culex pipiens 7 or 10 days after sequentially feeding to repletion on RVF virus immune hamsters and RVF viremic hamsters, or after feeding on a mixture of RVF virus immune sheep serum and RVF viremic hamster blood through a pledget. No significant differences in infection or dissemination rates were detected among Ae. fowleri and Cx. pipiens feeding to repletion on immune hamsters before or after feeding to repletion on a viremic hamster. Similarly, no significant differences in infection, dissemination, or transmission rates were observed among Ae. fowleri and Cx. pipiens feeding to repletion on immune hamsters or nonimmune (control) hamsters 0 or 24 hr after inoculation with RVF virus. Infection rates were significantly higher for Ae. fowleri (56/66, 85%) and Cx. pipiens (123/148, 83%) fed only on viremic hamsters than for those interrupted to complete feeding on an immune hamster (Ae. fowleri [24/49, 59%], Cx. pipiens [66/131, 50%]) or a nonimmune hamster (Ae. fowleri [32/51, 63%], Cx. pipiens [69/127, 54%]). However, no significant differences were detected in infection, dissemination, or transmission rates among Ae. fowleri, Ae. mcintoshi or Cx. pipiens fed on a viremic hamster and interrupted to complete feeding on an immune vs. a nonimmune hamster. Results from interrupted feeding experiments were significantly different from pledget feeding experiments.(ABSTRACT TRUNCATED AT 250 WORDS)

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Infection of Aedes albopictus and Aedes aegypti mosquitoes with dengue parent and progeny candidate vaccine viruses: a possible marker of human attenuation.

Dengue (DEN-1) and DEN-4 parent (P) and progeny candidate vaccine (CV) viruses were compared in their abilities to infect and to replicate in Aedes aegypti and Aedes albopictus mosquitoes. The DEN CV clones were temperature sensitive (ts) and had small plaque morphology. The DEN-1 and DEN-4 CV viruses differed in their ability to infect, to replicate in, and to be transmitted by mosquitoes. The DEN-1 CV virus was not attenuated for the vector mosquitoes; oral infection rates with the CV virus were as high as or higher than the P virus, and the CV virus replicated efficiently in mosquitoes after oral infection. The DEN-4 CV virus was attenuated; it was less efficient than its P virus in infection and replication in mosquitoes. Thus, the ts phenotype and small plaque morphology are not reliable biological markers for prediction of vector attenuation. Similar results were reported by others for attenuation in man and monkeys. These studies with DEN-1 and DEN-4 viruses, and previously reported studies with DEN-2 virus and with DEN-3 virus suggest that vector and vertebrate host attenuation are genetically linked. Thus, vector attenuation may be a biological marker for human attenuation.

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Venereal and vertical transmission of the Aedes albopictus parvovirus in Aedes aegypti mosquitoes.

Following per oral infection of Aedes aegypti larvae with Aedes albopictus parvovirus (AaPV), infected males and females adults were tested for their ability to transmit the virus venereally and vertically, respectively. Both types of transmission were observed. A low percentage (2.2%) of AaPV-free females were found contaminated by the virus after mating with AaPV-infected males. Although no significant difference was observed in the fecundity of orally infected and virus-free females, 17.1% of infected ones died before egg laying, whereas no mortality occurred during the same period in virus-free females. There was a clear relationship between the virus titer in the orally infected females and both mortality and infection in their offspring. The virus titer averaged 10(6.2) 50% tissue culture infective doses (TCID50s) in F1 females and 10(3.3) TCID50 in F1 females. Nevertheless, AaPV did not persist in an experimentally infected population of mosquitoes beyond the second generation.

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Differentiation of Aedes triseriatus (Say) from Aedes hendersoni Cockerell (Diptera: Culicidae) by restriction fragment length polymorphisms of amplified ribosomal DNA.

Aedes triseriatus is the primary vector of LaCrosse (LAC) virus, which can cause encephalitis, especially in young children. Aedes hendersoni, a sibling species of Ae. triseriatus, has a salivary gland barrier to LAC virus and, therefore, is not considered a vector of this virus. Adults of Ae. triseriatus are morphologically indistinguishable from those of Ae. hendersoni, and the two species are sympatric in the eastern United States. A definitive method of identifying field specimens is an important part of any disease surveillance program, particularly in the case of LAC virus. This study identifies restriction enzymes that produce species-specific restriction fragment length polymorphisms (RFLPs) from amplified ribosomal (r) DNA. In addition, sequences of the internal transcribed spacers 1 and 2 and the 5.8S regions of the rDNA were used to confirm the RFLP patterns. This study is the first to compare nucleotide sequences from Ae. triseriatus and Ae. hendersoni.

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Growth characteristics of the chimeric Japanese encephalitis virus vaccine candidate, ChimeriVax-JE (YF/JE SA14--14--2), in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes.

The Japanese encephalitis (JE) virus vaccine candidate, ChimeriVax-JE, which consists of a yellow fever (YF) 17D virus backbone containing the prM and E genes from the JE vaccine strain JE SA14--14--2, exhibits restricted replication in non-human primates, producing only a low-level viremia following peripheral inoculation. Although this reduces the likelihood that hematophagous insects could become infected by feeding on a vaccinated host, it is prudent to investigate the replication kinetics of the vaccine virus in mosquito species that are known to vector the viruses from which the chimera is derived. In this study ChimeriVax-JE virus was compared to its parent viruses, as well as to wild-type JE virus, for its ability to replicate in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes. Individual mosquitoes were exposed to the viruses by oral ingestion of a virus-laden blood meal or by intrathoracic (IT) virus inoculation. ChimeriVax-JE virus did not replicate following ingestion by any of the three mosquito species. Additionally, replication was not detected after IT inoculation of ChimeriVax-JE in the primary JE virus vector, Cx. tritaeniorhynchus. ChimeriVax-JE exhibited moderate growth following IT inoculation into Ae. aegypti and Ae. albopictus, reaching titers of 3.6-5.0 log(10) PFU/mosquito. There was no change in the virus genotype associated with replication in mosquitoes. Similar results were observed in mosquitoes of all three species that were IT inoculated or had orally ingested the YF 17D vaccine virus. In contrast, all mosquitoes either IT inoculated with or orally fed wild-type and vaccine JE viruses became infected, reaching maximum titers of 5.4-7.3 log(10) PFU/mosquito. These results indicate that ChimeriVax-JE virus is restricted in its ability to infect and replicate in these mosquito vectors. The low viremia caused by ChimeriVax-JE in primates and poor infectivity for mosquitoes are safeguards against secondary spread of the vaccine virus.

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