Horizontal and vertical transmission of Japanese encephalitis virus by Aedes japonicus (Diptera: Culicidae)

Aedes japonicus (Theobald) was evaluated for competence as a vector of Japanese encephalitis virus and for its ability to transmit the virus vertically to its F1 larvae. Ae. japonicus supported the growth of the virus at 20 and 28 degrees C after feeding on a virus-blood mixture of 10(6.2) plaque forming unit (PFU)/ml. This species was able to transmit the virus to suckling mice after feeding on a virus-blood meal (10(6.2) PFU/ml) or a viremic chick (10(3.7) PFU/ml). Vertical transmission of the virus in Ae. japonicus was demonstrated with a minimum infection rate of 0.7%.     

Susceptibility of selected strains of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) to chikungunya virus.

The relative susceptibility of selected strains of Aedes aegypti (L.) and Aedes albopictus (Skuse) fed on a viremic monkey to infection with chikungunya virus was determined. Infection rates were consistently higher in 10 strains of Ae. albopictus tested than in 7 strains of Ae. aegypti tested, regardless of the geographic location from which the strains originated or the dose of virus ingested. Similarly, virus dissemination rates were higher in the Ae. albopictus strains compared with the Ae. aegypti strains. For nearly all (11 of 12) strains tested of both species, groups of mosquitoes with one or more females with a disseminated infection transmitted virus by bite to weanling mice. Based on these studies, Ae. albopictus appears to be a more competent laboratory vector of chikungunya virus than does Ae. aegypti.     

Sindbis virus-associated pathology in Aedes albopictus (Diptera: Culicidae)

Virus dissemination and associated pathology were examined in Aedes albopictus after intrathoracic inoculation of Sindbis virus (SIN), the prototypic Alphavirus. At 10 days postinfection, virus RNA was detected in all three-body segments of the insect. Colocalization of virus antigen with structural pathology was observed in mosquito salivary glands and midgut-associated visceral muscles, representing yet another example of arbovirus-associated pathology in a mosquito host. SIN antigen and gross pathology were detected in lateral lobes, but not the medial lobe of salivary glands, whereas virus antigen, vacuolated cytoplasm, and myofilament misalignment were detected in the visceral muscles at the midgut exterior surface. Early in the midgut infection, virus antigen was localized in small foci on the organ surface that progressed to a grate work-like banding pattern that eventually cleared. Both the salivary glands and the midgut are essential to insect survival and reproduction. Additionally, these organs provide a pathway for virus transmission in nature. Although SIN infection may not shorten the mosquito life span, persistent coexistence could permit survival of both host and microbe as well as contribute to alterations in insect behavior.     

A novel technique for removing Wolbachia infections from Aedes albopictus (Diptera: Culicidae).

Intracellular bacteria of the genus Wolbachia often behave as reproductive parasites by manipulating host reproduction to enhance the vertical transmission of infections. Wolbachia infections in Aedes albopictus (Skuse) cause a reproductive manipulation known as cytoplasmic incompatibility, which can reduce brood hatch. Because field populations of Ae. albopictus are naturally infected, studies of Wolbachia-induced effects on Ae. albopictus reproduction and fitness require that Wolbachia be artificially removed. Although simiple techniques for clearing Wolbachia infections from other host insects have been developed, removal of Wolbachia bacteria from Ae. abopictus is difficult. Here we describe an improved method for removing Wolbachia infections Ae. albopictus. This method differs from earlier techniques in that it relies upon the tetracycline treatment of adults instead of larvae. We demonstrate that tetracycline treatment of adult Ae. albopictus can predictably generate uninfected individuals, simplify the procedure required for Wolbachia removal, and reduce the level of inbreeding required to produce uninfected lines.     

Failure of dengue-2 virus antibody to interfere with the isolation of dengue-2 virus from Aedes aegypti (Diptera: Culicidae)

When isolating dengue virus (DEN) from mosquitoes collected in endemic areas, pools may contain both anti-dengue antibodies from freshly engorged females and virus from DEN infected females. To determine if these antibodies may interfere with virus isolation, we simulated the isolation procedure using Aedes aegypti (L.) that we infected with the 16,681 strain of dengue type 2 virus by intrathoracic inoculation. At 7 d postinfection, we allowed females to engorge on immunized or normal mouse blood. Virus in a mixture of anti-dengue-2 antibodies and dengue-2 virus became inactive after incubation at 37 degrees C for 1 h, but remained infective without incubation. Therefore, at ambient conditions antibodies would not interfere with virus isolation from field-collected Ae. aegypti from endemic areas. In addition, DEN antibodies enhanced virus replication when inoculated into Ae. aegypti, but not C6/36 cells. The mechanism for this in vitro antibody enhancement of infection remains unclear.     

Transmission of Venezuelan equine encephalomyelitis virus by strains of Aedes albopictus (Diptera: Culicidae) collected in North and South America

Experimental studies were undertaken to ascertain the vector potential of North American (Houston and Alsace) and South American (Sao Paulo and Santa Teresa) strains of Aedes albopictus (Skuse) for an epizootic (Trinidad donkey) strain of Venezuelan equine encephalomyelitis (VEE) virus. Infection rates were similar in all four strains of Ae. albopictus tested after ingestion of VEE virus from a viremic hamster. Virus disseminated from the midgut to the hemocoel in about 80% of infected mosquitoes, regardless of the dose ingested (10(4.6) to 10(5.7) plaque-forming units per mosquito) or the time of extrinsic incubation (7-35 d). Although all four strains of this mosquito transmitted VEE virus by bite to hamsters, transmission rates were significantly higher for the South American strains (24%, 40 of 170) than for the North American strains (5%, 9 of 165). Although VEE virus has never been isolated from Ae. albopictus, the introduction of this species into the Americas may allow it to serve as an amplification vector in areas where epizootic strains of VEE are found or introduced.     

Transmission of Venezuelan equine encephalomyelitis virus by Aedes sollicitans and Aedes taeniorhynchus (Diptera: Culicidae).

Experimental studies compared the vector competence of Aedes sollicitans (Skuse) and Ae. taeniorhynchus (Wiedemann) collected on Assateague Island, Va., for an epizootic strain (Trinidad donkey) of Venezuelan equine encephalomyelitis (VEE) virus. Infection rates were significantly higher in Ae. sollicitans (101/107, 94%) than in Ae. taeniorhynchus (103/175, 59%), even though both species fed concurrently on the same infected hamsters. Similarly, dissemination and transmission rates were significantly higher in the Ae. sollicitans population tested. Although both Ae. taeniorhynchus and Ae. sollicitans are natural vectors of VEE virus, the latter species should be considered a more efficient vector of VEE epizootic strains, based on its greater susceptibility to infection and higher transmission rates.     

Laboratory study of competition between United States strains of Aedes albopictus and Aedes aegypti (Diptera: Culicidae)

Mosquito abatement workers in Houston, Tex., and New Orleans, La., have observed that the recent introduction of Aedes albopictus (Skuse) has been accompanied by a decline and virtual disappearance of Ae. aegypti (L.). This suggests competitive displacement but contradicts the direction of displacement observed in native habitats of Ae. albopictus. The paper reports a series of laboratory studies on competition between U.S. strains of the two species. Larval survivorship, size, and development time were monitored in increasing densities of pure and mixed cultures of the two species under limited and optimal diets. Oviposition preferences were examined by offering gravid females cups containing first or fourth instars of either species. Ethological isolation was studied by simultaneously offering virgin females of one species to males of both species and monitoring courtship behavior as well as rates of oviposition, percentage of hatch, and embryonation. The results of the larval competition studies suggest that interspecific competition increased development time only slightly and had no effect on survival of Ae. albopictus. Competition increased survival in Ae. aegypti. Size of both species was influenced by interspecific competition, but the magnitude and direction of the effect was inconsistent between strains and diets such that neither species emerged consistently larger. The results of the oviposition study indicated a higher fecundity in Ae. aegypti but otherwise demonstrated that both species oviposit without regard to the presence, age, or species of larvae in the oviposition container. The ethological isolation tests demonstrated that Ae. aegypti males were more sexually aggressive than males or Ae. albopictus. However, Ae. albopitus females placed with increasing densities of Ae. aegypti males demonstrated no decrease in rates of oviposition, and eggs laid showed equal rates of hatch and embryonation. In general, the results do not suggest that the U.S Ae. albopictus population is inherently more competitive in the laboratory than Ae. aegypti. Other reasons for the observed decline in Ae. aegypti in the United States are discussed.     

Vector competence of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) for dengue virus in the Florida Keys

In 2009-2011, Monroe County in southern Florida experienced locally acquired and traveler-imported focal dengue outbreaks. Aedes aegypti (L.) is the primary vector of dengue virus (DENV) worldwide, is prevalent in Monroe County, and is the suspected vector in Florida. Ae. albopictus (Skuse) is also known to be an important vector of DENV and this species is ubiquitous in Florida; however, it is not yet established in Monroe County. Florida Ae. aegypti (Key West and Stock Island geographic colonies) and Ae. albopictus (Vero Beach geographic colony) were fed blood containing 3.7 Log10 plaque-forming unit equivalents of DENV serotype 1 isolated from a patient involved in the Key West, FL, outbreak in 2010. Mosquitoes were maintained at extrinsic incubation temperatures of 28 or 30 degrees C for an incubation period of 14 d. Vector competence was assessed using rates of infection (percent with virus-positive bodies), dissemination (percent infected with virus-positive legs), and transmission (percent infected with virus-positive saliva). No significant differences were observed in rates of infection or dissemination between Ae. aegypti or Ae. albopictus at either extrinsic incubation temperature. Transmission was observed only at 28 degrees C in both Ae. aegypti (Key West) and Ae. albopictus. The assessment of local mosquito populations for their DENV vector competence is essential and will aid mosquito control operators interested in pinpointing specific vector populations for control. The extent to which vector competence is affected by seasonal changes in temperature is discussed and provides baseline risk assessment data to mosquito control agencies.     

Reproductive and metabolic differences between Aedes aegypti and Ae. albopictus (Diptera: Culicidae).

Aedes albopictus (Skuse) females were more likely to develop eggs after they ingested small blood meals than were Aedes aegypti (L.) when both species were maintained under the same suboptimal adult nutritional regimen. The longevity of adult female Ae. albopictus under conditions of starvation was also significantly greater than that of Ae. aegypti. Analyses of total body proteins, lipids, and glycogen indicated that the increased reproductive efficiency of Ae. albopictus may be a result of its greater reserves accumulated during a longer larval period.     

Transmission of western equine encephalomyelitis virus by Argentine Aedes albifasciatus (Diptera: Culicidae).

Aedes albifasciatus (Macquart) has been considered a potential vector of western equine encephalitis (WEE) virus in Argentina because it has been found naturally infected, it is susceptible to infection per os, and its distribution and feeding habits are compatible with those of an epizootic and epidemic vector. Ae. albifasciatus females collected in the vicinity of Cordoba, Argentina, were highly susceptible to WEE virus perorally (ID50 less than 0.5 Vero cell plaque-forming units) when fed on viremic chicks. Virus transmission trials were done 9 to 16 d after infection by feeding potentially infected mosquitoes individually on chicks. Among 31 mosquitoes that survived the incubation period, six refed on susceptible chicks. Virus transmission by bite was demonstrated by five of six (83%) mosquitoes. Therefore, the final piece of evidence incriminating Ae. albifasciatus as a natural vector of WEE virus has been obtained. This is the first mosquito species to be incriminated as a vector of WEE virus in South America.     

Overwintering survival of Aedes albopictus (Diptera: Culicidae) eggs in Indiana

Eggs of 12 strains of Aedes albopictus (Skuse), two strains of A. aegypti L., and one strain of A. triseriatus (Say) were tested for ability to overwinter at three outdoor locations in Indiana. Some survival of A. albopictus eggs was observed during the winters of 1986-1987 and 1987-1988. A. triseriatus had greater overwintering ability than A. albopictus; A. aegypti did not survive at all locations. Prolonged cold-conditioning and photoperiodically induced diapause increased the overwintering ability of A. albopictus eggs. Strains of A. albopictus from northern Asia and North America showed higher overwintering survival rates than A. albopictus strains from tropical Asia, Hawaii, and Brazil; strains from Indiana had greater ability to overwinter than those from Texas, Louisiana, and Florida. Results are discussed in relation to the overwintering limits of this species in the United States.     

Cuticular hydrocarbon geographic variation among seven North American populations of Aedes albopictus (Diptera: Culicidae).

Cuticular hydrocarbons were used to differentiate among the following North American populations of Aedes albopictus: Chicago, Ill.; Milford, Del.; Jacksonville, Fla.; San Antonio, Tex.; New Orleans, La.; Houston, Tex.; and Lexington, Ky. Forty cuticular hydrocarbon peaks were identified from each population using gas chromatography electron impact-mass spectrometry: 16 n-alkanes, 21 monomethylalkanes, and 3 dimethylalkanes. Fourteen peaks occurred at equivalent chain lengths greater than 38.65 and consisted of complex mixtures of dimethyl- and trimethylalkanes. Quantitative analyses were performed on pooled samples of five females from each population using flame ionization gas chromatography. All populations contained the same cuticular hydrocarbons, but showed differences in percent composition. Five of seven populations were separable by discriminant analysis of hydrocarbon components.     

Food ingestion and digestive enzymes in larval Aedes aegypti and Ae. albopictus (Diptera: Culicidae).

Frequency of the labral brush movements of first, second, and fourth instars of Aedes aegypti (L.) and Aedes albopictus (Skuse) was studied comparatively in the laboratory. A frequency of 197 strokes per min for the first and second instars was observed in the former species compared to 118 strokes per min in the latter species. A faster ingestion rate of algal cells also was observed in first and second instars of Ae. aegypti (mean 57.5 cells per s) compared with first and second instars of Ae. albopictus (mean 22.4 cells per s). The digestive enzymes chymotrypsin (EC 3.4.21.1) and trypsin (EC 3.4.21.4) were more active in the peritrophic membrane (including food contents) than in the midgut epithelium of both species. Chymotrypsin activity in 11-d-old third and fourth instars of Ae. albopictus was 28 times higher than in the corresponding stadia of Ae. aegypti, indicating that the former species may have a superior enzymatic process for digesting food proteins.     

Experimental infection and transmission of Barmah Forest virus by Aedes vigilax (Diptera: Culicidae)

Aedes vigilax (Skuse) mosquitoes colonized from Townsville, Queensland, Australia, were fed on blood containing Barmah Forest virus (BF) isolated from the same species. The colony was susceptible to infection, with an ID50 of 10(2.6) CCID50 per mosquito. Infection and transmission rates for mosquitoes fed 10(3.5) CCID50 virus per mosquito varied from 58 to 100% and 36 to 100%, respectively, between days 3 and 13 after infection. Titers in infected mosquitoes were high by 5 d after infection and did not vary substantially thereafter. Virus transmission to suckling mice occurred from 3 d after infection. This work establishes that Ae. vigilax is a competent vector of BF.