Yukiko Higa

Dengue Vectors and their Spatial Distribution.

The distribution of dengue vectors, Ae. aegypti and Ae. albopictus, is affected by climatic factors. In addition, since their life cycles are well adapted to the human environment, environmental changes resulting from human activity such as urbanization exert a great impact on vector distribution. The different responses of Ae. aegypti and Ae albopictus to various environments result in a difference in spatial distribution along north-south and urban-rural gradients, and between the indoors and outdoors. In the north-south gradient, climate associated with survival is an important factor in spatial distribution. In the urban-rural gradient, different distribution reflects a difference in adult niches and is modified by geographic and human factors. The direct response of the two species to the environment around houses is related to different spatial distribution indoors and outdoors. Dengue viruses circulate mainly between human and vector mosquitoes, and the vector presence is a limiting factor of transmission. Therefore, spatial distribution of dengue vectors is a significant concern in the epidemiology of the disease. Current technologies such as GIS, satellite imagery and statistical models allow researchers to predict the spatial distribution of vectors in the changing environment. Although it is difficult to confirm the actual effect of environmental and climate changes on vector abundance and vector-borne diseases, environmental changes caused by humans and human behavioral changes due to climate change can be expected to exert an impact on dengue vectors. Longitudinal monitoring of dengue vectors and viruses is therefore necessary.

Effects of temperature and diet on development and interspecies competition in Aedes aegypti and Aedes albopictus

We asked whether climate change might affect the geographic distributions of Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae). We tested the effects of temperature, diet and the presence of congeneric species on the performance of immature stages of these two aedine species in the laboratory. Mosquitoes in three different species‐density combinations were reared at four constant temperatures (20 °C, 25 °C, 30 °C, 35 °C) on low‐ or high‐level diets. Of the four temperatures tested, mortality increased only at 35 °C in both species. Mortality was higher on the high‐level diet than on the low‐level diet at 35 °C, but not at other temperatures. The presence of congeneric species had a significant positive effect on mortality in Ae. albopictus, but not in Ae. aegypti. Both species developed more quickly at higher temperatures within the range of 20–30 °C; development was not enhanced at 35 °C. Population growth of Ae. albopictus was more stable, regardless of diet and temperature; that of Ae. aegypti varied more according to these two factors. These species‐specific attributes may help to explain the latitudinal distribution of the mosquitoes and degree of species dominance where they are sympatric.

Geographic Distribution of Aedes aegypti and Aedes albopictus Collected from Used Tires in Vietnam

Abstract The spatial distribution of Aedes aegypti and Aedes albopictus in environmental and geographical zones, e.g., urban-rural, coastal-mountainous, and north-south, was investigated throughout Vietnam. Immature stages were collected from used tires along roads. The effects of regions, seasons, and the degree of urbanization on the density and the frequency were statistically analyzed. Aedes aegypti predominated in the southern and central regions, while Ae. albopictus predominated in the northern region, which may be related to climatic conditions (temperature and rainfall). Larval collection from used tires may be suitable to assess rapidly the current distribution of dengue mosquitoes for estimating health risks and implementing vector control measures.

High potential risk of dengue transmission during the hot-dry season in Nha Trang City, Vietnam.

We visited houses and inspected water-holding containers to determine the potential risks of dengue transmission during different seasons. This survey was conducted in two neighbourhoods of Nha Trang City in July and December 2006, which correspond to the middle of the hot-dry season and the beginning of the cool-wet season, respectively. We inspected a total of 1438 wet containers in 196 premises during both survey periods; 20% of the containers were positive for Aedes aegypti larvae and 8% for A. aegypti pupae. Indoor water-holding containers, which were sparsely distributed, exhibited high pupal productivity and efficiency (pupal productivity of a type of container/prevalence of that type of container) in either the first survey conducted in July, or the second, conducted in December. Although rainfall may not influence the number and distribution of water-holding containers in the city, the high average temperature in the first survey period resulted in a higher potential risk of dengue transmission. Our analysis suggests that if intensive source reduction is conducted in summer and containers with high pupal productivity and efficiency are targeted, the risk of dengue transmission in the city could be effectively reduced.

Insecticide Resistance in Potential Vector Mosquitoes for West Nile Virus in Japan

Abstract Culex pipiens complex is the significant vector mosquito of West Nile virus. To take stock of the current situation of insecticide susceptibilities and design an ideal mosquito control strategy, we collected Culex pipiens pallens Coquillet, Culex pipiens form molestus Forskal, and Culex quinquefasciatus Say from fields in Japan and conducted bioassays for five larvicides (fenitrothion, temephos, etofenprox, diflubenzuron, and pyriproxyfen) by using a larval dipping method. Among five insecticides tested, obvious reduced susceptibilities were observed for etofenprox, which is the only pyrethroid compound registered as a larvicide in Japan. Twenty-two of 56 colonies exhibited a >10% survival rate at the etofenprox concentration of 5.7 μg/ml, which is a 10 times higher concentration of the working solution. The LC50 of a colony collected from Fukuoka prefecture for etofenprox exceeded 60 μg/ml (resistance ratio >2,307), and this colony also exhibited cross-resistance to other pyrethroids, permethrin (299-fold) and phenothrin (1,200-fold). The insect growth regulators diflubenzuron and pyriproxyfen were found to be sufficiently effective enough to control Culex larvae present, but decreased sensitivities to these insecticides were slightly detected in some colonies of Cx. p. form molestus collected from urban areas. Several etofenprox-resistant colonies of Cx. p. form molestus exhibited simultaneously decreased susceptibilities to other insecticides, including temephos, diflubenzuron, and pyriproxyfen.

The Effect of Pyriproxyfen as a “Population Growth Regulator” against Aedes albopictus under Semi-Field Conditions

An insect growth regulator, pyriproxyfen, has been used for the control of a range of pest insects, including mosquitoes. Pyriproxyfen is effective in inhibiting adult emergence and sterilizing adult females. The Asian tiger mosquito, Aedes albopictus (Skuse), is an important vector of dengue and chikungunya, and is expanding its distribution throughout Europe and the Americas. In the present study, we evaluated the impact of pyriproxyfen-treated bed nets on population growth of Ae. albopictus under semi-field conditions, using 6 small microcosms. We created microcosms containing breeding sites to simulate the natural ecosystem of vector mosquito and installing miniature bed net treated with 350 mg/m2 pyriproxyfen in Experiment I and 35 mg/m2 in Experiment II. For each experiment, we also established microcosms installing untreated polyethylene net (untreated control). The installing nets were provided with artificially torn holes, to simulate damage and allow mosquitoes to penetrate. We released 100 pairs of Ae. albopictus into each microcosm, and allowed them to feed on a mouse under the bed nets at approximately 1-week intervals. In comparison with the untreated control microcosms, the number of eggs laid by the released adults in the pyriproxyfen-treated microcosms was significantly lower in both Experiment I and II. Moreover, egg hatchability was significantly suppressed and pupal mortality was increased. Our results indicate that tarsal contact with pyriproxyfen has been shown to suppress egg production and hatchability in adult females and the auto-dissemination of pyriproxyfen into larval breeding sites by adult mosquitoes, through contact with pyriproxyfen-treated polyethylene bed nets, may suppress the mosquito population density.

Nationwide investigation of the pyrethroid susceptibility of mosquito larvae collected from used tires in Vietnam.

Pyrethroid resistance is envisioned to be a major problem for the vector control program since, at present, there are no suitable chemical substitutes for pyrethroids. Cross-resistance to knockdown agents, which are mainly used in mosquito coils and related products as spatial repellents, is the most serious concern. Since cross-resistance is a global phenomenon, we have started to monitor the distribution of mosquito resistance to pyrethroids. The first pilot study was carried out in Vietnam. We periodically drove along the national road from the north end to the Mekong Delta in Vietnam and collected mosquito larvae from used tires. Simplified susceptibility tests were performed using the fourth instar larvae of Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus. Compared with the other species, Ae. aegypti demonstrated the most prominent reduction in susceptibility. For Ae. aegypti, significant increases in the susceptibility indices with a decrease in the latitude of collection points were observed, indicating that the susceptibility of Ae. aegypti against d-allethrin was lower in the southern part, including mountainous areas, as compared to that in the northern part of Vietnam. There was a significant correlation between the susceptibility indices in Ae. aegypti and the sum of annual pyrethroid use for malaria control (1998–2002). This might explain that the use of pyrethroids as residual treatment inside houses and pyrethroid-impregnated bed nets for malaria control is attributable to low pyrethroid susceptibility in Ae. aegypti. Such insecticide treatment appeared to have been intensively administered in the interior and along the periphery of human habitation areas where, incidentally, the breeding and resting sites of Ae. aegypti are located. This might account for the strong selection pressure toward Ae. aegypti and not Ae. albopictus.

MOSQUITO BIODIVERSITY PATTERNS AROUND URBAN ENVIRONMENTS IN SOUTH-CENTRAL OKINAWA ISLAND, JAPAN

Abstract Okinawa is the largest, most urbanized, and densely populated island in the Ryukyus Archipelago, where mosquito species diversity has been thoroughly studied. However, the south-central Okinawa mosquito fauna has been relatively poorly studied. Here, we present results from a mosquito faunal survey in urban environments of Nishihara city, south-central Okinawa. Mosquitoes were sampled biweekly, from April 2007 to March 2008, at 3 different environments: a forest preserve, an animal farm, and a water reservoir. We employed 4 mosquito collection methods: 1) oviposition traps; 2) light traps; 3) sweep nets; and 4) larval surveys of tree holes, leaf axils, and artificial water containers. We collected a total of 568 adults and 10,270 larvae belonging to 6 genera and 13 species, including 6 species of medical importance: Aedes albopictus, Armigeres subalbatus, Anopheles Hyrcanus group, Culex bitaeniorhynchus, Cx. quinquefasciatus, and Cx. tritaeniorhynchus. Mosquito species composition was similar to data from previous studies in Okinawa Island. The flattening of the species accumulation curve suggests that our diversity sampling was exhaustive with light and oviposition traps, as well as the coincidence between the species richness we found in the field and estimates from the Chao2 index, a theoretical estimator of species richness based on species abundance. This study highlights the importance of combining several sampling techniques to properly characterize regional mosquito fauna and to monitor changes in the presence of mosquito species.

Effect of peridomestic environments on repeated infestation by preadult Aedes aegypti in urban premises in Nha Trang City, Vietnam.

To determine the effect of peridomestic environments on Aedes aegypti infestation in urban premises, we conducted two consecutive surveys in the hot-dry and cool-wet seasons. Most Ae. aegypti pupae (79%) were recovered from premises where preadult forms (larvae and/or pupae) had been detected in both surveys. Hence, repeated infestation appears to be a useful parameter to identify premises associated with a high potential risk of dengue transmission. Multivariate analysis revealed that not only the persistent presence of containers discarded outdoors, wells, large plastic buckets, jars, and concrete toilet basins in the premises (adjusted odds ratios [aORs] = 63.3, 23.3, 22.5, 6.6, and 5.6, respectively) but also the presence of six or more residents was significantly associated with repeated infestation (aOR = 6.1). Premises with six or more residents along with specific container types from which a large number of pupae were recovered should be targeted in dengue-control programs.

Surveillance of Japanese Encephalitis Virus Infection in Mosquitoes in Vietnam from 2006 to 2008

Japanese encephalitis virus (JEV) infection in mosquitoes was monitored in Vietnam from 2006 to 2008. A total of 15,225 mosquitoes, identified as 26 species in five genera were collected and 12,621 were grouped into 447 pools for examination of JEV infection by assays for cytopathic effects in C6/36 cells and by RT-PCR to detect flavivirus RNA. Three JEV strains were isolated from Culex tritaeniorhynchus Giles collected in northern and southern Vietnam and two JEV strains were isolated from Culex vishnui Theobald collected in the highlands of Vietnam. Genetic and phylogenetic analyses, based on complete E gene nucleotide sequences, revealed that the five JEV strains were classified into the genotype I group and six amino acid differences were found in these five strains. These results indicated that multiple JEV genotype I populations are circulating countrywide in Vietnam, transmitted by bites of their Cx. tritaeniorhynchus and Cx. vishnui.