Yoshio Tsuda

Different spatial distribution of Aedes aegypti and Aedes albopictus along an urban-rural gradient and the relating environmental factors examined in three villages in northern Thailand.

ABSTRACT A larval survey of dengue vectors was conducted from July to November 1996 and from May to November 1997 in Chiangmai Province, Thailand. Three villages in urban, transition, and rural areas were selected for the survey to clarify the spatial distribution of Ae. aegypti and Ae. albopictus along an urban–rural ecological gradient. The average number of Ae. aegypti larvae in larvitraps was higher in the urban area than in the rural area, as we expected, whereas the opposite was found for Ae. albopictus, rural area > urban area. A house survey of larvae-inhabiting containers showed significant differences in the number and composition of these containers among the study areas. Significant differences also were found in the average distance between houses, average tree height, and average percentage of vegetation cover for each house. The seasonal pattern of rainfall recorded in each study area did not show great differences among the study areas. The response of Ae. aegypti and Ae. albopictus to the urban–rural gradient is discussed in relation to the possibility of applying geographic information system techniques to plan the control strategy and surveillance of dengue vectors.

Heterogeneity in the host preference of Japanese encephalitis vectors in Chiang Mai, northern Thailand.

Experiments, using the capture-mark-release-recapture technique inside large nets, were carried out in Chiang Mai, northern Thailand, to examine heterogeneity in the host preference of Japanese encephalitis (JE) vectors. A significantly higher proportion of the vector species that were initially attracted to a cow fed when released into a net with a cow than when released into a net containing a pig. However, Culex vishnui individuals that had been attracted to a pig had a higher feeding rate in a net containing a pig rather than a cow. When mosquitoes were given a choice by being released into a net containing both animals, they exhibited a tendency to feed on the host to which they had originally been attracted. This feeding preference was, however, not shown by the offspring of pig-fed individuals. We have therefore shown evidence of physiological/behavioural conditioning in the host preference of JE vectors rather than genetic variability. Our results suggest that effective control of JE might be achieved by increasing the availability of cows (the dead-end hosts of JE virus) to deflect the vectors from pigs (the amplifying host). The behavioural imprinting which we have found would tend to re-inforce the initial tendency of the vectors to bite cows.

Bloodmeal Identification and Detection of Avian Malaria Parasite From Mosquitoes (Diptera: Culicidae) Inhabiting Coastal Areas of Tokyo Bay, Japan

ABSTRACT Bloodmeal identification and the detection of avian malaria parasite from mosquitoes (Diptera: Culicidae) were carried out by polymerase chain reaction-based methods for field samples collected in coastal areas of Tokyo Bay, Japan, from April to October 2007. The following seven mosquito species were collected: Aedes albopictus (Skuse), Culex pipiens pollens Coquillett, Culex pipiens form molestus Forskal, Culex tritaeniorhynchus Giles, Culex inatomii Kamimura & Wada, Culex bitaeniorhynchus Giles, and Lutzia vorax Edwards. Forty blood-fed mosquitoes were collected and 95% of bloodmeals of Cx. pipiens pallens were avian-derived, whereas only mammalian bloodmeals were identified for Ae. albopictus. Plasmodium DNA was amplified from 65% (15/23) of blood-fed Cx. pipiens pallens and unfed females of Cx. pipiens pallens and Cx. pipiens form molestus with a minimum infection rate of 29.9 and 13.5, respectively. One unfed female of Lt. vorax was also positive for Plasmodium parasites. Five genetically distinct lineages of Plasmodium were identified, with 0.21 to 5.86% sequence divergence. Rinshi-8, the most prevalent lineage at our study site, was identical to the published sequence of Plasmodium relictum-P5.

Host-Feeding Habits of Culex pipiens and Aedes albopictus (Diptera: Culicidae) Collected at the Urban and Suburban Residential Areas of Japan

ABSTRACT To evaluate the vectorial capacity of mosquitoes for viruses in Japan, the host-feeding habits of the mosquitoes were analyzed by sequencing polymerase chain reaction-amplified fragments of the cytochrome b and 16S ribosomal RNA regions of the mitochondrial DNA of 516 mosquitoes of 15 species from seven genera that were collected from residential areas during 2003–2006. Culex pipiens L. and Aedes albopictus Skuse were the most commonly collected species in urban and suburban residential areas. Anautogenous Culex pipiens pallens Coquillett was distinguished from the autogenous Cx. pipiens form molestus Forskal using a polymerase chain reaction-based identification method. Both Cx. p. pollens and Cx. p. form molestus exhibited similar host-feeding habits, broadly preferring avian (50.0 and 42.5% of avian, respectively) and mammalian (38.6 and 45.0% of avian, respectively) hosts, such as tree sparrows, ducks, and humans. Conversely, Ae. albopictus exhibited a highly mammalophilic and anthropophilic feeding pattern, with 84.2% feeding on mammalian hosts and 68.5% of these on humans. We concluded that in Japan, Cx. pipiens might play a significant role in the avian-to-mammal transmission of viruses, such as West Nile virus, whereas Ae. albopictus might play a role in the human-human transmission of dengue and Chikungunya viruses.

Prevalence of avian malaria parasite in mosquitoes collected at a zoological garden in Japan

Several species of captive birds at zoological gardens of Japan were found to be infected with avian Plasmodium. However, incriminated vector mosquito species have not been identified yet. To indicate the competent vectors of avian malaria parasite, we collected mosquitoes at a zoological garden in Japan and examined for the avian malaria parasite DNA. Totally, 1,361 mosquitoes of 11 species were collected in the zoological garden of Kanagawa, the south of Tokyo in Japan in 2005. Captured mosquitoes were pooled by each species, date collected, and location and used for DNA extraction. Eight out of 169 DNA samples were positive for the nested PCR of avian Plasmodium cyt b gene. Estimated minimum infection rates of mosquitoes were 5.9 per 1,000. The PCR positive mosquito species were Culex pipiens group and Lutzia vorax. Some DNA sequences amplified from collected mosquitoes were identical to avian Plasmodium lineages detected from captive birds in the same zoological garden studied. Our results suggest that C. pipiens group and L. vorax could be incriminated vectors of avian malaria parasite transmitting in captive birds kept in the zoological garden in Japan.

Identification and molecular characterization of a new nonsegmented double-stranded RNA virus isolated from Culex mosquitoes in Japan.

Two infectious agents were isolated from Culex species mosquitoes in Japan and were identified as distinct strains of a new RNA virus by a method for sequence-independent amplification of viral nucleic acids. The virus designated Omono River virus (OMRV) replicated in mosquito cells in which it produced a severe cytopathic effect. Icosahedral virus particles of approximately 40 nm in diameter were detected in the cytoplasm of infected cells. The OMRV genome was observed to consist of a nonsegmented, 7.6-kb double-stranded RNA (dsRNA) and contain two overlapping open reading frames (ORFs), namely ORF1 and ORF2. ORF1 was found to encode a putative dsRNA-binding protein, a major capsid protein, and other putative proteins, which might be generated by co- and/or post-translational processing of the ORF1 polyprotein precursor, and ORF2 was found to encode a putative RNA-dependent RNA polymerase (RdRp), which could be translated as a fusion with the ORF1 product by a -1 ribosomal frameshift. Phylogenetic analysis based on RdRp revealed that OMRV is closely related to penaeid shrimp infectious myonecrosis virus and Drosophila totivirus, which are tentatively assigned to the family Totiviridae. These results indicated that OMRV is a new member of the family of nonsegmented dsRNA viruses infecting arthropod hosts, but not fungal or protozoan hosts.

Survival and Development of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) Larvae Under a Seasonally Changing Environment in Nagasaki, Japan

Abstract Survival and development of larvae of Aedes aegypti (L.) and Aedes albopictus (Skuse) were examined under field conditions on the campus of Nagasaki University School of Medicine, Nagasaki, Japan, to explore the possibility of the establishment of Ae. aegypti in Kyusyu, southern Japan. Exposure experiments with hatched larvae were conducted 11 times at about 1-mo intervals from January 1998 to January 1999. In both species, larvae could develop to adults under field conditions from April to November, but not in January, February, and December 1998. In March 1998, only Ae. albopictus could develop to adults. The results suggest that there are differences in the resistance of hatched larvae to low temperatures between Ae. aegypti and Ae. albopictus and a threshold air temperature of larval development below 10°C for both species. To estimate the threshold temperatures of larval development, the relationships between the average air temperature and the developmental rate observed in March–November 1998 were analyzed by linear regression analysis. The estimated threshold air temperatures for Ae. aegypti were 8.52 and 9.45°C for females and males, respectively, and were higher than those of Ae. albopictus. A clear seasonal change was also observed in the body size of pupae. In both species, body size was largest at the beginning of the breeding season and continuously became gradually smaller until July. The results were compared with those from previous field studies in Nagasaki and the possibility of the establishment of Ae. aegypti in Nagasaki, Japan, is discussed from the viewpoint of adaptive life history to seasonally changing environments.

Characteristics of Anopheles minimus (Diptera: Culicidae) Larval Habitats in Northern Thailand

Abstract The effects of breeding habitat characteristics on the larval density of Anopheles minimus Theobald were studied in a perennial stream in the foothills of Chiang Mai province, northern Thailand. Data on 41 different variables related to plant cover and stream physical attributes in 200 sections, each 10 m long, were collected along with larval data during the dry and wet seasons of 1998 and 1999. Larval density was significantly higher in the dry season than in the wet season. In the dry season, An. minimus density was negatively correlated with water current velocity, height of aquatic large-leaved plants, and height of riparian small-leaved plants; and positively correlated with the cover of riparian ferns. The dry season prediction model, which explained 51% of the variation of An. minimus, was as follows: ymin = 0.1980–0.1733*water velocity – 0.0317*height of aquatic large-leaved plants – 0.0249*height of riparian small-leaved plants + 0.0192*cover of riparian ferns – 0.0170*height of stream banks. The influence of vegetation characteristics on larval density may not be as large as previously assumed. We conclude that factors other than those measured here may account for a large part of the variation in larval density.

Movement of Aedes aegypti (Diptera: Culicidae) Released in a Small Isolated Village on Hainan Island, China

Abstract A mark-release-recapture experiment was conducted in a small isolated village on Hainan Island, China, to examine the dispersal and movement of adult Aedes aegypti (L.). Two cohorts of mosquitoes marked with uniquely colored fluorescent dye were released at two different sites and recaptured for 6 d at every house in the village using human bait collections. The distribution pattern of houses around release site affected dispersal. The recapture rate of females released at the center of the village was higher (3.49%) than females released at the edge of the village (2.47%). The average day of recapture differed significantly between sexes, but not cohorts. The average day of recapture of females and males released at the center was 2.5 and 1.54 d, respectively. The total number of mosquitoes recaptured was the greatest at premises near the release site, and decreased at a constant rate of 0.43–0.48 with increasing distance from the release site. The proportion of nulliparous females decreased during the first 4 d and proportion of females with developing or mature ovaries increased during the latter half of the experiment. The daily survival rate for females and males released at the center of the village was estimated by log-regression to be 0.763 and 0.52, respectively.

A laboratory and field evaluation of Macrocyclops distinctus, Megacyclops viridis and Mesocyclops pehpeiensis as control agents of the dengue vector Aedes albopictus in a peridomestic area in Nagasaki, Japan

Abstract The use of the cyclopoid copepods Macrocyclops distinctus (Richard) Megacyclops viridis (Jurine) and Mesocyclops pehpeiensis Hu (Cyclopoida: Cyclopidae) as biological control agents against the dengue vector Aedes albopictus (Skuse) (Diptera: Culicidae) was evaluated. In the laboratory their predatory ability was highest against the younger instars of Ae. albopictus and none of the three copepods killed the fourth instar. Except for M. viridis, predatory ability was affected by the size of the container: the smaller the container, the higher the predation. A 4‐month field test was conducted to examine the impact of these predators on wild Ae. albopictus. Thirty artificial containers were placed in a peridomestic area to allow Ae. albopictus colonization. We showed continuous and similar oviposition responses in treated and control containers. The densities of Ae. albopictus showed considerable short‐term changes and were much reduced by the copepod species. Macrocyclops and the mixture of all three provided better Ae. albopictus control than either Megacyclops or Mesocyclops alone. When larval densities peaked in the control containers in August and September, the overall reduction due to the copepods was nearly complete. Mesocyclops inoculated alone had the highest population survival. However, the growth and survival of all the copepod species was poor when the three genera were mixed. Based on their performance and survival in the trial, Macrocyclops and Mesocyclops merit consideration as bio‐control agents of Ae. albopictus.