Detection of Avian Plasmodium spp. DNA Sequences from Mosquitoes Captured in Minami Daito Island of Japan
Hiroko EjiriYukita SatoEmi SasakiDaisuke SUMIYAMAYoshio TsudaKyoko SawabeShin MatsuiSayaka HorieKana AkataniMasaoki TakagiSumie OmoriKoichi MurataMasayoshi Yukawa
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Abstract:
Several species of birds in Minami Daito Island, an oceanic island located in the far south from the main islands of Japan, were found to be infected with avian Plasmodium. However, no vector species of the avian malaria in this island have been revealed yet. To speculate potential vectors, we collected mosquitoes there and investigated using a PCR procedure whether the mosquitoes harbor avian malaria or not. Totally 1,264 mosquitoes including 9 species were collected during March 2006 to February 2007. The mosquitoes collected were stored every species, sampled date and location for DNA extraction. Fifteen out of 399 DNA samples showed positive for the partial mtDNA cytb gene of avian Plasmodium. Estimated minimum infection rate among collected mosquitoes was 1.2% in this study. Four species of mosquitoes; Aedes albopictus, Culex quinquefasciatus, Lutzia fuscanus and Mansonia sp. had avian Plasmodium gene sequences. Detected DNA sequences from A. albopictus and L. fuscanus were identical to an avian Plasmodium lineage detected in bull-headed shrike (Lanius bucephalus) captured in the island. Different sequences were detected from C. quinquefasciatus, which were corresponding to an avian Plasmodium from a sparrow (Passer montanus) and Plasmodium gallinaceum. Our results suggest that A. albopictus, Lutzia fuscanus, C. quinquefasciatus, and Mansonia sp. could be potential vectors of avian malaria in Minami Daito Island. This study was the first report of molecular detection of avian Plasmodium from mosquitoes in Japan.Keywords:
Avian Malaria
Plasmodium gallinaceum
Plasmodium (life cycle)
Aedes albopictus
Culex quinquefasciatus
Autotroph
P-Aminobenzoic acid
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Several species of birds in Minami Daito Island, an oceanic island located in the far south from the main islands of Japan, were found to be infected with avian Plasmodium. However, no vector species of the avian malaria in this island have been revealed yet. To speculate potential vectors, we collected mosquitoes there and investigated using a PCR procedure whether the mosquitoes harbor avian malaria or not. Totally 1,264 mosquitoes including 9 species were collected during March 2006 to February 2007. The mosquitoes collected were stored every species, sampled date and location for DNA extraction. Fifteen out of 399 DNA samples showed positive for the partial mtDNA cytb gene of avian Plasmodium. Estimated minimum infection rate among collected mosquitoes was 1.2% in this study. Four species of mosquitoes; Aedes albopictus, Culex quinquefasciatus, Lutzia fuscanus and Mansonia sp. had avian Plasmodium gene sequences. Detected DNA sequences from A. albopictus and L. fuscanus were identical to an avian Plasmodium lineage detected in bull-headed shrike (Lanius bucephalus) captured in the island. Different sequences were detected from C. quinquefasciatus, which were corresponding to an avian Plasmodium from a sparrow (Passer montanus) and Plasmodium gallinaceum. Our results suggest that A. albopictus, Lutzia fuscanus, C. quinquefasciatus, and Mansonia sp. could be potential vectors of avian malaria in Minami Daito Island. This study was the first report of molecular detection of avian Plasmodium from mosquitoes in Japan.
Avian Malaria
Plasmodium gallinaceum
Plasmodium (life cycle)
Aedes albopictus
Culex quinquefasciatus
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Plasmodium gallinaceum
Avian Malaria
Culex quinquefasciatus
Plasmodium (life cycle)
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Development of effective strategies to control malaria transmission is an important issue in malaria research. This study aimed to investigate refractoriness of laboratory and field strains of the malaria vector Culex quinquefasciatus to the avian malaria parasite Plasmodium gallinaceum , a model of the human malaria disease. Transmission potential was determined by feeding batches of Cx. quinquefasciatus on P. gallinaceum infected chickens with 10% and 30% parasitemia. The mosquitoes were examined for percent infectivity and numbers of oocysts. Our study showed that when the mosquitoes were fed on the infected chickens with high parasitemia, the laboratory isolate of Cx. quinquefasciatus was more susceptible to P. gallinaceum transmission and produced significantly higher oocyst numbers than the field isolate. There were, however, no differences in term of the transmission potential and infectivity of the malaria parasite when the mosquitoes were allowed to feed on the P. gallinaceum -infected chickens with low parasitemia. In conclusion, our study demonstrated the differences in refractoriness of the natural vector Cx. quinquefasciatus to the transmission of the avian malaria P. gallinaceum , implying the existence of the natural pathogen resistance mechanisms in the Culex mosquito.
Plasmodium gallinaceum
Avian Malaria
Culex quinquefasciatus
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To identify potential vectors of avian malaria in Hawaiian native forests, the innate susceptibility of Aedes albopictus, Wyeomyia mitchellii, and Culex quinquefasciatus from 3 geographical sites along an altitudinal gradient was evaluated using local isolates of Plasmodium relictum. Mosquitoes were dissected 5–8 and 9–13 days postinfective blood meal and microscopically examined for oocysts and salivary-gland sporozoites. Sporogony was completed in all 3 species, but prevalence between species varied significantly. Oocysts were detected in 1–2% and sporozoites in 1–7% of Aedes albopictus that fed on infected ducklings. Wyeomyia mitchellii was slightly more susceptible, with 7–19% and 7% infected with oocysts and sporozoites, respectively. In both species, the median oocyst number was 5 or below. This is only the second Wyeomyia species reported to support development of a malarial parasite. Conversely, Culex quinquefasciatus from all 3 sites proved very susceptible. Prevalence of oocysts and sporozoites consistently exceeded 70%, regardless of gametocytemia or origin of the P. relictum isolate. In trials for which a maximum 200 oocysts were recorded, the median number of oocysts ranged from 144 to 200. It was concluded that Culex quinquefasciatus is the primary vector of avian malaria in Hawai'i.
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Culex quinquefasciatus
Avian Malaria
Plasmodium (life cycle)
Blood meal
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Aedes aegypti, infected with West Nile virus by imbibing suspension of the virus through membrane, developed virus concentrations up to 10?5 mouse i.c. LD5o. The virus had no effect on the susceptibility of the mosquitoes to subsequent infection with Plasmodium gallinaceum, or on the ability of the mosquitoes to transmit the malaria parasites by bite. Mosquitoes with an infection of P. gallinaceum oocysts on the gut wall were also capable of becoming infected by feeding on West Nile virus suspension and of developing high virus titers. However, in these experiments, fewer of the malariainfected mosquitoes became infected with or supported multiplication of virus than did mosquitoes which had not been infected 5 to 7 days previously with the malaria parasites. Malaria and many arboviruses are transmitted by mosquitoes, and the question arises as to whether certain combinations of virus and Plasmodium may interfere with one another in given host. Trager (1959) discovered virus which inhibited the development of Plasmodium lophurae Coggeshall in ducklings. Barnett (1956) observed that concurrent infection of canaries with P. relictum (Grassi and Feletti) and western equine encephalitis virus suppressed the level of viremia in these hosts. However, there was no apparent effect of the virus on the ability of Culex tarsalis Coq. to transmit the malaria parasite. More recently de Zulueta and coauthors (1962) postulated that interference between malaria parasites of man and the virus of O'Nyong-nyong, both transmitted by Anopheles funestus Giles and A. gambiae Giles, could have considerable epidemiological importance. They suggested the possibility that the extensive outbreaks of the viral disease in East Africa could have been responsible for great reduction in malaria The possibility that virus infection could suppress development of malaria parasites in mosquito was investigated extensively by Bertram, Varma, and Baker (1964), with Ae. aegypti, P. gallinaceum, and Semliki Forest virus as the experimental organisms. These authors showed that the mosquito could become doubly infected with the virus and the malaria parasite, and could transmit both Received for publication 20 January 1966. * This work was supported in part by research grant No. AI-00351-13 from the NIH, U. S. Public Health Service. simultaneously. However, they also observed sm ll but more or less consistently lower malaria indices in the doubly infected mosquitoes, and, in one experiment, there was an unusually high mortality in virus-infected mosquitoes short time after they had fed on malariapositive chick. The authors concluded that a malaria parasite can be, at least partially, suppressed in its intrinsic development by the presence of an arbovirus in vector. They point out, however, that such interference need not occur inevitably, nor on detectable scale, nor sufficiently to reduce malaria transmission. Aedes aegypti is an efficient host for P. gallinaceum and West Nile virus. The purpose of our study was to determine whether prior infection of the mosquito with the virus would have suppressant effect on subsequent infection with the malaria parasite, or, conversely, whether malaria-infected mosquitoes are less susceptible to subsequent infection with the virus. MATERIALS AND METHODS The Ae. aegypti were obtained from our laboratory colony; the P. gallinaceum has been maintained in our laboratory by blood transfer from chicken to chicken; the virus was strain of West Nile 101 in mouse brain suspension. In Experiment 2, the virus was brain suspension harvested from mice infected in the course of mosquito titrations; hence, this pool is first mouse brain passage following isolation from mosquitoes. Both pools were equally infectious to Ae. aegypti. The mosquitoes were infected with P. gallinaceum by allowing them to feed on chicken showing high gametocyte count. The fully engorged insects were selected for further use. They were infected with the virus by allowing them to feed through mem-
Plasmodium gallinaceum
Avian Malaria
West Nile virus
Plasmodium (life cycle)
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Avian Malaria
Plasmodium gallinaceum
Culex quinquefasciatus
Plasmodium (life cycle)
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Plasmodium gallinaceum
Artesunate
Avian Malaria
Culex quinquefasciatus
Plasmodium (life cycle)
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SummaryArmigeres and Aedes species of mosquitoes have been infected with, and have transmitted, P. gallinaceum of fowls very easily in our laboratory experiments. But it has not been possible to infect an anopheline, even when fed simultaneously on a fowl whose gametocytes were of a quantity and quality adequate to cause 100 per cent infections in Armigeres and Aedes. With the exception of oocysts in one Culex mimuloides the same difficulty in infecting Culex species has been encountered.It does not appear that local species of Anopheles or Culex genera are natural hosts for this plasmodium.
Plasmodium gallinaceum
Avian Malaria
Plasmodium (life cycle)
Mansonia
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SummaryIn a study to determine the natural vectors of avian Plasmodium infection in Kern County, California 3,364 mosquitoes were examined for parasites during the summers of 1946, 1947, 1949 and 1950. The following average infection rates were demonstrated: Culex tarsalis, 199 of 2,074 (9.6 per cent); Culex stigmatosoma, 28 of 180 (15.6 per cent); and Culex quinquefasciatus, 14 of 746 (1.9 per cent). These differences are believed to reflect variation in vector efficiency. Of 364 Anopheles and Aedes examined, only 2 Anopheles franciscanus were infected with what appeared to be young oocysts of unidentified parasites.The intensity of infection in the principal vector, Culex tarsalis, varied considerably in different years and in different areas. These variations are believed to be due to changeable factors such as climate, the effect of mosquito control on the vector population, and ecological differences between areas.Plasmodium relictum was isolated from naturally infected Culex tarsalis and Culex stigmatosoma.
Culex quinquefasciatus
Avian Malaria
Plasmodium (life cycle)
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