The first detection of avian haemosporidia from Culicoides biting midges in Japan, with notes on potential vector species and the transmission cycle.

BACKGROUND: Biting midges (Ceratopogonidae) are capable of transmitting a variety of pathogens including viruses, trypanosomes and haemosporidia. The majority of Haemoproteus parasites are transmitted by biting midges predominantly of the genus Culicoides and are known to cause significant physical and reproductive impacts on both wild and domestic birds. In Japan, Haemoproteus had been detected from various avian hosts, but not from arthropod vectors. In this study, we investigated the prevalence of avian haemosporidia at an educational forest in central Japan in attempt to reveal possible vector species of Haemoproteus, which would help to better understand the transmission cycle of Haemoproteus within Japan and to develop preventative measures for captive and domestic birds. METHODS: Biting midges were caught using UV light traps from 2016 to 2018. The collected samples were morphologically identified, and haemosporidian parasites were detected using PCR-based methods. The detected lineages were phylogenetically analyzed and compared with lineages previously detected from birds. Bloodmeal analyses were also carried out for part of the blood-fed individuals. RESULTS: Six Haemoproteus lineages were detected from 17 of 1042 female Culicoides (1.63%), including three species (C. sigaensis, C. arakawae, and C. pictimargo) in which Haemoproteus was detected for the first time. All detected lineages were placed in the subgenus Parahaemoproteus clade and were previously detected from crows of central Japan, strongly suggesting that parasites of these genetic lineages are transmitted between Culicoides and crows. Two Plasmodium lineages were also detected but are thought to be transmitted between Culex mosquitoes and birds of the educational forest based on previous detections. No amplifications were seen in bloodmeal analysis, possibly due to insufficient amount of blood, denaturation via digestion, or insufficient detectability of the used protocol. CONCLUSION: Haemoproteus DNA was detected from Culicoides for the first time in Japan, suggesting that transmission is possible within the country. These findings highlight the necessity to investigate Culicoides populations and Haemoproteus infections dynamics in Japan. However, vector competence could not be confirmed in this study and further studies are anticipated.

A long-term field study on mosquito vectors of avian malaria parasites in Japan.

Avian malaria is a mosquito-borne disease of birds caused by avian Plasmodium spp. in worldwide scale. Some naïve birds show serious symptoms which can result in death. Surveillance of vectors and parasites are important to understand and control this disease. Although avian malaria has been found in Japan, detailed prevalence and dynamics remained understudied. We aimed to observe annual changes in the abundance of mosquitoes and the prevalence of avian Plasmodium parasites in Japan. Mosquitoes were collected using dry ice traps over a 10-year period, at a fixed research area located in Kanagawa prefecture. Collected mosquitoes were investigated for the species composition, population size and prevalence of avian Plasmodium by PCR. Mosquitoes belonging to 13 species in 7 genera were collected (n=8,965). The dominant species were Aedes (Ae.) albopictus and Culex (Cx.) pipiens group (gr.). Seven avian Plasmodium lineages, all of which were previously known, were detected from Cx. pipiens gr., Ae. albopictus, and Tripteroides bambusa. Three genetic lineages were dominant and were probably transmitted by Cx. pipiens gr. whose could be the primary vector of these parasites. Annual variations in the seasonal prevalence of mosquitoes and avian Plasmodium were revealed for the first time during recent 10 years in Japan. Namely, avian Plasmodium occurrence in the vector population peaked often in June to July and September to October when the density of the vector population was presumably high enough for the transmission of avian Plasmodium upon appearance of infected birds.