Abstract
One of vector-borne avian protozoa, Leucocytozoon lovati, has been found in the Japanese rock ptarmigans (Lagopus mutus japonicus), the endangered bird species distributed in the alpine regions in Japan. Vector arthropod species of L. lovati has also been estimated as Simuliidae black flies distributed in the same habitat of the host bird, however, possible blood meals of the black flies were not identified yet. To reveal host animals of black flies, we estimated the blood resources by using molecular techniques. Black flies were collected at Mt. Chogatake, one of the alpine regions of Japan in which Japanese rock ptarmigans live in June 2005. The analyzed 144 specimens were morphologically identified into five species including Simulium japonicum (n = 87), Prosimulium hirtipes (n = 48), Prosimulium yezoense (n = 3), Twinnia japonensis (n = 3), and Cnephia mutata (n = 3). Individually extracted DNA from the black flies was subjected to polymerase chain reaction (PCR) amplification targeting the partial mitochondrial cytochrome b gene of birds or mammals to identify the blood meals. Of 144 black flies examined, 34 specimens were PCR positive for avian hosts (23.6%). No mammalian-derived bloods were detected from the samples studied through. Sequences amplified from 11 black flies consist of S. japonicum, P. hirtipes, and C. mutata showed high similarity to that of the Japanese rock ptarmigan. Therefore, present results conclusively suggest that these three species of black flies might suck the bloods of Japanese rock ptarmigans and could be the vector for L. lovati infection among this endangered bird species of Japan.
References
Akiba K (1960) Studies on the Leucocytozoon found in the chicken, in Japan: II. On the transmission of L. caulleryi by Culicoides arakawae. Jpn J Vet Sci 22:309–319
Bennett GF, Peirce MA, Earle RA (1994) An annotated checklist of the valid avian species of Haemoproteus, Leucocytozoon (Apicomplexa: Haemosporida) and Hepatozoon (Apicomplexa: Haemogregarinidae). Syst Parasitol 29:61–73
Clarke CHD (1935) Blood parasites of ruffed grouse (Bonase umbellus) and spruce grouse (Canachites canadensis), with description of Leucocytozoon bonase n. sp. Can J Res 12:646–650
Dunbar MR, Tornquist S, Giordano MR (2003) Blood parasites in sage-grouse from Nevada and Oregon. J Wildl Dis 39:203–208
Fukuda M, Bain O, Aoki C, Otsuka Y, Takaoka H (2005) Natural infection of Simulium (Nevermannia) with infective filarial larvae, probably from a bird, in Oita, Japan. Med Entomol Zool 56:93–98
Hagihara M, Yamaguchi T, Kitahara M, Murata K (2004) Leucocytozoon lovati infections in wild rock Ptarmigan (Lagopus mutus) in Japan. J Wildl Dis 40:804–807
Hellgren O, Bensch S, Malmqvist B (2008) Bird hosts, blood parasites and their vectors associations uncovered by molecular analyses of blackfly blood meals. Mol Ecol 17:1605–1613
Ito A, Gotanda T (2004) Field efficacy of recombinant R7 vaccine against chicken Leucocytozoonosis. J Vet Med Sci 66:483–487
Malmqvist B, Strasevicius D, Hellgren O, Aldler P, Bensch S (2004) Vertebrate host specificity of wild-caught blackflies revealed by mitochondrial DNA in blood. Proc Biol Sci 271:152–155
Massey B, Greeson M, Slaney D, Tompkins DM (2007) PCR detection of Plasmodium and blood meal identification in a native New Zealand mosquito. J Vector Ecol 32:154–156
Molaei G, Andreadis TG, Armstrong PM, Anderson JF, Vossbrinck CR (2006) Host feeding patterns of Culex mosquitoes and West Nile virus transmission, northeastern United States. Emerg Infect Dis 12:468–474
Murata K (2002) Prevalence of blood parasites in Japanese wild birds. J Vet Med Sci 64:785–790
Murata K, Tamada A, Ichikawa Y, Hagihara M, Sato Y, Nakamura H, Nakamura M, Sakanakura T, Asakawa M (2007) Geographical distribution and seasonality of the prevalence of Leucocytozoon lovati in Japanese rock ptarmigans (Lagopus mutus japonicus) found in the alpine regions of Japan. J Vet Med Sci 69:171–176
Norris K, Anwar M, Read AF (1994) Reproductive effort influences the prevalence of haematozoan parasites in great tits. J Anim Ecol 63:601–610
Sasaki H, Nishijima Y, Ono H (1985) Studies on the relation between blood sources and the shape of fore tarsal claw of the female black flies in Hokkaido. J Coll Dairying 11:187–192
Sasaki H, Nishijima Y, Ono H (1988) Blood sources of two major Simulium species in Hokkaido, Japan (Diptera, Simuliidae). Jpn J Sanit Zool 39:87–90
Sato Y, Tamada A, Mochizuki Y, Nakamura S, Okano E, Yoshida C, Ejiri H, Omori S, Yukawa M, Murata K (2009) Molecular detection of Leucocytozoon lovati from probable vectors, black flies (Simuliidae) collected in the alpine regions of Japan. Parasitol Res 104:251–255
Takaoka H, Okazawa T (1988) A revised list of Japanese blackflies (Simuliidae). Med Entomol Zool 39:97–103 in Japanese with English summary
Takaoka H, Baba M, Bain O (1989) Natural infection of Simulium bidentatum (Diptera: Simuliidae) with larvae of Onchocerca spp., in relation to a human zoonotic onchocerciasis in Oita, Japan. Jpn J Trop Med Hyg 17:279–284
Valkiunas G (2005) Avian malaria parasites and other Haemosporidia. CRC, Boca Raton
Acknowledgments
This study was partially supported by a Grant-in-Aid for Scientific Research (no. 21580406) from the Japan Society for the Promotion of Science and the Academic Frontier Project “Surveillance and control for zoonoses”, Strategic Research Base Development Program "International research on epidemiology of zoonoses and training for young researchers", and “High-Tech Research Center” Project for Private Universities: matching fund subsidy from Ministry of Education, Culture, Sports, Science and Technology of Japan, Global Environment Research Fund of the Ministry of the Environment of Japan (F-062), and Nihon University Research Grants.
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Imura, T., Sato, Y., Ejiri, H. et al. Molecular identification of blood source animals from black flies (Diptera: Simuliidae) collected in the alpine regions of Japan. Parasitol Res 106, 543–547 (2010). https://doi.org/10.1007/s00436-009-1667-7
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DOI: https://doi.org/10.1007/s00436-009-1667-7