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Synchronous Langat Virus Infection of Haemaphysalis longicornis Using Anal Pore Microinjection

Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan
Department of Pathological and Preventive Veterinary Science, The United Graduate School of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi 753-8515, Japan
Laboratory of Public Health, Faculty of Veterinary Medicine, Hokkaido University, Kita-ku Kita-18 Nishi-9, Sapporo, Hokkaido 060-0818, Japan
Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, Los Baños, Laguna 4031, Philippines
Department of Clinical and Population Health, College of Veterinary Medicine and Biomedical Sciences, Cavite State University, Cavite 4122, Philippines
National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan
Author to whom correspondence should be addressed.
Viruses 2017, 9(7), 189;
Received: 16 June 2017 / Revised: 6 July 2017 / Accepted: 6 July 2017 / Published: 17 July 2017
(This article belongs to the Section Animal Viruses)
The tick-borne encephalitis virus (TBEV) serocomplex of flaviviruses consists of arboviruses that cause important diseases in animals and humans. The transmission of this group of viruses is commonly associated with tick species such as Ixodes spp., Dermacentor spp., and Hyalomma spp. In the case of Haemaphysalis longicornis, the detection and isolation of flaviviruses have been previously reported. However, studies showing survival dynamics of any tick-borne flavivirus in H. longicornis are still lacking. In this study, an anal pore microinjection method was used to infect adult H. longicornis with Langat virus (LGTV), a naturally attenuated member of the TBEV serocomplex. LGTV detection in ticks was done by real-time PCR, virus isolation, and indirect immunofluorescent antibody test. The maximum viral titer was recorded at 28 days post-inoculation, and midgut cells were shown to be the primary replication site. The tick can also harbor the virus for at least 120 days and can successfully transmit LGTV to susceptible mice as confirmed by detection of LGTV antibodies. However, no transovarial transmission was observed from the egg and larval samples. Taken together, our results highly suggest that anal pore microinjection can be an effective method in infecting adult H. longicornis, which can greatly assist in our efforts to study tick and virus interactions. View Full-Text
Keywords: anal pore microinjection; Langat virus; Haemaphysalis longicornis; virus transmission anal pore microinjection; Langat virus; Haemaphysalis longicornis; virus transmission
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MDPI and ACS Style

Talactac, M.R.; Yoshii, K.; Hernandez, E.P.; Kusakisako, K.; Galay, R.L.; Fujisaki, K.; Mochizuki, M.; Tanaka, T. Synchronous Langat Virus Infection of Haemaphysalis longicornis Using Anal Pore Microinjection. Viruses 2017, 9, 189.

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