Abstract
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.
Highlights
Ticks are important vectors of disease-causing pathogens in domestic and wild animals and are considered to be second to mosquitoes in transmitting human diseases [1]
We initially observed that Langat virus (LGTV) injected ticks via anal pore microinjection remained positive to LGTV RNA even after 28 dpi (Table 2)
One of the most important determinants of vector competence is the susceptibility of midgut cells to virus infection [22], and anal pore microinjection method may prove to be an important technique in evaluating vector competency, since the virus will first come in contact with the tick gut
Summary
Ticks are important vectors of disease-causing pathogens in domestic and wild animals and are considered to be second to mosquitoes in transmitting human diseases [1]. TBEV serocomplex consists of arboviruses that cause important diseases in animals and humans. The transmission of this group of viruses is commonly associated with tick species that include hard ticks Ixodes ricinus, I. persulcatus, Dermacentor spp., and Hyalomma spp. Synchronization of tick infection with a defined viral inoculum is a notable limitation in this method [2]. Another method of tick infection is through percoxal microinjection, it bypasses the midgut barrier which makes it non-representative of natural route of infection and may not ensure consistent infection rates among fed ticks [11]. This infection method is simpler and relatively inexpensive; generating cohorts of infected ticks with equal pathogen burden is its major limitation [12]
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