Ixodid ticks and tick-borne encephalitis virus prevalence in the South Asian part of Russia (Republic of Tuva).

The most significant processes of arbovirus evolution can be expected to occur in the territories where ticks of different species cohabitate and at the boundaries of virus occurrence, where the probability of the appearance of new virus variants is high due to the possible shift in the main vectors and/or vertebrate hosts. One of the most interesting regions in this regard is the Republic of Tuva. Since most of its territory is covered by mountain ranges and intermountain basins, we were able to study the distribution of vectors and viruses in geographically isolated areas at different altitudes and in various landscapes. From 2008 to 2017, we conducted six expeditions to Tuva and collected 3,077 adult ticks and 24 nymphs. The distribution of tick species was confined to specific landscapes, as follows: Dermacentor nuttalli occurred in steppes, D. silvarum inhabited forest-steppe areas, and Ixodes persulcatus inhabited mixed forests. All three species of ticks were collected on plains and mountain slopes. The range of D. silvarum was shown to be lower than 1300 m above sea level (a.s.l.). Only D. nuttalli and I. persulcatus were collected at higher altitudes. According to our observations, single nymphs of D. nuttalli appear on animals one month before larvae appear. This finding confirms the hypothesis that the immature forms of D. nuttalli are able to overwinter under favourable conditions. We isolated 9 strains and 3 isolates of tick-borne encephalitis virus (TBEV) from I. persulcatus, one strain from D. nuttalli and one strain from D. silvarum. The TBEV strain from D. nuttalli was isolated from the territory inhabited only by Dermacentor ticks. All isolated strains belong to the Siberian subtype of TBEV. TBEV was detected in ticks from all the investigated altitudes. There were no statistically significant differences in the virus prevalence between the Dermacentor and Ixodes ticks. The results of our work provide additional support for the hypothesis of the existence of TBEV foci in areas with an absolute dominance of D. nuttalli.

Kholodilov I ，Belova O ，Burenkova L ，Korotkov Y ，Romanova L ，Morozova L ，Kudriavtsev V ，Gmyl L ，Belyaletdinova I ，Chumakov A ，Chumakova N ，Dargyn O ，Galatsevich N ，Gmyl A ，Mikhailov M ，Oorzhak N ，Polienko A ，Saryglar A ，Volok V ，Yakovlev A ，Karganova G ... -  《-》

Tick-borne encephalitis virus in arthropod vectors in the Far East of Russia.

Isolates of tick-borne encephalitis virus (TBEV) from arthropod vectors (ticks and mosquitoes) in the Amur, the Jewish Autonomous and the Sakhalin regions as well as on the Khabarovsk territory of the Far East of Russia were studied. Different proportions of four main tick species of the family Ixodidae: Ixodes persulcatus P. Schulze, 1930; Haemaphysalis concinna Koch, 1844; Haemaphysalis japonica douglasi Nuttall et Warburton, 1915 and Dermacentor silvarum Olenev, 1932 were found in forests and near settlements. RT-PCR of TBEV RNA in adult ticks collected from vegetation in 1999-2014 revealed average infection rates of 7.9 ± 0.7% in I. persulcatus, of 5.6 ± 1.0% in H. concinna, of 2.0 ± 2.0% in H. japonica, and of 1.3 ± 1.3% in D. silvarum. Viral loads varied in a range from 102 to 109 TBEV genome-equivalents per a tick with the maximal values in I. persulcatus and H. japonica. Molecular typing using reverse transcription with subsequent real time PCR with subtype-specific fluorescent probes demonstrated that the Far Eastern (FE) subtype of TBEV predominated both in mono-infections and in mixed infection with the Siberian (Sib) subtype in I. persulcatus pools. TBEV strains of the FE subtype were isolated from I. persulcatus, H. concinna and from a pool of Aedes vexans mosquitoes. Ten TBEV strains isolated from I. persulcatus from the Khabarovsk territory and the Jewish Autonomous region between 1985 and 2013 cluster with the TBEV vaccine strain Sofjin of the FE subtype isolated from human brain in 1937. A TBEV strain from H. concinna collected in the Amur region (GenBank accession number KF880803) is similar to the vaccine strain 205 isolated in 1973 from I. persulcatus collected in the Jewish Autonomous region. The TBEV strain Lazo MP36 of the FE subtype isolated from a pool of A. vexans in the Khabarovsk territory in 2014 (KT001073) differs from strains isolated from 1) I. persulcatus (including the vaccine strain 205) and H. concinna; 2) mosquitoes [strain Malishevo (KJ744034) isolated in 1978 from Aedes vexans nipponii in the Khabarovsk territory]; and 3) human brain (including the vaccine strain Sofjin). Accordingly, in the far eastern natural foci, TBEV of the prevailing FE subtype has remained stable since 1937. Both Russian vaccines against TBE based on the FE strains (Sofjin and 205) are similar to the new viral isolates and might protect against infection.

Pukhovskaya NM ，Morozova OV ，Vysochina NP ，Belozerova NB ，Bakhmetyeva SV ，Zdanovskaya NI ，Seligman SJ ，Ivanov LI ... -  《-》

Influence of altitude on tick-borne encephalitis infection risk in the natural foci of the Altai Republic, Southern Siberia.

The Altai Republic is a highly endemic area as far as tick-borne encephalitis (TBE) is concerned. The aim of the research was to study the effect of altitude on the risk of tick-borne encephalitis infection in the Altai Republic. The paper analyzes the following data: the study of ixodid ticks collected from the vegetation in 116 sites at the 200-2383m elevation above sea level in 2012-2014, TBE virus prevalence of these vectors, tick-bite incidence rate, and TBE incidence rate of the population. Species identification of 4503 specimens has shown that the most common species are Dermacentor nuttalli (45.3%), Ixodes persulcatus (33.1%), Dermacentor silvarum (9.4%), Dermacentor reticulatus (8.9%), and Haemaphysalis concinna (5.0%). A total of 2997 adult ixodid ticks were studied for the presence of the TBE virus; 2163 samples were examined by ELISA, while 834 specimens were tested by PCR. The TBE virus prevalence of Dermacentor spp. ticks in both reactions was significantly higher than of Ixodes persulcatus ticks (p<0.001). The work shows that the altitude is an important factor in the development of the epidemiological situation of tick-borne encephalitis: the higher the elevation of the area above sea level, the smaller the range of vectors. There is also a change of a leading species: in middle altitude (800-1700m above sea level) the virus is transmitted by ticks of D. nuttalli along with I. persulcatus, and in high mountains (above 1700m above sea level) D. nuttalli becomes an absolute dominant species. However, these species of ticks are less effective vectors than I. persulcatus. With the increase of altitude the tick-bite incidence rate decreases (r=-0.78, p<0.05), and TBE incidence also reduces (r=-0.67, p<0.05).

Shchuchinova LD ，Kozlova IV ，Zlobin VI 《-》

Dermacentor reticulatus is a vector of tick-borne encephalitis virus.

Tick-borne encephalitis virus (TBEV; family Flaviviridae) is the most medically important tick-borne virus in Europe and Asia. Ixodes ricinus and I. persulcatus ticks are considered to be the main vector ticks of TBEV in nature due to their specific ecological associations with the vertebrate hosts. Nevertheless, recent TBEV prevalence studies in ticks suggest that Dermacentor reticulatus ticks might play a relevant role in the maintenance of TBEV in nature. The goal of this study was to evaluate the vector competency of D. reticulatus for TBEV through experimental tick infections and comparative in vivo transmission studies involving D. reticulatus and I. ricinus ticks. We observed that after a transcoxal micro-capillary inoculation, adult female D. reticulatus ticks efficiently replicated TBEV during the observed period of 21 days. The mean virus load reached up to 2.5 × 105 gene copies and 6.4 × 104 plaque forming units per tick. The infected D. reticulatus ticks were able to transmit the virus to mice. The course of infection in mice was comparable to the infection after a tick bite by I. ricinus while the virus spread and clearance was slightly faster. Moreover, D. reticulatus ticks were capable of tick-to-tick non-viraemic transmission of TBEV to the Haemaphysalis inermis nymphs during co-feeding on the same animal. The co-feeding transmission efficiency was overall slightly lower (up to 54 %) in comparison with I. ricinus (up to 94 %) and peaked 1 day later, at day 3. In conclusion, our study demonstrated that D. reticulatus is a biologically effective vector of TBEV. In line with the recent reports of its high TBEV prevalence in nature, our data indicate that in some endemic foci, D. reticulatus might be an underrecognized TBEV vector which contributes to the expansion of the TBEV endemic areas.

Ličková M ，Fumačová Havlíková S ，Sláviková M ，Slovák M ，Drexler JF ，Klempa B ... -  《-》

Identification of tick-borne encephalitis virus in ticks collected in southeastern Hungary.

Tick-borne encephalitis virus (TBEV) is an arthropod-borne viral pathogen causing infections in Europe and is responsible for most arbovirus central nervous system infections in Hungary. Assessing the TBEV prevalence in ticks through detection of genomic RNA is a broadly accepted approach to estimate the transmission risk from a tick bite. For this purpose, 2731 ticks were collected from the neighboring area of the town of Dévaványa, located in southeastern Hungary, which is considered a low-risk-transmission area for TBEV. Altogether, 2300 ticks were collected from the vegetation, while 431 were collected from rodents. Samples were pooled and then screened for TBEV with a newly designed semi-nested RT-PCR (RT-snPCR) targeting the NS1 genomic region. PCR results were confirmed by direct sequencing of the second round amplicons. Among the 3 different collected tick species (Ixodes ricinus, Haemaphysalis concinna, Dermacentor marginatus), I. ricinus was the only species that tested positive for TBEV. TBEV-positive ticks were collected from small mammals or from the vegetation. One nymphal pool and 4 larval pools tested positive for TBEV. The only positive nymphal pool was unfed and came from vegetation, while ticks of the 4 positive larval pools were collected from rodents. Minimal TBEV prevalence in ticks was 0.08% for unfed nymphs and 0.78% for feeding larvae. Our results indicate that further long-term investigations on the occurrence of TBEV are needed to better describe the geographic distribution and the prevalence of infected ticks in Hungary.

Pintér R ，Madai M ，Vadkerti E ，Németh V ，Oldal M ，Kemenesi G ，Dallos B ，Gyuranecz M ，Kiss G ，Bányai K ，Jakab F ... -  《-》