Tick-borne Encephalitis Virus in Ixodes Ticks and Wild Birds in the Russia Komi Republic
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1 Tick-borne Encephalitis Virus in Ixodes Ticks and Wild Birds in the Russia Komi Republic State Research Center for Virology and Biotechnology Vector, Koltsovo, Russia Bangkok December, 2014
2 Family Flaviviridae Genus Flavivirus Pestivirus Hepacivirus unassigned viruses Number: species, viruses, candidate Flavivirus /2 Pestivirus /1 Hepacivirus /2 unassigned viruses 1 3
3 Flavivirus Genome ss (+) RNA genome Approximately 11 kb 5 -m7gpppamp cap Lacks 3 -polya tail Codes for 3 structural proteins Capsid (C), membrane (prm/m), envelope (E) 7 non-structural proteins NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5
4 The main human flaviviral infections: Dengue Yellow fever Japanese encephalitis West Nile fever Tick-borne encephalitis
5 27 viruses were detected in ticks in Europe?! Flaviviruses: TICK-BORNE ENCEPHALITIS, LOUPING-ILL, Tyuleniy and Meaban; Orthobunyaviruses: Bahig, Matruh; Phleboviruses: Grand Arbaud, Ponteves, Uukuniemi, Zaliv Terpeniya and St. Abb's Head; Nairoviruses: Soldado, Puffin Island, Avalon, Clo Mor, CRIMEAN- CONGO HEMORRHAGIC FEVER; Bunyavirus : Bhanja; Coltivirus : Eyach; Orbiviruses: Tribec, Okhotskiy, Cape Wrath, Mykines, Tindholmur and Bauline Thogotoviruses : Thogoto, Dhori; Asfivirus : AFRICAN SWINE FEVER VIRUS Hubálek Z, Rudolf I. Tick-borne viruses in Europe. Parasitol Res Apr 18. In addition: Such viruses were detected also in Russia: OMSK HEMORRHAGIC FEVER, WEST NILE VIRUS, POWASSAN and Kemerovo
6 Main tick-borne infections in Russia: TICK-BORNE ENCEPHALITIS; BORRELIOSIS (Lyme diseases); Rickettsia infections (Spotted fever group); Powassan Encephalitis; Omsk haemorrhagic fever; Anaplasmosis; Ehrlichiosis; Bartonellosis; West Nile fever!? Kemerovo!? The number of tick attacks are between human cases per year in Russia! And it is only official data!
7 1937 discovery of TBEV, putative place of field laboratory in Obor, Khabarovsk kray
8 Tick borne encephalitis in Russia Number cases of TBEV on of population Vector, Koltsovo Sites of study
9 The incidence of tick-borne encephalitis (TBE) has been reported for more than 25 European and Asian countries. This region is a home for over 700 million people, even not including China. Systematic data on the number of TBE cases in China are small.
10 Tick borne encephalitis foci in Europe. European distribution of natural foci of tick-borne encephalitis (CEE and RSSE) and louping ill (asterisks). Explanation: black dots and black areas, TBE virus isolation or the virus disease. The dotted line shows the limits of the Ixodes ricinus plus I.persulcatus area
11 Tick borne encephalitis in Europe.
12 Primorye-270 SALEM 263 HF-Bogolubovka Primorye Primorye-212 Primorye-253 Primorye-69 Far Eastern TBEV Neudoerfl Oshima5-10 European genotype TBEV Spanish sheep Hypr 205 Dalnegorsk Louping ill Sofjin-HO Turkish sheep Glubinnoe/2004 Greek goat Kolarovo-2008 Vasilchenko? Siberian genotype TBEV Zausaev EK SENZHANG MDJ-01 Kavalerovo
13 European genotype Siberian genotype Far Eastern genotype TBEV and molecular hours А О 0.02 Б 2750 years 2250 years 100 Л 1770 yrs. П М K 100 C И 100 Н В Г years. 590 yr. 650 yr. Д З Ж Е years 250 yr AB022291, Oshima3-6, Japan,1995 AB022292, OshimaI-1, Japan, 1996 AB237187, Oshima A-1, Japan,1995 AB001026, Kik629/97, Japan, 1997 AB022293, Oshima5-10, Japan, 1993 AB022294, OshimaC-1, Japan, 1996 AB237189, Oh696/97, Japan,1997 AB237184, Miz416/97, Japan, 1997 AB237188, Miz660/97, Japan, 1997 AF091008, Crimea, Ukraina, 1987 AF091013, N132, Vladivostok, yr. AF091016, RK1424, Latvia, yr. AF091019, T-blood, Perm, 1939 AB237192, Kita987/99, Japan, 1999 AB022703, Sofjin-HO, 1937 AB049345,VL-99-m11, Vladivostok, 1999 AB049346, KH99-m9, Khabarovsk, 1999 AB022295, KH98-2, Khabarovsk, 1998 AB022296, KH98-5, Khabarovsk, 1998 AB022297, KH98-10, Khabarovsk, 1998 AB237185, Kam586/97, Japan, AB237186, Kam588/97, Japan, yrs. AB049347, D1283, Khabarovsk, 1998 DQ862460, Glubinnoe/ yrs. AY174188, Senzhang, China, EU444077, Yar71, Yaroslavl, EU444078, Yar114, Yaroslavl, yrs. EU444079, Yar46-2, Yaroslavl, EU444080, Yar48, Yaroslavl, yrs. 100 AB049349, IR99-1m4, Irkutsk, AB049351, IR99-2m7, Irkutsk, yrs. 100 AF527415, Zausaev, Tomsk, 1985 AF091006, Aina, Irkutsk, 1963 AF069066, Vasilchenko, Novosibirsk, yrs. 99 AB049348, IR99-1m1, Irkutsk, 1999 AB049350, IR99-2m3, Irkutsk, 1999 Y07863, Louping ill, Scotland, 1929 AF091015, Pan, Moscow, yrs. AF091017, Scharl, Austria, 1956 AF091007, Als.I, France, AF ZZ9, Austria, AF091009, Iso 40, Switzerland, 1975 AF091010, K23, Germany, 1975 M27157, Neudoerfl, Austria, yrs. X60286, A52, Finland, AF091005, Absetterov, Sankt-Petersburg, 1951 AF091011, Kem I, Hungary, 1952 WNVNY99, WNV, strain NY99
14 New variants of lethal human TBEV infection Hemorrhagic TBEV (Novosibirsk, 1999) Age: Incubation time (after tick attack) Fever (start of disease) First small hemorrhagic CNS damage Massive hemorrhagic Death years 12,8 days 1 day 7 day 10 day days 16 day
15 Place of isolation of TBEV, 1937 Human TBEV lethal infection, strain, Glubinnoe/2004
16 Human lethal TBEV infection, Age of patient: strain Glubinnoe/ year Date of tick attack (2 ticks) 30 may 2004 Incubation time Clinical symptoms: 8 days Days Heavy fever 1 First CNS symptoms 2 CNS lesion (Air evacuation, Vladivostok TBEV center, active reanimation) Coma and apnoea 5 Death (heart failure) 10 4
17 Optical density at 492 nm Cultivation Glubinnoe/2004 of TBEV on PEG cells Virus yield, log TCID 50 ml Hours after infection 0
18 Amino acid substitutions for Glubinnoe/2004 in comparison with 205 and Sofjin strains C CTHDpr M E NS1 NS2a NS2b NS3 NS4a 2K NS4b NS5
19 Hypothesis. Mechanisms and pathways of tick-borne infections? Main hypotheses - wild birds. West Nile virus is a typical example. What evidence can provide research: tick-borne encephalitis virus, Omsk hemorrhagic fever virus, Powassan virus
20 Tomsk and tick-borne encephalitis The average incidence of TBEV infection in Russia varied from 1.9 to 4.4 cases per per year between 2001 and 2011 (State report of RF). Tomsk Region, the officially reported incidence of the infection was from 15.5 to 72.5 per in last decade (State report of Tomsk, 2011). The majority of these cases (74%) were within the Tomsk city the largest city in the region. Novosibirsk region located only 200 km to the south of Tomsk has been lower cases for the last three years (State report, Novosibirsk). Even lower incidence (0.36 to 1.0 cases per ) has been reported in Far Eastern region, where TBEV was originally discovered in 1937 (State report, Khabarovsk). Tomsk region Novosibirsk region
21 Tomsk, places for ticks collection N S
22 Tomsk, places for ticks collection
23 Tick-borne encephalitis viruses are genetically different!!! Strains of Kolarovo-2008 (Tomsk ) and Vasylchenko (Novosibirsk), Siberian genotype: The level of genetic difference reaches 10% in nucleotide sequence and 7% of its amino acid sequence. Zausaev (Tomsk) Kolarovo (Tomsk), Siberian genotype: Amino acid substitutions - 124, and the level of homology for 3 '-NCR is only 79%.
24 Phylogenetic tree of TBEV, Tomsk, Siberian genotype of TBEV - predominantly distributed genotype (89.5%) in suburban site, whereas in urban biotopes, 47% isolates of the TBEV were presented by Far Eastern genotype. Chronical TBE infection
25 Phylogenetic tree of TBEV, Tomsk, 2010 Only Far Eastern genotype of TBEV!!!
26 Wild birds and ixodid ticks in Tomsk The 736 wild birds representing 60 species were captured carrying a total of 804 I. pavlovskyi, I.persulcatus and I. plumbeus ticks. TBEV RNA and antigen were found in 9.7% and 22.8% samples collected from wild birds (40 species), respectively. TBEV markers were also detected in 9,8% I. persulcatus ticks, 4,7% - I. pavlovskyi and 4,2% - I. plumbeus ticks collected from wild birds.
27 Fieldfare - the principal disseminator (vector) for ixodid ticks!? Up to 70 ticks per birds (all stages), from these - 18 imago!!! Sand martin with Ixodes plumbeus
28 Ixodes ticks Tree Pipit (Anthus trivialis) - 13,25 ticks/bird Fieldfare (Turdus pilaris) - 5,73 ticks/bird
29 Ixodes ticks Redwing (Turdus iliacus) - 3,33 ticks/bird Common redstart (Phoenicurus phoenicurus) - 1,32 ticks/bird
30 Chaffinch (Fringilla coelebs) - 0,96 ticks/bird Ixodes ticks
31 Genotyping TBEV in birds and ixodid ticks collected from birds 25 isolates from Far Eastern genotype of TBEV, 9 isolates Siberian genotype of TBEV 2 viral strains FE genotype of TBEV were isolated from wild birds. I.pavlovskyi, I.persulcatus and I. plumbeus were collected from wild birds I.pavlovskyi - unusually widespread in urban biotopes of Tomsk and Novosibirsk!!!
32 Tick-borne encephalitis and Komi Republic?! 47 regions of RF Incidence of TBE infections in the Komi Republic (on 100 thousand of population) Russian Federation 1,98 2,62 2,19 Komi Republic 0,42 1,80 3,30
33 Taiga tick on the territory of the Komi Republic detected pathogens: Tick-borne encephalitis virus, West Nile virus, Borrelia spp., Ricketsia spp., Ehrlichia spp., Babesia spp., Bartonella spp., Anaplasma spp.
34 Phylogenetic tree of TBEV, Komi Republic, 2010 Direct genotyping of 16 variants of TBEV in ticks based on the direct sequencing of 5 -NCR showed that 3 isolates can be classified as Siberian genotype of TBEV, while the remaining 13 isolates of TBEV are the highly pathogenic variants of Far Eastern genotype.
35 Instead of conclusion: New flaviviruses, genetic variants, unusual disease outbreaks in recent years:
36 New Flaviviridae, genetic variants, unusual disease outbreaks 2006 Senegal New flavivirus - Ngoye 2007 India Presumably the new genotype of WNV, the level difference between 25-30% Japan New flavivirus - Culex flavivirus (CxFV), 2009 West Africa New flavivirus - Nounane (NOUV) 2009 Japan New flavivirus - Aedes flavivirus (AeFV) 2009 Mexico New flavivirus T Ho 2009 China Nanjianyin virus it is Kyasanur forest disease virus? now Nord Eurasia West Nile virus, genotype 2
37 New Flaviviridae, genetic variants, unusual disease outbreaks 2007 China Donggang virus - genotype 1 of Japanese encephalitis virus, for the first time in China China The outbreak of Duck virus in China, Tembusu virus (described by 1955, Malaysia, Culex mosquitoes) 2009 Finland Virus Lammi new mosquito flavivirus, is close to the Nounane virus 2007, , 2011 Senegal, Israel China, Kenya, Korea Virus Barkedji new mosquito flavivirus, the owner is not known, is close to the Nounane virus Chaoyang virus, discovered in China, then in Kenya and then in the Republic of Korea Portugal Finland Virus Hanko, isolated from mosquitoes, the Negev-like virus is homologous virus Hanko
38 New Flaviviridae, genetic variants, unusual disease outbreaks 2011, 2013 Senegal, Polynesia Virus Zika, southeastern Senegal; French Polynesia, Southern Europe?? 2014 Italy AeVF, Aedes flavivirus in north-eastern Italy 2014 South America New strains of Culex flavivirus isolated in Argentina
39 Acknowledgments: SRC VB Vector, Koltsovo Ivanova A.V. Subbotina E.l. Kazachinskaya E.I. Konovalova S.N. Kononova Yu.V. Tupota N.L. Protopopova E.V. Razumov I.A. Riabchikova E.I. Sviatchenko V.A. Ternovoi V.A. Chausov E.V. Shvalov A.N. Mikrukova N.P. Kartashov M.Yu. Agulova L.P. Bolshakova N.P. Gashkov S.I. Ivanova, N.V. Kravchenko L.B. Korobitsyn I.G. Kuranova V.N. Moskvitin S.S. Moskvitina N.S. Romanenko V.N. Suchkova N.G. Tuten kov O.Yu. Komi Republic Glushkova L.I. Korabelnikov I.V. Galimov P.P. Yegorova Yu.I. Many thanks my colleagues who have worked and helped us in these studies
40 Thank you for your attention!
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