J.E. ECHEVARRÍA 1, S. VÁZQUEZ-MORÓN 1, C. IBÁÑEZ 2, J.C. AZNAR 1, E. RUIZ 3, J. JUSTE 2 1...
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Transcript of J.E. ECHEVARRÍA 1, S. VÁZQUEZ-MORÓN 1, C. IBÁÑEZ 2, J.C. AZNAR 1, E. RUIZ 3, J. JUSTE 2 1...
J.E. ECHEVARRÍA1, S. VÁZQUEZ-MORÓN1, C. IBÁÑEZ2, J.C. AZNAR1, E. RUIZ3, J. JUSTE2
1 National Center of Microbiology, Carlos III Institute of Health, Majadahonda, Madrid2 Doñana Biological Station, CSIC, Seville3 Laboratorio Central Veterinario, Santa Fe, Granada
Spain
ASYMPTOMATIC RHABDOVIRUS INFECTION IN MERIDIONALSEROTINE BATS (Eptesicus isabellinus) FROM SPAIN
Número de individuos
Localidad añoExposición humana
Murciélago Virus
1 VALENCIA 1987 SI UNKNOWN“DUVENHAGE
LIKE”
1 GRANADA 1987 SIEptesicus serotinus
EBLV1
5 HUELVA 1989 NOEptesicus serotinus
EBLV1
1 GRANADA 1994 SIEptesicus serotinus
EBLV1
3 SEVILLA 19991 SI
2 NOEptesicus serotinus
EBLV1
1 MURCIA 1999 SIEptesicus serotinus
EBLV1
4 SEVILLA 2000 NOEptesicus serotinus
EBLV1
1 MURCIA 2002 SIEptesicus
serotinusEBLV1
Lyssavirus-infected bats detected by passive surveillance in Spain
Genes: CytB, ND1, RAG22900 bp
57Ese SY
90Ebtur IR
93Ebtur IR
92Ebtur IR
91Ebtur IR
94Ebsp IR
95Ebsp IR
61Ese SY
60Ese SY
43Ese CY
16Ebo SY
19Ebo SY
14Ebo JO
97Ebog IR
98Ebog IR
54Ese NorthSpain
55Ese North Spain
50Ese IT
44Ese CZ
64Ese UK
46Ese DE
65Ese UK
45Ese CZ
66Eni GE
51Ese LA
52Ese LA
42Ese CH
7Ean SY
11Ean TU
8Ean SY
10Ean TU
21Eho SA
22Eho SA
33Eis MO
36Eis South Spain
35Eis South Spain
34Eis MO
23Eis LI
28Eis LI
Eptesicus Eptesicus serotinusserotinus
Eptesicus Eptesicus isabellinusisabellinus
> 1
6 %
> 1
6 %
cytb
!!
cytb
!!
Filogeny of Eptesicus in the Paleartic
Europe (including north Spain)
South SpainNorth Africa
E. serotinus or E. Isabellinus?
Lyssaviruses are not pathogenic for bats (1)
-Prevalence of VRAB antibodies above 60% in a healthy colony of tailed bat (Tadarida brasiliensis) in USA (Steece –1989- Journal of Wildlife Diseases 25: 490-496 )
-Prevalence of EBLV1 antibodies above 20% in several colonies of serotine bats (Eptesicus isabellinus) in Spain. One seropositive individual captured healthy one year later(Pérez Jordá –1994- Journal of Wildlife Diseases 3: 372-377)
- Presence of EBLV1 in healthy Rousettus aegyptiacus months after an outbreak in a captive colony in The Netherlands. Virus is detected by PCR and IF on brain and other organs. (Wellenberg –2002- Archives of Virology 147:349-361)
- Presence of EBLV1 ARN on oro-pharyngeal swabs of healthy serotine bats in Spain, recovery of some positive individuals one year after.(Echevarría –2001- Journal of Clinical Microbiology 39(10): 3678-3683)
Lyssaviruses are not pathogenic for bats (2)
- Detection of EBLV2 specific antibodies on Daubenton´sBats from Scotland.(Brookes -2005- Emerging Infectious Diseases 11(4): 572-578).
Lyssaviruses are not pathogenic for bats (3)
- Detection of EBLV1 specific antibodies and RNA on blood pellets of different bat species other than serotine bats from healthy colonies in Spain. (Serra-Cobo –2002- Emerging Infectious Diseases 8 (4): 413-420)
- Survival of Tadarida brasiliensis after aerosol exposition to RABV. (Davis –2007- Journal of infectious Diseases195:1144-1150)
- Survival of Eptesicus fuscus after inoculation of RABV(Shankar –2004- Journal of Wildlife Diseases 40(3): 403-413)
Lyssaviruses are not pathogenic for bats (3)
SEVILLA (SECRETARIA)
SEVILLA (CATEDRAL)
TRAJANO
ALCALA DEL RIOSOTIEL CORONADAAZNALCOLLAR (Puente Cañaveroso)
MOLINO DEL DUENDEMOLINO DE NIEBLA
MOLINO DEL RINCON
TUNEL DEL MANZANO
PUENTE GADEA
LA ARADILLA PUENTE TAMUJOSO
ORJIVA (TUNEL GUADALFEO)
ORJIVA (REVOLCADERO)
TUNEL DEL PICOTE
TUNEL SALOMON
50 km
UMBRETE(Colegio)
SEVILLA (ALCAZAR)
-19 COLONIES OF Eptesicus isabellinus-1080 INDIVIDUALS-1227 CAPTURES (1998-2003)
STUDY OF EBLV1 INFECTION ON BAT COLONIES
- O-PH SWAB (PCR) - SERUM (RFFIT) - IDENTIFIED - DATA COLLECTED
GENERAL RESULTS
- 1 individual PCR positive and 5 antibody positive captured one year later.- Only one individual simultaneously carrying RNA and antibodies.
RTRT--PCR PCR RFFITRFFIT12271227 6266261919 1313
34 (2.8%) 51 (8.1%)NNúúmero de capturasmero de capturas 164164 151151PositivosPositivos 4 (2.4%)4 (2.4%) 10 (6.6%)10 (6.6%)NNúúmero de capturasmero de capturas 161161 9090PositivosPositivos 4 (2.5%)4 (2.5%) 5 (5.6%)5 (5.6%)NNúúmero de capturasmero de capturas 204204 128128PositivosPositivos 16 (7.8.%)16 (7.8.%) 12 (9.4%)12 (9.4%)NNúúmero de capturasmero de capturas 209209 9696PositivosPositivos 00 4 (4.2%)4 (4.2%)NNúúmero de capturasmero de capturas 287287 100100PositivosPositivos 10 (3.4%)10 (3.4%) 5 (5%)5 (5%)NNúúmero de capturasmero de capturas 201201 5454PositivosPositivos 00 00
NUMERO DE COLONIASNUMERO DE COLONIASTOTAL CAPTURASTOTAL CAPTURAS
20002000
20012001
20022002
20032003
19981998
19991999
TOTAL POSITIVOSTOTAL POSITIVOS
RTRT--PCR PCR RFFITRFFIT12271227 6266261919 1313
34 (2.8%) 51 (8.1%)NNúúmero de capturasmero de capturas 164164 151151PositivosPositivos 4 (2.4%)4 (2.4%) 10 (6.6%)10 (6.6%)NNúúmero de capturasmero de capturas 161161 9090PositivosPositivos 4 (2.5%)4 (2.5%) 5 (5.6%)5 (5.6%)NNúúmero de capturasmero de capturas 204204 128128PositivosPositivos 16 (7.8.%)16 (7.8.%) 12 (9.4%)12 (9.4%)NNúúmero de capturasmero de capturas 209209 9696PositivosPositivos 00 4 (4.2%)4 (4.2%)NNúúmero de capturasmero de capturas 287287 100100PositivosPositivos 10 (3.4%)10 (3.4%) 5 (5%)5 (5%)NNúúmero de capturasmero de capturas 201201 5454PositivosPositivos 00 00
NUMERO DE COLONIASNUMERO DE COLONIASTOTAL CAPTURASTOTAL CAPTURAS
20002000
20012001
20022002
20032003
19981998
19991999
TOTAL POSITIVOSTOTAL POSITIVOS
Different temporal pattern of circulation on each colony
Colonia Det. N. 1998 1999 2000 2001 2002 2003Puente Duende
PCR
Ab
137
107
1 (4.7)
0
0
2 (22.2)
1 (4.5)
1 (4.5)
2 (8.0)
0
Puente
Gadea
PCR
Ab
114
89
0
2(6.9)
1 (3.7)
2 (11.11)
0
3 (17.65)
0
3 (23)
0
1 (8.3)
Molino
Rincón
PCR
Ab
67
54
0
1 (12.5)
1 (4.76)
0
1 (4.76)
0
Tunel
Coronada
PCR
Ab
143
85
1 (5.0)
5 (25.0)
Tunel Salomón
PCR
Ab
20
18
2 (15.7)
1 (9.0)
La
Aradilla
PCR
Ab
7
7
0
1 (14.3)
Tunel del
Picote
PCR
Ab
51
23
1 (4.34)
1 (4.34)
Revolcad.Órgiva
PCR
Ab
71
27
0
1 (3.7)
Puente Cañave.
PCR
Ab.
138
108
7 (22.5)
4 (23.5)
Repetitive captures of Eptesicus isabellinus onthree colonies sampled six times
0
20
40
60
80
100
120
1 CAPTURE 2 CAPTURES 3 CAPTURES 1 CAPTURE 2 CAPTURES 3 CAPTURES 4 CAPTURES 5 CAPTURES 1 CAPTURE 2 CAPTURES 3 CAPTURES 4 CAPTURES
Puente Cañaveroso Puente Molino del Duende Túnel (tren) Coronada
79.0
18.8
2.2
40.9
26.3
8.8 5.8
18.2
52.5
25.4
15.3
6.8
Individuals captured ALWAYS in the same colony
Genetic isolation of colonies
Different haplotype distribution on each colony
Conclusion
Genetic isolation of Eptesicus isabellinus
natural colonies reflects the lack of interchange of
individuals that causes independent epidemiological
circulation of EBLV1.
321RT-PCR
10-1-2-3-4-5
BC
I
1: Brain neg., swab neg.
2: Brain pos, swab pos.
3: Brain neg, swab pos.
Male
Female
BODY CONDITION INDEX AND PATTERN OF INFECTION
Bats with EBLV1 RNA in oral cavity are healthy
Many infections are subclinical with asymptomatic
presence of virus in oro-pharyngeal cavity.
Conclusion
LYSSAVIRUSES ARE NOT THE ONLY BAT RHABDOVIRUSES
244 oro-pharyngeal swabs from healthy Eptesicus isabellinus captured in Spain. Two relevant sequences obtained.
GENERIC SEARCH OF DHIMARABDOVIRUSES
EBLV1 DHIMARHABDOINESPECIFIC AMPLIFICATION
50 BP 50 BP
Protein BLAST of a phragment of 39 aa
Protein BLAST of a phragment of 26 aa
SARS CORONAVIRUS EBOLA
HENIPAVIRUS
OTHER WELL-KNOWN BAT-ASSOCIATED HUMAN PATHOGENS
Familia Orthomyxoviridae: Gripe A (Nyctalus noctula) Familia Parayxoviridae:género Rubulavirus: Virus Mapuera (Sturnina lilium), Virus Menangle (Pteropus poliocephalus), Virus Tioman (Pteropus hypomelanus), Género no determinado (Rousettus leschenaultia).
Familia Togaviridae, Género Alphavirus: Virus Chikungunya (Scotophilus sp., Rousettus aegyptiacus, Hipposideros cafer, Chaerephon pumilus), Virus Sindbis (Rinnolophidae, Hipposeridae), Virus de la encefalitis equina venezolana (Desmodus rotundus, Uroderma bilibatum, Artibeus phaeotis).
Familia Flaviviridae, género Flavivirus,: Virus Bukalasa bat (Chaerophon pumilus), Virus Isla Carey (Cynopterus brachiotys), Virus Dakar bat(Cherephon pumilus, Taphozous perforatus, Scotophilus sp, Mops condylurus), Virus Entebbe bat (Chaerophon pumilus, Mops condylurus), Virus de la encefalitis japonesa (Hipposideros armiger, Miniopterus schreibersii, Rinolophus cornutus), Virus Jugra (Cynopterus brachiotis), Virus de la leucoencefalitis del Myotis de Montana (Myotis lucifugus), Virus Phom-Penh bat (Eonycteris spelaea, Cynopterus brachyotis), Virus Rio Bravo (Tadarida brasiliensis), Virus de la encefalitis de St. Louis (Tadarida brasiliensis), virus Saboya (Nycteris gambiensis), virus Sokuluk (Pipistrellus pipistrellus), Virus Tamana Bat (Pteronotus parnelli), virus Uganda (Rousettus sp, Tadarida sp), Virus Yokose (no identificado).
Familia Bunyaviridae, género Bunyavirus: Virus Catu (Molossus obscurus), Virus Guama (no identificado), virus Nepuyo (Artibeus jamaicensis). Género Hantavirus: Virus Hantaan (Eptesicus serotinus, Rhinolophus ferrumequinum). Género Phlebovirus, Virus de la fiebre del valle del Rif (Micropterus pusillus, Hipposideros abae, Hipposideros caffer, Miniopterus schreibersii, Epomops franqueti, Glauconycteris argentata), virus Toscana (Pipistrellus kulhii). Género desconocido, virus Kaeng Khoi (Cheropon plicatus), virus Bangi (Scotophilus sp, Pipstrellus sp, Tadarida sp.).
Familia Reoviridae, género Orbivirus: Virus Ife (Eidolum helvum), virus Japanaut (Syconicteris australis), virus Fomede (Nycteris nana). Género orthoreovirus, virus Nelson Bay (Pterupus poliocephalus), virus Palau (Pteropus hypomelanus), virus Broome (Pteropus alecto).
Familia arenaviridae, virus Tacaribe (Artibeus lituratus).
Familia picornaviridae, género indeterminado: virus Juruaca (no identificado)
No clasificados: virus Issyc-Kul (virus Keterah) (Nyctalus noctula, Eptesicus serotinus, Pipistrellus pipistrellus, Myotis blythii, Rinolophus ferrumequinum, Rinolophus lepidus, Rinolophus horsfeldi, Megaderma spasma, Scotophilus kulhii, Cynopterus branchyotis, Eonycteris spelea, Cherephon plicatus, Hipposideros diadema, Tazophous melanopogon, virus Mojui dos Campos (no identificado), virus Yoge (Rousettus aegyptiacus), virus Kasokero (Rousettus aegyptiacus).
OTHER BAT ASSOCIATED VIRUSES (Calisher et al. –2006- Clinical Microbiological Reviews 19(3):531-545)
1.- ON THE BAT SIDE + Estación Biológica de Doñana, Sevilla - Javier Juste - Carlos Ibáñez 2.- ON THE VIRUS SIDE + Centro Nacional de Microbiología, Majadahonda, Madrid - Juan E. Echevarría - Sonia Vázquez - Carolina Aznar - José Miguel Berciano 3.- PATHOLOGIST + Laboratorio Central Veterinario. Santa Fe, Granada - Eduardo Ruiz Villamor
TWO Eptesicus SPECIES TWO EBLV1 SUBTYPES?
87
7598
6275
8387
71
6988
88
946370
64
87
80
84
68
93
6899
65
Subtype 1a
Subtype 1b
The NetherlandsGermanyDenmarkFrancePolandRussiaUcrania
Spain FranceThe NetherlandsGermany
Not today, but more work needed on both sides
EBLV1subtypes