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Dirofilaria immitis in Bulgaria: the first genetic baseline data and an overview of the current status

M. Panayotova-Pencheva
M. Panayotova-Pencheva
, 
V. Šnábel
V. Šnábel
, 
V. Dakova
V. Dakova
, 
V. Čabanová
V. Čabanová
, 
S. Cavallero
S. Cavallero
, 
A. Trifonova
A. Trifonova
, 
R. Mirchev
R. Mirchev
, 
Z. Hurníková
Z. Hurníková
, 
Z. Vasilková
Z. Vasilková
 y 
M. Miterpáková
M. Miterpáková
   | 05 ago 2020
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Article Category: Research Article
Publicado en línea: 05 ago 2020
Páginas: 211 - 218
Recibido: 14 feb 2020
Aceptado: 27 mar 2020
DOI: https://doi.org/10.2478/helm-2020-0026
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© 2020 M. Panayotova-Pencheva, V. Šnábel, V. Dakova, V. Čabanová, S. Cavallero, A. Trifonova, R. Mirchev, Z. Hurníková, Z. Vasilková, M. Miterpáková, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1

Gel electrophoresis of PCR products in a 1.5% agarose gel using Dirofilaria immitis-specific primers for ITS2 region (1a) and pan-filarioid primers for ITS2 region (1b). Lane M: MW marker; lane 1, sample 1: D. immitis DNA (dog); lane 2, sample 2: D. immitis DNA (jackal); lane 3, sample 3: D. immitis DNA (fox), lane 4, sample 4: positive control (D. immitis DNA); lane 5, sample 5: negative control.
Gel electrophoresis of PCR products in a 1.5% agarose gel using Dirofilaria immitis-specific primers for ITS2 region (1a) and pan-filarioid primers for ITS2 region (1b). Lane M: MW marker; lane 1, sample 1: D. immitis DNA (dog); lane 2, sample 2: D. immitis DNA (jackal); lane 3, sample 3: D. immitis DNA (fox), lane 4, sample 4: positive control (D. immitis DNA); lane 5, sample 5: negative control.

Fig. 2

Neighbor-joining (NJ) phylogram generated from ITS2 and 5.8S (231 bp) sequences showing the relationships among the Dirofilaria immitis from Bulgaria and GenBank reference conspecific sequences. Onchocerca gibsonii (Og) was used as the outgroup. Numbers next to the branches indicate the bootstrap value calculated from 1,000 replicates. The evolutionary distances were computed using the Tamura 3-parameter method and are in the units of the number of base substitutions per site. Bulgarian isolates involved in the phylogram: D. immitis from domestic dog (DD-B), golden jackal (GJ-B), and red fox (RF-B). Geographical origins of the referenced isolates: POR - Portugal, TUR - Turkey, IRA - Iran, CHN - China, IND - India, TAI - Taiwan, BRA - Brazil, CHI - Chile. Hosts and vectors of the referenced isolates (their labels are in brackets following the country codes next to the phylogram): D - dog, RP - red panda, M - mosquito.
Neighbor-joining (NJ) phylogram generated from ITS2 and 5.8S (231 bp) sequences showing the relationships among the Dirofilaria immitis from Bulgaria and GenBank reference conspecific sequences. Onchocerca gibsonii (Og) was used as the outgroup. Numbers next to the branches indicate the bootstrap value calculated from 1,000 replicates. The evolutionary distances were computed using the Tamura 3-parameter method and are in the units of the number of base substitutions per site. Bulgarian isolates involved in the phylogram: D. immitis from domestic dog (DD-B), golden jackal (GJ-B), and red fox (RF-B). Geographical origins of the referenced isolates: POR - Portugal, TUR - Turkey, IRA - Iran, CHN - China, IND - India, TAI - Taiwan, BRA - Brazil, CHI - Chile. Hosts and vectors of the referenced isolates (their labels are in brackets following the country codes next to the phylogram): D - dog, RP - red panda, M - mosquito.

Fig. 3

Geographical distribution of Dirofilaria immitis in dogs (circles), red foxes (squares), golden jackals (rhombs) and wolf (triangle) recorded in the territory of Bulgaria. Orange marks represent D. immitis isolates analysed in the present study.
Geographical distribution of Dirofilaria immitis in dogs (circles), red foxes (squares), golden jackals (rhombs) and wolf (triangle) recorded in the territory of Bulgaria. Orange marks represent D. immitis isolates analysed in the present study.

Review on the occurrence of Dirofilaria immitis in dogs and wild-living carnivores in Bulgaria.

Study year(s)Host species (n)LocalityDetection methodPrevalence % (ex/pos)Reference
1991 – 1996Dogs (341)various regionsKnott´s test5.3Kanev et al. (1996)
1998Dogs (20)Stara ZagoraNecropsy10.0Gerogieva et al. (1999)
Dogs (258)Knott´s test7.4
HW antigen test7.4
Dogs (40)Stara ZagoraNecropsy12.0
1997 – 1999Red foxes (78)Necropsy5.2Georgieva et al. (2001)
Golden jackals (45)Necropsy4.4
Wolves (18)Necropsy5.5
Dogs (487)Knott´s test8.6
2001 – 2006Red foxes (113)various regionsHW Necropsy antigen test9.2 3.0Kirkova et al. (2007)
Golden jackals (56)Necropsy8.9
SofiaKnott´s test8.7
2011Dogs (240)HW antigen test8.7Kostadinov (2012)
RuseKnott´s test15.7
HW antigen test15.7
2012 – 2013Red foxes (87)various regionsNecropsy27.6Mirchev et al. (2013)
2015Dogs (167)Stara ZagoraHW antigen test16.2Pantchev et al. (2015)
Dogs (27)Burgas(8/4)
Pazardzhik(5/3)
Silistra(1/1)
Sofia(2/1)
Red foxes (113)Burgas26.7
Pazardzhik44.4
Plovdiv40.9
Silistra(5/2)
Sliven(9/3)
2012 – 2013Stara Zagora HaskovoNecropsy(8/2) (1/1)Panayotova-Pencheva et al. (2016)
Golden jackals (56)Burgas29.1
Pazardzhik58.7
Pleven27.8
Plovdiv48.1
Silistra44.4
Sliven18.2
Stara Zagora(5/3)
Haskovo(4/4)
Yambol14.3
2013 – 2014Dogs (33)SofiaKnott´s test HW antigen test15.2 15.2Radev et al. (2016)
2012 – 2017Dogs (367)Stara ZagoraHW antigen test34.3Iliev et al. (2017)
2017 – 2018Dogs (80)SofiaHW antigen test31.3Stoyanova et al. (2019)
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