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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

Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. G. Bonchev StrSofia, Bulgaria
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Panayotova-Pencheva, M.
, 
V. Šnábel

V. Šnábel

Institute of Parasitology, Slovak Academy of SciencesHlinkova, Slovakia
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Šnábel, V.
, 
V. Dakova

V. Dakova

Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. G. Bonchev StrSofia, Bulgaria
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Dakova, V.
, 
V. Čabanová

V. Čabanová

  • Institute of Parasitology, Slovak Academy of SciencesHlinkova, Slovakia
  • Biomedical Research Centre, Institute of Virology, Slovak Academy of SciencesBratislava, Slovakia
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Čabanová, V.
, 
S. Cavallero

S. Cavallero

Department of Public Health and Infectious Diseases, Section of Parasitology, Sapienza University of RomeRome, Italy
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Cavallero, S.
, 
A. Trifonova

A. Trifonova

Southwest State EnterpriseBlagoevgrad, Bulgaria
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Trifonova, A.
, 
R. Mirchev

R. Mirchev

Southwest State EnterpriseBlagoevgrad, Bulgaria
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Mirchev, R.
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Z. Hurníková

Z. Hurníková

Institute of Parasitology, Slovak Academy of SciencesHlinkova, Slovakia
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Hurníková, Z.
, 
Z. Vasilková

Z. Vasilková

Institute of Parasitology, Slovak Academy of SciencesHlinkova, Slovakia
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Vasilková, Z.
 und 
M. Miterpáková

M. Miterpáková

Institute of Parasitology, Slovak Academy of SciencesHlinkova, Slovakia
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Miterpáková, M.
  
05. Aug. 2020
Dirofilaria immitis in Bulgaria: the first genetic baseline data and an overview of the current status's Cover Image
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Artikel-Kategorie: Research Article
Online veröffentlicht: 05. Aug. 2020
Seitenbereich: 211 - 218
Eingereicht: 14. Feb. 2020
Akzeptiert: 27. März 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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