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Molecular screening of vector-borne pathogens in ectoparasites from bats in

Ukraine

Poster · June 2021

DOI: 10.13140/RG.2.2.10937.90728

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Cristian Raileanu Anton S. Vlaschenko

TRON gGmbH H.S. Skovoroda Kharkiv National Pedagogical University
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Denys Muzyka Valeria Bohodist

National Scientific Center Institute of Experimental and Clinical Veterinary Medicine H.S. Skovoroda Kharkiv National Pedagogical University
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 Molecular screening of vector-borne pathogens in ectoparasites
from bats in Ukraine
1 1 2,3 3,4 5 4
Cristian Răileanu
1,6
, Oliver Tauchmann , Anton Vlaschenko , Denys Muzyka , Valeria Bohodist , Serhii Filatov ,
Cornelia Silaghi
1
Institute of Infectology, Friedrich-Loeffler-Institut, Suedufer 10, 17492, Greifswald-Insel Riems, Germany
2
Bat Rehabilitation Center of Feldman Ecopark, 62340 Lesnoye, Kharkiv Region, Ukraine
3
Institute of Natural Sciences, Department of Zoology, H.S. Skovoroda Kharkiv National Pedagogical University, Valentynivska st., 2, Kharkiv, 61168, Ukraine
4
National Scientific Center “Institute of Experimental and Clinical Veterinary Medicine”, Pushkinska St., 83, Kharkiv, 61023, Ukraine.
5
Bila Tserkva National Agrarian University, Veterinary Medicine department, Stavishchanskaya st., 126, 09111, Bila Tserkva, Ukraine
6
Department of Biology, University of Greifswald, Domstraße 11, 17489, Greifswald, Germany

Context and Objective

Bats (Mammalia: Chiroptera) represent the second-most diverse order of mammals after rodents, being described as hosts for a wide range of
bloodsucking arthropods that are able to circulate bacterial, protozoal and viral agents. Bats join other mammals in facilitating the spread of
pathogens and are natural reservoir hosts of a large variety of zoonotic diseases with the ability to cross species barriers [1,2]. In addition, due to
their migratory habits, bats can facilitate the long-distance dispersal of pathogens.
The aim of this study was to perform a molecular screening for selected vector-borne pathogens in ectoparasites collected from bats in Ukraine.

Methodology and Results

1. COLLECTION OF ECTOPARASITES FROM BATS 2. SCREENING OF ECTOPARASITES FOR PATHOGENS

Bats were captured and checked for ectoparasites at three different locations in All ectoparasites were tested individually by:
Ukraine: • PCR for Rickettsia spp., Bartonella spp., Ehlichia/Anaplasma spp. and Babesia spp.
• A mine system (Liptsy mines) used by bats for swarming and hibernation. • Carios vespertilionis ticks were tested additionally by nested PCR for Borrelia spp.
• Urban area (Kharkiv city): bats were examined during hibernation season. The amplified PCR products were sequenced for species identification.
• Rural areas (Kharkiv region): the breeding colonies from buildings were checked.
From the total number of collected ectoparasites, so far 143 samples were mor-
phologically identified and screened for pathogens: 100 Carios vespertilionis (short-
legged bat tick) and 43 Nycteridopsylla eusarca (noctule bat flea) (Table below). Sequencing of Carios vespertilionis samples
Pathogen Target Positive Sequenced Identified species
gene samples samples
Bat host species Carios Nycteridopsylla Collection area Rickettsia spp. ompA 7 7 Rickettsia parkeri
vespertilionis eusarca
1 Candidatus E. shimanensis
Myotis dasycneme 28 0 Mine system
Anaplasma/ 16S rRNA 2
Myotis daubentonii 2 0 Mine system Ehrlichia spp. 1 Uncultured Anaplasma sp.

Nyctalus noctula 0 100 Urban area Carios vespertilionis Borrelia spp. 16S-23S 2 2 -
https://biology.stackexchange.com
IGS
Pipistrellus kuhlii 1 0 Rural area
Pipistrellus pygmaeus 11 0 Rural area
Plecotus auritus 1 0 Mine system
Total 43 100 Sequencing of Nycteridopsylla eusarca samples
Nycteridopsylla eusarca
http://britishfleas.myspecies.info Pathogen Target Positive Sequenced Identified species
gene samples samples

Myotis dasycneme Nyctalus noctula Bartonella spp. gltA 10 7 Uncultured Bartonella sp.
https://batslife.eu https://batslife.eu
Pipistrellus pygmaeus
https://batslife.eu
2 Uncultured bacterium
Anaplasma/ 16S rRNA 56
Ehrlichia spp. 6 Wolbachia endosymbiont

Pipistrellus kuhlii Plecotus auritus

Myotis daubentonii https://batslife.eu
https://batslife.eu https://batslife.eu

3. PHYLOGENETIC ANALYSIS
R. parkeri ompA - C. vespertilionis Uncultured Anaplasma sp. 16S rRNA C. Ehrlichia shimanensis 16S rRNA Uncultured Bartonella sp. gltA
C. vespertilionis C. vespertilionis N. eusarca

ML + Tamura 3-parameter ML + Hasegawa-Kishino-Yano ML + Kimura 2-parameter using a discrete Gamma distribution

ML + Tamura 3-parameter using a discrete Gamma distribution
The phylogenetic tree based on the ompA partial sequence demonstrates that all
the Rickettsia sequences detected in Carios vespertilionis in this study belong to
Rickettsia parkeri, clustering with species within the spotted fever group (SFG).
The phylogenetic tree based on the 16s rRNA partial sequence indicates
that the Anaplasma sequence detected in Carios vespertilionis in this study
clusters in a clade that includes Uncultured Anaplasma and Anaplasma ovis.
The phylogenetic analysis based on the 16s rRNA partial sequence indica-
tes that the Ehrlichia sequence detected in Carios vespertilionis clusters
in a clade that includes Candidatus Ehrlichia shimanensis, Uncultured Ehr-
lichia, Ehrlichia minasensis or Ehrlichia canis.
The phylogenetic analysis based on the gltA partial sequence
shows that three Bartonella sequences detected in Nycteri-
dopsylla eusarca cluster in a clade that includes Uncultured Bar-
tonella, Bartonella washoensis and Bartonella sp. The additional
four obtained sequences cluster in a separate clade.

Conclusions
• This study reports for the first time in Ukraine the molecular detection of several bacterial agents in two species of References
ectoparasites found on bats. 1. Calisher, C. H., Childs, J. E., Field, H. E., Holmes, K. V. & Schountz, T. Bats: im-
• Detection of Rickettsia parkeri in Carios vespertilionis corroborated with previous reports indicating that this soft portant reservoir hosts of emerging viruses. Clin Microbiol Rev. 19, 531–545 (2006).
tick can bite humans, suggests the role of this species as a potential source of pathogens for humans. 2.Hornok, S. et al. DNA of piroplasms of ruminants and dogs in Ixodid bat ticks. PLoS
• The data presented extend the knowledge on the distribution of ectoparasite species on bats and their involvement One 11, e0167735 (2016).
to potentially circulate pathogenic agents.
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