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FREE-RANGING UNGULATES AS HOSTS OF IXODID TICKS AND TICK-BORNE
AGENTS IN THE MALÉ KARPATY MTS (SOUTH-WESTERN SLOVAKIA)
Kocianová E.1, Špitalská E.1, Berthová L.1, Svitálková Z.2, Mahríková L.2, Kazimírová M.2, Schnittger L.3
Institute of Virology, SAS, Dúbravská cesta 9, 845 05 Bratislava, SR ,
2 Institute of Zoology, SAS, Dúbravská cesta 9, 845 06 Bratislava, SR
3Instituto de Patobiologia, CICVyA, INTA-Castelar, 1686 Hurlingham, Prov. de Buenos Aires, Argentina
1
Introduction: Free-ranging ungulates are obligate hosts for adult and subadult stages of Ixodes
ricinus ticks. In collaboration with local hunters, spleen and feeding ticks were collected from game
animals shot in deciduous forests in the Malé Karpaty Mts during 2011-2014.
Material and methods: Tissue samples (spleen, blood, skin) and ticks feeding on free-ranging ungulates were obtained from suburban (Bratislava forest park, Železná
studnička) and natural (Fúgelka, Harmónia) sites in the Malé Karpaty Mts during 2011-2014. Ticks were collected from all examined species, except wild boar, from May
to December. The following microorganisms were detected in the biological samples: rickettsiae (PCR, targeting the gltA gene followed by sequencing), boreliae (PCR, 5S23S rRNA, followed by RFLP), Coxiella burnetii (PCR, com1 gene), Anaplasma phagocytophilum (real-time PCR, msp2 gene), babesiae (PCR, 18S rRNA gene,
followed by sequencing) and Candidatus Neoehrlichia mikurensis (CNM, real-time PCR, groEL gene).
Results: Ninety-two ungulates of five species - roe deer (Capreolus
capreolus), red deer (Cervus elaphus), mouflon (Ovis aries musimon),
fallow deer (Dama dama) and wild boar (Sus scrofa) were screened for
tick-borne microorganisms. Ticks were obtained from all examined
species. In the natural site and suburban site, 51.4% (18/35) and 36.8%
(21/57) game were infested by ticks, respectively. Ixodes ricinus
dominated (1524 ind: 90.5% larvae, 8.2% nymphs, 1% females and 0.3%
males). Haemaphysalis concinna were also present (206 ind.: 82%
larvae and 17.9% nymphs). In feeding ticks, overall prevalence of
rickettsiae (R. helvetica, R. monacensis and Rickettsia sp.) was 13.2%.
R. helvetica was detected in all development stages of I. ricinus, R.
monacensis only in larvae.
Prevalence of A. phagocytophilum in adult and subadult I. ricinus
feeding on deer species was 78.0%, while the prevalence of Theileria sp.
and Babesia venatorum was 36.4%.
IR 1 M
0
IR 7 F
Bab. sp.
R. helvetica 25%
C. burnetii 25%
Bab. sp. 25%
IR 59 F
IR 11 M
IR 3 M
IR 4 F
C. burnetii was detected in 2.6% of I. ricinus removed from
mouflons, fallow deer and roe deer. Presence of CNM, B. garinii, B.
afzelii and B. valaisiana was sporadically detected in subadult and adult
ticks.
A. phagocytophilum was detected in spleen of wild boar (29.2%)
and ruminants (88.1%).
Deer were found to be infected with Babesia sp. / Theileria sp.
(31.8%). Phylogenetic analysis using partial sequences of the 18S rRNA
gene showed that the isolates from feeding ticks and deer spleen
clustered with previously described isolates of T. capreoli / Theileria sp.
Spleens and blood of game animals were negative for rickettsiae,
C. burnetii, CNM and borreliae.
A. phag. 87.5%
33.3%
A. phag. 33.3%
Bab.sp. 42.9%
IR 1 M
R. helvetica 13.1%
A. phag. 81.6%
Bab. sp. 63.1%
C. burnetii 2.6%
R. helvetica
9.1%
A. phag. 81.8%
Bab.sp. 72.7%
IR 3 F
Rick. sp. 100%
B.valaisiana
9.1%
Infested 5/9
HC 1L
Bab. sp.
1
Infested 5/8
Infested 17/22
HC 6L 0
A. phag. 33.3%
Bab. sp. 66.7%
N. mikurensis
66.7%
Infested 10/14
Infested 9/13
IR 45 N
IR 6 L
IR 6 L
0
IR 1 N
0
R. helvetica
16.7%
A. phag. 66.6%
Bab. sp. 16.7%
IR 15 N
Rick. sp.
6.7%
A. phag. 66.6%
Bab. sp. 6.7%
IR 28L
Rick. sp 3.6%
R. monacensis
7.1%
C. burnetii 3.6%
A. phag. 60.7%
Bab. sp. 32.1%
B. val. 3.6%
IR 68 N
R. helvetica
7.4%
B. garinii 1.5%
B. valaisiana
1.5%
A. phag. 69.1%
Bab. sp. 13.2%
IR L 32
Rick. sp. 43.8%
B. valaisiana 3.1%
A. phag. 3.1%
Bab. sp. 56.3%
R. helvetica 2.2%
Rick. sp. 4.4%
Bab. sp. 26.7%
C. burnetii 8.9%
B. afzelii 2.2%
B. garinii 2.2%
N. mikurensis
2.2%
Fig. 1 Occurrence and prevalence of tick-borne pathogens in ticks feeding on free-ranging ungulates in the Malé Karpaty Mts (2011-2014).
Conclusions: The results show that free-ranging ungulates are good sentinels for transport of ticks, infected with the screened microorganisms, but
are probably reservoirs only for A. phagocytophilum and Theileria spp., suggesting the occurrence of natural foci in the studied sites of Slovakia.
Acknowledgments: This study is financially supported by the FP7 project EDENext (No. 261504), grants APVV DO7RP-0014-11 and APVV 0280-12 and grant VEGA No. 2/0061/13.