Reports on non-invasive blood sampling are limited, and there are only a few studies on using kissing bugs (Reduviidae) and medicinal leeches (Hirudo medicinalis) for hematology and biochemistry testing in various zoo animal species. The aim of this study was to evaluate the usefulness of non-invasive blood sampling with medicinal leeches for arbovirus epidemiological investigations in various animal species from one zoo collection. Medicinal leeches were manually applied on 35 animals of 11 species. Control blood samples were obtained by venipuncture of the jugular vein. Antibodies to tick-borne encephalitic virus (TBEV) were detected by using the immunoenzymatic method or an immunofluorescent assay (IFAT), depending on the animal species. One of the 35 animals (2.9%) was seropositive (Ovis aries), whereas the rest of the samples were seronegative in both methods of sampling (non-invasive by leeches vs. invasive by venipuncture). Blood sampling using medicinal leeches showed promising results. It is likely a good alternative to other more complex and invasive methods, and it can provide significant advancement in blood sampling for preventive medicine and epidemiological studies in zoo animals.

Department of Biology and Wildlife Diseases Faculty of Veterinary Hygiene and Ecology University of Veterinary Sciences Brno Palackého tř 1946 1 612 42 Brno Czech Republic

Department of Virology and Serology Prague State Veterinary Institute Prague Sídlištní 136 24 165 03 Prague Czech Republic

Institute for Poultry Birds Small Mammals and Reptiles Faculty of Veterinary Medicine University of Ljubljana Gerbičeva 60 1000 Ljubljana Slovenia

Institute of Microbiology and Immunology Faculty of Medicine University of Ljubljana Zaloška 4 1000 Ljubljana Slovenia

Ljubljana Zoo Večna Pot 70 1000 Ljubljana Slovenia

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Validation of Medicinal Leeches (Hirudo medicinalis) as a Non-invasive Blood Sampling Tool for Hematology and Biochemistry Profiling in Mammals