Detection of Toxoplasma gondii in a Eurasian Badger (Meles meles) Living in Wildlife Areas of Izmir, Turkey/Izmir Dogal Hayatinda Yasayan bir Avrasya Porsugunda (Meles meles) Toxoplasma gondii'nin Saptanmasi.
Toxoplasma gondii is an obligatory intracellular protozoon parasite that causes toxoplasmosis in humans and all warm-blooded animals (1). T. gondii has a complex life cycle including sexual and asexual reproductive phases. Felines are the definitive host that oocyst formation occurs through sexual reproduction. Also, tachyzoite and tissue cyst (bradyzoites) production occurs through asexual reproduction in felines and other intermediate hosts such as mammals and birds (1). Oocysts which are highly resistant to environmental conditions and meat containing tissue cysts are the main factors that spread the infection among wild animals (2, 3). The Eurasian badger (Meles meles) belonging to the family Mustelidae lives in Europe and Eurasia located in the northern hemisphere. Although the Eurasian badger is classifed as a carnivore; these animals also feed on a wide variety of plants (4). In some previous studies conducted in Europe, toxoplasmosis was detected in Eurasian badger using serological methods and/or conventional or nested polymerase chain reaction (PCR) methods (5-10). In this study, using real-time PCR, we aimed to investigate the presence of T. gondii in a Eurasian badger that was found dead in the wildlife area of Izmir located in the Aegean region of Turkey.
A Eurasian badger was found dead in wild life of Izmir located in the Aegean region of Turkey and was brought to Izmir Natural Life Park. On external examination, no pathological sign was detected on the body, fur, nails, and eyes. Only ticks were detected on the skin of the animal. Furthermore, no macroscopic abnormality was observed on necropsy of the abdomen and chest cavity. All experiments were performed under the instructions and approval of the Institutional Animal Care and Use Committee of Ege University for animal ethical norms (Permit number: 2014-16).
To investigate the presence of T. gondii in the Eurasian badger, brain tissue was homogenized as previously described (11). Briefly, the brain was removed by opening the skull using a costatome and was homogenized in sterile 0.9% NaCl using a blender (Waring, USA). During the homogenization process, 125 mL 0.9% NaCl was used for 10 g tissue. After homogenization, 0.5 g of trypsin (BD sciences, USA) was added to the homogenate and incubated at 37[degrees]C for 75 min using an incubator shaker (New Brunswick, USA). Later, the homogenate was fltered into a 50 ml falcon tube using two-layered sterile gauze and was centrifuged for 15 min at 910 g. After this process, the supernatant was discarded and the pellet was washed two more times with 0.9% NaCl. Following the last centrifugation, the supernatant was discarded, and DNA extraction from the pellet was performed using QIAamp DNA mini kit (Qiagen, USA).
T. gondii REP gene (Genebank Accession no: AF146527) was amplified using real-time PCR as previously described (11). To amplify a 134 bp gene fragment, the primers (F: 5'-AGGCGAGGGTGAGGATGA-3' and R: 5'-TCGTCTCGTCTGGATCGCAT-3') and hybridization probes [5'-GCCGGAAACATCTTCTCCCTCTCC-3'-FL and 5'-640-CTCTCGTCGCTTCCCAACCACG-3' (Metabion, Germany)] were used. Real-time PCR was performed using LightCycler 1.5 Real Time instrument with LightCycler software, version 4.1 (Roche, Germany). T. gondii genomic DNA, which was diluted from [10.sup.6]/mL to [10.sup.1]/mL, and distilled water were used as positive and negative controls, respectively. According to the quantification and melting curve analysis (Figure 1), T. gondii REP gene was positive in the Eurasian badger brain homogenate.
In Turkey, it is well known that badgers particularly inhabit the regions of Adana, Adapazari, Ankara, Aydin, Balikesir, Bingol, Burdur, Denizli, Eskisehir, Isparta, Kahramanmaras, Konya, Mersin, Mugla, Ordu, Trabzon, and Zonguldak. Furthermore, badgers live throughout Anatolia as well as Thrace region of Turkey and can live up to an altitude of 2000 m (12). In this study, a dead Eurasian badger was found in the wildlife area of Bornova-Izmir located in the Aegean region of Turkey. It was thought that investigations on this animal may provide new data for those who work in the field of animal ecology. On the other hand, the presence and/or frequency of T. gondii in various wild animals is currently being investigated to reveal the animal species that helps spread the parasite and to determine the potential source of toxoplasmosis in nature. The presence of T. gondii has been detected in the Eurasian badger in previous studies conducted in UK, Spain, Portugal, Slovakia, and Poland; these studies have shown the presence of anti-Toxoplasma antibodies and/or used conventional or nested PCR methods to show the presence of T. gondii (5-10). The presence of T. gondii has been reported in various wild animals belonging to the family Mustelidae, such as mink (Mustela vison), river otters (Lontra canadensis), southern sea otters (Enhydra lutris nereis), black-footed ferrets (Mustela nigripes), stone marten (Martes foina), pine marten (Martes martes), ferret (Mustela putorius furo), Eurasian otter otters (Lutra lutra), and polecat (Mustela putorius), using microscopy, in vitro cultivation, and serological methods (6, 13-16).
In this study, T. gondii REP gene was detected for the first time in a Eurasian badger that is found in Turkey using real-time PCR. This result showed that T. gondii is capable of infecting the Eurasian badger and causing chronic infection. Since chronically infected animals are accepted as a potential source of toxoplasmosis, the Eurasian badger can also be a source for the transmission of toxoplasmosis in the natural setting of Izmir, Turkey.
Peer-review: Externally peer-reviewed.
Author Contributions: Concept--M.D., M.K.; Design--M.K., M.D., H.C.; Supervision--M.D., A.D.D., Y.G.; Resources--M.K., E.A.S., H.G.O., D.A.; Materials--M.K., M.D., H.G.O., D.A.; Data Collection and/or Processing--M.K., H.G.O., D.A., H.C., A.D.D.; Analysis and/or Interpretation--M.D., H.C., Y.G., A.D.D.; Literature Search--M.K., E.A.S., H.C., A.D.D.; Writing Manuscript--M.K., H.C., M.D.; Critical Review--Y.G., A.D.D., H.C., M.D.; Other--M.K., E.A.S., D.A., H.G.O.
Conflict of Interest: Authors have no conflicts of interest to declare.
Financial Disclosure: The authors declared that this study has received no financial support.
Hakem Degerlendirmesi: Dis bagimsiz.
Yazar Katkilari: Fikir--M.D., M.K.; Tasarim--M.K., M.D., H.C.; Denetleme--M.D., A.D.D., Y.G.; Kaynaklar--M.K., E.A.S., H.G.O., D.A.; Malzemeler--M.K., M.D., H.G.O., D.A.; Veri Toplanmasi ve/veya Islemesi--M.K., H.G.O., D.A., H.C., A.D.D.; Analiz ve/veya Yorum--M.D., H.C., Y.G., A.D.D.; Literatur Taramasi--M.K., E.A.S., H.C., A.D.D.; Yaziyi Yazan--M.K., H.C., M.D.; Elestirel Inceleme--Y.G., A.D.D., H.C., M.D.; Diger--M.K., E.A.S., D.A., H.G.O.
Cikar Catismasi: Yazarlar cikar catismasi bildirmemislerdir.
Finansal Destek: Yazarlar bu calisma icin finansal destek almadiklarini beyan etmislerdir.
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Muhammet Karakavuk (1)[iD], Duygu Aldemir (2)[iD], Esra Atalay Sahar (1)[iD], Huseyin Can (3)[iD], Huseyin Gokhan Ozdemir (4)[iD], Aysu Degirmenci Doskaya (1)[iD], Adnan Yuksel Guruz (1)[iD], Mert Doskaya (1)[iD]
(1) Department of Parasitology, Ege University School of Medicine, Izmir, Turkey
(2) Department of Internal Medicine, Uludag University Institute of Health Sciences, Faculty of Veterinary Medicine, Bursa, Turkey
(3) Department of Biology, Molecular Biology Section, Ege University Faculty of Science, Izmir, Turkey
(4) The Protection and Development, Union of Izmir Bird Paradise, Izmir, Turkey
Address for Correspondence / Yazisma Adresi: Huseyin Can E.mail: email@example.com
Gelis Tarihi: 08.02.2018
Kabul Tarihi: 10.04.2018
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|Title Annotation:||Olgu Sunumu/Case Report|
|Author:||Karakavuk, Muhammet; Aldemir, Duygu; Can, Huseyin; Ozdemir, Huseyin Gokhan; Doskaya, Aysu Degirmenci|
|Publication:||Turkish Journal of Parasitology|
|Article Type:||Author abstract|
|Date:||Sep 1, 2018|
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