Monitoring of infectious diseases is one of the most important pillars of preventive medicine in zoos. Screening for parasitic and bacterial infections is important to keep animals and equipment safe from pathogens that may pose a risk to animal and human health. Zoos usually contain many different animal species living in proximity with people and wild animals. As an epidemiological probe, 188 animals (122 mammals, 65 birds, and one reptile) from a zoo in Slovenia were examined for selected pathogens. Antibodies to Toxoplasma gondii and Neospora caninum were detected by ELISA in 38% (46/122) and 3% (4/122) of mammals, and in 0% (0/64) and 2% (1/57) of birds, respectively; the reptile (0/1) was negative. A statistically significant difference in T. gondii prevalence was found in Carnivora compared to Cetartiodactyla and primate antibodies to Encephalitozoon cuniculi were detected by IFAT in 44% (52/118) of mammals and 20% (11/56) of birds, respectively; the reptile (0/1) was negative. Herbivores had a higher chance of being infected with E. cuniculi compared to omnivores. Antibodies to Chlamydia abortus and Coxiella burnetii were not detected in any of the 74 tested zoo animals. The sera of 39 wild rodents found in the zoo were also examined; they were negative for all three parasites. The parasite T. gondii was detected by PCR in the tissue of two mute swans (Cygnus olor), three eastern house mice (Mus musculus), one yellow-necked field mouse (Apodemus flavicollis), and one striped field mouse (A. agrarius). Positive samples were genotyped by a single multiplex PCR assay using 15 microsatellite markers; one sample from a mute swan was characterized as type II. This micro-epidemiological study offers a better understanding of pathogens in zoo animals and an understanding of the role of zoos in biosurveillance.

Biological Resource Center for Toxoplasma National Reference Centre for Toxoplasmosis Limoges University Hospital 87100 Limoges France

Department of Biology and Wildlife Diseases Faculty of Veterinary Hygiene and Ecology University of Veterinary and Pharmaceutical Sciences Brno 60200 Brno Czech Republic

Department of Mathematics and Statistics Faculty of Science Masaryk University 60200 Brno Czech Republic

Department of Virology and Serology State Veterinary Institute Prague Sídlištní 136 24 16503 Prague 6 Czech Republic

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

Ljubljana Zoo 1000 Ljubljana Slovenia

Tropical Neuroepidemiology Unit University of Limoges 87100 Limoges France

See more in PubMed

McNamara T. The role of zoos in biosurveillance. Int. Zoo Yearb. 2007;41:12–15. doi: 10.1111/j.1748-1090.2007.00019.x. DOI

Bártová E., Lukášová R., Vodička R., Váhala J., Pavlačík L., Budíková M., Sedlák K. Epizootological study on Toxo-plasma gondii in zoo animals in the Czech Republic. Acta Trop. 2018;187:222–228. doi: 10.1016/j.actatropica.2018.08.005. PubMed DOI

Camps S., Dubey J.P., Saville W.J.A. Seroepidemiology of Toxoplasma gondii in zoo animals in selected zoos in the mid-western United States. J. Parasitol. 2008;94:648–653. doi: 10.1645/GE-1453.1. PubMed DOI

Pas A., Dubey J.P. Fatal Toxoplasmosis in Sand Cats (Felis margarita) J. Zoo Wildl. Med. 2008;39:362–369. doi: 10.1638/2007-0149.1. PubMed DOI

Lobato J., Silva D.A., Mineo T.W., Amaral J.D., Segundo G.R.S., Costa-Cruz J.M., Mineo J.R. Detection of immuno-globulin G antibodies to Neospora caninum in humans: High seropositivity rates in patients who are infected by human im-munodeficiency virus or have neurological disorders. Clin. Vaccine Immunol. 2006;13:84–89. doi: 10.1128/CVI.13.1.84-89.2006. PubMed DOI PMC

Barratt J.L.N., Harkness J., Marriott D., Ellis J.T., Stark D. Importance of nonenteric protozoan infections in immuno-compromised people. Clin. Microbiol. Rev. 2010;23:795–836. doi: 10.1128/CMR.00001-10. PubMed DOI PMC

Reichel M.P., Ayanegui-Alcérreca M.A., Gondim L.F., Ellis J.T. What is the global economic impact of Neospora caninum in cattle—The billion dollar question. Int. J. Parasitol. 2013;43:133–142. doi: 10.1016/j.ijpara.2012.10.022. PubMed DOI

Donahoe S.L., Lindsay S.A., Krockenberger M., Phalen D., Šlapeta J. A review of neosporosis and pathologic findings of Neospora caninum infection in wildlife. Int. J. Parasitol. Parasites Wildl. 2015;4:216–238. doi: 10.1016/j.ijppaw.2015.04.002. PubMed DOI PMC

Sangster C., Bryant B., Campbell-Ward M., King J.S., Šlapeta J. Neosporosis in an Aborted Southern White Rhinoceros (Ceratotherium simum simum) Fetus. J. Zoo Wildl. Med. 2010;41:725–728. doi: 10.1638/2009-0250.1. PubMed DOI

Guscetti F., Mathis A., Hatt J.-M., Deplazes P. Overt fatal and chronic subclinical Encephalitozoon cuniculi microsporidiosis in a colony of captive emperor tamarins (Saguinus imperator) J. Med. Primatol. 2003;32:111–119. doi: 10.1034/j.1600-0684.2003.00016.x. PubMed DOI

Juan-Sallés C., Garner M.M., Didier E.S., Serrato S., Acevedo L.D., Ramos-Vara J.A., Nordhausen R.W., Bowers L.C., Parás A. Disseminated Encephalitozoonosis in Captive, Juvenile, Cotton-top (Saguinus oedipus) and Neonatal Emperor (Saguinus imperator) Tamarins in North America. Vet. Pathol. 2006;43:438–446. doi: 10.1354/vp.43-4-438. PubMed DOI

Asakura T., Nakamura S., Ohta M., Une Y., Furuya K. Genetically unique microsporidian Encephalitozoon cuniculi strain type III isolated from squirrel monkeys. Parasitol. Int. 2006;55:159–162. doi: 10.1016/j.parint.2006.02.002. PubMed DOI

Davis M.R., Kinsel M., Wasson K., Boonstra J., Warneke M., Langan J.N. Fatal disseminated encephalitozoonosis in a captive, adult Goeldi’s Monkey (Callimico goeldii) and subsequent serosurvey of the exposed conspecifics. J. Zoo Wildl. Med. 2008;39:221–227. doi: 10.1638/2007-0114R.1. PubMed DOI

Kitz S., Grimm F., Wenger S., Hatt J., Kipar A., Hetzel U. Encephalitozoon cuniculi infection in Barbary striped grass mice (Lemniscomys barbarus) Schweiz. Arch. Tierheilkd. 2018;160:394–400. doi: 10.17236/sat00166. PubMed DOI

Berri M., Bernard F., Lécu A., Ollivet-Courtois F., Rodolakis A. Molecular characterisation and ovine live vaccine 1B evaluation toward a Chlamydophila abortus strain isolated from springbok antelope abortion. Vet. Microbiol. 2004;103:231–240. doi: 10.1016/j.vetmic.2004.08.009. PubMed DOI

Salinas J., Caro M.R., Vicente J., Cuello F., Reyes-Garcia A.R., Buendía A.J., Rodolakis A., Gortázar C. High preva-lence of antibodies against Chlamydiaceae and Chlamydophila abortus in wild ungulates using two “in house” blocking-ELISA tests. Vet. Microbiol. 2009;135:46–53. doi: 10.1016/j.vetmic.2008.10.001. PubMed DOI

Cutler S.J., Bouzid M., Cutler R.R. Q fever. J. Infect. 2007;54:313–318. doi: 10.1016/j.jinf.2006.10.048. PubMed DOI

Agerholm J.S. Coxiella burnetii associated reproductive disorders in domestic animals-a critical review. Acta Vet. Scand. 2013;55:13. doi: 10.1186/1751-0147-55-13. PubMed DOI PMC

Ruiz-Fons F., Rodríguez Ó., Torina A., Naranjo V., Gortázar C., De La Fuente J. Prevalence of Coxiella burnetti infection in wild and farmed ungulates. Vet. Microbiol. 2008;126:282–286. doi: 10.1016/j.vetmic.2007.06.020. PubMed DOI

Clemente L., Fernandes T.L., Barahona M.J., Bernardino R., Botelho A. Confirmation by PCR of Coxiella burnetii infection in animals at a zoo in Lisbon, Portugal. Vet. Rec. 2008;163:221–222. doi: 10.1136/vr.163.7.221. PubMed DOI

Dorko E., Rimarova K., Pilipcinec E., Travnicek M. Prevalence of Coxiella burnetii antibodies in wild ruminants in Ka-vecany zoo, Kosice, Eastern Slovakia. Ann. Agric. Environ. Med. 2009;16:321–324. PubMed

Lloyd C., Stidworthy M.F., Wernery U. Coxiella burnetii Abortion in Captive Dama Gazelle (Gazella dama) in the United Arab Emirates. J. Zoo Wildl. Med. 2010;41:83–89. doi: 10.1638/2009-0005.1. PubMed DOI

Bartova E., Sedlak K., Nagl I., Vodicka R., Racka K., Slezakova R., Ajzenberg D. Genotyping of Toxoplasma gondii iso-lated from Pallas’s cats with fatal toxoplasmosis; Proceedings of the Infectious and Parasitic Diseases of Animals, 5th International Scientific Conference; Košice, Slovakia. 4–9 September 2014.

Sós E., Szigeti A., Fok É., Molnár V., Erdélyi K., Perge E., Biksi I., Gál J. Toxoplasmosis in Tammar wallabies (Macropus eugenii) in the Budapest Zoo and Botanical Garden (2006–2010) Acta Vet. Hung. 2012;60:361–370. doi: 10.1556/avet.2012.031. PubMed DOI

Epiphanio S., Sinhorini I., Catão-Dias J. Pathology of Toxoplasmosis in Captive New World Primates. J. Comp. Pathol. 2003;129:196–204. doi: 10.1016/S0021-9975(03)00035-5. PubMed DOI

Minervino A.H.H., Soares H.S., Barrêto-Júnior R.A., Neves K.A.L., Pena H.F.D.J., Ortolani E.L., Dubey J.P., Gennari S.M. Seroprevalence of Toxoplasma gondii Antibodies in Captive Wild Mammals and Birds in Brazil. J. Zoo Wildl. Med. 2010;41:572–574. doi: 10.1638/2010-0046.1. PubMed DOI

Sedlák K., Bártová E. Seroprevalences of antibodies to Neospora caninum and Toxoplasma gondii in zoo animals. Vet. Parasitol. 2006;136:223–231. doi: 10.1016/j.vetpar.2005.11.021. PubMed DOI

Kim K., Weiss L.M. Toxoplasma gondii: The model apicomplexan. Int. J. Parasitol. 2004;34:423–432. doi: 10.1016/j.ijpara.2003.12.009. PubMed DOI PMC

Costache C.A., Colosi H.A., Blaga L., Györke A., Paştiu A.I., Colosi I.A., Ajzenberg D. First isolation and genetic characterization of a Toxoplasma gondii strain from a symptomatic human case of congenital toxoplasmosis in Romania. Parasite. 2013;20:11. doi: 10.1051/parasite/2013011. PubMed DOI PMC

De Sousa S.R., Ajzenberg D., Canada N., Freire L., Da Costa J.M.C., Dardé M.-L., Thulliez P., Dubey J.P. Biologic and molecular characterization of Toxoplasma gondii isolates from pigs from Portugal. Vet. Parasitol. 2006;135:133–136. doi: 10.1016/j.vetpar.2005.08.012. PubMed DOI

Dubey J.P., Schares G. Neosporosis in animals—The last five years. Vet. Parasitol. 2011;180:90–108. doi: 10.1016/j.vetpar.2011.05.031. PubMed DOI

Mineo T.W., Carrasco A.O., Raso T.F., Werther K., Pinto A.A., Machado R.Z. Survey for natural Neospora caninum infection in wild and captive birds. Vet. Parasitol. 2011;182:352–355. doi: 10.1016/j.vetpar.2011.05.022. PubMed DOI

de Barros L.D., Miura A.C., Minutti A.F., Vidotto O., Garcia J.L. Neospora caninum in birds: A review. Parasitol. Int. 2018;67:397–402. doi: 10.1016/j.parint.2018.03.009. PubMed DOI

Jordan C.N., Zajac A.M., Lindsay D.S. Encephalitozoon cuniculi infection in rabbits. Compend. Contin. Educ. Pract. Vet. N. Am. Ed. 2006;28:108.

Webster J.D., Miller M.A., Vemulapalli R. Encephalitozoon cuniculi-Associated Placentitis and Perinatal Death in an Alpaca (Lama pacos) Vet. Pathol. 2008;45:255–258. doi: 10.1354/vp.45-2-255. PubMed DOI

Hinney B., Sak B., Joachim A., Kváč M. More than a rabbit’s tale—Encephalitozoon spp. in wild mammals and birds. Int. J. Parasitol. Parasites Wildl. 2016;5:76–87. doi: 10.1016/j.ijppaw.2016.01.001. PubMed DOI PMC

Cristina N., Oury B., Ambroise-Thomas P., Santoro F. Restriction-fragment-length polymorphisms among Toxoplasma gondii strains. Parasitol. Res. 1991;77:266–268. doi: 10.1007/BF00930870. PubMed DOI

Ajzenberg D., Collinet F., Mercier A., Vignoles P., Dardé M.L. Genotyping of Toxoplasma gondii isolates with 15 mi-crosatellite markers in a single multiplex PCR assay. J. Clin. Microbiol. 2010;48:4641–4645. doi: 10.1128/JCM.01152-10. PubMed DOI PMC

Muller N., Zimmermann V., Hentrich B., Gottstein B. Diagnosis of Neospora caninum and Toxoplasma gondii infection by PCR and DNA hybridisation immunoassy. J. Clin. Microbiol. 1996;34:2850–2852. doi: 10.1128/JCM.34.11.2850-2852.1996. PubMed DOI PMC

Valencáková A., Bálent P., Novotný F., Cisláková L. Application of specific primers in the diagnosis of Encephalitozoon spp. Ann. Agric. Environ. Med. 2005;12:321–323. PubMed

Statsoft, Inc. STATISTICA (Data Analysis Software System), Version 12. [(accessed on 8 November 2020)]; Available online: www.statsoft.com.