Seroprevalences of antibodies to Neospora caninum and Toxoplasma gondii in zoo animals

Veterinary Parasitology 136 (2006) 223–231 www.elsevier.com/locate/vetpar

Seroprevalences of antibodies to Neospora caninum and Toxoplasma gondii in zoo animals K. Sedla´k a, E. Ba´rtova´ b,* a

Department of Virology and Serology, State Veterinary Institute Prague, Sı´dlisˇtnı´ 136/24, 165 03 Prague 6, Czech Republic b Department of Biology and Wildlife Diseases, University of Veterinary and Pharmaceutical Sciences, Palacke´ho 1/3, 612 42 Brno, Czech Republic Received 27 July 2005; received in revised form 26 October 2005; accepted 22 November 2005

Abstract Neospora caninum is an apicomplexan parasite that causes neuromuscular disease in dogs and abortions in cattle. Little is known about the prevalence of antibodies to this parasite in zoo animals. Sera from 556 animals, from 13 Czech and Slovak zoos were tested for antibodies to N. caninum and Toxoplasma gondii by indirect fluorescent antibody test. Antibodies to N. caninum were found in 31 of 556 zoo animals (5.6%), representing 18 of 114 species tested: Eurasian wolf (Canis lupus lupus), Maned wolf (Chrysocyon brachyurus), fennec (Vulpes zerda), cheetah (Acinonyx jubatus), jaguarundi (Herpailurus yaguarondi), Eurasian lynx (Lynx lynx), Indian lion (Panthera leo goojratensis), fisher (Martes pennanti), blackbuck (Antilope cervicapra), European bison (Bison bonasus), lechwe (Kobus leche), African buffalo (Syncerus caffer caffer), eland (Taurotragus oryx), sitatunga (Tragelaphus spekei gratus), Thorold’s deer (Cervus albirostris), Eastern elk (C. elaphus canadensis), Vietnam sika deer (C. nippon pseudaxis) and Pe`re David’s deer (Elaphurus davidianus). Titres ranged from 1:40 to 1:2560. The highest prevalence 50% was found in family mustelidae of the order carnivora. Antibodies to T. gondii were detected in 193 of 556 zoo animals (34.7%) representing 72 of 114 species tested, with titres ranging from 1:40 to 1:40960. The highest prevalence 100% was found in families: hyaenidae, mustelidae, ursidae and viveridae of the order carnivora. The results of this study indicate that zoo animals have more exposure to T. gondii than to N. caninum. It is the first report of seroprevalence of antibodies to N. caninum in European zoo animals. # 2005 Elsevier B.V. All rights reserved. Keywords: Neosporosis; Toxoplasmosis; Zoo animals; Serologic survey; IFAT

1. Introduction Neospora caninum and Toxoplasma gondii are apicomplexan parasites that may cause clinical * Corresponding author. Tel.: +420 541562633. E-mail addresses: [email protected], [email protected] (E. Ba´rtova´).

disease in domestic and wild animals (Dubey and Beattie, 1988; Dubey, 2003). Little is known about the pathogenicity of N. caninum infection in wildlife, however, fatal neosporosis was reported in two black tailed deer (Odocoileus hemionus columbianus) from California, one full-term stillborn Elds’ deer (Cervus eldi siamensis) from Paris zoo, two full-term stillborn

0304-4017/$ – see front matter # 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2005.11.021

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twin calves of lesser kudu (Tragelaphus imberbis) from German zoo, one 16-day-old white rhinoceros calve (Ceratotherium simium) from the National zoological gardens of South Africa and juvenile fallow deer (Dama dama) from Switzerland (Woods et al., 1994; Dubey et al., 1996; Peters et al., 2001; Williams et al., 2002; Soldati et al., 2004). Antibodies to N. caninum were found in 6 wildlife species (white-tailed deer, moose, raccoon, gray fox, coyot, wolf) from USA (Lindsay et al., 1996, 2001a, 2001b, 2002; Dubey et al., 1999; Gondim et al., 2004), 10 wildlife species (opossum, 5 species of brocket, Maned wolf, Chiloes fox, crab-eating fox, pampas fox) from South America (Patitucci et al., 2001; Melo et al., 2002; Yai et al., 2003; Cano´n-Franco et al., 2004; Tiemann et al., 2005), 7 wildlife species (European bison, Alpine ibex, chamois, red deer, roe deer, red fox, brown hare) from Europe (Buxton et al., 1997; Simpson et al., 1997; Wolfe et al., 2001; Ferroglio et al., 2001a; Ferroglio and Rossi, 2001b; Ferroglio and Trisciuoglio, 2003; Cabaj et al., 2005), 9 wildlife species (zebra, eland, Thompson gazelle, impala, buffalo, warthog, lion, cheetah, spotted hyena) from Africa (Cheadle et al., 1999; Ferroglio et al., 2003) and 1 free-ranging species (dingo) from Australia (Barber et al., 1997). Dubey et al. (2003) reported N. caninum antibodies in 7 marine mammals species (sea otter, Pacific harbour seal, sea lion, ringed seal, bearded seal, Atlantic bottlenose dolphin, walrus). Since reports of prevalence of N. caninum in zoo animals are limited to two studies in felids from South Africa and North America (Cheadle et al., 1999; Spencer et al., 2003), one report in canids from South America (Vitaliano et al., 2004) and one report in cervids from South America (Tiemann et al., 2005), prevalence of neosporosis in other mammal species is largely unknown. To date, no study reported seroprevalence of N. caninum in animals from European zoos. There are many reports on toxoplasmosis in zoo animals. For example, fatal toxoplasmosis was recorded in captive dik-dik and manul from zoo in USA (Dubey et al., 2002; Riemann et al., 1974a, 1974b), in lions from zoo in Africa (Ocholi et al., 1989) and in Siberian tiger from zoo in Belgium (Dorny and Fransen, 1989). In the Czech Republic, fatal toxoplasmosis was recorded in saiga and nilgais antelope from Prague and Chomutov zoos (Sedla´k et al., 2004).

Antibodies to T. gondii were found for example in 6 felids from zoo in Brazil (Silva et al., 2001) and in nine felids and other 104 exotic mammals from zoo in USA (Lappin et al., 1991; Riemann et al., 1974a, 1974b). In the Czech Republic, serologic assay in zoo animals was not done yet. The present study was done to determine the prevalence of antibodies to N. caninum in zoo animals in Czech and Slovak zoos. Additionally, we determined the prevalence of antibodies to another apicomplexan parasite T. gondii in these animals.

2. Materials and methods Sera from 556 zoo animals (114 mammal species or subspecies) were obtained from 12 Czech zoos, 1 Slovak zoo and 4 small private exotic breeds in the Czech Republic. Samples were collected by private veterinaries for State Veterinary Institute Prague during years 1995 till February 2005. The majority of sera samples had been collected from clinically healthy zoo animals. Sera were centrifuged and frozen at 20 8C until they were examined in the year 2005. Serum samples were examined for antibodies against N. caninum and T. gondii by an indirect fluorescence antibody test (IFAT) using a commercially available NC-1 tachyzoite slides (VMRD, Pullman, USA) and Sevatest toxoplasma NIFR (Sevapharma, Prague, Czech Republic). Species-specific conjugate were used: anti-monkey IgG (Sigma-Aldrich s.r.o., Prague, Czech Republic) for cercopithecidae, antihuman IgG (Sevapharma) for hominidae, anti-dog IgG (Sigma) for canidae, anti-cat IgG (Sigma) for felidae, hyaenidae, mustelidae, otariidae, ursidae and viveridae, anti-horse IgG (VMRD) for equidae, anti-llama IgG (VMRD) for camelidae, anti-swine IgG (Sigma) for suidae, anti-deer IgG (KPL) for cervidae, anti-bovine IgG (VMRD) for bovidae except genus Ammotragus, Budorcas, Capra, Hemitragus, Nemorhaedus, Oreamnos, Ovibos, Ovis and Pseudois, in which anti-goat IgG (VMRD) was used. The sera were diluted in a two-fold series starting at 1:20 as a basic dilution. The titre 1:40 was considered positive. Continuous peripheral fluorescence was considered specific (Conrad et al., 1993; Pare´ et al., 1995). N. caninum positive and negative dog and cattle sera (VMRD) were used as controls. Sera from

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Table 1 Results of indirect fluorescence tests for antibody to N. caninum and T. gondii in zoo animals by species Order and family

Species or subspecies

N. caninum No. positive/ tested

Primates Cercopithecidae

T. gondii Range of titers

No. positive/ tested

Range of titers

Rhesus monkey (Macaca mulatta) Barbary macaque (M. sylvanus)

0/1 0/10

0/1 2/10

1:640

Gorilla (Gorilla gorilla) Chimpanzee (Pan troglodytes) Orangutan (Pongo pygmaeus)

0/5 0/1 0/5

4/5 1/1 3/5

1:320–1:1280 1:80 1:320–1:640

Dingo (Canis lupus dingo) Eurasian wolf (C. lupus lupus) Maned wolf (Chrysocyon brachyurus) African wild dog (Lycaon pictus) Bush dog (Speothos venaticus) Fennec (Vulpes zerda)

0/2 2/10 1/6 0/9 0/3 2/2

1/2 10/10 6/6 6/9 3/3 2/2

1:160 1:40–1:80 1:320–1:2560 1:40–1:80 1:40–1:80 1:320–1:2560

Cheetah (Acinonyx jubatus) Jaguarundi (Herpailurus yaguarondi) Ocelot (Leopardus pardalis) Little spotted cat (L. tigrinus) Serval (Leptailurus serval) Eurasian lynx (Lynx lynx) Pallas’s cat (Otocolobus manul) Clouded leopard (Neofelis nebulosa) African lion (Panthera leo) Indian lion (P. l. goojratensis) Jaguar (P. onca) Amur leopard (P. pardus orientalis) Siberian tiger (P. tigris altaica) Sumatran tiger (P. t. sumatrae) Bengal tiger (P. t. tigris)

2/15 1/1 0/1 0/1 0/1 1/2 0/2 0/1 0/2 1/2 0/1 0/3 0/2 0/6 0/1

13/15 1/1 1/1 1/1 1/1 2/2 2/2 0/1 2/2 2/2 1/1 3/3 2/2 6/6 1/1

1:320–1:5120 1:5120 1:320 1:1280 1:320 1:2560–1:20480 1:5120–1:40960

Hyaenidae Mustelidae Otariidae

Brown hyena (Parahyaena brunnea) Fisher (Martes pennanti) South American sea lion (Otaria byronia)

0/3 1/2 0/2

3/3 2/2 0/2

1:320–1:1280 1:320–1:640

Ursidae

Red panda (Ailurus fulgens) American black bear (Ursus americanus) Polar bear (U. maritimus)

0/1 0/3 0/1

1/1 3/3 1/1

1:2560 1:80–1:320 1:320

Viverridae

Fossa (Cryptoprocta ferox)

0/2

2/2

1:320–1:640

Somali wild ass (Equus africanus somaliensis) Boehm’s zebra (E. burchellii boehmi) Chapman’s zebra (E. b. chapmanni) Grevy’s zebra (E. grevyi) Kulan (E. hemionus kulan) Tibetan wild ass (E. kiang) Przewalski’s horse (E. przewalskii) Hartmann’s mountain zebra (E. zebra hartmannae)

0/1 0/4 0/2 0/3 0/1 0/8 0/13 0/14

0/1 4/4 0/2 1/3 0/1 1/8 3/13 6/14

1:40 1:40–1:80 1:40–1:160

Blesbok (Damaliscus pygargus phillipsi)

0/11

3/11

1:160–1:320

Hominidae

Carnivora Canidae

Felidae

Perissodactyla Equidae

Artiodactyla Bovidae

1:80–1:320 1:640

1:320 1:80–1:160 1:80

1:40

1:40

1:40

1:40–1:80 1:640–1:1280 1:160 1:40–1:320 1:160–1:1280 1:160–1:10240 1:160

1:40–1:160 1:40

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Table 1 (Continued ) Order and family

Species or subspecies

N. caninum No. positive/ tested

Camelidae

White-tailed gnu (Connochates gnou) Blue wildebeest (C. taurinus taurinus) Roan antelope (Hippotragus equinus) Sable antelope (H. niger) Scimitar-horned oryx (Oryx dammah) Gemsbok (O. gazella) Addax (Addax nasomaculatus) Mountain reedbuck (Redunca fulvorufula) Lechwe (Kobus leche) Nile lechwe (K. megaceros) Impala (Aepyceros melampus) Springbok (Antidorcas marsupialis) Blackbuck (Antilope cervicapra) Slender-horned gazelle (Gazella leptoceros) Saiga antelope (Saiga tatarica) Barbary sheep (Ammotragus lervia) West Caucasian tur (Capra caucasica) East Caucasian tur (C. cylindricornis) Markhor (C. falconeri) Alpine ibex (C. ibex) Siberian ibex (C. sibirica) Grey goral (Nemorhaedus caudatus) Japanese serow (N. crispus) Rocky mountain goat (Oreamnos americanus) Himalayan tahr (Hemitragus jemlahicus) Californian bighorn sheep (Ovis canadensis californiana) Mouflon (O. orientalis musimon) Bharal (Pseudois nayaur) Golden takin (Budorcas taxicolor bedfordi) Mishmi takin (B. t. taxicolor) Muskox (Ovibos moschatus) American bison (Bison bison) European bison (B. bonasus) Gayal (Bos gaurus f. frontalis) Yak (B. mutus f. grunniens) Watussi cattle (B. primigenius f. taurus) African buffalo (Syncerus caffer caffer) Dwarf forest buffalo (S. c. nanus) Water buffalo (Bubalus arnee f. bubalis) Lowland anoa (B. depressicornis) Mountain anoa (B. quarlesi) Nilgai (Boselaphus tragocamelus) Eland (Taurotragus oryx) Nyala (Tragelaphus angasii) Bongo (T. eurycerus) Lesser kudu (T. imberbis) Sitatunga (T. spekei gratus) Greater kudu (T. strepsiceros)

0/3 0/2 0/17 0/17 0/9 0/6 0/8 0/5 1/4 0/7 0/4 0/2 2/9 0/1 0/11 0/20 0/5 0/3 0/1 0/1 0/3 0/1 0/1 0/3 0/13 0/3

Bactrian camel (Camelus ferus f. bactrianus) Guanaco (Lama guanicoe) Llama (L. glama)

0/10 0/2 0/3

0/1 0/5 0/2 0/3 0/1 0/9 1/4 0/1 0/10 0/3 1/5 0/2 0/4 0/6 0/1 0/2 1/12 0/10 0/4 0/2 1/7 0/1

T. gondii Range of titers

1:80

1:640–1:1280

1:80

1:40

1:80

1:160

No. positive/ tested 0/3 0/2 0/17 0/17 1/9 1/6 0/8 0/5 0/4 2/7 0/4 0/2 2/9 0/1 0/11 8/20 1/5 0/3 1/1 0/1 1/3 0/1 0/1 1/3 6/13 2/3 0/1 3/5 0/2 0/3 0/1 6/9 4/4 0/1 4/10 0/3 1/5 0/2 1/4 1/6 0/1 0/2 0/12 3/10 0/4 0/2 1/7 0/1 1/10 1/2 1/3

Range of titers

1:320 1:320

1:40–1:80

1:40

1:40–1:80 1:40 1:1280 1:40

1:40 1:80–1:1280 1:80–1:320

1:80–1:160

1:80–1:320 1:160–1:2560 1:320–1:640 1:80 1:640 1:80

1:40–1:5120

1:320 1:160 1:80 1:1280

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Table 1 (Continued ) Order and family

Species or subspecies

N. caninum No. positive/ tested

Cervidae

Suidae

T. gondii Range of titers

No. positive/ tested

Range of titers

Alpaca (L. pacos) Vicugna (Vicugna vicugna)

0/3 0/1

1/3 1/1

1:320 1:80

Chinese water deer (Hydropotes inermis) Reeves’s muntjac (Muntiacus reevesi) Thorold’s deer (Cervus albirostris) Prince Alfred’s spotted deer (C. alfredi) Eastern elk (C. elaphus canadensis) Manitoban elk (C. e. manitobensis) Eld’s deer (C. eldii) Vietnam sika deer (C. nippon pseudaxis) Timor deer (C. timorensis) Pe`re David’s deer (Elaphurus davidianus) Fallow deer (Dama dama) Persian fallow deer (D. mesopotamica) Moose (Alces alces) Reindeer (Rangifer tarandus) Roe deer (Capreolus capreolus) Southern pudu (Pudu puda)

0/2 0/6 4/7 0/3 1/1 0/14 0/7 1/3 0/4 7/28 0/3 0/1 0/13 0/9 0/4 0/5

1/2 3/6 3/7 1/3 0/1 7/14 0/7 0/3 0/4 3/28 0/3 0/1 8/13 1/9 2/4 1/5

1:80 1:320–1:640 1:160–1:320 1:160

Babirusa (Babyrousa babyrussa) Red river hog (Potamochoerus porcus pictus) Wild boar (Sus scrofa)

0/1 0/4 0/2

1:80–1:320 1:1280

1:160 1:320–1:2560

1:40–1:160

1:160–1:2560 1:80 1:80–1:320 1:1:80

0/1 2/4 0/2

1:80–1:320

193/556

1:40–1:40960

Total animals

31/556

domestic dog, domestic cat, brown hyena (Parahyaena brunnea), fisher (Martes pennanti), American black bear (Ursus americanus), fossa (Cryptoprocta ferox), domestic horse, yak (Bos mutus f. grunniens), cattle, domestic goat, llama, moose (Alces alces) and domestic pig, screened by IFAT (at dilutions 1:40, 1:80, 1:160, 1:320, 1:640), latex-agglutination test (LAT) (Pastorex Toxo, Bio-Rad, Marnes-la-Coquette, France) or Sabin Feldman dye test (SFR) (at dilutions 1:2, 1:4, 1:8, 1:16, 1:32, 1:64) served as T. gondii positive and negative controls. Additionally, T. gondii positive and negative human sera (Sevatest Toxoplasma set IFR, Sevapharma) were used as the control.

positive titres to T. gondii and N. caninum antibodies are given by species in Table 1. Seroprevalence of N. caninum among all mammals species tested was 5.6% (31 of 556) and seropositive animals were identified from 18 of 114 species tested. Seroprevalence among the different orders and families ranged from 0 to 12.6% and 0 to 50%, respectively. The highest prevalence was found in family mustelidae of the order carnivora (Table 2). The highest titre 1:2560 was found in Pe`re David´s deer. Seroprevalence of T. gondii among all mammals species tested was 34.7% (193 of 556) and seropositive animals were identified from 72 of 114 species tested. Seroprevalence among the different orders and families ranged from 0 to 89.7% and 0 to 100%, respectively. The highest prevalence was found in hyaenidae, mustelidae, ursidae and viveridae families of the order carnivora (Table 2). The highest titre 1:40960 was found in Pallas’s cat. Detailed serologic results of 31 mammals found to be positive for fluorescent antibodies to N. caninum

3. Results Serum samples taken from 556 individual animals representing 4 orders, 14 families and 114 species or subspecies were tested for the presence of N. caninum and T. gondii antibodies. The prevalence and range of

1:40–1:2560

1:40–1:160

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Table 2 Results of serologic testing by indirect fluorescence antibody test to N. caninum and T. gondii in zoo animals by mammal order and family Order

Number N. caninum T. gondii tested Number of positive Number of cases (%) positive cases (%)

Primates

22

Carnivora

87

Perissodactyla Artiodactyla

46 401

0 11 (12.6)

0 20 (5.0)

Family

Number N. caninum T. gondii tested Number of positive Number of positive cases (%) cases (%)

10 (45.5)

Cercopithecidae Hominidae

11 11

0 0

78 (89.7)

Canidae Felidae Hyaenidae Mustelidae Otariidae Ursidae Viverridae

32 41 3 2 2 5 2

5 (15.6) 5 (12.2) 0 1 (50.0) 0 0 0

28 38 3 2 0 5 2

15 (32.6)

Equidae

46

0

15 (32.6)

90 (22.4)

Bovidae Camelidae Cervidae Suidae

compared with antibody titres to T. gondii are summarized in Table 3. Fourteen animals had antibodies solely to N. caninum. In seventeen animals mixed infection was recorded, whereas four animals had the same titres to N. caninum and T. gondii, five animals had higher titres to N. caninum and eight animals had higher titres to T. gondii.

265 19 110 7

7 (2.6) 0 13 (11.8) 0

2 (18.2) 8 (72.7)

53 5 30 2

(87.7) (92.7) (100) (100) (100) (100)

(20.0) (26.3) (27.3) (28.6)

4. Discussion A serological survey in zoo animals has not been yet done in the Czech Republic; although clinical neosporosis was diagnosed in dog (Koudela et al., 1998) and antibodies to N. caninum were detected in aborting dairy cows (Va´clavek et al., 2003).

Table 3 Detailed serologic results from mammals found to be positive for fluorescent antibodies to N. caninum Species or subspecies

N. caninum titres in IFAT

T. gondii titres in IFAT

Eurasian wolf (Canis lupus lupus) no. 1, 2 Fennec (Vulpes zerda) no. 1, 2 Maned wolf (Chrysocyon brachyurus) no. 1 Cheetah (Acinonyx jubatus) no. 1, 2 Indian lion (Pantera leo goojratensis) no. 1 Jaguarundi (Herpailurus yaguarondi) no. 1 Eurasian lynx (Lynx lynx) no. 1 Fisher (Martes pennanti) no. 1 African buffalo (Syncerus caffer caffer) no. 1 Blackbuck (Antilope cervicapra) no. 1, 2 Eland (Taurotragus oryx) no. 1 European bison (Bison bonasus) no. 1 Lechwe (Kobus leche) no. 1 Sitatunga (Tragelaphus spekei gratus) no. 1 Eastern elk (Cervus elaphus canadensis) no. 1 Pe`re David’s deer (Elaphurus davidianus) no. 1, 2, 3, 4, 5, 6, 7 Thorold’s deer (Cervus albirostris) no. 1, 2, 3, 4 Vietnam sika deer (Cervus nippon pseudaxis) no. 1

1:80, 1:320 Both 1:320 1:640 1:80, 1:160 1:40 1:80 1:40 1:40 1:40 1:640, 1:1280 1:80 1:80 1:80 1:160 1:1280 1:640, 1:2560, others 1:320 1:80, 1:160, 1:320, 1:320 1:160

Both 1:80 1:320, 1:2560 1:320 Both 1:320 1:1280 1:5120 1:20480 1:640 <1:40 Both 1:40 <1:40 1:2560 <1:40 <1:40 <1:40 All <1:40 <1:40, 1:160, 1:320, 1:160 <1:40

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The results of this study indicate that zoo animals have more exposure to T. gondii (34.7%) than to N. caninum (5.6%). While antibodies to N. caninum were found only in mustelidae (50%), canidae (15.6%), felidae (12.2%) bovidae (2.6%) and cervidae (11.8%), all 14 families tested, except otariidae, had antibodies to T. gondii. More than 50% of samples with positive N. caninum antibody titres, had antibodies solely to N. caninum. Prevalence of antibodies to N. caninum was set out in animals, in case that five or more animals were tested and was following: Thorold’s deer (57%), Pe`re David’s deer (25%), blackbuck (22.2%), African buffalo (20%), Eurasian wolf (20%), maned wolf (16%), sitatunga (14.3%), cheetah (13.3%) and eland (8.3%). In our study, higher prevalence was recorded in maned wolf 16% compared to 8.5% in maned wolf from Brazil zoo (Vitaliano et al., 2004) and in cheetah 13.3% compared to 6.3% in cheetah from South Africa (Cheadle et al., 1999) or negative results in cheetah from USA zoo (Spencer et al., 2003). On the other hand, lower prevalence was recorded in wolf 20% compared to 39% in wolf from USA (Gondim et al., 2004), in eland 8.3% compared to 92.3% in eland from Africa (Ferroglio et al., 2003) and in African buffalo 20% compared to 50% in African buffalo from Africa (Ferroglio et al., 2003). All animals with antibody to N. caninum came from six czech zoos and from one private exotic breed. The most infected Thorold’s deer came from zoo in ´ stı´ nad Labem; Pe`re David’s deer came from zoo in U Chomutov; blackbuck came from zoo in Plzenˇ; African buffalo, cheetah and lechve came from zoo in Dvu˚r Kra´love´; Eurasian wolf, maned wolf, fennec, Indian lion, jaguarundi, Eurasian lynx, fisher, European bison and sitatunga came from Prague zoo; Eastern elk and Vietnam sika deer were kept in zoo in Ostrava; eland came from private exotic breed. We could not say what was the main source of infection, because zoos included in our study had different way of housekeeping. Since herbivores can be infected by food contaminated with N. caninum oocysts, we suppose that the potential source is hay, which has different origin, come from givers or from different farmers. On the other hand, carnivores can be infected by meat containing N. caninum cysts. For example in Prague zoo, feeding ration for carnivores is composed from 80% of beef meat with bone, and other

229

meat as lamb, pork and horse meat. So, we suppose that carnivores were infected by contaminated beef meat. Antibodies to T. gondii were found in 89.7% carnivora, 45.5% of primates, 32.6% perissodactyla and 22.4% artiodactyla. When compared to another similar study concerning zoo animals, T. gondii antibodies were found in 46.6% carnivora, 25.2% artiodactyla and 22.5% primates (Gorman et al., 1986). The definitive host of T. gondii are felids, with 92.7% prevalence in our study compared to 32, 64.9 and 75.8% in zoo from California (Riemann et al., 1974a, 1974b), Brazil (Silva et al., 2001) and Florida (Lappin et al., 1991), respectively. The higher prevalence in carnivora and primates shows to higher spreading of T. gondii infection in Czech and Slovak zoos compared to zoos in the world. The potential source of T. gondii infection for carnivores is meat, contaminated with T. gondii tissue cysts; herbivora can be infected by food contaminated with T. gondii oocysts and omnivorous animals by both ways. Contamination of zoo environments by oocysts must be prevented, but it is common practice that free-ranging felids have access to some zoos, resulting in high prevalence in some zoo animals. Although clinical disease in animals is not always apparent and is misdiagnosed, there are many reports of clinical toxoplasmosis from zoological parks. In the Czech Republic, there was recorded abortions in nilgais and fatal toxoplasmosis in adult saiga antelope from two zoos (Sedla´k et al., 2004). In our study, representatives of this two species were also tested, but with negative results. A disadvantage of IFAT, compared to another serologic surveys, is a necessary of using of speciesspecific conjugates. However, we used 10 commercially available conjugates for examination of 14 families of 4 mammal orders. To our knowledge, using of species-specific conjugate in mammals is mostly limited into family (i.e. anti-horse conjugate detected antibodies in all equidae, but not in rhinocerotidae) (Sedla´k unpubl. results). However, in the present study, serum samples of some non-feline carnivores (hyaenidae, mustelidae, otariidae, ursidae and viveriidae) reacted strongly with anti-cat conjugate. Function of IFAT with heterologous conjugates was confirmed on control sera by LAT and SFR.

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We report for the first time the presence of N. caninum and T. gondii antibodies in mammals from Czech and Slovak zoos. The results of this study indicate that while T. gondii is fairly common, N. caninum is not widespread among zoo animals, but some of them are potential hosts of N. caninum. To our knowledge, this is the first report of N. caninum antibodies in fennec, jaguarundi, Eurasian lynx, fisher, blackbuck, lechwe, sitatunga, Thorold’s deer, Eastern elk, Vietnam sika deer and Pe`re David’s deer.

Acknowledgements We thank Dr. R. Vodicˇka (Prague Zoo) and Dr. J. Va´hala (Dvu˚r Kra´love´ Zoo) for their assistance in the collection of the serum samples.

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