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Seroprevalence of equine babesiosis in the Black Sea region of Turkey
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Parasitology International 57 (2008) 198 – 200 www.elsevier.com/locate/parint
Seroprevalence of equine babesiosis in the Black Sea region of Turkey Mustafa Acici a,⁎, Sinasi Umur a , Tolga Guvenc b , H. Hilal Arslan c , Mithat Kurt d a
Department of Parasitology, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey Department of Pathology, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey Department of Internal Medicine, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey d Parasitology Laboratory, Samsun Veterinary Control and Research Institute, Samsun, Turkey b
c
Received 31 May 2007; received in revised form 13 December 2007; accepted 16 December 2007 Available online 27 December 2007
Abstract The prevalence of Theileria equi and Babesia caballi was determined in equid blood samples in five provinces of the Black Sea region of Turkey by using the indirect fluorescent antibody test (IFAT). Of 153 samples, 53 (34.6%) and 33 (21.5%) were seropositive to B. caballi and T. equi, respectively. In addition, 8 (5.2%) of samples were seropositive to both T. equi and B. caballi. Anti T. equi and B. caballi antibodies were detected in all five regions. The prevalence of B. caballi was higher than T. equi in all counties. Antibodies to T. equi and B. caballi were detected in horses of all ages, and there were no significant differences among age groups. Out of 84 horses, 32 (38.0%) were positive for B. caballi infection and 20 (23.8%) were positive for T. equi infection. Five horses (5.6%) were found to be seropositive to both B. caballi and T. equi. Of 38 donkeys, 14 (36.8%) were found to be positive for B. caballi infection and 5 (13.1%) positive for T. equi infection. In addition, 2 (5.2%) samples were seropositive for both T. equi and B. caballi infections. Out of 31 mules, 8 (25.8%) were positive for B. caballi infection and 8 (25 8%) positive for T. equi infection. One (3.2%) sample was seropositive for both T. equi and B. caballi infections. Of all the animals in this study, only 3 horses were infected by Rhipicephalus turanicus and Hyalomma detritum, and no haemoparasites were detected by microscopic examination. © 2007 Elsevier Ireland Ltd. All rights reserved. Keywords: Babesiosis; Equidae; IFAT; Turkey
1. Introduction Equine babesiosis, caused by the hemoprotozoans T. equi and B. caballi, is responsible for important economic losses in the equine industry of tropical and subtropical areas of the world [1–3]. Levine [4] stated that equine babesiosis is an acute, subacute or chronic disease of equids that is caused by the intraerytrocytic protozoans T. equi and B. caballi which share the same vectors. Of the estimated world population of 120 million horses, 90% are bred in areas with endemic babesiosis [1]. In Turkey, there have been reports of hemoprotozoal diseases [5,6], and more recently in Elazığ and Kayseri in Turkey [7,8]. Information on the distribution and prevalence of equine babesiosis is limited in Turkey. One of the ⁎ Corresponding author. Department of Parasitology, Faculty of Veterinary Medicine, University of Ondokuz Mayis 55139 Kurupelit, Samsun, Turkey. Tel.: +90 3623121919 2822; fax: +90 3624576000 2801. E-mail address: [email protected] (M. Acici). 1383-5769/$ - see front matter © 2007 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.parint.2007.12.009
objectives of this research was to perform an epidemiological survey of T. equi and B. caballi antibodies in equid animals of part of the Black Sea region of Turkey. 2. Materials and methods The research was carried out mainly in areas ranging from 50 m to 700 m altitude and covering parts of five northern Turkish provinces: Sinop (Sazli and Akbas); Samsun (Tekkekoy–Yeşil Alan, Yukarı Çinik; Terme–Ahmetbey; Çarşamba, Yamalı); Ordu (Perşembe–Yumrutaş, Yeniköy, Ramazan, Töngeldüzü, Boğazcık); Amasya (Direkli) and Tokat (Kadıvakfı). The study covered 13 villages and included 120 farms, each with one or more animals. 2.1. Blood and tick collection In May and June of 2004, blood samples were collected on each of the 120 selected farms from a total of 153 animals. The
M. Acici et al. / Parasitology International 57 (2008) 198–200
local crossbred animals included 84 horses, 38 donkeys and 31 mules. The blood samples were allowed to coagulate, after which the serum was removed and stored at −20 °C until further use. Thin, Giemsa-stained blood smears were prepared from fresh, periferal capillary blood and examined microscopically for the presence of parasites. Tick specimens were collected from all areas of animals' bodies, and preserved in 70% ethanol, until identified from the keys of Karaer et al. [9]. Tick smears were prepared and checked for Babesia vermicules in the eggs [10].
199
Table 2 Distribution of equine babesiosis and Babesia species in different age groups⁎ Age (year)
N 0–3 4–9 10–15 16 ≤
Babesiosis N
%
153 27 50 53 23
37.0 56.0 56.6 43.5
B. caballi χ2
3.79
N
%
153 27 50 53 23
25.9 42.0 37.7 30.4
T. equi χ2
2.35
N
%
153 27 50 53 23
18.5 18.0 26.4 17.4
χ2
1.49
⁎: p N 0.05.
2.2. IFAT analysis The T. equi and B. caballi antigens, 12-well IFA substrate slides (Lot BKG-120-040907) and the anti-horse IgG FITC conjugate (EG-20-040217) used in the Indirect Fluorescent Antibody Test (IFAT) were acquired from Fuller Laboratories, Fullerton, CA, USA. T. equi (BEP-031029) and B. caballi (BKP040219) were used as positive controls; T. equi and B. caballi (NE50-040325) was used as a negative control. Test procedures were performed according to the manufacturer's instructions for use. The antigen slides were stored at −20 °C until the analysis and they were brought to room temperature before opening slide envelopes. All sera were diluted 1:80 in PBS before the test. From each serum dilution (10 μl) was added to a slide well and incubated at 37 °C for 30 min. Then, the slides were washed three times with PBS (pH 7.2). Conjugate (10 μl, FITC-labelled rabbit anti-horse IgG with bovine serum albumin and Evans'blue counterstain) was added to a slide well. The slides were incubated for 30 min. at 37 °C. After five washings with PBS the glass slides were dried out and mounted in glycerol/PBS (1:1) for examination under an epifluorescence microscope. Buffer control, conjugate control and negative and positive control sera were included in each test. Titers of 1:80 or more that showed strong fluorescence were considered to be positive in diluted sera and IFAT was applied to the differential diagnosis of T. equi and B. caballi infections [11]. The chi-square test was used to test for significant difference between groups. 3. Results Out of 153 sampling, 33 (21.5%) and 53 (34.6%) were positive for T. equi and B. caballi infections, respectively, at a dilution ≥ 1:80. Among the sera collected from horses, donkeys and mules, 32 (38.0%), 14 (36.8%) and 8 (25.8%) were positive
Table 1 Prevalence of equine babesiosis and Babesia species in the Black Sea region Province
N Amasya Ordu Samsun Sinop Tokat ⁎: p b 0.01.
Babesiosis N
%
153 30 30 30 31 32
83.3 53.3 56.7 35.5 28.1
B. caballi χ
N
%
22.68⁎
153 30 30 30 31 32
60.0 36.7 46.7 32.3 6.3
2
T. equi χ
N
%
χ2
21.48⁎
153 30 30 30 31 32
33.3 20.0 23.3 3.2 25.0
9.10⁎
2
to B. caballi, and 20 (23.8%), 5 (13.1%) and 8 (25.8%) were positive to T. equi, respectively. Anti-Babesia antibodies were detected in every village and its prevalence is shown in Tables 1 and 2. B. caballi infections were more prevalent in horses than in donkeys and mules. The percentages were 38.0%, 36.8% and 25.8%, respectively. T. equi (13.1%) was least prevalent in donkeys. There were statistical differences (p b 0.01) among counties for Babesia prevalence (Table 1). However, there were no statistical differences (p N 0.05) between age groups for their prevalence (Table 2). Five Rhipicephalus turanicus were found in horses in Samsun and two Hyalomma detritum in Amasya and Tokat. No Babesia parasites were detected in periferal blood smears or in the ticks assayed. None of these ticks were infected with Babesia spp. 4. Discussion Several direct and indirect detection methods, including blood smears [12], in-vitro cultures [13], DNA probes [14] and serology [15,16], have been used for the diagnosis of B. caballi and T. equi infections. Standard serological tests for babesiosis are the complement fixation test and the indirect fluorescent antibody test [2]. However, it has been reported that because of its low sensitivity and specificity, the complement fixation test fails to discriminate accurately between negative and carrier animals [17]. Weiland [18] demonstrated the strong crossreactivity of anti-B. caballi horse serum with the lysate of T. equi-infected erythrocytes, using an Enzyme-Linked Immunosorbent Assay. In this study, 8 (5.2%) of samples in equines were seropositive for both B. caballi and T. equi. IFAT and ELISA tests have been used to detect equine babesiosis in Turkey [5,8]. According to Inci [7], B. caballi and T. equi infections were detected in 7.86% and 4.49% of horses in the Kayseri region, respectively. Balkaya and Erdogmus [8] researched babesiosis in Elazig and Malatya provinces of Turkey and found B. caballi in 2.5% and B. equi in 20.5% of horses, respectively. Ozcan [19] reported B. caballi (71.87%) and T. equi (28.12%) in horses and donkeys respectively in Ankara in Turkey. In this study, the prevalence of B. caballi was 38.0% in horses, 36.8% in donkeys, and 25.8% in mules. T. equi was detected in 23.8% of horses, 13.1% of donkeys, and 25.8% of mules. On the other hand, no infections were found in donkeys less than one year of age. Equine babesiosis is endemic in many tropical and subtropical areas and is transmitted by ticks, including the genera Boophilus, Hyalomma, Dermacentor and Rhipicephalus [20]. In this study, H. detritum and R.
200
M. Acici et al. / Parasitology International 57 (2008) 198–200
turanicus were found on equines. However, not Babesia spp. was detected in them. In conclusion, this study showed a high prevalence of equine babesiosis seropositivity in equids in the northern region of Turkey. The results presented are in agreement with the ELISA study reported by Balkaya and Erdogmus [8] which showed no significant differences between age groups in the prevalence of equine babesiosis. Although no parasites were detected in Giemsa-stained blood smears, indirect IFAT results showed that T. equi and B. caballi are endemic in the Black Sea region. Acknowledgements The authors are grateful for the support of the Scientific Research Committee of Ondokuz Mayis University, Samsun, Turkey (Project No. Vet. 008) and to Gregory Thomas Sullivan of the same university for his assistance in English editing. References [1] Schein E. Equine babesiosis. In: Ristic M, editor. Babesiosis of Domestic Animals and Man. Boca Raton, FL: CRC Press; 1988. p. 197–209. [2] Friedhoff KT. The piroplasms of equidae significance fort he international equine trade. Berl Münch Tierarztl Wochenschr 1982;95:368–74. [3] Kuttler KL. Chemotherapy of babesiosis. In: Ristic M, editor. Babesiosis of Domestic Animals and Man. Boca Raton, FL: CRC Press; 1988. p. 10–2. [4] Levine ND. Veterinary Protozoology. Ames. Iowa: Iowa State University Press; 1985. [5] Inci A. Gemlik Askeri Harası atlarında Babesia caballi (Nuttal, 1910) ve Babesia equi (Laveran, 1901)'nin mikroskobik muayeneyle saptanması. Tr J Vet Anim Sci 1997;21:43–6. [6] Aktas M, Dumanlı N. Malatya Sultansuyu Tarım Isletmesi atlarında subklinik Babesia equi (Laveran, 1901) ve Babesia caballi (Nuttal, 1910) enfeksiyonları. Turk Parasitol Derg 2000;24:55–6.
[7] Inci A. Kayseri yöresinde tek tırnaklılarda Babesia equi (Laveran, 1901) ve Babesia caballi (Nuttal, 1910) yaygınlığının mikroskobik muayeneyle araştırılması. Fırat Üniv Sağ Bil Derg 2002;16:85–8. [8] Balkaya I, Erdogmus SZ. Investigation of prevalence of Babesia equi (Laveran, 1901) and Babesia caballi (Nuttal, 1910) in horses by serological methods in Elazığ and Malatya province. FU Sağlık Bil Derg 2006;20:61–3. [9] Karaer Z, Yukari BA, Aydin L. Türkiye keneleri ve vektörlükleri. In: Özcel A, Daldal N, editors. Parazitoloji'de Artropod Hastalıkları ve Vektörler. Türkiye Parazitoloji Derneği Yayını, İzmir; 1997. No:13. [10] Holbrook AA, Anthony DW, Johnson AJ. Observations on the development of Babesia caballi (Nuttall) in the tropical horse tick Dermacentor nitens Neumann. J Protozool 1968;15:391–6. [11] Madden PA, Holbrook AA. Equine piroplasmosis: indirect fluorescent antibody test for Babesia caballi. Am J Vet Res 1968;29:117–23. [12] Retief GB. A comparison of equine piroplasmosis in South Africa and the United States. J Am Vet Med Assoc 1964;145:912–6. [13] Holman PJ, Freriches WM, Chieves L, Wagner GG. Culture confirmation of the carrier status of Babesia caballi infected horses. J Clin Microbiol 1993;31:698–701. [14] Posnett ES, Ambrosio RE. DNA probes fort he detection of Babesia caballi. Parasitology 1991;102:365–7. [15] Bose R, Peymann B. Diagnosis of Babesia caballi infections in horses by Enzyme-Linked Immunosorbent Assay (ELISA) and Western blot. Int J Parasitol 1994;24:341–6. [16] Brüning A, Phipps P, Possnett E, Canning EU. Monoclonal antibodies against Babesia caballi and Theileria equi and their application in serodiagnosis. Vet Parasitol 1997;58:11–26. [17] Tenter AM, Friedhoff KT. Serodiagnosis of experimental and natural Babesia equi and B. caballi infections. Vet Parasitol 1986;20:49–61. [18] Weiland G. Species-specific serodiagnosis of equine piroplasma infections by means of complement fixation test, immunofluorescence and enzymelinked immunosorbent assay. Vet Parasitol 1986;20:43–8. [19] Ozcan CH. Ankara ve civarında evcil hayvanlarda görülen piroplasmose vak'aları ve tedavileri üzerinde araştırmalar. Ankara Univ Vet Fak Yayın. 143, Ankara Univ Basımevi; 1961. [20] Rampersad J, Cesar E, Campbell MD, Samlal M, Ammons D. A field evaluation of PCR for the routine detection of Babesia equi in horses. Vet Parasitol 2003;114:81–7.
Parasitology International 57 (2008) 198 – 200 www.elsevier.com/locate/parint
Seroprevalence of equine babesiosis in the Black Sea region of Turkey Mustafa Acici a,⁎, Sinasi Umur a , Tolga Guvenc b , H. Hilal Arslan c , Mithat Kurt d a
Department of Parasitology, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey Department of Pathology, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey Department of Internal Medicine, Faculty of Veterinary Medicine, Ondokuz Mayis University, Kurupelit, Samsun, Turkey d Parasitology Laboratory, Samsun Veterinary Control and Research Institute, Samsun, Turkey b
c
Received 31 May 2007; received in revised form 13 December 2007; accepted 16 December 2007 Available online 27 December 2007
Abstract The prevalence of Theileria equi and Babesia caballi was determined in equid blood samples in five provinces of the Black Sea region of Turkey by using the indirect fluorescent antibody test (IFAT). Of 153 samples, 53 (34.6%) and 33 (21.5%) were seropositive to B. caballi and T. equi, respectively. In addition, 8 (5.2%) of samples were seropositive to both T. equi and B. caballi. Anti T. equi and B. caballi antibodies were detected in all five regions. The prevalence of B. caballi was higher than T. equi in all counties. Antibodies to T. equi and B. caballi were detected in horses of all ages, and there were no significant differences among age groups. Out of 84 horses, 32 (38.0%) were positive for B. caballi infection and 20 (23.8%) were positive for T. equi infection. Five horses (5.6%) were found to be seropositive to both B. caballi and T. equi. Of 38 donkeys, 14 (36.8%) were found to be positive for B. caballi infection and 5 (13.1%) positive for T. equi infection. In addition, 2 (5.2%) samples were seropositive for both T. equi and B. caballi infections. Out of 31 mules, 8 (25.8%) were positive for B. caballi infection and 8 (25 8%) positive for T. equi infection. One (3.2%) sample was seropositive for both T. equi and B. caballi infections. Of all the animals in this study, only 3 horses were infected by Rhipicephalus turanicus and Hyalomma detritum, and no haemoparasites were detected by microscopic examination. © 2007 Elsevier Ireland Ltd. All rights reserved. Keywords: Babesiosis; Equidae; IFAT; Turkey
1. Introduction Equine babesiosis, caused by the hemoprotozoans T. equi and B. caballi, is responsible for important economic losses in the equine industry of tropical and subtropical areas of the world [1–3]. Levine [4] stated that equine babesiosis is an acute, subacute or chronic disease of equids that is caused by the intraerytrocytic protozoans T. equi and B. caballi which share the same vectors. Of the estimated world population of 120 million horses, 90% are bred in areas with endemic babesiosis [1]. In Turkey, there have been reports of hemoprotozoal diseases [5,6], and more recently in Elazığ and Kayseri in Turkey [7,8]. Information on the distribution and prevalence of equine babesiosis is limited in Turkey. One of the ⁎ Corresponding author. Department of Parasitology, Faculty of Veterinary Medicine, University of Ondokuz Mayis 55139 Kurupelit, Samsun, Turkey. Tel.: +90 3623121919 2822; fax: +90 3624576000 2801. E-mail address: [email protected] (M. Acici). 1383-5769/$ - see front matter © 2007 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.parint.2007.12.009
objectives of this research was to perform an epidemiological survey of T. equi and B. caballi antibodies in equid animals of part of the Black Sea region of Turkey. 2. Materials and methods The research was carried out mainly in areas ranging from 50 m to 700 m altitude and covering parts of five northern Turkish provinces: Sinop (Sazli and Akbas); Samsun (Tekkekoy–Yeşil Alan, Yukarı Çinik; Terme–Ahmetbey; Çarşamba, Yamalı); Ordu (Perşembe–Yumrutaş, Yeniköy, Ramazan, Töngeldüzü, Boğazcık); Amasya (Direkli) and Tokat (Kadıvakfı). The study covered 13 villages and included 120 farms, each with one or more animals. 2.1. Blood and tick collection In May and June of 2004, blood samples were collected on each of the 120 selected farms from a total of 153 animals. The
M. Acici et al. / Parasitology International 57 (2008) 198–200
local crossbred animals included 84 horses, 38 donkeys and 31 mules. The blood samples were allowed to coagulate, after which the serum was removed and stored at −20 °C until further use. Thin, Giemsa-stained blood smears were prepared from fresh, periferal capillary blood and examined microscopically for the presence of parasites. Tick specimens were collected from all areas of animals' bodies, and preserved in 70% ethanol, until identified from the keys of Karaer et al. [9]. Tick smears were prepared and checked for Babesia vermicules in the eggs [10].
199
Table 2 Distribution of equine babesiosis and Babesia species in different age groups⁎ Age (year)
N 0–3 4–9 10–15 16 ≤
Babesiosis N
%
153 27 50 53 23
37.0 56.0 56.6 43.5
B. caballi χ2
3.79
N
%
153 27 50 53 23
25.9 42.0 37.7 30.4
T. equi χ2
2.35
N
%
153 27 50 53 23
18.5 18.0 26.4 17.4
χ2
1.49
⁎: p N 0.05.
2.2. IFAT analysis The T. equi and B. caballi antigens, 12-well IFA substrate slides (Lot BKG-120-040907) and the anti-horse IgG FITC conjugate (EG-20-040217) used in the Indirect Fluorescent Antibody Test (IFAT) were acquired from Fuller Laboratories, Fullerton, CA, USA. T. equi (BEP-031029) and B. caballi (BKP040219) were used as positive controls; T. equi and B. caballi (NE50-040325) was used as a negative control. Test procedures were performed according to the manufacturer's instructions for use. The antigen slides were stored at −20 °C until the analysis and they were brought to room temperature before opening slide envelopes. All sera were diluted 1:80 in PBS before the test. From each serum dilution (10 μl) was added to a slide well and incubated at 37 °C for 30 min. Then, the slides were washed three times with PBS (pH 7.2). Conjugate (10 μl, FITC-labelled rabbit anti-horse IgG with bovine serum albumin and Evans'blue counterstain) was added to a slide well. The slides were incubated for 30 min. at 37 °C. After five washings with PBS the glass slides were dried out and mounted in glycerol/PBS (1:1) for examination under an epifluorescence microscope. Buffer control, conjugate control and negative and positive control sera were included in each test. Titers of 1:80 or more that showed strong fluorescence were considered to be positive in diluted sera and IFAT was applied to the differential diagnosis of T. equi and B. caballi infections [11]. The chi-square test was used to test for significant difference between groups. 3. Results Out of 153 sampling, 33 (21.5%) and 53 (34.6%) were positive for T. equi and B. caballi infections, respectively, at a dilution ≥ 1:80. Among the sera collected from horses, donkeys and mules, 32 (38.0%), 14 (36.8%) and 8 (25.8%) were positive
Table 1 Prevalence of equine babesiosis and Babesia species in the Black Sea region Province
N Amasya Ordu Samsun Sinop Tokat ⁎: p b 0.01.
Babesiosis N
%
153 30 30 30 31 32
83.3 53.3 56.7 35.5 28.1
B. caballi χ
N
%
22.68⁎
153 30 30 30 31 32
60.0 36.7 46.7 32.3 6.3
2
T. equi χ
N
%
χ2
21.48⁎
153 30 30 30 31 32
33.3 20.0 23.3 3.2 25.0
9.10⁎
2
to B. caballi, and 20 (23.8%), 5 (13.1%) and 8 (25.8%) were positive to T. equi, respectively. Anti-Babesia antibodies were detected in every village and its prevalence is shown in Tables 1 and 2. B. caballi infections were more prevalent in horses than in donkeys and mules. The percentages were 38.0%, 36.8% and 25.8%, respectively. T. equi (13.1%) was least prevalent in donkeys. There were statistical differences (p b 0.01) among counties for Babesia prevalence (Table 1). However, there were no statistical differences (p N 0.05) between age groups for their prevalence (Table 2). Five Rhipicephalus turanicus were found in horses in Samsun and two Hyalomma detritum in Amasya and Tokat. No Babesia parasites were detected in periferal blood smears or in the ticks assayed. None of these ticks were infected with Babesia spp. 4. Discussion Several direct and indirect detection methods, including blood smears [12], in-vitro cultures [13], DNA probes [14] and serology [15,16], have been used for the diagnosis of B. caballi and T. equi infections. Standard serological tests for babesiosis are the complement fixation test and the indirect fluorescent antibody test [2]. However, it has been reported that because of its low sensitivity and specificity, the complement fixation test fails to discriminate accurately between negative and carrier animals [17]. Weiland [18] demonstrated the strong crossreactivity of anti-B. caballi horse serum with the lysate of T. equi-infected erythrocytes, using an Enzyme-Linked Immunosorbent Assay. In this study, 8 (5.2%) of samples in equines were seropositive for both B. caballi and T. equi. IFAT and ELISA tests have been used to detect equine babesiosis in Turkey [5,8]. According to Inci [7], B. caballi and T. equi infections were detected in 7.86% and 4.49% of horses in the Kayseri region, respectively. Balkaya and Erdogmus [8] researched babesiosis in Elazig and Malatya provinces of Turkey and found B. caballi in 2.5% and B. equi in 20.5% of horses, respectively. Ozcan [19] reported B. caballi (71.87%) and T. equi (28.12%) in horses and donkeys respectively in Ankara in Turkey. In this study, the prevalence of B. caballi was 38.0% in horses, 36.8% in donkeys, and 25.8% in mules. T. equi was detected in 23.8% of horses, 13.1% of donkeys, and 25.8% of mules. On the other hand, no infections were found in donkeys less than one year of age. Equine babesiosis is endemic in many tropical and subtropical areas and is transmitted by ticks, including the genera Boophilus, Hyalomma, Dermacentor and Rhipicephalus [20]. In this study, H. detritum and R.
200
M. Acici et al. / Parasitology International 57 (2008) 198–200
turanicus were found on equines. However, not Babesia spp. was detected in them. In conclusion, this study showed a high prevalence of equine babesiosis seropositivity in equids in the northern region of Turkey. The results presented are in agreement with the ELISA study reported by Balkaya and Erdogmus [8] which showed no significant differences between age groups in the prevalence of equine babesiosis. Although no parasites were detected in Giemsa-stained blood smears, indirect IFAT results showed that T. equi and B. caballi are endemic in the Black Sea region. Acknowledgements The authors are grateful for the support of the Scientific Research Committee of Ondokuz Mayis University, Samsun, Turkey (Project No. Vet. 008) and to Gregory Thomas Sullivan of the same university for his assistance in English editing. References [1] Schein E. Equine babesiosis. In: Ristic M, editor. Babesiosis of Domestic Animals and Man. Boca Raton, FL: CRC Press; 1988. p. 197–209. [2] Friedhoff KT. The piroplasms of equidae significance fort he international equine trade. Berl Münch Tierarztl Wochenschr 1982;95:368–74. [3] Kuttler KL. Chemotherapy of babesiosis. In: Ristic M, editor. Babesiosis of Domestic Animals and Man. Boca Raton, FL: CRC Press; 1988. p. 10–2. [4] Levine ND. Veterinary Protozoology. Ames. Iowa: Iowa State University Press; 1985. [5] Inci A. Gemlik Askeri Harası atlarında Babesia caballi (Nuttal, 1910) ve Babesia equi (Laveran, 1901)'nin mikroskobik muayeneyle saptanması. Tr J Vet Anim Sci 1997;21:43–6. [6] Aktas M, Dumanlı N. Malatya Sultansuyu Tarım Isletmesi atlarında subklinik Babesia equi (Laveran, 1901) ve Babesia caballi (Nuttal, 1910) enfeksiyonları. Turk Parasitol Derg 2000;24:55–6.
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