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Research Article

volume 54 issue 8 (august 2020) : 1034-1038,   Doi: 10.18805/ijar.B-1051

Molecular Detection of Neospora caninum from Naturally Infected Dogs in Van Province, East Turkey

A
Ali Bilgin Yilmaz
Y
Yaşar Göz
&
Özlem Orunç Kilinç
V
Vural Denizhan
1Faculty of Health Sciences, Van Yuzuncu Yil University, 65100, Turkey.
Cite article:- Yilmaz Bilgin Ali, Göz Yaşar, Kilinç Orunç Özlem, Denizhan Vural (2020). Molecular Detection of Neospora caninum from Naturally Infected Dogs in Van Province, East Turkey. Indian Journal of Animal Research. 54(8): 1034-1038. doi: 10.18805/ijar.B-1051.

ABSTRACT

Neospora caninum is a coccidian parasite causing abortion in cattle and neurological problems in horses. Dogs are definitive hosts of N. caninum. Polymerase chain reaction is the most specific method used for the detection of N. caninum oocytes. In the present study, a total of 100 fecal samples were collected from naturally infected dogs. Of the 100 samples analyzed, 11 of them were detected with Hammondia/Neospora-like oocytes. Genomic DNA was isolated using a commercially available DNA extraction kit. The Nc5 gene specific to N. caninum was amplified by PCR and two of the eleven samples with Hammondia/Neospora-like oocytes formed ~337 bp repeatable band. In conclusion, N. caninum, which has been shown to cause neurological disorders in dogs and to abortion in cattle, was detected in naturally infected dogs in Van Province in Polymerase chain reaction. 

REFERENCES

  1. Al Warid, H.S. (2014). The effect of freezing-thawing on DNA extraction from Cryptosporidium oocysts ýn fecal samples. International Journal of Pharmacy and Biological Sciences. 1:14-18.
  2. Anderson, M.L., Blanchard, P.C., Barr, B.C., Dubey, J.P., Hoffman, R.L., Conrad, P.A. (1991). Neospora- like protozoan infection as a major cause of abortion in California dairy cattle. Journal of Veterinary Diagnostic Investigation.198: 241-244. 
  3. Björkman, C., Uggla, A. (1999). Serological diagnosis of Neospora caninum infection. International journal for parasitology. 29: 1497-1507. 
  4. Björkman, C., Lunden, A., Ugla, A. (1994). Prevalence of antibodies to Neospora caninum and Toxoplasma gondii in Swedish dogs. Acta veterinaria Scandinavica. 35: 45–47. 
  5. Barber, J.S. Gasser, R.B. Ellis, J. Reichel, M.P. McMillan, D. and Trees, A.J. (1997). Prevalence of antibodies to Neospora caninum in different canid populations. The Journal of Parasitology. 83: 1056-1058. 
  6. Coskun, S.Z., Aydin, L., Bauer, C. (2000). Seroprevalence of Neospora caninum infection in domestic dogs in Turkey. The Veterinary Record. 146:649. 
  7. Dubey, J.P., Carpenter, J.L., Speer, C.A., Topper, M.J., Uggla, A. (1988). Newly recognized fatal protozoan disease of dogs. Journal of the American Veterinary Medical Association. 192:1269-1285. 
  8. Dubey, J.P and Lindsay, D.S. (1996). A review of Neospora caninum and Neosporosis. Veterinary parasitology. 67: 1-59. 
  9. Dubey, J.P., Schares, G., Ortega-Mora, L.M. (2007). Epidemiology and control of Neosporosis and Neospora caninum. Clinical microbiology reviews. 20:323-367. 
  10. Dubey, J.P. (1999). Recent advances in Neospora and neosporosis. Veterinary parasitology. 84: 349-367. 
  11. Dubey, J.P, (2003). Review of Neospora caninum and neosporosis in animals. The Korean journal of parasitology. 41:1-16. 
  12. Dubey, J.P., Sreekumar, C., Knickman, E., Miska, K.B., Vianna, M.C., Kwok, O.C., Hill, D.E., Jenkins, M.C., Lindsay, D.S., Greene, C.E (2004). Biologic, morphologic and molecular characterization of Neospora caninum isolates from littermate dogs. International Journal for Parasitology. 34:1157-1167.
  13. Düzlü, Ö., Ýnci, A., Yýldýrým, A., Önder, Z., Çiloðlu, A. (2014). Köpek periferal kan örneklerinde Neospora caninum ve Toxoplasma gondii takizoitlerinin TaqMan prob bazlý Real Time PCR ile araþtýrýlmasý. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 61:283-288. 
  14. Gondim, L.F., McAllister, M.M., Pitt, W.C., Zemlicka, D.E, (2004). Coyotes (Canis latrans) are definitive hosts of Neospora caninum. International Journal for Parasitology. 34:159- 161. 
  15. Hill, D.E., Liddell, S., Jenkins, M.C., Dubey, J.P, (2001). Specific detection of Neospora caninum oocyst in fecal samples from experimentally-infected dogs using the polymerase chain reaction. Journal of Parasitology. 87:395-398.
  16. King, J.S., Brown, G.K., Jenkins, D.J., Ellis, J.T., Fleming, P.J., Windsor, P.A., Slapeta, J. (2012). Oocyst and high seroprevalence of Neospora caninum in dogs living in remote Aboriginal communities and wild dogs in Australia. Veterinary Parasitology. 187:85-92. 
  17. King, J.S., Slapeta, J., Jenkins, D.J., Al-Qassab, S.E., Ellis J.T., Windsor, P.A. (2010). Australian dingoes are definitive hosts of Neospora caninum. International journal for parasitology. 40: 945–950.
  18. Langoni, H., Matteucci, G., Medici, B., Camossi, L.G., Richini-Pereira, V.B., Silva, R.C. (2012). Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders. Revista da Sociedade Brasileria de Medicina Tropical. 45:365-368. 
  19. Leon, A., Richard, E., Fortier, C., Laugier, C., Fortier, G., Pronost, S. (2012). Molecular detection of Coxiella burnetii and Neospora caninum in equine aborted foetuses and neonates. Preventive veterinary medicine. 104: 179-183. 
  20. Malmasi, A., Hosseininejad, M., Haddadzadeh, H., Badii, A., Bahonar, A. (2007). Serologic study of anti-Neospora caninum antibodies in household dogs and dogs living in dairy and beef cattle farms in Tehran, Iran. Parasitology Research. 100:1143-1145. 
  21. Naguleswaran, A., Hemphill, A., Rajapakse, R.P., Sager, H. (2004). Elaboration of a crude antigen ELISA for serodiagnosis of caprine neosporosis: validation of the test by detection of Neospora caninum-specific antibodies in goats from Sri Lanka. Veterinary parasitology. 126:257-262. 
  22. Osawa, T., Wastling, J., Maley, S., Buxton, D., Innes, E.A. (1998). A multiple antigen ELISA to detect Neospora-specific antibodies in bovine sera, bovine foetal fluids, ovine and caprine sera. Veterinary parasitology. 79:19-34. 
  23. Palavicini, P., Romero, J.J., Dolz, G., Jimènez, A.E., Hill, D.E., Dubey, J.P. (2007). Fecal and serological survey of Neospora caninum in farm dogs in Costa Rica. Veterinary Parasitology. 149:265-270. 
  24. Razimi, G. (2009). Fecal and molecular survey of Neospora caninum in farm and household dogs in Mashhad area, Khorasan province, Iran. Korean Journal of Parasitology. 47:417-420. 
  25. Reichel, M.P., Ellis, J.T., Dubey, J.P. (2007). Neosporosis and Hammondiosis in dogs. The Journal of Small Animal Practice. 48:308-312. 
  26. Santos, S.L., de Souza Costa, K., Gondim, L.Q., da Silva, M.S., Uzêda, R.S., Abe-Sandes, K., Gondim, L.F. (2010). Investigation of Neospora caninum, Hammondia sp. and Toxoplasma gondii in tissues from slaughtered beef cattle in Bahia, Brazil. Parasitology Research. 106:457-461. 
  27. Sheather, A. L. (1923). The detection of intestinal and manage parasites by a floatation technique. Journal of Comparative Pathology. 36: 266-275.
  28. Silva, M.S., Uzèda R.S., Costa, K.S., Santos SL, Macedo, A.C., Abe-    Sandes, K., Gondim, L.F. (2009). Detection of Hammondia heydorni and related coccidia (Neospora caninum and Toxoplasma gondii) in goats slaughtered in Bahia, Brazil. Veterinary parasitology. 162:156-159.
  29. Sevgili, M., ªahin, T., Çimtay, Ý., Çetin, H., Keskin, O., Gökçen, O. (2003). Þanlýurfa Yöresi Safkan Arap Kýsraklarýnda Neospora caninum Antikorlarýnýn Belirlenmesi.Yüzüncü Yýl Üniversitesi Veteriner Fakültesi Dergisi. 14:15-17. 
  30. Trees, A.J., Guy, F., Tennant, B.J., Balfour, A.H., Dubey, J.P. (1993). Prevalence of antibodies to Neospora caninum in a population of urban dogs in England. The Veterinary record. 132: 125-126. 
  31. Yildiz, K., Yasa Duru, S., Yagcý, B.B., Babur, C., Ocal, N., Gurcan, S., Karaca, S. (2009). Seroprevalence of Neospora caninum and Coexistence with Toxoplasma gondii in Dogs. Türkiye Parazitoloji Dergisi. 33: 116-119.
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Indian Journal of Animal Research
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    Research Article

    volume 54 issue 8 (august 2020) : 1034-1038,   Doi: 10.18805/ijar.B-1051

    Molecular Detection of Neospora caninum from Naturally Infected Dogs in Van Province, East Turkey

    A
    Ali Bilgin Yilmaz
    Y
    Yaşar Göz
    &
    Özlem Orunç Kilinç
    V
    Vural Denizhan
    1Faculty of Health Sciences, Van Yuzuncu Yil University, 65100, Turkey.
    Cite article:- Yilmaz Bilgin Ali, Göz Yaşar, Kilinç Orunç Özlem, Denizhan Vural (2020). Molecular Detection of Neospora caninum from Naturally Infected Dogs in Van Province, East Turkey. Indian Journal of Animal Research. 54(8): 1034-1038. doi: 10.18805/ijar.B-1051.

    ABSTRACT

    Neospora caninum is a coccidian parasite causing abortion in cattle and neurological problems in horses. Dogs are definitive hosts of N. caninum. Polymerase chain reaction is the most specific method used for the detection of N. caninum oocytes. In the present study, a total of 100 fecal samples were collected from naturally infected dogs. Of the 100 samples analyzed, 11 of them were detected with Hammondia/Neospora-like oocytes. Genomic DNA was isolated using a commercially available DNA extraction kit. The Nc5 gene specific to N. caninum was amplified by PCR and two of the eleven samples with Hammondia/Neospora-like oocytes formed ~337 bp repeatable band. In conclusion, N. caninum, which has been shown to cause neurological disorders in dogs and to abortion in cattle, was detected in naturally infected dogs in Van Province in Polymerase chain reaction. 

    REFERENCES

    1. Al Warid, H.S. (2014). The effect of freezing-thawing on DNA extraction from Cryptosporidium oocysts ýn fecal samples. International Journal of Pharmacy and Biological Sciences. 1:14-18.
    2. Anderson, M.L., Blanchard, P.C., Barr, B.C., Dubey, J.P., Hoffman, R.L., Conrad, P.A. (1991). Neospora- like protozoan infection as a major cause of abortion in California dairy cattle. Journal of Veterinary Diagnostic Investigation.198: 241-244. 
    3. Björkman, C., Uggla, A. (1999). Serological diagnosis of Neospora caninum infection. International journal for parasitology. 29: 1497-1507. 
    4. Björkman, C., Lunden, A., Ugla, A. (1994). Prevalence of antibodies to Neospora caninum and Toxoplasma gondii in Swedish dogs. Acta veterinaria Scandinavica. 35: 45–47. 
    5. Barber, J.S. Gasser, R.B. Ellis, J. Reichel, M.P. McMillan, D. and Trees, A.J. (1997). Prevalence of antibodies to Neospora caninum in different canid populations. The Journal of Parasitology. 83: 1056-1058. 
    6. Coskun, S.Z., Aydin, L., Bauer, C. (2000). Seroprevalence of Neospora caninum infection in domestic dogs in Turkey. The Veterinary Record. 146:649. 
    7. Dubey, J.P., Carpenter, J.L., Speer, C.A., Topper, M.J., Uggla, A. (1988). Newly recognized fatal protozoan disease of dogs. Journal of the American Veterinary Medical Association. 192:1269-1285. 
    8. Dubey, J.P and Lindsay, D.S. (1996). A review of Neospora caninum and Neosporosis. Veterinary parasitology. 67: 1-59. 
    9. Dubey, J.P., Schares, G., Ortega-Mora, L.M. (2007). Epidemiology and control of Neosporosis and Neospora caninum. Clinical microbiology reviews. 20:323-367. 
    10. Dubey, J.P. (1999). Recent advances in Neospora and neosporosis. Veterinary parasitology. 84: 349-367. 
    11. Dubey, J.P, (2003). Review of Neospora caninum and neosporosis in animals. The Korean journal of parasitology. 41:1-16. 
    12. Dubey, J.P., Sreekumar, C., Knickman, E., Miska, K.B., Vianna, M.C., Kwok, O.C., Hill, D.E., Jenkins, M.C., Lindsay, D.S., Greene, C.E (2004). Biologic, morphologic and molecular characterization of Neospora caninum isolates from littermate dogs. International Journal for Parasitology. 34:1157-1167.
    13. Düzlü, Ö., Ýnci, A., Yýldýrým, A., Önder, Z., Çiloðlu, A. (2014). Köpek periferal kan örneklerinde Neospora caninum ve Toxoplasma gondii takizoitlerinin TaqMan prob bazlý Real Time PCR ile araþtýrýlmasý. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 61:283-288. 
    14. Gondim, L.F., McAllister, M.M., Pitt, W.C., Zemlicka, D.E, (2004). Coyotes (Canis latrans) are definitive hosts of Neospora caninum. International Journal for Parasitology. 34:159- 161. 
    15. Hill, D.E., Liddell, S., Jenkins, M.C., Dubey, J.P, (2001). Specific detection of Neospora caninum oocyst in fecal samples from experimentally-infected dogs using the polymerase chain reaction. Journal of Parasitology. 87:395-398.
    16. King, J.S., Brown, G.K., Jenkins, D.J., Ellis, J.T., Fleming, P.J., Windsor, P.A., Slapeta, J. (2012). Oocyst and high seroprevalence of Neospora caninum in dogs living in remote Aboriginal communities and wild dogs in Australia. Veterinary Parasitology. 187:85-92. 
    17. King, J.S., Slapeta, J., Jenkins, D.J., Al-Qassab, S.E., Ellis J.T., Windsor, P.A. (2010). Australian dingoes are definitive hosts of Neospora caninum. International journal for parasitology. 40: 945–950.
    18. Langoni, H., Matteucci, G., Medici, B., Camossi, L.G., Richini-Pereira, V.B., Silva, R.C. (2012). Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders. Revista da Sociedade Brasileria de Medicina Tropical. 45:365-368. 
    19. Leon, A., Richard, E., Fortier, C., Laugier, C., Fortier, G., Pronost, S. (2012). Molecular detection of Coxiella burnetii and Neospora caninum in equine aborted foetuses and neonates. Preventive veterinary medicine. 104: 179-183. 
    20. Malmasi, A., Hosseininejad, M., Haddadzadeh, H., Badii, A., Bahonar, A. (2007). Serologic study of anti-Neospora caninum antibodies in household dogs and dogs living in dairy and beef cattle farms in Tehran, Iran. Parasitology Research. 100:1143-1145. 
    21. Naguleswaran, A., Hemphill, A., Rajapakse, R.P., Sager, H. (2004). Elaboration of a crude antigen ELISA for serodiagnosis of caprine neosporosis: validation of the test by detection of Neospora caninum-specific antibodies in goats from Sri Lanka. Veterinary parasitology. 126:257-262. 
    22. Osawa, T., Wastling, J., Maley, S., Buxton, D., Innes, E.A. (1998). A multiple antigen ELISA to detect Neospora-specific antibodies in bovine sera, bovine foetal fluids, ovine and caprine sera. Veterinary parasitology. 79:19-34. 
    23. Palavicini, P., Romero, J.J., Dolz, G., Jimènez, A.E., Hill, D.E., Dubey, J.P. (2007). Fecal and serological survey of Neospora caninum in farm dogs in Costa Rica. Veterinary Parasitology. 149:265-270. 
    24. Razimi, G. (2009). Fecal and molecular survey of Neospora caninum in farm and household dogs in Mashhad area, Khorasan province, Iran. Korean Journal of Parasitology. 47:417-420. 
    25. Reichel, M.P., Ellis, J.T., Dubey, J.P. (2007). Neosporosis and Hammondiosis in dogs. The Journal of Small Animal Practice. 48:308-312. 
    26. Santos, S.L., de Souza Costa, K., Gondim, L.Q., da Silva, M.S., Uzêda, R.S., Abe-Sandes, K., Gondim, L.F. (2010). Investigation of Neospora caninum, Hammondia sp. and Toxoplasma gondii in tissues from slaughtered beef cattle in Bahia, Brazil. Parasitology Research. 106:457-461. 
    27. Sheather, A. L. (1923). The detection of intestinal and manage parasites by a floatation technique. Journal of Comparative Pathology. 36: 266-275.
    28. Silva, M.S., Uzèda R.S., Costa, K.S., Santos SL, Macedo, A.C., Abe-    Sandes, K., Gondim, L.F. (2009). Detection of Hammondia heydorni and related coccidia (Neospora caninum and Toxoplasma gondii) in goats slaughtered in Bahia, Brazil. Veterinary parasitology. 162:156-159.
    29. Sevgili, M., ªahin, T., Çimtay, Ý., Çetin, H., Keskin, O., Gökçen, O. (2003). Þanlýurfa Yöresi Safkan Arap Kýsraklarýnda Neospora caninum Antikorlarýnýn Belirlenmesi.Yüzüncü Yýl Üniversitesi Veteriner Fakültesi Dergisi. 14:15-17. 
    30. Trees, A.J., Guy, F., Tennant, B.J., Balfour, A.H., Dubey, J.P. (1993). Prevalence of antibodies to Neospora caninum in a population of urban dogs in England. The Veterinary record. 132: 125-126. 
    31. Yildiz, K., Yasa Duru, S., Yagcý, B.B., Babur, C., Ocal, N., Gurcan, S., Karaca, S. (2009). Seroprevalence of Neospora caninum and Coexistence with Toxoplasma gondii in Dogs. Türkiye Parazitoloji Dergisi. 33: 116-119.
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