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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 3 (march 2019) : 382-387,   Doi: 10.18805/ijar.B-3507

A comparative analysis of microscopy and PCR based detection methods for Babesia and Trypanosoma infecting bovines and assessment of risk factors

B
B.R. Maharana
B
B. Kumar
J
J.P. Joseph
T
T.K. Patbandha
1Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Junagadh Agricultural University, Junagadh-362 001, Gujarat, India.
Cite article:- Maharana B.R., Kumar B., Joseph J.P., Patbandha T.K. (2018). A comparative analysis of microscopy and PCR based detection methods for Babesia and Trypanosoma infecting bovines and assessment of risk factors. Indian Journal of Animal Research. 53(3): 382-387. doi: 10.18805/ijar.B-3507.

ABSTRACT

The present study was carried out to evaluate and compare the status of infection and assessment of risk factors in 353 blood samples (144 cattle and 209 buffaloes) of bovines by PCR assay along with microscopic examinations (ME). ME revealed prevalence of Babesia bigemina and Trypanosoma evansi to be 22.91% and 0.69%, respectively in cattle and 12.44% and 0.95%, respectively in buffaloes. Conversely, PCR assay was able to detect 40.97% and 3.47 % prevalence of B. bigemina and T. evansi in cattle and 23.92% and 6.69% in buffaloes, respectively. The result revealed that the PCR assay was 100% sensitive and 82.9% specific when compared with ME for babesiosis and 100% sensitive and 95.42% specific for trypanosomosis. Multivariate logistic regression models showed that risk of babesiosis was significantly higher in cattle (Odds ratio (OR) =2.207, P=0.001) compared to buffaloes. The risk for surra in male buffaloes increased by 6.37 times (OR= 6.375, P=0.013). Conversely, risk of babesiosis was significantly lower in male cattle than females (OR= 0.467, P=0.044). 

REFERENCES

  1. Almeria, S., Castella, J., Ferrer, D., Ortuno, A., Estrada-Pena, A. and Gutierrez, J. F. (2001). Ovine piroplasms in Minorca (Balearic Islands, Spain): a comparison of PCR-based and light microscopy detection. Veterinary Parasitology, 99: 249-259.
  2. Clausen, P.H, Wiemann, A., Patzelt, R., Kakaire, D., Poetzsch, C., Peregrine, A. and Mehlitz, D. (1998). Use of a PCR assay for the specific and sensitive detection of Trypanosoma spp. in naturally infected dairy cattle in peri-urban Kampala,Uganda. Annals of the New York Academy of Sciences, 849: 21–31.
  3. Criado-Fornelio, A. (2007). A review of nucleic acid-based diagnostic tests for Babesia and Theileria, with emphasis on bovine piroplasms. Parassitologia (Rome), 49: 39–44.
  4. Dávila, A.M., Herrera, H.M., Schlebinger, T., Souza, S. S. and Traub-Cseko, Y. M. (2003). Using PCR for unraveling the cryptic epizootiology of livestock trypanosomosisin the Pantanal, Brazil. Veterinary Parasitology, 117: 1–13. 
  5. Desquesnes, M. and D´avila, A. M. R. (2002). Applications of PCR-based tools for detection and identification of animal trypanosomes: a review and perspectives. Veterinary Parasitology, 109(3-4): 213–231.
  6. Figueroa, J.V., Chieves, L.P., Johnson, G.S. and Buening, G. M. (1992). Detection of Babesia bigemina-infected carriers by polymerase chain reaction amplification. Journal of Clinical Microbiology, 30: 2576–2582. 
  7. Ganguly, A., Bisla, R.S., Singh, H., Bhanot, V., Kumar, A., Kumari, S., Maharana, B.R. and Ganguly, I. (2017). Prevalence and hematobiochemical changes of tickborne haemoparasitic diseases in crossbred cattle of Haryana, India. Indian Journal of Animal Sciences, 87(5):552-557.
  8. Kumar, B., Maharana, B. R., Prasad, A., Joseph, J. P., Patel, B. and Patel, J.S. (2016). Seasonal incidence of parasitic diseases in bovines of south western Gujarat (Junagadh), India. Journal of Parasitic Disease, 40(4):1342-1346.
  9. Maharana, B.R., Kumar, B., Prasad, A., Patbandha, T.K., Sudhakar, N.R., Joseph, J.P. and Patel, B.R. (2016). Prevalence and assessment of risk factors for haemoprotozoan infections in cattle and buffaloes of South-West Gujarat, India. Indian Journal of Animal Research, 50 (5): 733-739.
  10. McLeod, R. and Kristjanson, P. (1999). Final Report of Joint ESYS/International Livestock Research Institute/Australian Centre for International Agricultural Research Tick Cost Project-Economic Impact of Ticks and Tick-Borne Diseases to Livestock in Africa, Asia and Australia, International Livestock Research Institute, Nairobi, Kenya.
  11. Olmeda, A.S., Armstrong, P.M., Rosenthal, B.M., Valladares, B., del Castillo, A., de Armas, F., Miguelez, M., Gonzalez, A., Rodriguez., J.A. and Spielman, A. (1997. Telford 3rd SR. A subtropical case of human babesiosis. Acta Tropica, 67: 229–234.
  12. Passos, L.M., Bell-Sakyi, L. and Brown, C.G. (1998). Immunochemical characterization of in vitro culture-derived antigens of Babesia bovis and Babesia bigemina. Veterinary Parasitology, 76: 239–249.
  13. Roy, S., Tiwari, A., Galdhar, C.N., Upadhyay, S.R., Ratre, H.K., Sahu, S.K. and Maiti, S.K. (2004). Indian Journal of Veterinary Medicine, 24:5-7.
  14. Shams, S., Ayaz, S., Ali, I., Khan, S., Gul, I., Naila, Gul. and Khan, S. N. (2013). Sensitivity and Specificity of PCR & Microscopy in detection of Babesiosis in domesticated cattle of Khyber Pakhtunkhwa, Pakistan. International Journal of Advancement in Research and Technology, 5(2):37-41.
  15. Sharma, A., Singla, L.D., Tuli, A., Kaur, P., Batth, B.K., Javed, M. and Juyal, P.D. (2013). Molecular Prevalence of Babesia bigemina and Trypanosoma evansi in Dairy Animals from Punjab, India, by Duplex PCR: A Step Forward to the Detection and Management of Concurrent Latent Infections. BioMED Research International, doi: 10.1155/2013/893862. 
  16. Sharma, A., Singla, L.D., Tuli, A., Kaur, P. and Bal, M.S. (2015). Detection and assessment of risk factors associated with natural concurrent infection of Trypanosoma evansi and Anaplasma marginale in dairy animals by duplex PCR in eastern Punjab. Tropical Animal Health and production, 47:251–257.
  17. Soulsby, E. J. L. (1982). Helminths, Arthropods and Protozoa of Demesticated Animals. 7th ed. ELBS, Bailliers Tindall and Cassel, London.
  18. Vohra, S.P., Patel, J.V., Patel, B.B., Patel, S.B. and Umale, R.H. (2012). Seasonal incidence of haemoprotozoan diseases in cross bred cattle and buffaloes in Kaira and Anand districts of Gujarat, India. Veterinary World, 5: 223-224.
  19. Wuyts, N., Chokesajjawatee, N., Sarataphan, N. and Panyim, S. (1995). PCR amplification of crude blood on microscopic slides in diagnosis of Trypanosoma evansi infection in dairy cattle. Annales De La Societe Belge De Medecine Tropicale. 75: 229-37.
  20. Yadav, C.L., Gupta, R.P. and Ruprah, N.S. (1985). The prevalence of haemoprotozoan infections in cattle and buffaloes. Journal of Indian Veterinary Medicine, 9: 205–209. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 3 (march 2019) : 382-387,   Doi: 10.18805/ijar.B-3507

    A comparative analysis of microscopy and PCR based detection methods for Babesia and Trypanosoma infecting bovines and assessment of risk factors

    B
    B.R. Maharana
    B
    B. Kumar
    J
    J.P. Joseph
    T
    T.K. Patbandha
    1Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Junagadh Agricultural University, Junagadh-362 001, Gujarat, India.
    Cite article:- Maharana B.R., Kumar B., Joseph J.P., Patbandha T.K. (2018). A comparative analysis of microscopy and PCR based detection methods for Babesia and Trypanosoma infecting bovines and assessment of risk factors. Indian Journal of Animal Research. 53(3): 382-387. doi: 10.18805/ijar.B-3507.

    ABSTRACT

    The present study was carried out to evaluate and compare the status of infection and assessment of risk factors in 353 blood samples (144 cattle and 209 buffaloes) of bovines by PCR assay along with microscopic examinations (ME). ME revealed prevalence of Babesia bigemina and Trypanosoma evansi to be 22.91% and 0.69%, respectively in cattle and 12.44% and 0.95%, respectively in buffaloes. Conversely, PCR assay was able to detect 40.97% and 3.47 % prevalence of B. bigemina and T. evansi in cattle and 23.92% and 6.69% in buffaloes, respectively. The result revealed that the PCR assay was 100% sensitive and 82.9% specific when compared with ME for babesiosis and 100% sensitive and 95.42% specific for trypanosomosis. Multivariate logistic regression models showed that risk of babesiosis was significantly higher in cattle (Odds ratio (OR) =2.207, P=0.001) compared to buffaloes. The risk for surra in male buffaloes increased by 6.37 times (OR= 6.375, P=0.013). Conversely, risk of babesiosis was significantly lower in male cattle than females (OR= 0.467, P=0.044). 

    REFERENCES

    1. Almeria, S., Castella, J., Ferrer, D., Ortuno, A., Estrada-Pena, A. and Gutierrez, J. F. (2001). Ovine piroplasms in Minorca (Balearic Islands, Spain): a comparison of PCR-based and light microscopy detection. Veterinary Parasitology, 99: 249-259.
    2. Clausen, P.H, Wiemann, A., Patzelt, R., Kakaire, D., Poetzsch, C., Peregrine, A. and Mehlitz, D. (1998). Use of a PCR assay for the specific and sensitive detection of Trypanosoma spp. in naturally infected dairy cattle in peri-urban Kampala,Uganda. Annals of the New York Academy of Sciences, 849: 21–31.
    3. Criado-Fornelio, A. (2007). A review of nucleic acid-based diagnostic tests for Babesia and Theileria, with emphasis on bovine piroplasms. Parassitologia (Rome), 49: 39–44.
    4. Dávila, A.M., Herrera, H.M., Schlebinger, T., Souza, S. S. and Traub-Cseko, Y. M. (2003). Using PCR for unraveling the cryptic epizootiology of livestock trypanosomosisin the Pantanal, Brazil. Veterinary Parasitology, 117: 1–13. 
    5. Desquesnes, M. and D´avila, A. M. R. (2002). Applications of PCR-based tools for detection and identification of animal trypanosomes: a review and perspectives. Veterinary Parasitology, 109(3-4): 213–231.
    6. Figueroa, J.V., Chieves, L.P., Johnson, G.S. and Buening, G. M. (1992). Detection of Babesia bigemina-infected carriers by polymerase chain reaction amplification. Journal of Clinical Microbiology, 30: 2576–2582. 
    7. Ganguly, A., Bisla, R.S., Singh, H., Bhanot, V., Kumar, A., Kumari, S., Maharana, B.R. and Ganguly, I. (2017). Prevalence and hematobiochemical changes of tickborne haemoparasitic diseases in crossbred cattle of Haryana, India. Indian Journal of Animal Sciences, 87(5):552-557.
    8. Kumar, B., Maharana, B. R., Prasad, A., Joseph, J. P., Patel, B. and Patel, J.S. (2016). Seasonal incidence of parasitic diseases in bovines of south western Gujarat (Junagadh), India. Journal of Parasitic Disease, 40(4):1342-1346.
    9. Maharana, B.R., Kumar, B., Prasad, A., Patbandha, T.K., Sudhakar, N.R., Joseph, J.P. and Patel, B.R. (2016). Prevalence and assessment of risk factors for haemoprotozoan infections in cattle and buffaloes of South-West Gujarat, India. Indian Journal of Animal Research, 50 (5): 733-739.
    10. McLeod, R. and Kristjanson, P. (1999). Final Report of Joint ESYS/International Livestock Research Institute/Australian Centre for International Agricultural Research Tick Cost Project-Economic Impact of Ticks and Tick-Borne Diseases to Livestock in Africa, Asia and Australia, International Livestock Research Institute, Nairobi, Kenya.
    11. Olmeda, A.S., Armstrong, P.M., Rosenthal, B.M., Valladares, B., del Castillo, A., de Armas, F., Miguelez, M., Gonzalez, A., Rodriguez., J.A. and Spielman, A. (1997. Telford 3rd SR. A subtropical case of human babesiosis. Acta Tropica, 67: 229–234.
    12. Passos, L.M., Bell-Sakyi, L. and Brown, C.G. (1998). Immunochemical characterization of in vitro culture-derived antigens of Babesia bovis and Babesia bigemina. Veterinary Parasitology, 76: 239–249.
    13. Roy, S., Tiwari, A., Galdhar, C.N., Upadhyay, S.R., Ratre, H.K., Sahu, S.K. and Maiti, S.K. (2004). Indian Journal of Veterinary Medicine, 24:5-7.
    14. Shams, S., Ayaz, S., Ali, I., Khan, S., Gul, I., Naila, Gul. and Khan, S. N. (2013). Sensitivity and Specificity of PCR & Microscopy in detection of Babesiosis in domesticated cattle of Khyber Pakhtunkhwa, Pakistan. International Journal of Advancement in Research and Technology, 5(2):37-41.
    15. Sharma, A., Singla, L.D., Tuli, A., Kaur, P., Batth, B.K., Javed, M. and Juyal, P.D. (2013). Molecular Prevalence of Babesia bigemina and Trypanosoma evansi in Dairy Animals from Punjab, India, by Duplex PCR: A Step Forward to the Detection and Management of Concurrent Latent Infections. BioMED Research International, doi: 10.1155/2013/893862. 
    16. Sharma, A., Singla, L.D., Tuli, A., Kaur, P. and Bal, M.S. (2015). Detection and assessment of risk factors associated with natural concurrent infection of Trypanosoma evansi and Anaplasma marginale in dairy animals by duplex PCR in eastern Punjab. Tropical Animal Health and production, 47:251–257.
    17. Soulsby, E. J. L. (1982). Helminths, Arthropods and Protozoa of Demesticated Animals. 7th ed. ELBS, Bailliers Tindall and Cassel, London.
    18. Vohra, S.P., Patel, J.V., Patel, B.B., Patel, S.B. and Umale, R.H. (2012). Seasonal incidence of haemoprotozoan diseases in cross bred cattle and buffaloes in Kaira and Anand districts of Gujarat, India. Veterinary World, 5: 223-224.
    19. Wuyts, N., Chokesajjawatee, N., Sarataphan, N. and Panyim, S. (1995). PCR amplification of crude blood on microscopic slides in diagnosis of Trypanosoma evansi infection in dairy cattle. Annales De La Societe Belge De Medecine Tropicale. 75: 229-37.
    20. Yadav, C.L., Gupta, R.P. and Ruprah, N.S. (1985). The prevalence of haemoprotozoan infections in cattle and buffaloes. Journal of Indian Veterinary Medicine, 9: 205–209. 
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