NAAS Score 2019

                   5.36

Declaration Format

Please download DeclarationForm and submit along with manuscript.

UserOnline

Free counters!

Previous Next

Seroprevalence of Brucellosis in Buffaloes of Malwa Region of Madhya Pradesh

Sachin Verma G. P. Jatav Supriya Shukla A. K. Jayraw H. C. Chauhan Nidhi Shrivastava
Vol 9(3), 256-262
DOI- http://dx.doi.org/10.5455/ijlr.20181114062927

Brucellosis is one of the five main notifiable bacterial diseases of zoonotic importance in the world. The most widely used serological tests for diagnosis of brucellosis in animals are Rose Bengal plate test, standard tube agglutination test and enzyme linked immunosorbent assay. The present study was undertaken to assess the seroprevalence of brucellosis in 150 buffaloes belonging to different regions of Malwa tract and nearby areas of Indore district by using Rose Bengal plate test (RBPT), indirect enzyme linked immunosorbent assay (i-ELISA), Milk ring test (MRT) and California mastitis test (CMT) for detecting anti-Brucellaantibodies. Sero-prevalence of brucellosis in buffaloes by using RBPT, i-ELISA, MRT and CMT was recorded as 13.33% (20/150), 00% (00/150), 38.88% (7/18) and 55.55% (10/18), respectively.


Keywords : Brucellosis CMT ELISA MRT RBPT Seroprevalence

Brucellosis in buffaloes is the best known as one of the main reproductive disease, capable of causing abortion storms in the breeding season during the last trimester of pregnancy, retention of the fetal membranes, still births and reduction in milk yield resulting in great economic losses (Refai, 2003). In ruminants, organism has a marked affinity for lymphoid and reproductive organs, since the organism replicates to a high numbers in the gravid uterus. In non-pregnant and persistently infected cows, the udder and supramammary lymph nodes are the most common sites for localization (Meador, 1989). It was reported that after abortion up to 80% of infected cows develop chronic infection localized in mammary glands and supramammary lymph node, and marnmary gland infection may persist throughout the lifetime of the cow (Beytutet al., 2009; Xavieret al., 2009; Hamdy and Amin, 2002).Accurate diagnosis of brucellosis is essential for institution of control strategies, either disease as a whole or as species-specific. The most widely used serological tests for diagnosis of brucellosis in animals are Rose Bengal plate test (RBPT), standard tube agglutination test (STAT) and enzyme linked immunosorbentassay (ELISA).

Materials and Methods

The present study was conducted in the Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Mhow (M.P). Samples were collected from 150 buffaloes (both lactating and non-lactating) from Cantonment Board slaughter house, Mhow and nearby areas of Indore. Milk and blood without anticoagulant (for serum) were collected for the present investigation.

Serum samples and milk samples were examined by RBPT, i-ELISA and MRT, respectively for detecting anti-Brucella antibodies. The collected milk samples were also screened by California mastitis test (OIE, 2012). The reagents like, RBPT antigen and Milk Ring Test (MRT) / Abortus Bang ring test (ABR) – ABR-antigen were procured from the Institute of Animal Health and Veterinary Biologicals (IAH and VB), Hebbal, Bengaluru, Karnataka-560024. Whereas, the indirect-enzyme linked Immunosorbent assay –Brucella Antibody Test Kit, ELISA along with the user’s manual was procured from National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI), Bengaluru. While the California mastitis test (CMT) reagents were prepared according to the method described by Schalm et al. (1971).

Results and Discussion

The study was undertaken to assess the sero-prevalence of brucellosis in buffaloes belonging to different regions of Malwa tract and nearby areas of Indore district. The serum samples were collected from buffaloes before slaughter from Cantonment Board slaughter house, Mhow. For detection of  anti-Brucella antibodies in serum and milk samples of buffaloes, the prescribed tests, viz. RBPT, i-ELISA (NIVEDI, Bengaluru) and MRT and CMT, respectively were used as per the standard protocol for detecting anti-Brucella antibodies and the findings of which are shown in Table 1, 2, 3 and 4 and Plate 1, 2 and 3.

Seroprevalence of Brucellosis in Buffaloes

Detection of Anti-Brucella Antibodies in Buffaloes by using RBPT

For this study a total of 150 serum samples were examined for the presence of anti-Brucella antibodies in buffaloes (both lactating and non-lactating) using the RBPT. Out of 150 serum samples, 20 (13.33 %) serum samples were found positive for anti-Brucella antibodies by Rose Bengal Plate Test (RBPT).

 

Table 1: Prevalence of anti-Brucella antibodies in serum samples using the Rose Bengal Plate Test (RBPT)

S. No. Particulars Number of Animals Prevalence (%)
1 Positive 20 13.33
2 Negative 130 86.67

Table 2: Prevalence of brucellosis by using the indirect enzyme linked immunosorbent assay (i-ELISA)

S. No. Particulars Number of Animals Prevalence (%)
1 Positive 0 0
2 Negative 150 100

 

Plate 1: Photograph showing positive and negative RBPT for brucellosis

The findings of the present study are in close proximity with the findings of Lodhi et al. (1995) with 12.98 % seropositive cases, Sandhuet al. (2001) with 9.33% infection rate, Varasada (2003) with 16.80 % rate of infection, Nasiret al. (2004) with 15.38% buffaloes at government livestock farms, Bhattacharya et al. (2005) with 10.27 %, Dinka and Chala (2009) with 11.2 % infection in cattle, Saha et al. (2010) with 12.02% serum samples in organized herds in West Bengal, Karthik et al. (2014) with 16.49 % infection in cattle and Gogoi et al. (2017) with 12.69 % prevalence of brucellosis in bovines. The higher seroprevalence recorded in the present study is attributable to the fact that the present survey was carried out in slaughtered buffaloes which are being culled owing to their unproductivity, as culling of unproductive buffaloes is the routine practice of farmers of Malwa region of Madhya Pradesh.

Detection of Anti-Brucella Antibodies in Buffaloes by using i-ELISA

Findings of the present study are in contrast with the observations of Varasada (2003) with 20.00% infection, Muniret al. (2008) with 78% rate of infection, Grushina et al. (2010) with 100% positive serum samples in Kazakhstan, Kaleem et al.  (2016) with 57.58 % infection and Gogoi et al. (2017) with 13.84% rate of infection when compared with findings of the present study which revealed all the animals were sero-negative by  i-ELISA.

Plate 2: Photograph showing negative i- ELISA in walls of microtitre plate

 

Detection of Anti-Brucella Antibodies in Milk Samples of Buffaloes by MRT

Out of 150 buffaloes (18 lactating and 132 non-lactating), 18 milk samples were collected for detection of anti-Brucella antibodies by the MRT. Out of 18 milk samples, 07 samples were found positive for MRT with a prevalence of 38.88% (Table 3).

Table 3: Prevalence of brucellosis by using the Milk Ring Test (MRT)

S. No. Particulars Number of Animals Prevalence (%)
1 Positive 7 38.88
2 Negative 11 61.55

The MRT is an agglutination test conducted on fresh milk collected from dairy animals, but it does not work on pasteurised or homogenized milk (Fleischhauer, 1937). This test detectsIgM and IgA antibodies bound to the fat globules which has wide acceptability as it is cost effective, easy to perform and can cover a large population in a short time (Cadmus et al., 2008). Findings of this study are in consonance with the findings of Soomro et al. (2014) with 47.19% prevalence of brucellosis in aborted cattle and buffaloes in district of Hyderabad, Pakistan. Whereas the findings are in contrast with the findings of Shafee et al. (2011) with 4.6 and 1.7 % infection from cattle and buffaloes, Mohamand et al. (2014) with 18.35 % positive cases in dairy cow for anti-Brucella antibodies, Gogoi et al. (2017) with 10.53 % prevalence of brucellosis in bovines and Dalal et al. (2017) with 21.73% in pooled milk samples from Kaladera and Manpura Machedi.

Plate 3: Photograph showing positive and negative milk ring test for brucellosis

Detection of Mastitis by the California Mastitis Test

In the present study, 18 milk samples were also examined for mastitis by the CMT. Out of 18 milk samples, 10 cases were found positive for CMT with a prevalence of 55.55% (Table 4).

Table 4: Prevalence of mastitis by using the California mastitis test (CMT)

S. No. Particulars Number of Animals Prevalence (%)
1 Positive 10 55.55
2 Negative 8 44.45

Findings of the present study are in consonance with the findings of Mdegelaa et al. (2009) with 51.6 % infection in Tanzania and Gitau et al. (2014) with 56% cases of clinical mastitis.

Conclusion

Serological examination of 150 serum samples revealed 20 (13.33%) serum samples positive for anti-Brucella antibodies by RBPT. Whereas blood samples of all the animals (n=150) were found negative by i-ELISA. Overall 38.88% samples were recorded as positive by MRT and 55.55% cases were recorded as positive for mastitis by the Callifornia mastitis test that was more sensitive than the other for mastitis.

Acknowledgment

The authors are highly thankful to Head of Institute (Dean), College of Veterinary Science and Animal Husbandry, Mhow (NDVSU, Jabalpur) and the Dean, CVS & AH, SDAU, SK Nagar (Gujarat) for supporting this study.

References

  1. Alton, G., Jones, L.M., Angus, R.D. and Verger, J. M. (1988).Techniques for the Brucellosis Laboratory. INRA, Paris.
  2. Beytut, E., Sahin, M., Erginoy, S. and Sozmen, M. (2009).Pathological, immunohistochemical and bacteriogical findings in the mammary glands and supramammary lymph nodes of cows with a history of abortion due to Brucellaabortus. Turkish Journal of Veterinary and Animal Sciences, 33: 37-43.
  3. Bhattacharya, D.K., Ahmed, K. and Rahman, H. (2005).Studies on seroprevalence of bovine brucellosis by different tests.Journal of Veterinary Public Health, 3: 131-133.
  4. Cadmus S.I.B., Adesokan H.K. and Stack J. (2008). The use of the milk ring test and Rose Bengal test in brucellosis control and eradication in Nigeria. Journal of the South African Veterinary Association, 79: 113-115.
  5. Dalal, S., Bhatt, L., Shrimali, S. and Agrawal, S. (2017). Screening of Milk Samples for Brucellosis around Jaipur. Haryana Veterinarian, 56(1): 102-103.
  6. Dinka, H. and Chala, R. (2009).Seroprevalence Study of Bovine Brucellosis in Pastoral and Agro-Pastoral Areas of East Showa Zone, Oromia Regional State. Ethiopia American-Eurasian Journal of Agriculture & Environmental Science, 6(5): 508-12.
  7. Fleischhauer, G. (1937). Die Abortus-Bang-Ring-probe (ABR) zurFestellung von bangverdächtigenVollmilchproben. BerlTierarztlWochenschr, 53: 527-528.
  8. Gitau, G.K., Royford, M. Bundi, R.M., Vanleeuwen, J. and Mulei, C.M. (2014). Mastitogenic bacteria isolated from dairy cows in Kenya and their antimicrobial sensitivity. Journal of the South African Veterinary Association, 85(1): 8.
  9. Gogoi, S.B., Hussain, P., Sarma P.C., Barua A.G., Mahato G., Bora, D.P., Konch, P. and Gogoi P. (2017). Prevalence of bovine brucellosis in Assam, Indian Journal of Entomology and Zoology Studies, 5(4): 179-185.
  10. Grushina, T., Atshabara, B., Syzdykova, M., Daulbaevaa, S., Tserelsona, L., Kuznetsova, A., Baramovab, S., Seidakhmetovab, R., Sultanovb, A., Ospanovb, Y., Mikhalevb, A., Amireeva, S., Ospanovc, K., Kazakovc, S., Mizanbayevac, S., Myrzabekovc, A., Rementsovaa, M., Berezovskiya, D., Akashevac, R., Khasenovc, M., Nussipovac, Zh.,Yud, W. and Nielsend, K. (2010). Universal indirect enzyme-linked immunosorbent assay for monitoring of human and animal brucellosis. Kazakhstan Vaccine, 28: 46-48.
  11. Hamdy, M.E. and Amin, A.S. (2002).Detection of Brucella species in the milk of infected cattle, sheep, goats and camels by PCR. The Veterinary Journal, 163: 299-305.
  12. Kaleem, M., Durrani, A.Z., Rizwan, M.A., Arain, M.A., Saeed, M., Bhutto, Z.A., Khan Kasi, K.K. and Bacha, U. (2016).Epidemiological Investigation of Outbreak of Brucellosis at Private Dairy Farm, Central Punjab-Pakistan. Indian Veterinary Research Institute, Uttar Pradesh, 4(8) 395.
  13. Karthik, K., Rathore, R., Thomas, P., Arun, T.R., Viswas, K.N., Agarwal, R.K., Manjunathachar H.V. and Dhama, K. (2014).Loop-mediated isothermal amplification (LAMP) test for specific and rapid detection of Brucella abortus in cattle.US National Library of Medicine National Institutes of Health. 34(4):174.
  14. Lodhi, L.A., Jamil, H., Qureshi, Z.I. and Ahmad, I. (1995).Sero-surveillance of brucellosis in buffaloes in and around Faisalabad. Pakistan Veterinary Journal, 15: 127-28.
  15. Mdegelaa, R.H., Ryobab, R., Karimuriboa, E.D., Phiria, E.J., Løkenc, T., Reksenc, O., Mtengetib, E. and Urio, N.A. (2009). Journal of the South African Veterinary Association, 80(3): 163–168.
  16. Meador, V.P., Deyoe, B.L. and cheville, N.F. (1989).Pathogenesis of Brucellaabortus infection of the mammary gland and supramammary lymph node of the goats. Veterinary Pathology, 26: 357-368.
  17. Mohamand, N., Gunaseelan, L., Sukumar, B. and Porteen, K. (2014). Milk Ring Test for spot identification of Brucellaabortusinfection in single cow herds. Journal of Advanced Veterinary and Animal Research, 1(2): 70-72.
  18. Munir, R., Rehman, S. T. Kausar, U. R., Saqlan, R., Naqvi, S. and Farooq, M. U. (2008). Indirect Enzyme Linked Immunosorbent Assay for Diagnosis of Brucellosis in Buffaloes ActaVeterinaria Brno, 77: 401-06.
  19. Nasir, A.A., Parveen, Z., Shah, M.A. and Rashid, M. (2004). Seroprevalence of brucellosis in animals at Government and private livestock farms in Punjab. Pakistan Veterinary Journal, 24(3): 144-46.
  20. OIE (2012).Manual of Diagnostic Tests and Vaccines for Terrestrial Animals online http://oie.int/manual-of-diagnostic-tests-and-vaccines-for-terrestrial-animal.
  21. Refai, M.K. (2003). Brucellosis in animals and man in Egypt. Egyptian Journal of Veterinary Science, 37: 1-31.
  22. Saha, T., Guha, C., Chakraborty, D., Biswas, U., Pal, B., Sarkar, M. and Chatterji, A. (2010). Seroprevalence of brucellosis and isolate on and identification of Brucella in cattle in West Bengal. Indian Journal of Animal Sciences. 80(11): 1087–1088.
  23. Sandhu, K. S., Filia, G., Sharma, D. R., Dhand, N.K., Singh, J. and Saini, S.S. (2001). Prevalence of brucellosis among dairy animals of Punjab. Indian Journal of Comparative Microbiology Immunology and Infectious Diseases, 22: 160-161.
  24. Schalm, O.W., Carroll, E.J. and Jain, N.C. (1971).Bovine Mastitis, 1stedition Lea Febiger, Philadelphia.
  25. Shafee, M., Rabbani, M., Sheikh, A.A., Ahmad, M.D. and Razzaq, A. (2011).Prevalence of Bovine Brucellosis in Organized Dairy Farms, Using Milk ELISA, in Quetta City, Baluchistan. Pakistan Veterinary Medicine International, Article 34: 1-3.
  26. Soomro, A. H., Kamboh, A.A., Rind, R., Dawani, P., Sarwar, M., Abro, S.H. and Awais, M. (2014).A Study on Prevalence and Risk Factors of Brucellosis in Cattle and Buffaloes in District Hyderabad, Pakistan. Journal of Animal Health and Production, 2(3): 33 – 37.
  27. Varasada R.N., (2003). Seroprevalence of brucellosis in cattle, buffalo and human being in central Gujarat. M.V.Sc. Thesis, Gujarat Agricultural University, Sardar Krushi Nagar, India.
  28. Xavier, M.N., Paixao, T.A., Poester, F.P., Lage, A.P. and Santos, R.L. (2009). Pathological, immunohistochemical and bacteriological study of tissues and milk of cows and fetuses experimentally infected with Brucellaabortus. Journal of Comparative Pathology, 140: 149-157.
Abstract Read : 70 Downloads : 18
Previous Next
Close