Detection of animal brucellosis by cultural method

M V HEMADE; R S GANDGE; A S BANNALIKAR

doi:10.56093/ijans.v85i6.49282

Authors

M V HEMADE Livestock Development Officer, Dhule district
R S GANDGE Associate Professor, Bombay Veterinary College, Parel, Mumbai, Maharashtra 400 012 India
A S BANNALIKAR Professor, Bombay Veterinary College, Parel, Mumbai, Maharashtra 400 012 India

https://doi.org/10.56093/ijans.v85i6.49282

Keywords:

Animal brucellosis, Brucella abortus, Brucella melitensis, Cultural identification, Cultural isolation, PCR

Abstract

Brucellosis, an infectious disease of many vertebrate species, is an important re-emerging zoonosis with a worldwide distribution. The present research was aimed for detection of brucellosis in animals by isolation and identification of Brucella species isolates by conventional method and PCR. Clinical specimens (61) which included vaginal swabs (48), foetal membranes (08), semen samples (03) and abomasal contents of aborted foetuses (02) were subjected for isolation on Brucella Agar Medium (BAM), wherein 15 Brucella spp. isolates were recovered with isolation rate of 24.59%. All 15 conventionally identified Brucella abortus species isolates were further confirmed by 2 different PCR assays using genus specific BCSP31 and species specific IS711 (AB and BM) primers. The BCSP31 (223 bp) PCR assay was simple, effective and sensitive in detection of Brucella isolates at genus level, whereas, IS711/AB PCR was useful to detect isolates at species level. In IS711/AB PCR, abortus specific amplification product of 498 bp was produced in all 15 isolates confirming their identity as B. abortus, whereas none of isolate showed B. melitensis specific amplification in IS711/BM PCR.

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References

Aher A S. 2010. ‘Evaluation of BCSP31 and IS711 PCR assays in detection of bovine and human brucellosis’. M.V.Sc. Thesis submitted to MAFSU, Nagpur.

Al-Farwachi, M I, Al-Badrani B A and Al-NimaTh M. 2010. Detection of Brucella antigen in the aborted ovine fetal stomach contents using a modified ELISA test. Iraqi Journal of Veterinary Sciences 24 (1): 1–4. DOI: https://doi.org/10.33899/ijvs.2010.5577

Al-Shamahy H A. 1999.Seropositivity for brucellosis in a sample of animals in the republic of yemen. Eastern Mediteranean Health Journal 5 (5): 1035–41. DOI: https://doi.org/10.26719/1999.5.5.1042

Alton G, Jones L M, Angus R D and Verger J M. 1988. Techniques for the Brucellosis Laboratory. INRA, Paris.

Amin A S, Hamdy M E and Ibrahim A K. 2001.Detection of Brucellamelitensisin semen using the polymerase chain reaction assay.Veterinary Microbiology 83: 37–44. DOI: https://doi.org/10.1016/S0378-1135(01)00401-1

Baily G C, Kraahn J B, Drasar B S and Stokeer N G. 1992. Detection of Brucella melitensis and Brucella abortus by DNA amplification. The Journal of Tropical Medicine and Hygiene. 95: 271–75.

Bayatti S A and Thwani A N. 2009. DNA polymorphism investigation among local Brucella isolates. Iraqi Journal of Veterinary Sciences 23 (II): 213–18.

Bhiogade P P. 2012. ‘Molecular characterization of outer membrane protein (omp) gene of Brucella abortus.’ M.V.Sc. Thesis submitted to MAFSU, Nagpur.

Bricker B J and Halling S M. 1994. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis and Brucella suis bv. 1 by PCR. Journal of Clinical Microbiology 32: 2660–66. DOI: https://doi.org/10.1128/jcm.32.11.2660-2666.1994

Bricker B J. 2002. PCR as diagnostic tool for Brucellosis. Veterinary Microbiology 90 (1–4): 435–46. DOI: https://doi.org/10.1016/S0378-1135(02)00228-6

Brown T A. 2007. Essential Molecular Biology. Oxford University Press. 2nd edn. 1: 96–100.

Casanas M C, Queipo-Ortuno M I, Rodriguez-Torres A, Orduna A, ColmeneroJ D and Morata P. 2001. Specificity of a polymerase chain reaction assay of a target sequence on the 31–kilodalton Brucella antigen DNA used to diagnose human brucellosis. European Journal of Clinical Microbiology and Infectious Diseases 20: 127–31. DOI: https://doi.org/10.1007/PL00011242

Chahotal R, Sharmal M, Katochl R C, Verma S, Singh M, Kapoorl V and AsraniR K. 2003.Brucellosis outbreak in an organized dairy farm involving cows and in contact human beings, in Himachal Pradesh, India. Veterinarski Arhiv.73: 95–102.

Corbel M J and Brinley-Morgan W J. 1984. Genus Brucella Meyer and Shaw, 1920,173AL. Bergey’s Mannual of Systematic Bacteriology. (Eds) Krieg N R and Holt J G. Willians and Wilkins, Baltimore.

Das A M, Paranjape V L and Corbel M J. 1990. Investigation of brucellosis associated diseases in dairy buffaloes and cattles. Indian Journal of Animal Sciences 60: 1193–94.

Farrel I D and Robertson L. 1972. Journal of Applied Bacteriology 35: 625. DOI: https://doi.org/10.1111/j.1365-2672.1972.tb03744.x

Geering W A, Forman J A and Nunn M J. 1995. Exotic Diseases of Animals. Australian Government Publishing Service, Canberra, Australia, 301–06.

Guarino A, Serpe L, Fusco G, Scaramuzzo A and Gallo P. (2000) Detection of Brucellaspecies in buffalo whole blood by genespecific PCR. Veterinary Record 147: 634–36. DOI: https://doi.org/10.1136/vr.147.22.634

Herman L and Herman De. 1992. Identification of Brucellaspp.by using the polymerase chain reaction. Applied and Environmental Microbiology 58: 2099–101. DOI: https://doi.org/10.1128/aem.58.6.2099-2101.1992

Kanani A. 2007. ‘Serological, cultural and molecular detection of Brucella infection in breeding bulls.’ PhD thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand.

Kollannur J D, RathoreR and Chauhan R S. 2007. Epidemiology and economics of brucellosis in animals and its zoonotic significance.Animal health, animal welfare and biosecurity. Proceedings of 13th International Congress in Animal Hygiene, Tartu, Estonia.1: 466–68.

Leal-Klevezas D S, Martinez V I O, Garcia C J, Lopez M A and P.Martinez S J. 2000. Use of polymerase chain reaction to detect Brucella abortus biovar1 in infected goats. Veterinary Microbiology 75: 91–97. DOI: https://doi.org/10.1016/S0378-1135(00)00200-5

Londhe S P. 2009. ‘Molecular detection and characterization of Brucella abortus from bovines.’ M.V.Sc thesis submitted to Maharashtra Animal & Fishery Sciences University, Nagpur.

Madale D. 2011. ‘Detection of Brucella spp. in animals by conventional and molecular techniques. ’ M.V.Sc thesis submitted to MAFSU, Nagpur.

Mehra K N, Dhanesar N S and Chaturvedi V K. 2000. Seroprevalence of brucellosis in bovines of Madhya Pradesh. Indian Veterinary Journal 77: 571–73.

Mukherjee F, Jain J, Patel V and Nair M. 2007. Multiple genusspecific markers in PCR assays improve the specificity and sensitivity of diagnosis of brucellosis in field animals. Journal of Medical Microbiology 56: 1309–16. DOI: https://doi.org/10.1099/jmm.0.47160-0

Musa M T, Jahan K L and Shigidi M T A. 2000. Brucella organism isolated from the sudan I: Species biovars associated with indigenous animals. Sudan Journal of Veterinary Research 16: 23–28.

OIE. 2004. Bovine brucellosis, chapter 2.3.1 Manual of Standard for Diagnostic Test and Vaccines. 5th edn OIE. Paris on 16 March 2006, Paris. OIE. 2009. OIE Terrestrial Manual: Bovine Brucellosis. Version adopted by the World Assembly of Delegates of the OIE in May 2009.

Quinn P J, Carter M E, Markey B and Carter G R. 1994. Clinical Veterinary Microbiology. Wolfe publisher, England.21–66.

Radostits M O, Gay C C, Hinchcliff K W and Constable P D. 2008. Veterinary Medicine Textbook of Diseases of Cattle, Sheep, Goats, Pig and Horses. Vol. 10th, p.967 London. Bailliere, Tindall.

Rathore B S, Barman T K, Singh K P, Singh R and Mehrotra M L. 2002. Microbiological and epidemiological studies on brucellosis in an organized herd and rural cattle and buffaloes of Uttar Pradesh. Indian Journal of Comparative Microbiology, Immunology and Infectious Diseases 23:195–96.

Renukaradhya G J, Isloor S and Rajsekhar M. 2002. Epidemiology, zoonotic aspect, vaccination and control / eradication of Brucellosis in India. Veterinary Microbiology 90: 183–95. DOI: https://doi.org/10.1016/S0378-1135(02)00253-5

Romero C, Gamezo C, Pardo M and Lopez-GoniI. 1995. Specific Detection of Brucella DNA by PCR. Journal of Clinical Microbiology 33: 615–17. DOI: https://doi.org/10.1128/jcm.33.3.615-617.1995

Sarumathi C, Reddy T V, Sreedevi B and Rao U V N M. 2003.Comparison of avidin-biotin ELISA with RBPT and STAT for screening of antibodies to bovine brucellosis. Indian Veterinary Journal 80: 1106–08.

Shin S J, Drazek F J, Fairbrother J M, Kelly M S, McDonough P L and Rogers J D. 1978. Recent developments in the culturing of Brucella abortus: methods of increasing the isolation rate from various specimens. Proceedings of the American Association of Veterinary Laboratory Diagnosticians 21: 297– 308.

Singh S P, Kumar M and Kumar A. 2002. Advance in veterinary public health. First annual conference IAVPH. Pantnagar.

Suryawanshi A A. 2011. ‘Detection of Brucella spp. in occupationally exposed humans by serological, bacteriological and molecular techniques.’ M.V.Sc. Thesis submitted to MAFSU, Nagpur.

Trujillo I Z, Zavala A N, Cacres J G and Miranda C Q. 1994. Brucellosis. Infectious disease clinics of North America 8: 225– 41. DOI: https://doi.org/10.1016/S0891-5520(20)30581-X

Tuba I C A, Fuat A Y D I N, Semih K G, Duygu P. Sumerkan A B, Fulya O C A K, Secil A H, Okan D O G A N, Arzu F I N D I K and Ciftci A. 2012. Conventional and molecular biotyping of Brucella strains isolated from cattle, sheep and human. Ankara Üniversitesi Veteriner Fakültesi Dergisi 59: 259–64. DOI: https://doi.org/10.1501/Vetfak_0000002536