Molecular characterization of Brucella species detected from clinical samples of cattle and buffaloes
Abstract views: 156 / PDF downloads: 130 / PDF downloads: 19
Keywords:Brucella abortus, B. abortus 544, BCSP 31, IS711 PCR assays
The present study was undertaken for molecular characterization of Brucella species of cattle and buffaloes. Clinical samples (1145) of unvaccinated cattle and buffaloes (200 blood samples, 710 sera, 190 vaginal swabs, 20 abomasal contents of foetus, 25 foetal tissues) and 146 blood samples of vaccinated animals were collected from dairy farms in and around Mumbai and Pune region. These samples were processed for isolation of Brucella organisms and further characterized by PCR and sequencing. A total of 26 (11.06%) Brucella isolates were recovered from 235 samples. Also, 5 isolates received from human cases were included in the study. BCSP 31 PCR showed an amplicon of 223 bp in all 31 isolates, 123 (61.5%) blood samples, 123 (64.73%) vaginal swabs and 27 (60%) aborted foetal material. IS711/AB and BM PCR showed an amplicon of 498 bp and 731 bp in 17 and 14 isolates, 42 (21%) and 38 (19%) blood samples, 43 (22.63%) and 34 (17.89%) vaginal swabs, while 7(15.55%) and 6 (13.33%) aborted foetal material, respectively. The phylogenetic analysis detected the ancestral origin of the organism. Rapid and correct diagnosis of brucellosis and vaccination is important to eradicate the disease. The molecular methods used in the present study speed up the diagnosis of the disease.
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.
Ahmed R, Muhammad K, Rabbani M, Khan M S, Ali M A, Naureen S, Kanwal F, Raza S, Aqib A I, Sadia H and Chang Y F. 2017. Phylogenetic analysis of soil borne Brucella species by targeting insertion sequence 711 element in Punjab, Pakistan. International Journal of Agriculture and Biology 19(6): 1457–62.
Al-Attas R A, Al-Khalifa M and Alqurashi A R. 2000. Evaluation of PCR, culture and serology for the diagnosis of acute human brucellosis. Annual Saudi Medicine. 20 (3-4): 224–28. DOI: https://doi.org/10.5144/0256-4947.2000.224
Al-Dahouk S, Tomaso H, Nockler K, Neubauer H and Frangoulidis D. 2003. Laboratory-based diagnosis of brucellosis: A review of the literature. Part II: Serological tests for brucellosis. Clinical Laboratory 49(11-12): 577–89.
Awwad E, Adwan K, Farraj M, Essawi T, Rumi I, Manasra A, Baraitareanu S, Gurau M R and Danes D. 2015. Cell envelope virulence genes among field strains of Brucella melitensis isolated in West Bank part of Palestine. Agriculture and Agricultural Science Procedia 6: 281–86. DOI: https://doi.org/10.1016/j.aaspro.2015.08.073
Barua A, Kumar A, Thavaselvam D, Mangalgi S, Prakash A, Tiwari S, Arora S and Sathyaseelan K. 2016. Isolation and characterization of Brucella melitensis isolated from patients suspected for human brucellosis in India. The Indian Journal of Medical Research 143(5): 652–58. DOI: https://doi.org/10.4103/0971-5916.187115
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(11): 2660–66. DOI: https://doi.org/10.1128/jcm.32.11.2660-2666.1994
Das V M, Paranjape V L and Corbel M J. 1990. Investigation of brucellosis-associated abortion in dairy buffaloes and cows in Bombay. Indian Journal of Animal Sciences 60(10): 1193–94.
Deka R P, Magnusson U, Grace D and Lindahl J. 2018. Bovine brucellosis: prevalence, risk factors, economic cost and control options with particular reference to India-A review. Infection Ecology and Epidemiology 8(1): 1556548. DOI: https://doi.org/10.1080/20008686.2018.1556548
Ilhan Z, Solmaz H, Aksakal A, Gulhan T, Hakkl Ekin I. and Oynukara B. 2007. Comparison of PCR assay and bacteriological culture method for the detection of Brucella melitensis in stomach content samples of aborted sheep fetuses. Deutsche Tierarztliche Wochenschrift 114: 459–63.
Jeyaprakash C, Ranjitsingh A J A and Amuthan A. 1999. Isolation of Brucella spp. from indigenous and cross-bred cows and evaluation of their antibiogram. Indian Journal of Animal Sciences 33: 99–103.
Kala M, Sankhala L, Kant L, Kumar L, Puniya S and Shah N. 2018. Isolation, identification and molecular detection of Brucella abortus from bovines of North Gujarat. Journal of Entomology and Zoology Studies 6(3): 1523–27.
Kaur P, Sharma N S and Arora A K. 2018. Investigation of brucellosis in cattle and buffaloes by conventional and molecular assays. Indian Journal of Animal Research 52(10): 1482–87. DOI: https://doi.org/10.18805/ijar.B-3375
Leal-Klevezas D S, Martinez-Vazquez I O, Lopez-Merino A and Martinez-Soriano J P. 1995. Single-step PCR for detection of Brucella spp. from blood and milk of infected animals. Journal of Clinical Microbiology 33: 3087–90. DOI: https://doi.org/10.1128/jcm.33.12.3087-3090.1995
Londhe S P, Aher A S, Jagadale S D, Dighe S D and Bannalikar A S. 2013. Evaluation of BCSP 31 kDa and IS711 PCR assays in detection of bovine brucellosis. Indian Journal of Veterinary Research 22(1): 8–14.
Mittal M, Sharma V, Nehra K, Chakravarti S, Kundu K, Bansal V K, Churamani C P and Kumar A. 2018. Abortions in an organized dairy farm from North India reveal the possibility of breed susceptibility to Bovine Brucellosis. One Health, 5: 1–5. DOI: https://doi.org/10.1016/j.onehlt.2017.11.001
Mukherjee F, Jain J, Patel V and Nair M. 2007. Multiple genus- specific markers in PCR assays improve the specificity and sensitivity of diagnosis of brucellosis in field animals. Journal of Medical Microbiology 56(10): 1309–16. DOI: https://doi.org/10.1099/jmm.0.47160-0
O’Leary S, Sheahan M and Sweeney T. 2006. Brucella abortus detection by PCR assay in blood, milk and lymph tissue of serologically positive cows. Research in Vveterinary Sscience 81(2): 170–76. DOI: https://doi.org/10.1016/j.rvsc.2005.12.001
OIE Terrestrial Manual 2009. Bovine brucellosis. Retrieved February 02, 2015 from http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.04.03.
Romero C, Gamazo C, Pardo M and Lopez-Goni I. 1995. Specific detection of Brucella DNA by PCR. Journal of Clinical Microbiology 33(3): 615–17. DOI: https://doi.org/10.1128/jcm.33.3.615-617.1995
Seleem M N, Boyle S M and Sriranganathan N. 2010. Brucellosis: A re-emerging zoonosis. Veterinary Microbiology 140(3-4): 392–98. DOI: https://doi.org/10.1016/j.vetmic.2009.06.021
Shakuntala I, Milton A A P, Sanjukta R K, Kakoty K, Karam A, Dutta A, Puro K, Sen A, Das S and Ghatak S. 2021. Isolation and sero-genomo-epidemiological studies on Brucella infection in dairy cattle in Meghalaya, India. Comparative Immunology, Microbiology and Infectious Diseases 78: 101694. DOI: https://doi.org/10.1016/j.cimid.2021.101694
Shome R, Kalleshamurthy T, Nagaraj C, Rathore Y, Ramanjinappa K D, Skariah S, Mohandoss N, Shome B R, Chanda M M and Hemadri D. 2022. Countrywide cross-sectional study of swine brucellosis sero-prevalence in Indian subcontinent during 2018–2019. Tropical Animal Health and Production 54(2): 1–10. DOI: https://doi.org/10.1007/s11250-022-03107-9
Surucuoglu S, Ural S E R A P, Gazi H O R U, Kurutepe S E M R A, Taskiran P and Yurtsever S G. 2009. Evaluation of real-time PCR method for rapid diagnosis of brucellosis with different clinical manifestations. Polish Journal of Microbiology 58(1): 15–19.
Suryawanshi A and Gandge R. 2017. Detection and antibiotic sensitivity testing of brucella species. World Journal of Pharmacy and Pharmaceutical Sciences 6(7): 1817–29. DOI: https://doi.org/10.20959/wjpps20177-9621
Thenamutha M, Zakiah M D, Azizul O and Maswati M A. 2017. Isolation and molecular characterization of Brucella abortus and Brucella melitensis from samples received by the regional veterinary laboratory, Bukit Tengah, Malaysia. Malaysian Journal of Veterinary Research 8: 79–87.
Tyasningsih W. 2015. ‘Potensi Outer Membrane Protein 36 kDa Brucella abortus Isolat Lokal Sebagai Kandidat Vaksin Brucellosis MelaluI Aktivitas Respon Imun Seluler Dan Humoral.’ (Doctoral dissertation, Universitas Airlangga).
How to Cite
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.