ASSESSMENT OF BRUCELLOSIS IN MILK OF COWS AND BUFFALOES USING BACTERIOLOGICAL AND SEROLOGICAL METHODS

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Background: Brucellosis is a zoonotic disease caused by Brucella species, gram-negative intracellular bacteria that primarily infect food-producing animals such as cattle and buffaloes. The infection poses a serious public health and economic threat in developing countries due to its impact on livestock productivity and its potential for human transmission through unpasteurized milk. The Milk Ring Test (MRT) serves as a rapid, cost-effective, and indirect method for detecting anti-Brucella antibodies in milk samples. Objective: This study aimed to determine the frequency of brucellosis among milk samples of cows and buffaloes in Tehsil Bahrain, Swat, using serological and bacteriological methods. Methods: The study was conducted in the Department of Microbiology, Government College Madyan, Swat, from July to December 2024 after ethical approval from the institutional research committee. A total of 160 raw milk samples were collected—100 from cows and 60 from buffaloes—from both individual animals and bulk milk tanks. Samples were aseptically collected in sterile containers, refrigerated at 4°C, and analyzed using the MRT. For each test, 1 mL of milk was mixed with 0.03 mL of hematoxylin-stained antigen and incubated at 37°C for one to three hours. The formation of a blue ring indicated a positive result. Biochemical identification of Brucella species was performed according to standard protocols, and data were analyzed using SPSS version 16. Results: Out of 160 milk samples, 20 (12%) tested positive for Brucella antibodies. Among these, 14 (14%) were from cows and 6 (10%) from buffaloes. The predominant species isolated were Brucella melitensis (64.2% in cows; 66.6% in buffaloes) and Brucella abortus (35.7% in cows; 33.3% in buffaloes). Seasonal variation analysis revealed the highest seropositivity in December and the lowest in July, suggesting environmental influence on disease occurrence. Conclusion: Brucellosis remains a persistent zoonotic and economic concern in Tehsil Bahrain, Swat, with higher seroprevalence observed during the rainy season. MRT proved to be a reliable and practical diagnostic method for rapid surveillance of Brucella infection in dairy herds, underscoring the need for routine screening and preventive measures.

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