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Asian Journal of Biological Sciences
Year: 2017  |  Volume: 10  |  Issue: 2  |  Page No.: 47 - 55

Establishment and Application of the Resazurin Micro-plate Method for Rapid Diagnosis of Bovine Mastitis-causing Staphylococci

Lubna Hassan and Fahim Ullah    

Abstract: Background and Objective: Staphylococci strains causing bovine mastitis, has produced serious therapeutic problems. However, the underlying mechanisms are poorly understood and it is necessary to develop new effective method for accurate diagnosis. Resazurin micro-plate method was used for the diagnosis of bovine mastitis caused by staphylococci. The objective of this study was to discuss about the method of resazurin micro-plate for the diagnosing of bovine mastitis which caused by staphylococci. Methodology: While for testing the infected mastitis milk samples were inoculated in the selection of the Staphylococcus-mBP medium with 96-well micro-plate and pure cultures of bacteria and after 2 h of the incubation the resazurin indicator was added to that solution up to 0.4 g L–1 and then continued the cultivation for 10 h, depending on whether the medium color changes of bacterial growth. For statistical analysis chi-square test was performed using SPSS Software version 22 (IBM SPSS Statistics for Windows, Armonk, NY, USA: IBM Corp.). Results: The results from the bacterial pure culture plate showed that the resazurin micro-plate indicated the growth of S. aureus i.e., S. epidermidis, S. chromogens, S. haemolyticus, S. simulans , S. xylosus and E. coli did not appear on the culture plates. Further from the investigated results showed that the color changing in the medium was due to the staphylococcal concentration and time incubation. However, the results of the infection milk samples showed that the resazurin micro-plate had a detection sensitivity of 1×103 CFU mL–1 and in 12 h the diagnosis could be obtained. Conclusion: Furthermore from the results of the mastitis milk samples, the positive culture plate showed that the resazurin micro-diagnosis of S. aureus-positive and negative milk sample at the rate of 98.3% (n = 60) and 95.0% (n = 40) respectively. Similarly, from all the investigated results it is concluded that the resazurin micro-plate can replace the conventional method for rapid diagnosis of Staphylococcus milk samples.

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