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Evaluation of real-time PCR based on SYBR Green I fluorescent dye for detection of Bacillus anthracis strains in biological samples
2018
Kędrak-Jabłońska, Agnieszka | Budniak, Sylwia | Szczawińska, Anna | Reksa, Monika | Krupa, Marek | Szulowski, Krzysztof
Introduction: The aim of the study was the application and evaluation of real-time PCRs based on the fluorescence of SYBR Green I intercalating dye for the detection of three Bacillus anthracis genes in contaminated liver and blood samples. The goals for detection were rpoB gene as a chromosomal marker, pag gene located on plasmid pXO1, and capC gene located on plasmid pXO2. Material and Methods: Five B. anthracis strains were used for the experiments. Additionally, single strains of other species of the genus Bacillus, i.e. B. cereus, B. brevis, B. subtilis, and B. megaterium, and strains of six other species were used for evaluation of the specificity of the tests. Three SYBR Green I real-time PCRs were conducted allowing confirmation of B. anthracis in the biological samples. Results: The observation of amplification curves in real-time PCRs enabled the detection of the chromosomally encoded rpoB gene, pag gene, and capC gene of B. anthracis. The specificity of the tests was confirmed by estimation of the melting temperature of the PCR products. The sensitivity and linearity of the reactions were determined using regression coefficients. Strains of other microbial species did not reveal real-time PCR products. Conclusion: All real-time PCRs for the detection of B. anthracis in biological samples demonstrated a significant sensitivity and high specificity.
Mostrar más [+] Menos [-]Influence of incorporating dried fruits on dairy drinks characteristics focusing on their antimicrobial effects
2024
Neveen S. M. Soliman | Ayah B. Abdel-Salam | Shimaa R. Emam | Ahmed Orabi | Sara M. Nader | Mena Saad
Objective: The study was designed to show the effect of adding different levels of dried fruit extracts for 14 days on sensory and chemical parameters in dairy drinks. The survival of Pseudomonas aeruginosa and Bacillus cereus in artificially contaminated dairy drinks fortified with these extracts was also studied. Materials and Methods: The freshly watery extracts and nonaqueous extracts of dried fruits were prepared by rotary evaporators and solvents, respectively. The determination of the minimum inhibitory concentration of dried fruit extracts was achieved using the disc diffusion test. The sensory evaluation of samples was done, while the chemical parameters of the examined samples were determined by the calibrated analyzer. In addition, the degree of survival of P. aeruginosa and B. cereus in inoculated milk samples was also estimated. Results: In pasteurized and Rayeb milk samples, the water extract of carob and all alcoholic dried fruit extracts had a significant effect on compositional parameters in comparison to control samples. At day 14 of pasteurized milk storage, the watery (20.0%) and alcoholic (10.0%) extracts of carob significantly improved its sensory parameters. Conclusion: Based on the survival results, all utilized dried fruit extracts had a significant inhibitory effect on P. aeruginosa and B. cereus growth in the fortified milk samples at the end of storage. This trial of the survival of these new dairy drinks is the first investigation, particularly in the Middle East. Extracts of utilized dried fruits have prospective functions that enhance dairy drink characteristics. [J Adv Vet Anim Res 2024; 11(1.000): 55-64]
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