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.

The present investigation was undertaken to study the occurrence of enterotoxigenic B. cereus from 200 samples including 100 raw milk, 50pasteurized milk and 50 ice cream samples sold in Puducherry. The results revealed that 68 samples (34 %) belonged to B. cereus group which included 36 from raw milk (36%), 9 from pasteurized milk (18%) and 23 from ice cream samples (46 %). On PCR analysis, 64 out of 68 isolates showed the presence of gyrB gene which differentiates B. cereus from B. cereus group isolates. Multiplex PCR revealed the presence of four enterotoxigenic genes hblA, nheA, cytK and entF Min 32.35 %, 77.94 %>, 54.41 % and 94.12% of the 64 isolates. On biofilm production assay, out of 64 enterotoxigenic B. cereus isolates, 60 (93.75 %) isolates showed the ability to form biofilm in which 13 (20.31 %) were strong, 21 (32.81 %) were moderate and26 (40.63 %) were weak biofilm producers. All the isolates were resistant to ampicillin and penicillin G and sensitive to ciprofloxacin, gentamicin, vancomycin, streptomycin and norfloxacin. The current study directs the need for proper hygienic measures during collection, production, processing and storage of milk to reduce the contamination level. It also indicates the need for surveillance and action to reduce the contamination of enterotoxigenic B. cereus in milk and milk products in Puducherry.

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]