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.
It is well known in cattle that reproductive disorders are intimately associated with low or high body condition score (BCS). However, little is known concerning the relationship between BCS and oxidative stress, particularly in the reproductive tract. Thus, the aim of the present study was to investigate the oxidative stress biomarkers according to cows BCS values in the plasma, ovaries, oviductal, follicular and uterine fluids. The study was conducted on 58 cows classified in four classes (1, 2, 3 and 4) of BCS varying from1-1.5, 2-2.5, 3-3.5 and 4-4.5, respectively. The genital tracts and plasma were collected from slaughtered cows and processed within 5h after slaughtering. The number of ovarian follicles was measured using ultrasonography and the oxidative stress was assessed by considering total antioxidant status (TAS), catalase activity (CAT), and thiobarbituric acid-reactive substances (TBARS). The results showed that follicles number was significantly affected by BCS; cows with medium BCS (3) presented higher number of follicles than cows with low or high BCS (<3 or >3). Overall, BCS affects significantly the oxidative stress status at different levels of the reproductive tract. Cows with medium BCS (3) showed the best oxidative status than those with low or high values (BCS <3 or >3). In conclusion, the current results suggest that reproductive disorders observed in cows with low or high BCS values could be mediated through oxidative stress affecting consequently the reproductive tract and finally compromising fertility outputs.
First insemination success is considered as good indicator of heifer fertility to ensure renewal of dairy herds. The objective of this study was to determine the relationship between first insemination success and biomarkers of oxidative stress (OS). A total of 25 heifers were divided into two groups according to their success at first insemination: group FS+ (heifers that were pregnant at first service, n = 14) and group FS- (heifers that were not pregnant at first service, n = 11). The serum of these two groups were analyzed for malondialdehyde (MDA), glutathione S-transferase (GST), reduced glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT), myeloperoxidase (MPO), nitric Oxide (NO) and superoxide dismutase (SOD) as oxidative stress biomarkers and biochemical parameters. Heifers in the group FS+ showed no significant difference in all OS parameters compared to heifers in the group FS-. The OS parameters showed almost similar values in both groups except for GST and CAT where the difference was at the limit of significance. The plasma concentration of OS biomarkers assessed in our study were not related to first service success in heifers. Further studies are needed to clarify the role of oxidative status in the reproductive performance of heifers.
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