OPTIMIZATION AND VALIDATION OF HPLC METHOD FOR TETRAMETHRIN DETERMINATION IN HUMAN SHAMPOO FORMULATION.

High-performance liquid chromatography (HPLC) method with diode array detection (DAD) were optimized and validated for separation and determination of tetramethrin in an antiparasitic human shampoo. In order to optimize separation conditions, two different columns, different column oven temperatures, as well as mobile phase composition and ratio, were tested. Best separation was achieved on the Supelcosil TM LC-18- DB column (4.6 x 250 mm), particle size 5 jim, with mobile phase methanol : water (78 : 22, v/v) at a flow rate of 0.8 mL/min and at temperature of 30°C. The detection wavelength of the detector was set at 220 nm. Under the optimum chromatographic conditions, standard calibration curve was measured with good linearity [r2 = 0.9997]. Accuracy of the method defined as a mean recovery of tetramethrin from shampoo matrix was 100.09%. The advantages of this method are that it can easily be used for the routine analysis of drug tetramethrin in pharmaceutical formulas and in all pharmaceutical researches involving tetramethrin.

Clinical Evidence on Expansion of Essential Oil-Based Formulation's Pharmacological Activity in Bovine Mastitis Treatment: Antifungal Potential as Added Value.

Bovine mastitis, as a significant and prevalent health problem in dairy herds, is primarily attributed to bacterial infections. Yeasts, although considered relatively rare causative agents, have also been associated with mastitis in dairy cattle. Current mastitis treatment predominantly relies on antibiotics, with limited emphasis on antifungal treatment. However, mycotic mastitis is challenging to treat, since these fungi are often resistant to antibiotics and may even utilize them for energy. In the current research, the in vivo antimicrobial activity of the essential oil-based formulation (Phyto-Bomat), as a possible alternative mastitis treatment associated with yeasts, was studied. This study involved a total of 68 animals from two dairy farms with diagnosed mastitis, and three treatment groups were established: conventional antibiotic treatment, Phyto-Bomat treatment, and the combination of both. The findings suggest significant variations in the presence of Candida samples based on the treatment administered, and the most significant difference was noted in cows treated with the combination (Phyto-Bomat and antibiotics). Yet, it is important to note that the results reveal that, regardless of the treatment type, there are statistically significant differences in the presence of Candida samples across the examined time points. These results aim to provide valuable insights into the potential of EOs as an alternative therapy in bovine mastitis, specifically targeting yeasts. Such findings could offer new strategies in the mycotic mastitis control and reducing the occurrence of secondary infections following antibiotic treatment.

Environmental Bovine Mastitis Pathogens: Prevalence, Antimicrobial Susceptibility, and Sensitivity to Thymus vulgaris L., Thymus serpyllum L., and Origanum vulgare L. Essential Oils.

Mastitis is considered to be one of the most important diseases of dairy cows in terms of health, production, and economy. Being the most common cause of antibiotic consumption in dairy cows, treatment of this disease is one of the biggest challenges in the veterinary profession as an increasing number of pathogens develop resistance to antibiotics used in the treatment. Therefore, new alternative approaches for limiting the use of antibiotics in livestock are required. For this reason, our study aimed to investigate prevalence of environmental mastitis associated bacterial strains, as well as the sensitivity of isolated strains to different antibiotics. Additionally, the therapeutic potential of three essential oils (EOs) was tested against bovine Serratia spp. and Proteus spp. mastitis pathogens, based on their chemical composition, as well as antibacterial potential. The study was carried out on 81 milk samples collected from dairy cows with mastitis. In order to determine prevalence of S. marcescens and P. mirabilis, microbiological isolation and identification were performed. Antimicrobial susceptibility testing was performed by disk diffusion method and the microdilution method was used to determine the antibacterial activity of selected EOs. In the oregano EO, a total of 23 compounds were detected, with carvacrol as a dominant component (78.94%). A total of 26 components were present in the EO of common thyme, where thymol was the most abundant compound (46.37%). Thymol also dominated (55.11%) the wild thyme EO. All tested EOs displayed antibacterial activity against all strains to different extents, while wild and common thyme EOs were the most effective. It could be concluded that the tested EOs represent promising therapeutic candidates for effective non-antibiotic treatment of mastitis.

Effect of an Antioxidant Supplement Combination on Boar Sperm.

The study was conducted on a commercial pig farm located in Serbia. Thirty Duroc or Landrace breed boars were randomly selected for this study. The experimental group was fed a compound feed with added organic selenium and Oxynat 3D. The antioxidant status parameters of boar seminal plasma were evaluated using a biochemical analyzer and commercial Randox kits. The sperm chromatin structure assay (SCSA) using flow cytometry (FC) provided information about spermatozoa's DNA status. Additionally, the total number of motile spermatozoa and spermatozoa kinematic parameters were measured using the computer-assisted sperm analysis (CASA) system. The aim of this study was to improve the parameters of semen by combining two preparations that have a potential antioxidant effect, but also to establish the level of various antioxidant enzymes in native sperm. There was no statistically significant difference in total antioxidant capacity and glutathione peroxidase activity in the seminal plasma obtained from the experimental and control groups of boars. Regarding the superoxide dismutase activity, the research results showed a difference in the control group compared to the experimental one. Moreover, spermatozoa DNA fragmentation and the total number of motile spermatozoa showed statistically significant lower and higher values, respectively, in experimental compared to the control groups. The combination of these two preparations shows significantly enhanced vital parameters of semen. To the best of our knowledge, this study is the first in which the ejaculate parameters were examined after the application of a combination of these two antioxidant supplements.