Artur Burmańczuk, Cezary Kowalski, Zbigniew Roliński, Rafał Zań and Dorota Krasucka
Introduction: The objective of this study was to determine the current profile of bacteria responsible for the infection of the mammary gland and to assess their sensitivity to selected β-lactam antibiotics.
Material and Methods: The study was conducted on 119 (n = 119) dairy cows of the Polish Black-White breed aged 4 to 10 years with inflammation of the mammary gland. The cows came from different farms: smallholder farms and large dairy cattle farms in the Lublin and Bialystok Provinces. Before the process of collection of milk samples, the teats were cleaned and immersed in a liquid disinfectant. The first streams were collected into containers which were subsequently utilised. Afterwards, 2-4 mL of milk or secretions was milked into sterile disposable tubes. The milk samples were collected into plastic bottles and kept in a cooler with ice until transportation to the laboratory. Tests of resistance to β-lactam antibiotics were performed by disc diffusion.
Results: Contagious and environmental bacteria were isolated from all dairy barns. In the group of contagious bacteria, the presence of typical pathogens responsible for the mammary gland infections, i.e. Staph. aureus, Str. agalactiae, and C. bovis, was detected. A relatively broad group of the isolates was formed by environmental bacteria responsible for inflammation of the mammary gland: Str. dysgalactiae, Str. uberis, Staph. chromogenes, Staph. hyicus, Staph. warneri, and E. coli. Among the environmental organisms, streptococci constituted the largest percentage (23%), followed by staphylococci (13.2%), and E. coli (8.8%). The largest group of infectious pathogens comprised Str. agalactiae (29.6%) and Staph. aureus (20.8%).
Conclusion: Our investigation of the current profile of the isolates responsible for mastitis in the Lublin and Bialystok Provinces showed that environmental bacteria are the major cause of the disease. In view of the substantially varying degrees of sensitivity of the microorganisms isolated from cases of mastitis to β-lactam antibiotics, each therapeutic treatment should be preceded by susceptibility testing.
Wojciech Jerzy Pietro, Aneta Woźniak, Katarzyna Pasik, Wojciech Cybulski and Dorota Krasucka
A liquid chromatography-ultraviolet detection method for the determination of florfenicol (FF) and thiamphenicol (TAP) in feeds is presented. The method comprises the extraction of analytes from the matrix with a mixture of methanol and acetonitrile, drying of the extract, and its dissolution in phosphate buffer. The analysis was performed with a gradient programme of the mobile phase composed of acetonitrile and buffer (pH = 7.3) on a Zorbax Eclipse Plus C18 (150 × 4.6 mm, 5 μm) analytical column with UV (λ = 220 nm) detection. The analytical procedure has been successfully adopted and validated for quantitative determination of florfenicol and thiamphenicol in feed samples. Sensitivity, specificity, linearity, repeatability, and intralaboratory reproducibility were included in the validation. The mean recovery of amphenicols was 93.5% within the working range of 50-4000 mg/kg. Simultaneous determination of chloramphenicol, which is banned in the feed, was also included within the same procedure of FF and TAP stability studies. Storing the medicated feed at room temperature for up to one month decreased concentration in the investigated drugs even by 45%. These findings are relevant to successful provision of therapy to animals.
Wojciech Jerzy Pietroń, Wojciech Cybulski, Dorota Krasucka, Agata Mitura, Katarzyna Kos and Maja Antczak
A fast and reliable method of liquid chromatography and ultraviolet detection of sulfaguanidine, sulfadiazine, sulfamethazine, sulfamethizole, and sulfamethoxazole in feedingstuffs was described. The method involves THE procedure of preparation of spiked samples, and extraction of sulphonamides from the matrix using a mixture of methanol and acetonitrile, followed by drying the extract and dissolving it in a phosphate buffer. The analysis uses octadecyl (C18) analytical column with UV detection at λ = 260 nm and a gradient programme of mobile phase composition. The analytical procedure has been successfully adopted and validated for quantitative determination of the sulfonamides in feedingstuff samples. Validation included sensitivity, specificity, linearity, repeatability, and intra-laboratory reproducibility. The mean recovery of sulfonamides was 84%, within the working range of 200-2000 mg/kg. Direct, simple sample preparation and HPLC-UV analysis allow the method to be successfully included in the scope of routine analyses. The presented results could be an answer to a need of simple and easy method for sulfonamide determination applicable in medicated feedingstuffs analysis.