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Open access

Artur Burmańczuk, Zbigniew Roliński, Cezary Kowalski and Rafał Zań


Introduction: The objective of this study was to describe a pharmacokinetic–pharmacodynamic (PK/PD) approach for determination of a rational dosage of ampicillin (AMP) and depletion of the antibiotic residues in milk after intramammary administration to cows.

Material and Methods: The cows came from different farms from the Lublin Province area. They (n = 9) received 5 g of the drug, containing 75 mg of AMP sodium in physiological solution, through a syringe tube by intramammary administration. Following single intramammary administration, the milk samples (5 mL) were collected after 2, 4, 6, 8, 10, 24, 36, 48, and 60 h. The liquid chromatography-mass spectrometry analysis was performed on the Agilent 1200 system connected to an AB Sciex API 4000™ mass spectrometer. The pharmacokinetic analysis of the concentrations of the antibiotic in milk was performed using software Phoenix® WinNonlin® 6.4. Calculations were made in non-compartmental (slopes, highest, amounts, and moments) and compartmental analysis.

Results: The pharmacokinetic characteristics of AMP after intramammary administration indicate rapid elimination of the drug from milk. The mean residence time had a several-fold lower value than the designated elimination half-life and amounts to only 3.4 h. The concentration of the drug in the milk dropped relatively quickly and the process was very dynamic.

Conclusion: The conducted research confirms the rationale of using the PK/PD model in order to verify the dosing regimen for other antibiotic groups and various indicators of the applied PK/PD model.

Open access

Artur Burmańczuk, Grabowski Tomasz, Małgorzata Gbylik-Sikorska, Anna Gajda and Cezary Kowalski


Introduction: There are many veterinary products containing β-lactam antibiotics which are used for mastitis treatment in cows. The aim of the study was to determine whether mastitis could have any effect on amoxicillin (AMX) or penicillin G procaine (PEN) withdrawal period from milk, in the context of current maximum residue limits established by the European Commission.

Material and Methods: The study was conducted on 17 dairy Black and White cows with clinical mastitis during the lactation period. The first group (n = 8) received 200 mg of amoxicillin (AMX), whereas the second group (n = 9) received 200,000 IU/mg of penicillin G procaine (PEN) by intramammary administration. For the measurement of AMX and PEN concentrations in milk, the liquid chromatography tandem mass spectrometry method was applied. Pharmacokinetic calculations were performed using Phoenix WinNonlin 6.4 software.

Results: The determined AMX and PEN half-life values in the mammary gland suggest that the drug withdrawal is at a level of 99.9% within 81 h (≈3.5 days) and 116 h (≈5 days) after administration of AMX and PEN, respectively. The present research indicates that, at 60 h after administration, the average PEN concentration in the milk from cows with clinical signs of mastitis may still reach 4.96 g/kg and that of AMX can even be 6.92 g/kg.

Conclusion: The results obtained confirm that, in mastitis cases, a 72-h withdrawal period is sufficient for elimination of AMX to a lower level than the established maximum residue limit (MRL) values. However, in the case of PEN, at 69 h after administration, the drug concentration may be close to that of the determined MRL.

Open access

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.

Open access

A. Burmańczuk, T. Grabowski, T. Błądek, C. Kowalski and P. Dębiak


The aim of the study was to carry out retrospective and prospective comparative analyses of the pharmacokinetics of CEF after single intramammary (IMM) administration in cows. The prospective study (study A) was conducted on 9 dairy cows of the Polish Black-White race with clinical mastitis during the lactation period. Milk samples were collected at 2, 4, 6, 8, 10, 24, 36, 48, 72 and 84 h after single IMM administration of 250 mg of CEF to one quarter. Drug concentrations in milk samples were determined by HPLC-MS/MS technique and the results of the pharmacokinetic analysis were compared to those obtained in previous studies based on the microbiological (study B) and HPLC-UV methods (study C and D). Pharmacokinetic parameters were calculated based on adapted two-compartment model of drug distribution. One of the findings of the comparison of the analysed investigations is that the CEF kinetics determined with the microbiological method is consistent with the results obtained by the authors of this paper. Both studies yielded similar results of the key pharmacokinetic parameters related to the level of the drug distribution to tissues and elimination half-life. In the pharmacodynamic analysis, the observations in all four studies were entirely consistent and have shown lower values of T>MIC90 in healthy animals and significantly higher values in infected dairy cows. The comparison of studies A, B, C, and D revealed that the time of complete CEF wash-out of 90.90% varied and amounted to 5.7, 8.0, 2.2, and 2.2 days after administration of the drug, respectively.

It was confirmed that not only the type of the analytical method but also correct sampling have a significant impact on determination of the correct value of the drug half-life after IMM administration. The comparative analysis of studies in which the milk yield was high and low allows a conclusion that this parameter in the case of CEF has no significant effect on T>MIC90.

Open access

Małgorzata Gbylik-Sikorska, Anna Gajda, Artur Burmańczuk, Tomasz Grabowski and Andrzej Posyniak


Introduction: Quercetin is a polyphenolic flavonoid which has been used in traditional Chinese medicine as a natural therapeutic agent with a broad spectrum of activities (antioxidant, anticancer, neuroprotective, anti-inflammatory, antiviral and antibacterial). The aim of this study was to develop and validate a rapid and simple ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method for the determination of quercetin in milk.

Material and Methods: Sample preparation was based on a liquid-liquid extraction with 0.5% formic acid in acetonitrile. The chromatographic separation was performed on a ZORBAX SB-C18 column with methanol and 0.5% formic acid as a mobile phase.

Results: The procedure was successfully validated. The mean recovery of the analyte was 98%, with the corresponding intra- and inter-day variation less than 10% and 15%, respectively, and the repeatability and reproducibility were in the range of 3%–7.2% and 6.1%–12%, respectively. The lowest level of quantification was 1.0 μg/kg.

Conclusion: The proposed method was successfully applied in evaluating the pharmacokinetics of quercetin in milk obtained from dairy cows with clinical mastitis after intramammary administration.

Open access

Renata Klebaniuk, Ewa Tomaszewska, Piotr Dobrowolski, Małgorzata Kwiecień, Artur Burmańczuk, Dmytro Yanovych, Zvenyslava Zasadna, Sylwia Szymańczyk, Natalia Burmańczuk and Siemowit Muszyński


An effect of the exposure to chloramphenicol (CAP) at doses used therapeutically was studied in pigs at the age of slaughter. Pigs were treated with CAP intramuscularly (20 mg/kg b.w. two times every 24 hours). Histomorphometrical and immunohistochemical analyses of small intestine and liver were done. CAP increased the thickness of myenteron and submucosa, and the length of villi; decreased the depth of crypts in the duodenum and jejunum. CAP influenced the Auerbach plexus. A decrease in cell proliferation, an increase in the number of apoptotic cells and T lymphocytes in the CAP-treated pigs were observed. CAP induces hepatotoxicity, neurotoxicity and disturbed intestinal epithelium. It can be concluded that short exposure of pigs to CAP at doses used therapeutically results in disturbed digestion and absorption process in the intestine.