Identification of flunixin glucuronide and depletion of flunixin and its marker residue in bovine milk.

Residues of flunixin [and its marker residue 5-hydroxyflunixin (5OHFLU)] were determined in milk from cows that intravenously received therapeutic doses of the drug. The samples were collected during each milking (every 12 h) for six consecutive days, and concentrations of flunixin and its metabolites were determined by the method with and without enzymatic hydrolysis (beta-glucuronidase). The highest flunixin concentration in milk was observed 12 h after dosing (2.4 ± 1.42 µg/kg, mean ± SD). Flunixin concentrations in the samples determined with enzymatic hydrolysis were significantly higher (P < 0.05), which suggests the transfer of flunixin glucuronide to the milk. Additionally, unambiguous identification of flunixin glucuronide in the bovine milk was performed with linear ion-trap mass spectrometry. The 5OHFLU concentrations analyzed without enzymatic hydrolysis (22.3 ± 16.04 µg/kg) were similar to this obtained with enzymatic hydrolysis. Flunixin and 5OHFLU concentrations dropped below the limits of detection at 48 h after last dosing.

Etiological agents of dairy cows' mastitis in western part of Poland.

The purpose of the study was to determine the etiological mastitis agents in cows from herds located in the western part of Poland in years 2003-2005. Bacteriological examinations of 18,713 samples taken from California Mastitis Test (CMT) positive quarters were performed according to standard methods. Some Gram-negative bacilli and Gram-positive cocci were examined with the use of API tests (Analytical Profile Index). It was found that 32.7% of samples that were CMT positive were culture negative and 9.3% of samples were contaminated. Streptococcus species (15.7%), coagulase negative staphylococci (CNS) (14.6%), Staph. aureus (8.6%), Gram-negative bacilli (4.0%) and Corynebacterium species (3.8%) were most frequently isolated. Escherichia coli (52.3%) dominated among Gram-negative bacilli followed by Klebsiella pneumoniae (4.1%), Pseudomonas aeruginosa (3.6%), Enterobacter cloacae (3.6%), Serratia marcescens (3.1%), Pasteurella multocida (3.1%), Acinetobacter lwofii (3.1%), and 26 other bacteria species. Stahylococcus xylosus (31.5%), Staph. hyicus (12.2%), Staph. haemolyticus (12.2%), Staph. sciuri (11.6%), Staph. chromogenes (8.8%), Staph. epidermidis (8.3%) and Staph. simulans (6.1%) were the most frequent CNS. Streptococcus uberis (50%), Str. dysgalactiae (19.7%), Str. acidominimus (6.6%) and Enterococcus faecalis (5.3%) were mostly found among CAMP-negative streptococci. An increase in frequency of mastitis caused by non-agalactiae streptococci, Staph. aureus, A. pyogenes and yeast-like fungi and a decrease in that produced by Str. agalactiae in 2005 as compared to years 2003 or 2004 were observed.

Influence of various treatment methods on bacteriological findings in cows with puerperal endometritis.

The aim of the study was to identify the species of microorganisms isolated from the uterus of healthy cows (control group) and cows affected with puerperal metritis (PM) before and after an experimental therapy with an immunomodulator and antibiotics versus commonly applied methods (antibiotic + beta-blocker or antibiotic + PGF2alpha). Examinations were carried out on 110 cows with PM in three farms with similar system of rearing and nutrition. The control group consisted of 21 cows without postpartum disturbances. Smears from the uterus were taken before treatment and then at 21st day of observation. Escherichia coli and other species of Enterobacteriaceae family were isolated from 48.2% of PM cows and 47.6% of healthy cows. The degree of Arcanobacterium pyogenes infection was statistically lower in healthy than in sick cows (9.5% versus 30.0%). Streptococcus sp. was isolated from 13.6% of PM cows and from 16% of control ones. Staphylococci were isolated from 10% of PM and from 30% of control cows. Other bacteria species were isolated from about 10% of the examined cows. The best elimination of infections (66%) was noted in cows treated with the intrauterine antibiotic in combination with PGF2alpha i.m. injection. Examination showed that species of bacteria in the postparturient uterus were similar in healthy and sick cows. However, Arcanobacterium pyogenes was isolated 3 times more often from the sick animals. This pathogen was identified in 11 cows out of 19 (57.9%) culled subsequently because of infertility after the metritis puerperalis was clinically cured.

The effect of some drugs injection to pregnant heifers on blood antioxidant status.

The purpose of the trial was to establish the effect of the injection of the lysozyme dimer or vitamins connected with Se on the activity of chosen antioxidant enzymes and the total antioxidant status in pregnant heifers. Examinations were carried out during winter season in one farm on 21 heifers aged 22-24 months. Between the 21st and 14st day before expected parturition, seven heifers were once i.m. injected with antioxidants (Vitamin A-600 000 i.u.; Vitamin D3-200 000 i.u.; Vitamin E-1.5 mg/kg b.w., Selenium-0.022 mg/kg b.w.), and the next seven animals with lysozyme dimer (Lydium-KLP) at a dose of 0.02 mg/kg b.w. versus 7 non-treated control animals. Blood samples were taken before injection and then in hour 24 and 72 after injection, and between, the 7th and 14th day after calving. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSHpx), glutathione reductase (GSHred) and total antioxidant status (TAS) were measured by colorimetric method with the use of Randox kits. The mean value of SOD activity 21-14 days before expected calving was 704.8 +/- 294.6 U/ml of whole blood, GSHpx 59222 +/- 23699 U/l of whole blood, GSHred 110.8 +/- 22.5 U/l and TAS 0.33 +/- 0.15 mmol/l of serum. These indicators did not change in the control group with the exception of a statistically insignificant decrease in SOD activity after parturition. Statistically significant increase in blood SOD activity was noted only in the first day after injection of vitamins combined with selenium. These antioxidants also caused an insignificant increase in blood GSHpx activity in 72 hour following the injection, and in the second week after calving (statistically significant). The injection of antioxidants or lysozyme dimer did not change the activity of blood GSHred. However, an increase in the TAS was found in hour 24 (non significant) and 72 (statistically significant) following the single injection of lysozyme dimer.

Field trials on the prophylaxis of intramammary infections in pregnant heifers.

The study was carried out in 5 farms on 174 pregnant heifers. Clinical examination of the udder and bacteriological tests of quarter secretion were performed between the 8th and 3rd week before parturition, and then the animals were divided into a control group (64 heifers) and 3 experimental groups and immediately treated. A group of 32 experimental heifers was injected once with antioxidants (Vitamin A--600,000 i.u.; Vitamin D3--200,000 i.u.; Vitamin E--1.5 mg/kg b.w., Selenium--0.022 mg/kg b.w., i.m.). The next group (26 heads) was intramammary infused with antibiotic DC product (cloxacillin). Heifers from last experimental group (52) were injected with lysosyme dimer in a single dose of 0.02 mg/kg b.w. Clinical and bacteriological examinations were made during the first week after calving. The presence of bacteria was found in secretion of 22.6-38.9% udder quarters in 56.2-71.2% of pregnant heifers. The number of infected quarters (cows) did not change distinctly in the first week after calving except the lysozyme dimer group, where a decrease by 30% was noted. The percentage of quarters with elevated somatic cell count was higher in antibiotic DC group and closely similar in the other groups. None of examined methods showed an acceptable prophylactic effect. Clinical mastitis cases during first week after parturition were mostly caused by Escherichia coli, Staph. chromogenes, Staph. simulans, Staph. aureus, Staph. hyicus, Str. uberis, Str. acidominimus and Enterococcus faecalis.