The incidence of milk leakage (ML) after dry-off (DO) and related risk factors was studied in 1,175 dairy cows from 41 commercial herds in 8 European countries: Belgium, Czech Republic, Denmark, France, Germany, Italy, the Netherlands, and Spain. Milk leakage was assessed twice for 30 s each during 3 visits at 20 to 24 h, 30 to 34 h, and 48 to 52 h after DO. Information related to dry-cow management and udder health was collected at herd and cow level, including individual somatic cell count (ISCC) from test-day controls and occurrence of clinical mastitis cases from DO until 30 d in lactation. Mixed-effect logistic regression analyses were used to identify possible risk factors for ML and to study the association between ML and new intramammary infections. Intramammary infections were defined as clinical mastitis cases during the dry period and in the first 30 d in lactation or a rise in ISCC from before to after the dry period (threshold: 200,000 cells/mL) or both. Milk leakage was observed in 24.5% of the cows between 20 and 52 h after DO, where the herd incidence varied between 0.0 and 77.8%. The reduction in number of milkings in the weeks before DO had statistically significant effect on the ML incidence. When the milking frequency was reduced from 3 times/d to 2 or maintained at twice a day, cows had 11 (95% CI = 3.43-35.46) or 9 (95% CI = 1.85-48.22) times higher odds of leaking milk, respectively, compared with cows where the milking frequency was reduced from twice to once a day. Also, the milk production 24 h before DO was associated with ML incidence. Hence, cows with a milk production between 13 and 21 L or above 21 L had 2.3 (95% CI = 1.48-3.53) and 3.1 (95% CI = 1.79-5.3) times higher odds of leaking milk, respectively, compared with cows with a milk production below 13 L. A higher ML incidence was present in the group of cows with an average ISCC in the last 3 mo before DO ≥200,000 cells/mL (odds ratio = 1.7; 95% CI = 1.13-2.41) compared with cows with an average ISCC <100,000 cells/mL. Quarters with ML tended to have 2.0 times higher odds of developing clinical mastitis compared with quarters not leaking milk. Cows with ML tended to have 1.5 times higher odds of intramammary infections (i.e., an increase of ISCC or clinical mastitis) compared with cows without ML.

• Klíčová slova
• clinical mastitis, dry-off, intramammary infection, milk leakage, risk factor,
• MeSH
• incidence MeSH
• laktace MeSH
• mastitida skotu epidemiologie   patofyziologie   MeSH
• mléčné žlázy zvířat patofyziologie   MeSH
• mlékárenství * MeSH
• mléko cytologie   MeSH
• nemoci skotu epidemiologie   patofyziologie   MeSH
• počet buněk veterinární   MeSH
• rizikové faktory MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

The goal of the present study was to develop a new intramammary combination preparation for veterinary use based on silver citrate, arginine and dexpanthenol for the treatment and prophylaxis of subclinical mastitis in cattle. When creating combined medicines for veterinary use, attention must be paid to the factors that influence their stability in a combined presence, and the composition of drugs, as well as the technology for their preparation should be determined. In the current study, a qualitative and quantitative control of model mixtures of active ingredients, as well as of a silver citrate-based combined preparation was carried out. The following parameters that define intramammary drug stability were studied: clarity, pH, and the quantitative content of the active ingredient. Based on the experimental studies, optimal pH limits for a new preparation were determined ranging from 6.4 to 6.7. Considering the instability of solutions based on silver citrate, studies on the stabilization of the preparation were conducted at the obtained pH values. As the result, we used povidone, which possessed the stabilizing properties. The amount of arginine that provides the optimum pH level for the selected combination of active ingredients was also determined. In conclusion, the optimal composition of an intramammary preparation for veterinary medicine in 10 ml ampoules with a shelf life of 6 months was developed.Key words: silver citrate arginine dexpanthenol intramammary dosage form stability.

• MeSH
• arginin terapeutické užití   MeSH
• citráty terapeutické užití   MeSH
• kyselina pantothenová analogy a deriváty   terapeutické užití   MeSH
• mastitida skotu farmakoterapie   MeSH
• skot MeSH
• stabilita léku MeSH
• stříbro terapeutické užití   MeSH
• veterinární léky terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• arginin MeSH
• citráty MeSH
• dexpanthenol MeSH Prohlížeč
• kyselina pantothenová MeSH
• stříbro MeSH
• veterinární léky MeSH

A sterile polyethylene device was introduced in the milk cistern of two mammary quarters of forty dairy cows. The cows were divided into three groups according to the length of exposure of the device in the cistern (3 days, 14 days, 365 days). The somatic cell counts were studied for a year in the first 10 ml fraction of milk and an increase in the somatic cell counts was found in the quarters having the intramammary device, as compared with the control quarters (having no devices). This difference was marked during lactation and prior to the onset of drying off. For the reduction in the frequency of occurrence of new natural intramammary infections, the activity of the device against S. aureus was limited and no activity against S. agalactiae was proved. The proportions of polymorphonuclears, round-cell elements and macrophages were histologically studied and compared for the udder quarters with and without the intramammary body in correlation with the time of exposure of the device to the milk cistern milieu. The most marked differences in favour of the udder quarters with the intramammary device were recorded in the alveoli containing more cells with a significant proportion of polymorphonuclear leucocytes. Small differences were found in the interstitial and subepithelial zone of the milk cistern. The activity of acid and alkaline phosphatase and adenosine triphosphatase was histochemically determined in the tissue structures of the udder and was found not to change under the influence of the device. Leucocytes and macrophages adhering to the surface of the body were observed under scanning electron microscope.

• MeSH
• buněčná imunita MeSH
• imunoterapie přístrojové vybavení   metody   MeSH
• mastitida skotu imunologie   prevence a kontrola   MeSH
• mléčné žlázy zvířat MeSH
• mléko cytologie   mikrobiologie   MeSH
• polyethyleny MeSH
• přirozená imunita MeSH
• protézy a implantáty MeSH
• skot MeSH
• stafylokokové infekce prevence a kontrola   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• polyethyleny MeSH

BACKGROUND: Bovine mastitis is one of the main causes of reduced production in dairy cows. The infection of the mammary gland is mainly caused by the bacterium Staphylococcus aureus, whose resistant strains make the treatment of mastitis with conventional antibiotics very difficult and result in high losses. Therefore, it is important to develop novel therapeutic agents to overcome the resistance of mastitis-causing strains. In this study, novel selenium-tellurium based nanoparticles (SeTeNPs) were synthesized and characterized. Their antibacterial activity and biocompatibility were evaluated both in vitro and in vivo using a bovine model. A total of 10 heifers were divided into experimental and control groups (5 animals each). After intramammary infection with methicillin resistant S. aureus (MRSA) and the development of clinical signs of mastitis, a dose of SeTeNPs was administered to all quarters in the experimental group. RESULTS: Based on in vitro tests, the concentration of 149.70 mg/L and 263.95 mg/L of Se and Te, respectively, was used for application into the mammary gland. Three days after SeTeNPs administration, MRSA counts in the experimental group showed a significant reduction (P < 0.01) compared to the control group. The inhibitory effect observed within the in vitro experiments was thus confirmed, resulting in the suppression of infection in animals. Moreover, the superior biocompatibility of SeTeNPs in the organism was demonstrated, as the nanoparticles did not significantly alter the inflammatory response or histopathology at the site of application, i.e., mammary gland, compared to the control group (P > 0.05). Additionally, the metabolic profile of the blood plasma as well as the histology of the main organs remained unaffected, indicating that the nanoparticles had no adverse effects on the organism. CONCLUSIONS: Our findings suggest that SeTeNPs can be used as a promising treatment for bovine mastitis in the presence of resistant bacteria. However, the current study is limited by its small sample size, making it primarily a proof of the concept for the efficacy of intramammary-applied SeTeNPs. Therefore, further research with a larger sample size is needed to validate these results.

• Klíčová slova
• Antibacterial, Biocompatibility, Heifer, Intramammary, MRSA, Mammary gland, Nanomaterial, Resistance, SeTe,
• Publikační typ
• časopisecké články MeSH

Streptococcus uberis is one of the most important mastitis-causing pathogens. Although the pathogenesis and virulence factors required for the intramammary infection development are not yet well established, several putative virulence-associated genes have been described. This work aimed to investigate the presence of ten known and putative virulence-associated genes in S. uberis isolated from subclinical or clinical mastitis and its closely related species Streptococcus parauberis in 135 dairy farms in the Czech Republic. The PCR analysis detected that all the examined isolates possessed at least four virulence genes and most isolates carried eight out of ten virulence genes. All S. uberis isolates were positive for the oppF, gapC and sua genes. Among the most prevalent virulence-associated genes skc (98%) and pauA (97%) were also found. The hasA and hasB genes were always present together in 94% of the isolates. The genes cfu and lbp were detected in 6% and 2%, respectively. In the S. uberis isolates, 14 different virulence gene profiles were observed. The most frequent profile was hasA + hasB + sua + skc + pauA + gapC + oppF with variable hasC, observed in 86% of the tested isolates, occurring in 127 out of 135 farms. S. parauberis was identified very sporadically and, although it is closely related to S. uberis, only a rare occurrence of the examined virulence-associated genes was found.

• Klíčová slova
• cows, intramammary infections, mammary gland, pathogens, virulence factors,
• Publikační typ
• časopisecké články MeSH

The ubiquitous occurrence and high heterogeneity of Streptococcus uberis strains cause difficulties in the development and implementation of effective control strategies in dairy herds. In this study, S. uberis strains from 74 farms, obtained predominantly from subclinical, acute, and chronic recurrent mastitis, as well as from udder surface swabs and milk from healthy udders, were analysed for their genetic diversity using multilocus sequence typing (MLST). Isolates were tested for the presence of the genes encoding the virulence factors using polymerase chain reaction. Antibiotic susceptibility testing was performed using a microdilution assay including 14 antimicrobials. The virulence profiles and antimicrobial (AMR) profiles of the isolates were assembled and the overall heterogeneity was evaluated. Among the 124 isolates, 89 MLST genotypes, 7 different virulence profiles, and 12 AMR profiles were identified. The large number of different MLST allelic profiles in this study points to the high heterogeneity of strains in dairy herds in the Czech Republic. Isolates of a certain MLST genotype may possess a different set of virulence factor genes. We detected up to three different resistance profiles within a single MLST genotype. The results of our study showed that fully susceptible isolates coexisted with resistant or even multiresistant isolates in the same herd. Multiple genotypes within a herd were detected on many farms (up to seven MLST genotypes and four AMR profiles in one herd). This heterogenic population structure might suggest that environmental transmission is the predominant route of infection in herds in the Czech Republic.

• Klíčová slova
• MLST, antimicrobial resistance, cows, intramammary infection, virulence factors,
• Publikační typ
• časopisecké články MeSH

Streptococcus uberis is one of the primary causative agents of mastitis, a clinically and economically significant disease that affects dairy cattle worldwide. In this study, we analyzed 140 S. uberis strains isolated from mastitis milk samples collected from 74 cow herds in the Czech Republic. We employed whole-genome sequencing to screen for the presence of antimicrobial resistance (AMR) genes and genes encoding virulence factors, and to assess their genetic relationships. Our analysis revealed the presence of 88 different sequence types (STs), with 41% of the isolates assigned to global clonal complexes (GCCs), the majority of which were affiliated with GCC5. The STs identified were distributed across the major phylogenetic branches of all currently known STs. We identified fifty-one putative virulence factor genes, and the majority of isolates carried between 27 and 29 of these genes. A tendency of virulence factors and AMR genes to cluster with specific STs was observed, although such clustering was not evident within GCCs. Principal component analysis did not reveal significant diversity among isolates when grouped by GCC or ST prevalence. The substantial genomic diversity and the wide array of virulence factors found in S. uberis strains present a challenge for the implementation of effective anti-mastitis measures.

• Klíčová slova
• MLST, antimicrobial resistance genes, intramammary infection, mastitis, phylogenetic tree, sequence type, virulence genes,
• Publikační typ
• časopisecké články MeSH

Streptococcus uberis is one of the most important causative agents of mastitis and is a common reason for the use of antimicrobials in dairy cows. In this study, we assessed the antimicrobial susceptibility of 667 S. uberis isolates originating from 216 Czech dairy farms collected between 2019 and 2023 using the broth microdilution method. We tested 140 of the isolates for the presence of antimicrobial genes using whole-genome sequencing and evaluated their relationship with phenotypic resistance. Streptococcus uberis isolates showed high levels of resistance to tetracycline (59%), followed by streptomycin (38%) and clindamycin (29%). Although all of the isolates were susceptible to beta-lactams, a relatively high percentage of intermediately susceptible isolates was recorded for ampicillin (44%) and penicillin (18%). The isolates were mainly resistant to tetracycline alone (31.3%); the second most frequent occurrence of the phenotypic profile was simultaneous resistance to tetracycline, streptomycin, and clindamycin (16.6%). The occurrence of antibiotic resistance genes did not always match the phenotypic results; in total, 36.8% of isolates that possessed the ant(6)-Ia gene did not show phenotypic resistance to streptomycin. To a lesser extent, silent genes were also detected in clindamycin and tetracycline. This study confirmed the high susceptibility of S. uberis to penicillins used as first-line antimicrobials for S. uberis mastitis treatment.

• Klíčová slova
• Streptococcus uberis, antimicrobial resistance gene, antimicrobial resistence, bovine mastitis, intramammary infection, whole-genome sequencing,
• Publikační typ
• časopisecké články MeSH

During two years in a herd of dairy cows with an average number of 94 cows the effects on infection level were investigated of single nonselective antibiotics treatment of cow's udder in the dry period. The treatment influenced favourably the results of complex periodic three-months examinations: --level of infection with infectious mastitis was reduced from 38.3% to 9.8%; --occurrence of clinical forms of mastitis dropped from 17.4% to 5.9%;--occurrence of NK-test positive reactions decreased from 55.8% to 21.8%. An investigation into therapeutic efficiency of three intramammary preparations revealed the highest total therapeutic effect of Stapenor retard (Bayer, FRG)--93.2%; Syntarpene 500 (Polfa, Poland) had the total therapeutic efficiency of 92.6% and the efficiency of Oxaclene foam (Spofa, CSSR) made 86.5%. The results demonstrate good efficiency of the method because the levels of mastitis infection in the herd were positively influenced, and excellent therapeutic efficiency of semisynthetic penicillins containing intramammary preparations used for mastitis treatment of dairy cows in the dry period.

• MeSH
• bakteriální infekce farmakoterapie   veterinární   MeSH
• mastitida skotu farmakoterapie   MeSH
• oxacilin terapeutické užití   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• oxacilin MeSH

The objective of this study was to determine the oxytetracycline residues in milk from cows with clinical mastitis dosed with two extra-label routes of oxytetracycline administration not only during antibiotic's treatment (5 days), but also six days after treatment by use of a liquid chromatography method of testing with a detection limit of 20 ppb. Both groups of animals were treated once daily for five milkings at 24-hour intervals following morning milkings. Composite milk samples (equal volumes of foremilk from each quarter) were collected during morning and afternoon milkings, mixed together (1:1), and stored until analyzed. Milk samples were analyzed just before the first treatment (0 hour) and ten times at 24-hour intervals. Residue studies in milk cows indicate that oxytetracycline passes into milk. Residues in milk were higher for the cows receiving oxytetracycline by intramammary route (Tab. I) than for the cows receiving oxytetracycline intramuscularly (Tab. II). The highest mean data were 195.68 mg/kg after intramammary infusion (Fig. 2) and 2.74 mg/kg after intramuscular injection (Fig. 3) on the 5th day of the treatment beginning. The analysis data showed that oxytetracycline persisted in milk for as long as two days after both treatments at the concentration 0.03 mg/kg versus 0.02 mg/kg, respectively. No residues were detected in milk of any animal from the 4th day of the cessation of the therapy (Fig. 1) as detected by the HPLC method.

• MeSH
• antibakteriální látky analýza   terapeutické užití   MeSH
• laktace metabolismus   MeSH
• mastitida skotu farmakoterapie   metabolismus   MeSH
• mléko chemie   MeSH
• oxytetracyklin analýza   terapeutické užití   MeSH
• rezidua léčiv analýza   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• oxytetracyklin MeSH