Search Results (68)

Background/Objectives: Streptococcus uberis is a Gram-positive bacterium and a major cause of bovine mastitis. The use of antimicrobial treatments raises concerns about resistance. This study aimed to characterize S. uberis isolates from one of the ten largest milk-producing regions in Europe. Methods: Thirty-six isolates from 36 cows with mastitis were identified using MALDI-TOF and VITEK^®MS. Susceptibility to 9 antibiotics (penicillin G, ampicillin, tetracycline, erythromycin, clindamycin, cefotaxime, ceftriaxone, levofloxacin, and moxifloxacin) was determined with VITEK^®2. Whole-genome sequencing was performed using MinION Mk1C. Results: Alleles were identified for 7 loci: arcC, ddl, gki, recP, tdk, tpi, and yqiL. Only 10 isolates had alleles for all the loci. The loci with the highest number of alleles were ddl and tdk (33/36 strains), while arcC had the fewest (19/36). Four isolates were assigned to known sequence types (ST6, ST307, and ST184), and novel alleles were detected in 32 of the 36 isolates. Twelve isolates showed phenotypic resistance to one or more of the following antibiotics: tetracycline, erythromycin, clindamycin, and ceftriaxone. The lnu was the most frequently detected resistance gene (27 out of 102 total gene appearances). A total of 19 virulence factors were identified. All strains were predicted to be capable of infecting human hosts. Conclusions: Streptococcus uberis is a potential reservoir of antimicrobial resistance genes. The use of antimicrobials to treat bovine mastitis has reduced the susceptibility of this microorganism to several antibiotics, underscoring the importance of monitoring antimicrobial use in veterinary practice. The results also highlight the high genetic diversity of the isolates, suggesting a strong capacity to adapt to different environmental conditions. Full article

This cross-sectional study aimed to (a) determine the apparent prevalence of mastitis pathogens and (b) to identify risk factors for intramammary infections (IMIs) at the herd level in dairy herds in Bavaria, Germany. A stratified random sample of 305 herds was selected based on herd size, administrative district, and season. During the farm visits between July 2023 and July 2024, management data were recorded, quarter milk samples (QMSs) from 14,700 lactating cows were collected aseptically and analyzed, and the somatic cell count (SCC) at the quarter level was determined. Risk factors for the within-herd prevalence of Staphylococcus (S.) aureus, Streptococcus (Strep.) uberis, Strep. dysgalactiae, and non-aureus staphylococci (NAS) were analyzed by negative binomial regression, while risk factors for the presence of Escherichia (E.) coli and Strep. agalactiae IMIs on dairy farms were identified by logistic regression. The most frequently detected pathogens were NAS, found in 5.0% of all QMSs (n = 57,251), followed by Strep. uberis (1.9%) and S. aureus (1.8%), Strep. agalactiae (0.2%), and E. coli (0.1%). At the herd level, NAS, Strep. uberis, S. aureus, and Strep. dysgalactiae were found in 92%, 69%, 67%, and 57% of farms, respectively. Risk factors for increased within-herd prevalence included automated milking systems (NAS), organic production (Strep. uberis, S. aureus), straw bedding (Strep. uberis), and lack of bedding or mattress cubicles (Strep. dysgalactiae). The odds for a herd to be positive were increased with audible liner slips (E. coli) and the irregular cleaning of water troughs (Strep. agalactiae), and without a maintenance agreement for milking equipment (Strep. agalactiae). These results provide valuable insights into options for the targeted prevention of IMI. Full article

The rise in antimicrobial resistance and strict milk withdrawal regulations drive the search for safe, non-antibiotic intramammary therapies. This pilot field study focused on clinical parameters, including the somatic cell count (SCC) and the assessment of changes, as well as overall safety, which together enabled a prospective evaluation of whether the substance exerted any therapeutic effect. In this study, 48 Holstein–Friesian cows with naturally occurring clinical mastitis (somatic cell count > 400,000 cells/mL; single quarter) were randomized to receive either seven daily infusions of 10% pectin (n = 24) or two standard intramammary doses of a licensed multi-component antibiotic formulation (n = 24). The clinical severity scores (0–3) and SCC were monitored from 72 h before to 168 h after treatment initiation; the bacteriological cultures, milk TNF-α, milk yield, and blood hematology/biochemistry were also assessed. Both groups exhibited comparable and significant reductions in the mastitis scores and log₂-transformed SCC by 48 h post-treatment, with equivalent bacteriological cure rates and pathogen profiles (predominantly Streptococcus uberis, coagulase-negative staphylococci, and Escherichia coli) and no local irritation, systemic adverse effects, or alterations in the milk yield, TNF-α, or blood parameters. These findings indicate that intramammary pectin at a 10% concentration is safe and well tolerated and that it provides efficacy equivalent to standard antibiotic therapy, supporting its potential as an alternative mastitis treatment that avoids antibiotic residues and contributes to antimicrobial stewardship. Full article

The objective of this study was to evaluate the effects of feeding a combination of chicory–plantain silage and supplementing Se yeast on the response of early-lactating ewes to induce subclinical mastitis. Polypay ewes (n = 32) were fed either chicory–plantain silage or grass silage and supplemented with 3.6 mg Se yeast/ewe/day for approximately 2 months prior to the infusion of S. uberis into both mammary glands (i.e., intramammary infection or IMI). The ewes had a typical subclinical mastitis response with an 8-fold increase in milk somatic cell count within 24 h post-IMI, a decrease in milk yield, and changes in all milk components measured. The ewes experienced a mild systemic inflammation post-IMI as determined by an increase in rectal temperature and decrease in feed and water intake and, in blood, by an increase in the concentration of ceruloplasmin, haptoglobin, and myeloperoxidase and a decrease in paraoxonase, Zn, advanced oxidation protein products, and hematocrit with no effect on pro-inflammatory cytokines. No effect of silage type, likely due to a low concentration of secondary compounds, or Se supplementation was detected in response to IMI. In summary, the subclinical mastitis model used was effective in mounting an inflammatory response, although this was mild; however, feeding chicory–plantain silage with a low concentration of secondary compounds and supplementing Se yeast had no significant effect on the response of ewes to mammary infection. Full article

This study aimed to evaluate the antibacterial activity of several technical lignins against major environmental bacteria that cause mastitis in dairy cattle. The efficacy of four types of technical lignins against environmental mastitis pathogens was evaluated using MIC and MBC assays. The best candidate, sodium lignosulfonate (NaL-O), was further tested using sawdust bedding substrates. Substrates were prepared in different cleanliness conditions: sawdust only, sawdust plus urine, sawdust plus feces, or sawdust plus a combination of both. The antimicrobial activity of NaL-O against the mixture of environmental mastitis-causing pathogens was determined on days 0, 2, and 6 of incubation. In addition, the components of bedding substrates were analyzed to help understand the dynamics of pathogen loads. In the MIC and MBC assays, NaL-O showed the best antimicrobial performance against all pathogens except Escherichia coli. When testing in the bedding substrates, the addition of NaL-O decreased the concentration of Staphylococcus chromogenes, Streptococcus uberis, and Pseudomonas aeruginosa across all bedding cleanliness levels at d 0, 2, and 6 of incubation. As the incubation time increased, the antimicrobial effect decreased. NaL-O also lowered the counts of E. coli and Klebsiella pneumoniae across all incubation times, but to a lesser extent. The presence of feces significantly reduced the antibacterial effects of NaL-O for these two bacteria. Among the technical lignins tested, NaL-O showed the broadest antibacterial activity against the mastitis pathogens tested. This study suggests that NaL-O has promising potential as a bedding conditioner to control environmental pathogens on dairies due to its low cost, ready availability, and compatibility with sustainable livestock practices. Combined with bedding cleanliness, bedding conditioner application may play a crucial role in reducing the growth of EM pathogens and subsequent mastitis incidence. Full article

Introduction: Streptococcus uberis is a poorly controlled cause of bovine intramammary infections and a common motivation for the use antibiotics in dairy farms worldwide. Therefore, studying the genomic characteristics of this pathogen is fundamental to understand its complex epidemiology and behavior against antimicrobials. Methods: A comparative genomic analysis of 10 S. uberis strains was performed and their antimicrobial susceptibility was assessed. Results: Ten different novel sequence types were found, and genes (tetM, tetO, patB, lnuC, lnuA, lsaE, ermB, ANT(6)-la) and mobile genetic elements previously associated with antimicrobial resistance (repUS43, ISSag2, and ISEnfa4) and virulence (315.2 phage) were detected. Additionally, our strains had the highest relative rate of recombination to mutation (8.3) compared to other S. uberis strains isolated from different continents (America: 7.7, Asia: 2.9, Europe: 5.4, and Oceania: 6.6). Most of the strains (80%) tested showed phenotypic resistance to clindamycin and 70% exhibited intermediate susceptibility to penicillin. Conclusions: The high heterogeneity of strains observed and the presence of genetic factors linked to antimicrobial resistance represent a challenge for the implementation and surveillance of measures focused on the control and elimination of this pathogen. Full article

Background/Objectives: Bovine mastitis remains a challenge for the Irish dairy industry. This study aimed to explore the seasonality and antimicrobial resistance of mastitis pathogens obtained by the regional laboratories (RVL) of the Department of Agriculture, Food and the Marine. Methods: Seasonality of isolation of the most common bacterial species and antimicrobial resistance of those species repeatedly obtained in the same herds in different years were explored using the RVL diagnostic data. Additionally, whole genome sequencing (WGS) was employed to establish the persistency of Staphylococcus aureus strains within the same herd. Results: A clear seasonality was observed in the isolation of Staphylococcus aureus, Streptococcus uberis, and Escherichia coli from milk. Seasonal differences were statistically significant within and between bacterium. Persistence of resistance within herds was highest in S. aureus against penicillin (35.5% of herds) and in S. uberis against pirlimycin (14% of herds), while E. coli did not show persistence of resistance to any antimicrobials. Sequencing of S. aureus isolates revealed that the strains causing mastitis in ten out of twenty-one herds were similar genetically in different years. In seven of these herds, S. aureus was persistently resistant to penicillin. Isolates from two different herds were practically identical and carried the human immune evasion cluster genes (IEC, scn, sak, chp and sea) suggesting a recent human-bovine host switch event. Conclusions: These findings underscore the importance of implementing targeted biosecurity measures and monitoring programs to mitigate the spread of mastitis-causing pathogens and enhance antimicrobial stewardship in the Irish dairy industry, while it also highlights the significance of including a One Health perspective in surveillance programs. Full article

In this study, we sought to evaluate the prevalence of bacterial pathogens of mastitis in dairy cattle in the Inner Mongolia Autonomous Region, China. The study was conducted from 2015 to 2024 using a total of 12,053 clinical mastitis (CM) and sub-clinical mastitis (SCM) samples. The pathogens were isolated and identified by standard bacteriological and mycological methods. The most common pathogens isolated were Escherichia coli (13.82%), Staphylococcus aureus (10.28%), Klebsiella spp. (8.96%), Streptococcus agalactiae (7.45%), Streptococcus uberis (6.60%), coagulase-negative staphylococci (5.84%), and Streptococcus dysgalactiae (4.21%). From 2015 to 2017, the primary pathogens responsible for causing mastitis in cows were Staphylococcus aureus and Streptococcus agalactiae. In 2018, the most frequently isolated pathogen was Staphylococcus aureus. Notably, the isolation rate of Escherichia coli increased from 12.31% to 21.72%, and the isolation rate of Klebsiella spp. increased from 7.52% to 14.01% from 2019–2024. Mycoplasma was only detected in clinical mastitis cases, with a separation rate as high as 6.95%. In summary, the isolation rate of environmental pathogens is gradually increasing, while that of contagious pathogens has been continuously declining. This indicates that the current prevention strategies for infectious pathogens are effective. As a next step, it will be important to develop new strategies specifically targeting environmental pathogenic microorganisms. Full article

Despite the impact of mastitis on sheep production worldwide, the pathogenesis and host response to bacterial infection of the ovine mammary gland are poorly characterized. Studies in cattle highlight the significance of the mammary epithelium in pathogen recognition and the subsequent host response. The objective of this study was to assess bacterial adherence, invasion, and Toll like receptor (TLR) gene expression in primary sheep mammary epithelial cells (pMEC) following co-culture with the three principal mastitis pathogens of sheep, Mannheimia haemolytica, Staphylococcus aureus, and Streptococcus uberis. S. aureus was 140-fold more adherent than S. uberis and 850-fold more adherent than M. haemolytica. However, only S. aureus was internalized after 3 h of co-culture. TLR1, 2, 3, 4, 6, and 9 were shown to be constitutively transcribed by pMEC. M. haemolytica induced upregulation of transcription of TLR1, 2, 3, and 4. By contrast, S. uberis and S. aureus induced concentration-dependent transcription of TLR2 and TLR4 with a higher level of transcription in cells stimulated with the bacteria at a multiplicity of infection (MOI) of 200 compared to cells stimulated with a MOI of 20. These experiments define the range of TLR genes constitutively transcribed in sheep pMEC and show that bacterial infection has the capacity to regulate transcription in a species-specific and concentration-dependent manner. Full article

The objective of this study was to investigate the distribution of mastitis pathogens in quarter milk samples (QMSs) submitted to the laboratory of the Bavarian Animal Health Service (TGD) between 2014 and 2023 in general, in relation to the clinical status of the quarters, and to analyze seasonal differences in the detection risk. Each QMS sent to the TGD during this period was analyzed and tested using the California Mastitis Test (CMT). Depending on the result, QMSs were classified as CMT-negative, subclinical, or clinical if the milk character showed abnormalities. Mastitis pathogens were detected in 19% of the QMSs. Non-aureus staphylococci (NAS) were the most common species isolated from the culture positive samples (30%), followed by Staphylococcus (S.) aureus (19%), Streptococcus (Sc.) uberis (19%), and Sc. dysgalactiae (9%). In culture-positive QMSs from CMT-negative and subclinically affected quarters, the most frequently isolated pathogens were NAS (44% and 27%, respectively), followed by S. aureus (25% and 17%, respectively) and Sc. uberis (8% and 22%, respectively). In QMSs from clinically affected quarters, the most frequently isolated pathogens were Sc. uberis (32%), S. aureus (13%), Sc. dysgalactiae (11%), and Escherichia (E.) coli (11%). The distribution of NAS and Sc. uberis increased throughout the study period, while that of S. aureus decreased. From June to October, QMSs from subclinically affected quarters increased and environmental pathogens, such as Sc. uberis, were detected more frequently. In conclusion, this study highlights the dynamic nature of the distribution of mastitis pathogens, influenced by mastitis status and seasonal factors. Environmental pathogens still play an important role, especially in clinical mastitis and seasonal dependency, with the number of positive samples continuing to increase. It is therefore essential to continue mastitis control measures and to regularly monitor the spread of mastitis pathogens in order to track trends and adapt targeted prevention measures. Full article

The objectives of these studies were to identify associations between udder half defects (hard or lump) and bacteria isolated from milk or mammary tissue swabs, to compare with samples from normal udder halves at different physiological time points and to compare bacterial species isolated via milk and swabs of mammary tissue from within the same udder halves. A total of 1054 samples were aseptically collected from each udder half of 199 non-dairy breed (Romney) ewes from three different studies (Study A, n = 77; Study B, n = 74; and Study C, n = 48). Conventional bacterial culture and MALDI-ToF mass spectrometry were used for bacterial identification. Of the 225 samples from which bacteria were isolated, Mannheimia haemolytica and Streptococcus uberis were the dominantly identified species from defective udder halves, whereas coagulase-negative staphylococcus (CNS) species, mostly Staphylococcus simulans and Staphylococcus chromogenes, were more frequently isolated from normal udder halves. The ongoing presence of bacterial species over time was variable, although less frequently identified species showed less stability over time. A very high agreement (91.5%) of bacterial species identified was observed between the mammary tissue swab and udder half milk samples during post-weaning. In summary, palpable udder half defects were associated with bacterial positivity, and the ongoing presence of the bacteria over time was dependent on the species involved. Hence, culling ewes with palpable udder half defects that had more stable bacterial species could contribute to reducing the recurrence of palpable defects or mastitis. Full article

An LC–Orbitrap–HRMS method was developed and validated for the simultaneous determination of four tetracyclines—oxytetracycline (OTC), tetracycline (TC), doxycycline (DC), and chlortetracycline (CTC)—in milk. This method involves sample extraction with McIlvaine–EDTA buffer solution (pH 4) and solid-phase extraction (SPE) with Oasis HLB cartridges, followed by the evaporation of the extract and its reconstitution with a 14% methanol aqueous solution before injection into the instrumental system. This method has been validated in terms of linearity, sensitivity, selectivity, precision, and accuracy, in accordance with Commission Decision 2002/657/EC requirements. Compared to existing methods, this approach optimally combines a quantitative procedure for extracting analytes from the milk of different species, including sheep, bovines, and goats, with a very short LC–Orbitrap–HRMS instrumental analysis time (only 8 min), simultaneously ensuring high precision, sensitivity, and applicability as a rapid confirmation method in official food control laboratories. The proposed method was applied to determine the concentration levels of OTC in milk samples derived from healthy Sardinian sheep and those naturally infected with Streptococcus uberis, after the intramuscular administration of an antibiotic, in order to evaluate how much of the drug was “subtracted” during penetration from blood into milk, with a potential effect of reducing its therapeutic efficacy. Full article

This article focuses on the eco-friendly (green) synthesis of silver nanoparticles (AgNPs) and their incorporation into a polymer matrix. For AgNPs synthesis, Lavandula angustifolia (lavender) leaf extract was used as a reducing and stabilizing agent, and as a silver precursor, AgNO₃ solution with different concentrations of silver (50, 100, 250, and 500 mg/L) was used. Prepared AgNPs colloids were characterized using UV–vis spectrophotometry, transmission electron microscopy (TEM), and X-ray diffraction (XRD). The spherical morphology of AgNPs with an average size of 20 nm was confirmed across all samples. Further, the antimicrobial properties of the AgNPs were evaluated using the disk diffusion method on algae (Chlorella kessleri) and the well diffusion method on bacteria (Staphylococcus chromogenes, Staphylococcus aureus, and Streptococcus uberis), along with root growth inhibition tests on white mustard (Sinapis alba). Polymer composite (PVA–AgNPs) was prepared by incorporation of AgNPs into the polymer matrix. Subsequently, non-woven textiles and thin foils were prepared. The distribution of AgNPs within the nanocomposites was observed by scanning electron microscopy (SEM). Antibacterial properties of PVA–AgNPs composites were analyzed on bacteria Streptococcus uberis. It was found that not only AgNPs showed good antimicrobial properties, but toxic properties were also transferred to the PVA–AgNPs nanocomposite. Full article

Streptococcus uberis is a common bovine mastitis pathogen in dairy cattle. The rapid identification and characterization of antimicrobial resistance (AMR) in S. uberis plays an important role in its diagnosis, treatment, and prevention. In this study, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to identify S. uberis and screen for potential AMR biomarkers. Streptococcus uberis strains (n = 220) associated with bovine mastitis in northern Thailand were identified using the conventional microbiological methods and compared with the results obtained from MALDI-TOF MS. Streptococcus uberis isolates were also examined for antimicrobial susceptibility using a microdilution method. Principal component analysis (PCA) and the Mann–Whitney U test were used to analyze the MALDI-TOF mass spectrum of S. uberis and determine the difference between antimicrobial-resistant and -susceptible strains. Using MALDI-TOF MS, 73.18% (161/220) of the sampled isolates were identified as S. uberis, which conformed to the identifications obtained using conventional microbiological methods and PCR. Using PCR, antimicrobial-resistant strains could not be distinguished from antimicrobial-susceptible strains for all three antimicrobial agents, i.e., tetracycline, ceftiofur, and erythromycin. The detection of spectral peaks at 7531.20 m/z and 6804.74 m/z was statistically different between tetracycline- and erythromycin-resistant and susceptible strains, respectively. This study demonstrates a proteomic approach for the diagnosis of bovine mastitis and potentially for the surveillance of AMR among bovine mastitis pathogens. Full article