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The objective of this cross-sectional study was to determine the minimal inhibitory concentrations of cephalosporins of the first (cefalonium and cefapirin) and fourth generation (cefquinome) against bacteria isolated from intramammary infections in dairy cows with elevated somatic cell counts in Germany. Additionally, possible regional differences of the minimal inhibitory concentrations within Germany should be evaluated. In total, 6936 quarter milk samples from cows with a somatic cell count >200,000 cells/ml were taken in 43 herds. The concentrations of the first generation cephalosporins inhibiting at least 90% of the isolates of a pathogen (MIC 90 ) were ≥64 μg/ml against Gram-negative bacteria and enterococci whereas the respective MIC 90 against the other Gram-positive bacteria were ≤4 μg/ml. The MIC 90 of cefquinome were ≥16 μg/ml against Gram-negative bacteria, bacilli and enterococci, and ≤2 μg/ml against the other Gram-positive bacteria. Only the minimal inhibitory concentrations against coagulase-negative staphylococci differed significantly between regions in parametric survival models with shared frailties for the herds. However, the minimal inhibitory concentrations of cefquinome against staphylococci were higher than the minimal inhibitory concentrations of the tested cephalosporins of the first generation. Therefore, cefquinome should not be the first choice to treat staphylococcal mastitis in dairy cows. [ABSTRACT FROM AUTHOR]

To comply with antibiotic restriction policies in the European Union, internal teat sealants (TS) are increasingly used at dry off (DO) in selective dry cow treatment protocols to maintain udder health. Postcalving TS residue attachment to milking equipment and associated cleaning difficulties is a reason some farmers stay away from blanket TS use. Our objective was therefore to improve insight into TS excretion visibility and to compare quantity, pattern, and presence versus absence of TS excretion postcalving between the typical 2 cow categories at DO: high (H)- and low (L)-SCC cows, treated with antibiotic (AB) plus TS (H-ABTS) or TS only (L-TS), respectively. In herds in the Netherlands (n = 3), and Germany (n = 4), cows were enrolled at DO, and categorized as H-ABTS (n = 93), or L-TS (n = 99). Postcalving, quarter-level TS visibility, quantities, patterns, and percentage of TS infused and excreted postcalving were recorded from 50 mL of premilk of every quarter at each of the first 15 or 16 milkings. Udder quarter health status was determined by bacteriological culture and SCC of quarter milk samples taken at DO and at d 3 postcalving and by clinical mastitis incidence from DO until 30 DIM. Univariable and multivariable models were created to explore associations of TS excretion presence versus absence at the first 3 milkings. Irrespective of SCC category, both laboratory personnel and farmers saw TS residues at the first milking in an equal 72% of quarters. Compared with laboratory as the gold standard, farmer sensitivity to spotting TS in premilk was 74.5% at the first milking and decreased to a maximum of 8.3% at the last 3 milkings. At the first milking, TS excretion quantities showed a bimodal distribution pattern and the mean percentage of TS infused (3.83 g) that was excreted in premilk at the first milking, was higher in the L-TS cows (45.5%) compared with the H-ABTS cows (32%). At the second and third milking, mean-adjusted TS percentage excreted was higher in the H-ABTS cows (8.5% and 1.8%, respectively) compared with the L-TS cows (4.6% and 0.4%, respectively). The multivariable model of the first 3 milkings showed parity at both the first and second milking, and the study group at both the second and third milking was significantly associated with TS presence. The univariable model showed no association between TS presence at the first milking and udder health. In conclusion, in premilk of the first milking, TS residue excretion was bimodal, higher in L-TS cows, more likely to be present in multiparous cows, and not associated with udder health. At the second and third milking, excretion was higher in H-ABTS cows and TS presence was only more likely in multiparous cows at the second milking., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Mastitis-causing pathogens are shed from infected mammary gland quarters and thus contribute to an increased risk of new intramammary infections. The objective of the current study was to investigate the shedding characteristics of various mastitis-causing pathogens and associated animal-specific (somatic cell score and parity) and environmental (heat stress) factors. In a longitudinal study, infected udder quarters were sampled consecutively on 5 dairy farms in Germany. To capture climatic factors, temperature-humidity index (THI) was calculated. In the laboratory analysis, the pathogens and their counts in the milk samples were determined. A generalized linear mixed model with gamma link was used to evaluate the factors influencing pathogen-shedding characteristics. The variables somatic cell count, pathogen, parity, and THI had significant influence on pathogen shedding. Staphylococci were shed in lower values than streptococci. The pathogen shedding from mammary gland quarters with intramammary infections was higher in the first and second lactation than in higher lactations. Exceeding the THI threshold 60 resulted in higher pathogen counts on the same day. This was only caused by the pathogens yeasts and Streptococcus uberis. Possible mechanisms causing differences in pathogen shedding are changes in the counts due to influenced milk quantities, better growth conditions at higher temperatures, or altered immunological reactions. The mechanisms often remain speculative and require further investigation. The study underlines the contribution of cows with high somatic cell counts regarding the transmission of mastitis pathogens within a herd. Furthermore, it becomes clear that heat stress in Germany influences udder health and that prevention measures are useful., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Recurrent clinical mastitis contributes to around half of all infections having an economic impact in the dairy industry. It leads to milk yield reduction, increased risk of mortality, and culling, and may be caused by new infections or a persistent infection after previous treatment. Disease management is dependent on the infecting species, necessitating accurate identification of the pathogen in the range of persistent and reinfection cases among recurrent infections using culture and molecular biological analysis. Milk samples from diagnosed clinical mastitis cases were collected from three Northern German dairy farms between 2011 and 2015. Totally, 2043 diagnosed mastitis cases were examined at quarter level (1598 (78.2 %) first and 445 (21.8 %) recurrent mastitis cases in lactation). Among the recurrent cases, 145 (32.6 %) cases were confirmed to harbor the same pathogenic species as previous infections. RAPD PCR confirmed the same species strain in 49 (11 %) of the recurrent infections. The contribution of new infections as compared to persistent infections in cases of clinical mastitis is clear from the data. Future studies in recurrent clinical mastitis control should be focused on influencing factors to prevent new infections in addition to therapeutic intervention and bacteriological cure., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Staphylococcus aureus is one of the most common pathogens associated with bovine mastitis in Germany and Denmark. Successful therapy is strongly linked to the susceptibility of the pathogen to the administered antimicrobial. An increase in resistant pathogens in human and veterinary medicine has become a concern worldwide and hampers therapy due to reduced susceptibility. In the present study, susceptibility testing was performed for 85 and 93 S. aureus isolates originating from mastitis cases on 12 German and 8 Danish dairy farms, respectively. Phenotypic examination was performed by detection of minimal inhibitory concentration (MIC) values using the broth microdilution method, followed by genotypic investigations of the blaZ and mecA resistance genes via PCR. The tested antimicrobials were the most frequently used β-lactams in German and Danish dairy farms, including cefquinome, cefoperazone, cephapirin, penicillin, oxacillin, cloxacillin, amoxicillin-clavulanic acid, and cephalexin-kanamycin. Special attention was paid to varying therapy concepts because, in Germany, third- and fourth-generation cephalosporins have been predominantly used in mastitis therapy, whereas in Denmark, restrictive use of penicillin is followed by a general avoidance of cephalosporins. Differences in MIC values were mainly based on determined MIC 90 values (MIC at which 90% of isolates are inhibited). In general, Danish S. aureus isolates were inhibited at comparatively lower MIC 90 values than S. aureus isolated from German dairy farms for most β-lactams. No differences were observed regarding cefquinome, because both German and Danish isolates exhibited MIC 50 and MIC 90 values of 0.5 and 1 µg/mL, respectively. In contrast, the MIC 90 for penicillin against German and Danish S. aureus were 0.5 and ≤0.06 µg/mL, respectively. Resistance genes (blaZ, mecA) were only detected in German S. aureus isolates on 3 dairy farms in Germany. A total of 5 isolates tested positive for both blaZ and mecA, whereas 1 isolate carried the blaZ resistance gene only. A direct correlation between frequently used antimicrobials and reduced susceptibility could not be determined based on results of the present study. In addition to further research to determine factors associated with resistance development, we emphasize the urgent need for internationally standardized clinical breakpoints to assess resistance situations more accurately., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Non-S. aureus staphylococci (NAS) are the most frequently isolated pathogens from bovine milk and can cause intramammary infections (IMI). They can also be found in teat canals, on bovine skin and in cows' environment, which may lead to unnoticed contamination of milk samples. The aim of this study was to investigate the role of NAS species as mastitis-causing pathogens or contaminants, and to identify possible differences between NAS species. A longitudinal study was conducted with consecutive milk sampling in five German dairy herds. Species identification was performed using matrix-assisted laser desorption ionization-time of flight mass spectrometry. Infections were distinguished from contaminations using two different definitions based on the repeated detection of an NAS species. Of 15 NAS species found, eight and ten, respectively, were associated with an IMI. Staphylococcus simulans and S. chromogenes were associated with IMI in more than 90 % of the findings. S. warneri, S. xylosus, S. microti, S. haemolyticus, and S. succinus seem to be frequent causes of IMI as well as contaminants. If a species-differentiation is available after cultivating NAS, the findings should be interpreted in consideration of the observations made in this study, whether it is more likely a question of a contaminant or a cause of intramammary infection. The bacteria shedding intensity of the NAS species with a more substantially adverse effect on udder health seems to be higher than that of the less important NAS pathogens., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The objective of the present study was to investigate the association between the milk fat-protein ratio and the incidence rate of clinical mastitis including repeated cases of clinical mastitis to determine the usefulness of this association to monitor metabolic disorders as risk factors for udder health. Herd records from 10 dairy herds of Holstein cows in Saxony, Germany, from September 2005-2011 (36,827 lactations of 17,657 cows) were used for statistical analysis. A mixed Poisson regression model with the weekly incidence rate of clinical mastitis as outcome variable was fitted. The model included repeated events of the outcome, time-varying covariates and multilevel clustering. Because the recording of clinical mastitis might have been imperfect, a probabilistic bias analysis was conducted to assess the impact of the misclassification of clinical mastitis on the conventional results. The lactational incidence of clinical mastitis was 38.2%. In 36.2% and 34.9% of the lactations, there was at least one dairy herd test day with a fat-protein ratio of <1.0 or >1.5, respectively. Misclassification of clinical mastitis was assumed to have resulted in bias towards the null. A clinical mastitis case increased the incidence rate of following cases of the same cow. Fat-protein ratios of <1.0 and >1.5 were associated with higher incidence rates of clinical mastitis depending on week in milk. The effect of a fat-protein ratio >1.5 on the incidence rate of clinical mastitis increased considerably over the course of lactation, whereas the effect of a fat-protein ratio <1.0 decreased. Fat-protein ratios <1.0 or >1.5 on the precedent test days of all cows irrespective of their time in milk seemed to be better predictors for clinical mastitis than the first test day results per lactation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The internal teat sealant OrbeSeal® (Zoetis, Berlin, Germany) is intended to prevent new intramammary infection (IMI) throughout the dry period. The aim of this field study was to determine new infection rates in udder quarters applied exclusively with the sealant at dry-off and untreated ones (control group). For that, the new infection rate of udder quarters treated with the sealant was evaluated in a split-udder design (front right and rear left udder quarters treated, the other ones untreated) in 128 cows from nine dairy farms in North Germany that were healthy in all four udder quarters (i.e. <100 000 cells/ml, bacteriologically negative). After calving, duplicate quarter milk samples were collected twice at DIM (days in milk) 5-12 and 7 d later for culture. The new infection rates for treated and untreated groups were 3·4 and 10·5%, respectively. The results showed that applying a standardised study design with very few confounders under field conditions, using internal teat sealants reduced the new infection rate on quarter level to a third of that of the control group. Thus, internal teat sealants represent a viable option for dry-off treatment of udder-healthy cows.

It was the objective of the study to estimate the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in bulk tank milk from German dairy herds and to characterize isolates from bulk tank milk with respect to their Staph. aureus protein A (spa) and staphylococcal cassette chromosome mec (SCCmec) type, their phenotypic antimicrobial resistance and resistance- resp. virulence-associated genes using broth microdilution and a microarray for Staph. aureus. Bulk tank milk samples (25 mL) were tested for MRSA using a 2-step selective enrichment protocol. Presumptive MRSA were confirmed by PCR. Thirty-six isolates collected from bulk tank milk of dairy herds in 2009 and 2010 were included in the characterization. All isolates displayed spa-types assigned to the clonal complex CC398. Based on the epidemiological cut-off values for the interpretation of minimum inhibitory concentrations isolates were resistant to tetracycline (100%), clindamycin (58%), erythromycin (52%), quinupristin/dalfopristin (36%), and kanamycin (27%). Isolates did not carry genes associated with typical virulence factors for Staph. aureus such as the Panton-Valentine leukocidin. However, they did carry hemolysin genes. Livestock-associated MRSA of CC398 does occur in German dairy herds and the strains have similar properties as described for strains from pigs., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The internal teat sealant OrbeSeal (Pfizer Animal Health) should prevent new IMI throughout the dry period. The aim of our field studies was to determine the efficacy of infusion with an internal teat seal at dry off in reducing new intramammary infections during the dry period in relation to an untreated control. In experiment 1 128 cows were enrolled from 24 dairy herds in Lower Saxony. Prior to final milking, all quarters were sampled for bacteriological culture and SCC analysis in three weekly intervals. After final milking, all 4 quarters of test group animals were infused with a commercially available internal teat seal. Following calving quarter milk samples were collected between 1 to 3 DIM and again between 8 to 10 and 15 to 17 DIM for culture and SCC analysis. The rate of new IMI during the dry period was 7.4% of quarters in the OrbeSeal-group and 12.7% in the control group, resp. In experiment 2 the new infection rate of udder quarters treated with OrbeSeal was evaluated in a split udder design with 93 cows healthy on all four udder quarters from six dairy farms in North Germany. In this study the front right and rear left udder quarters were treated, while both other quarters served as control. Applied methods occurred like in experiment 1. Here a new infection rate of 2.8% (OrbeSeal group) and in the control group a new infection rate of 11.5% was reached (P < 0.05).