SummaryEnvironmental mastitis is the most common and costly form of mastitis in modern dairy herds where contagious transmission of intramammary pathogens is controlled through implementation of standard mastitis prevention programmes. Environmental

The farmer field school (FFS) is a concept for farmers' learning, knowledge exchange, and empowerment that has been developed and used in developing countries. In Denmark, a research project focusing on explicit nonantibiotic strategies involves farmers who have actively expressed an interest in phasing out antibiotics from their herds through promotion of animal health. One way of reaching this goal was to form participatory focused farmer groups in an FFS approach, which was adapted to Danish conditions and named "stable schools." Four stable schools were established and went through a 1-yr cycle with 2 visits at each of the 5 or 6 farms connected to each group. A facilitator was connected to each group whose role was to write the meeting agenda together with the host farmer, direct the meeting, and write the minutes to send to the group members after the meeting. Through group focus interviews and individual semistructured qualitative interviews of all participants, the approach of the farmers' goal-directed work toward a common goal was judged to be very valuable and fruitful and based on a common learning process. Complex farming situations were the focus of all groups and in this context, problems were identified and solutions proposed based on each farmer's individual goals. In this article, we describe the experiences of 4 stable school groups (each comprising farmers and a facilitator), and the common process of building a concept that is suitable for Danish organic dairy farming.

Promotion of animal health and well-being at the individual animal and herd level is an important goal in organic farming. At the same time, chemical products affecting the natural balance among living organisms are prohibited in all areas of the organic farm. From an animal welfare point of view, however, no animal must suffer. Therefore, veterinary drugs are allowed under the European Union's regulations for organic farming, despite the fact that they are powerful cell toxins affecting both pathogenic and necessary bacteria, and as such in organic terminology, are regarded as "chemical" or "artificial" products. In this article, we present and discuss interviews with 12 Danish organic dairy producers who claim that minimized use or nonuse of antimicrobial drugs is an explicit goal. The dairy producers were at different levels with regard to reduced antimicrobial treatment. An explicit strategy of no antimicrobial treatments is based primarily on a long-term effort to improve herd health, and secondarily, on finding alternative treatments for diseased animals. Improved hygiene, outdoor access, use of nursing cows, and blinding of chronic mastitis quarters were the main techniques in developing a strategy of not using antimicrobial treatments in the herd by dairy producers. Producers' perception of disease changed from something unavoidable to a disturbing break in the daily rhythm that often could have been avoided. Change toward a nonantimicrobial strategy was gradual and stepwise. All dairy producers in this study desired to preserve the possibility of using antimicrobial drugs in emergencies.

With the aim of phasing out the use of antimicrobials 23 Danish organic dairy producers from the same organic dairy company participated in Stable School farmer groups from February 2004 to March 2005 in order to go through a common learning and development process towards their common goal. Data on production and herd health were evaluated from two years before to three years after the start of the Stable Schools. For comparison, data was collected from the remaining 35 herds delivering to the same dairy company, 118 organic dairy herds delivering to other dairies and 115 conventional herds. On average, the project herds were smaller with lower production and had half the incidence rate of mastitis treatment than the organic herds from other dairies before the start of the project. The incidence rate of mastitis treatments was reduced considerably from 20 treatments per 100 cow years to 10 treatments per 100 cow years after the project period. Somatic cell count (SCC) and scores for acute and chronic intramammary infections did not change significantly during the study period, and milk production increased at the same rate as in the other herd groups. The incidence rate of mastitis treatments or the reduction of the incidence rate could be related to the herd SCC or the prevalence of blind quarters. The incidence rates of locomotive disorders and reproductive disorders were lower in the project herds compared with herds from other dairies before the project start, and the differences increased during the project period though the reduction of the incidence rates in the project herds was not statistically significant. It is concluded that the farmers participating in the Stable Schools managed to reduce the use of antimicrobials in their herds also after the project period without apparent negative effects on production and udder and herd health.

Staphylococcus aureus is one of the most common pathogens that cause mastitis in dairy cows. Various subtypes, virulence genes and mobile genetic elements have been associated with isolates from bulk tank milk and clinical mastitis. So far, no Danish cattle associated S. aureus isolates have been whole-genome sequenced and further analyzed. Thus, the main objective was to investigate the population structure and genomic content of isolates from bulk tank milk and clinical mastitis, using whole-genome sequencing. This may reveal the origin of strains that cause clinical mastitis. S. aureus isolates from bulk tank milk (n = 94) and clinical mastitis (n = 63) were collected from 91 and 24 different farms, respectively and whole-genome sequenced. The genomic content was analyzed and a phylogenetic tree based on single nucleotide polymorphisms was constructed. In general, the isolates from both bulk tank milk and clinical mastitis were of similar genetic background. This suggests that dairy cows are natural carriers of the S. aureus subtypes that cause clinical mastitis if the right conditions are present and that a broad range of subtypes cause mastitis. A phylogenetic cluster that mostly consisted of ST151 isolates carried three mobile genetic elements that were primarily found in this group. The prevalence of resistance genes was generally low. However, the first ST398 methicillin resistant S. aureus isolate from a Danish dairy cow with clinical mastitis was detected.

Milk leakage in dairy cows is a symptom of impaired teat sphincter function. Milk leakage is related to an increased risk of mastitis in heifers and cows, and causes hygiene problems. The aim of our study was to assess whether teat shape, condition of teat orifice, and peak milk flow rate are risk factors for milk leakage. We conducted a longitudinal observational study in 15 German dairy farms in which cows were maintained in loose housing. The farms were visited monthly at 2 consecutive milkings. During the evening milking, milk flow curves were measured with the LactoCorder. Milk leakage was recorded during the subsequent morning milking, when cows entered the milking parlor. Immediately after detachment of the milking cluster, teat shape, teat end shape, and condition of the teat orifice of cows were assessed between 9 and 100 d in milk (DIM) and during late lactation (>250 DIM). Data from 1600 cows were analyzed. Milk leakage was treated as the binary response variable in a logistic regression model with herd as a random effect. Primiparous cows with high peak milk flow and teat canal protrusion were at greater risk of milk leakage. High peak milk flow rate, short teats, teat canal protrusion, inverted teat ends, and early lactation increased the risk of milk leakage in multiparous cows. Random herd effects accounted for only 10% of the total variation, indicating that the impact of management or other herd-level factors on the occurrence of milk leakage is virtually negligible for practical purposes.

Matrix-assisted laser desorption/ionization timeof-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method to identify the most common pathogenic bacteria in humans and animals. The goals of this study were to amend a commercial database with additional species, evaluate the amended database for identification of bacterial genera and species causing bovine mastitis, and describe the plethora of species involved. In total, 500 udder pathogenic isolates were subjected to MALDI-TOF MS using bacterial or fungal colony material; 93.5% could be identified to the species level, and 6.5% were identified only to the genus level. Isolates identified to the genus level required further identification to the species level by conventional methods or 16S rDNA sequencing. Mass spectra from verified species were used to expand the MALDI-TOF MS database to improve future identification ability. A total of 24 genera and 61 species were identified in this study. Identified isolates were mainly staphylococci, streptococci, Enterobacteriaceae, and coryneforme bacteria. In conclusion, MALDI-TOF MS is a powerful, rapid, and reliable technique to identify the most common microorganisms causing bovine mastitis, and the database can be continuously expanded and improved with additional species.

The role of non-aureus staphylococci (NAS) in the risk of acquisition of intramammary infections with Staphylococcus aureus is vague and still under debate. The objectives of this study were to (1) investigate the distribution patterns of NAS species from milk and teat skin in dairy herds with automatic milking systems, and (2) examine if the isolated NAS influences the expression of S. aureus virulence factors controlled by the accessory gene regulator (agr) quorum sensing system. In 8 herds, 14 to 20 cows with elevated somatic cell count were randomly selected for teat skin swabbing and aseptic quarter foremilk samples from right hind and left front quarters. Teat skin swabs were collected using the modified wet-dry method and milk samples were taken aseptically for bacterial culture. Colonies from quarters with suspicion of having NAS in milk or teat skin samples (or both) were subjected to MALDI-TOF assay for species identification. To investigate the interaction between S. aureus and NAS, 81 isolates NAS were subjected to a qualitative β-galactosidase reporter plate assay. In total, 373 NAS isolates were identified representing 105 from milk and 268 from teat skin of 284 quarters (= 142 cows). Sixteen different NAS species were identified, 15 species from teat skin and 10 species from milk. The most prevalent NAS species identified from milk were Staphylococcus epidermidis (50%), Staphylococcus haemolyticus (15%), and Staphylococcus chromogenes (11%), accounting for 76%. Meanwhile, the most prevalent NAS species from teat skin were Staphylococcus equorum (43%), S. haemolyticus (16%), and Staphylococcus cohnii (14%), accounting for 73%. Using reporter gene fusions monitoring transcriptional activity of key virulence factors and regulators, we found that out of 81 supernatants of NAS isolates, 77% reduced expression of hla, encoding a-hemolysin, 70% reduced expression of RNAIII, the key effector molecule of agr, and 61% reduced expression of spa encoding protein A of S. aureus, respectively. Our NAS isolates showed 3 main patterns: (1) downregulation effect such as S. chromogenes (milk) and Staphylococcus xylosus (milk and teat), (2) no effect such as Staphylococcus sciuri (teat) and S. vitulinus (teat), and the third pattern (c) variable effect such as S. epidermidis (milk and teat) and S. equorum (milk and teat). The pattern of cross-talk between NAS species and S. aureus virulence genes varied according to the involved NAS species, habitat type, and herd factors. The knowledge of how NAS influences S. aureus virulence factor expression could explain the varying protective effect of NAS on S. aureus intramammary infections.