District-Wide Herd Sanitation and Eradication of Intramammary Staphylococcus aureus Genotype B Infection in Dairy Herds in Ticino, Switzerland.

The present study demonstrates successful herd sanitation and eradication of contagious mastitis caused by Staphylococcus aureus genotype B (S. aureus GTB) in an entire Swiss district (Ticino) including 3,364 dairy cows from 168 farms. Herd sanitation included testing of all cows using a highly GTB specific and sensitive qPCR assay, implementation of related on-farm measures, appropriate antibiotic therapy of GTB-positive cows and culling of therapy-resistant animals, respectively. A treatment index was used as an objective criterion to select GTB-positive cows eligible for culling and replacement payment. 62 herds (37%) were initially GTB-positive with a cow prevalence between 10% and 100% and were submitted to sanitation. Twenty mo after the start of the campaign, all these herds were free from S. aureus GTB, whereby 73% of them were sanitized during the first 7 mo. At the cow level, a total of 343 animals were infected. 50 of them were immediately culled and financially compensated based on their treatment index value. The remaining 293 cows were intramammarily treated with antibiotics either during lactation using the combination of cephalexin-kanamycin or penicillin-gentamicin or at dry-off using cloxacillin. Out of these cows, 275 (93.9%) were treated successfully meaning that their milk was twice GTB-negative by qPCR after therapy. For lactational treatment, control samples were taken ≥10 and ≥20 d after treatment, for dry off treatment ≥14 and ≥24 d after parturition. Neither lactation number nor SCC before treatment of the cow nor the type of therapy were associated with therapeutic cure. Using data of 30 GTB-positive and 71 GTB-negative herds (1855 observations), the impact of GTB sanitation on bulk tank milk SCC (BTSCC) was evaluated applying a linear mixed statistical model. In the year before sanitation, BTSCC was always higher in GTB positive than in GTB negative herds. After the start of the campaign, BTSCC declined rapidly in the herds under GTB sanitation and achieved values that no longer differed statistically from those of GTB-free herds after only 2 mo, remaining very similar for the rest of the campaign. The farmers were very satisfied with the outcome of the campaign as all GTB positive herds could be sanitized rapidly, sanitation was sustainable, and milk quality increased.

Sanitation of Staphylococcus aureus genotype B-positive dairy herds: A field study.

Staphylococcus aureus is one of the most widespread mastitis pathogens infecting dairy cattle worldwide. In Switzerland, different bovine genotypes of Staph. aureus have been identified, and genotype B (GTB) was demonstrated to be a highly contagious subtype, causing herd problems in cattle. As the efficacy of antibiotic therapy against Staph. aureus is not satisfactory, the most promising strategy for controlling this udder pathogen is the implementation of specific sanitation programs for affected farms. The aim of the present longitudinal study was the field evaluation of 2 analytical approaches for the sanitation of Staph. aureus GTB-positive dairy herds. We compared a new real-time quantitative PCR (qPCR) assay based on the detection of the unique target gene adlb with classical bacteriology. Sanitation was successfully achieved using both analytical methods, but the qPCR approach showed some main advantages, namely the use of clean (instead of aseptically collected) milk samples facilitates sample collection in terms of time and costs, enabling the sampling of entire herds during a regular milking procedure and by the farm staff. The high inclusivity and exclusivity of the new target gene adlb enable very specific detection of only the genotype of interest. Because of the very high diagnostic sensitivity of qPCR, each GTB-positive cow can be correctly identified at any time point during lactation, allowing farmers to continuously update milking groups to prevent transmission during milking. Milk sample analysis becomes more objective, faster, less expensive, and more suitable for routine application, enabling the sanitation of even big herds in short time.

[AntibioticScout.ch: Decision support for the prudent use of antimicrobials: Application in cattle]. / AntibioticScout.ch: Entscheidungshilfe für den umsichtigen Einsatz von antimikrobiellen Wirkstoffen: Anwendung beim Rind.

INTRODUCTION: The administration of antibiotics in livestock has been criticized for many years, in particular because of an inappropriate use and the appearance of antibiotic residues in the environment, which can promote the emergence and spread of resistant bacteria. However, antibiotics are essential for the successful and sustainable control of bacterial pathogens. With the aim of optimizing the use of antibiotics in food animals and minimizing the prevalence of resistant bacteria, AntibioticScout. ch provides a decision aid for the prudent use of antimicrobial drugs. This approach emphasizes the importance of supportive therapy and the hallmarks of preventive concepts. Procedures to improve animal health and animal welfare in accordance with the principles of good veterinary practice are primary and effective tools to reduce the use of antimicrobial drugs. The necessary reduction in the use of antibiotics must, therefore, be accompanied by appropriate management strategies in animal husbandry. In particular, hygiene, animal welfare and biosecurity measures are crucial to ensure an optimal health status in farm animals.

INTRODUCTION: On discute depuis des années de l'usage des antibiotiques dans l'élevage des animaux de rente, en particulier en ce qui concerne leur utilisation incorrecte et la charge environnementale liée à des résidus d'antibiotiques susceptibles de favoriser l'apparition et la propagation de résistances. Toutefois les antibiotiques sont essentiels pour assurer une lutte efficace et durable contre les maladies d'origine bactérienne. Dans le but d'optimiser l'usage des antibiotiques dans l'élevage des animaux de rente et, par conséquence, de réduire le développement de résistances, AntibioticScout.ch propose une aide à la décision pour un usage prudent de ces substances ("prudent use"). Parallèlement, on attire l'attention sur les traitements adjuvants et sur les mesures de prévention. Des mesures visant à améliorer la santé et le bien-être des animaux en tenant compte des fondements d'une bonne pratique vétérinaire sont des instruments efficaces pour réduire l'usage des antibiotiques. Cette réduction indispensable doit donc être combinée avec des mesures de gestion adéquates dans les élevages. Ce sont en particulier l'hygiène et les conditions d'élevage correctes ainsi que la mise en place de mesures de biosécurité qui sont décisives pour l'optimisation de la santé des troupeaux.

Quarter- and cow-level risk factors for intramammary infection with coagulase-negative staphylococci species in Swiss dairy cows.

Bacteriological status, evaluation of udder symmetry, udder hygiene, and teat end scores of 92 dairy cows were assessed on 3 Swiss dairy farms in a longitudinal 1-yr study to determine risk factors for intramammary infection (IMI) with coagulase-negative staphylococci (CNS) species. Farm visits were performed monthly including sterile quarter milk sampling and udder evaluation of all lactating cows. Milk samples were evaluated for the presence of staphylococci using selective agar plates. Species identification was performed using MALDI-TOF mass spectrometry. Intramammary infection was defined as milk samples having ≥100 cfu per mL of milk according to culture results. Overall, 3,151 quarter samples were included in the statistical analysis. Staphylococcus chromogenes, Staphylococcus haemolyticus, Staphylococcus xylosus, and a Staphylococcus warneri-like species were the 4 most prevalent CNS species found. Hierarchical multivariable logistic regression models were built to evaluate risk factors for species-specific CNS IMI. Risk factors for Staph. chromogenes IMI were presence in herd B, the period from June 2014 to August 2014 and December 2014 to February 2015, and presence of udder edema. For Staph. haemolyticus, the relevant risk factor included coinfection with Staph. xylosus coinfection with other than the above-mentioned CNS species ("others") and the period from June 2014 to November 2014. Coinfection with Staph. haemolyticus and "others," the periods from June 2014 to August 2014 and December 2014 to February 2015, early phase of lactation (1-60 d in milk), and belonging to herd B were significantly associated with Staph. xylosus IMI. Mid and late lactation, coinfection with Staph. xylosus, and the period September 2014 to May 2015 were identified as significant risk factors for Staph. warneri-like IMI. For Staph. chromogenes, 60.6 and 26% of the variance was observed at the quarter and cow level, respectively, whereas for the other investigated species the highest variance was observed at the sample level. The predominant species within herds differed and was most pronounced for the Staph. warneri-like species.

Short communication: Role of Streptococcus pluranimalium in Mediterranean buffaloes (Bubalus bubalis) with different udder health statuses.

The aims of the current study were to describe presence and clinical role over time of Streptococcus pluranimalium isolated in milk samples of Mediterranean buffalo (MB). Two hundred composite milk samples originating from 40 primiparous MB were collected at 10, 30, 60, 90, and 150d in milk (DIM) and from 20 pluriparous MB at 77 to 120 DIM. Milk samples were used for analysis of somatic cell counts, bacteriological cultures, and identification (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). Nine of 200 (4.5%) samples of primiparous MB and 3 of 20 (15%) samples of pluriparous MB were positive for Strep. pluranimalium. The prevalence of the bacterium in primipari was 0% (0/40) at 10, 30, and 150 DIM, whereas it was 5 (2/40) and 17.5% (7/40) at 60 and 90 DIM, respectively. Eight primipari were positive only once, whereas 1 was positive at 2 different samplings. Mono-infection was not detected in any of the age categories or udder health status. Infections were transient in primipari. Clinical mastitis was observed in primipari once at 90 DIM, subclinical mastitis detected twice in the same animals at 60 and 90 DIM, and intramammary infections were diagnosed 1 and 5 times at 60 and 90 DIM in primipari, respectively, whereas 3 infections were diagnosed in pluripari. The clinical reflections demonstrate for the first time the presence of Strep. pluranimalium in MB and its association with different udder health status. Nevertheless, it cannot be excluded that the bacterium may simply follow a pattern of commensal or opportunistic behavior, taking advantage of a preexisting bacterial udder infection.

A multiarm randomized field trial evaluating strategies for udder health improvement in Swiss dairy herds.

The aims of this study were to quantify the effectiveness of specialist advice about udder health in Swiss dairy herds and to compare 3 different udder health improvement strategies against a negative control group. In 2010, 100 Swiss dairy herds with a high (between 200,000 and 300,000 cells/mL) yield-corrected bulk milk somatic cell count (YCBMSCC) were recruited for a 1-yr multiarm randomized field trial. The herds were visited between September and December 2011 to evaluate udder health-management practices and then randomly allocated into 1 of 4 study arms containing 25 herds each. The negative control study arm received neither recommendations for improving udder health nor any active support. The remaining 75 farmers received a herd-specific report with recommendations to improve udder health management. The positive control study arm received no further active support during 2012. The veterinarian study arm received additional support in the form of monthly visits by their herd veterinarian. Finally, the study group study arm received support in the form of bimonthly study group meetings where different topics concerning udder health were discussed. One year later, implementation of recommendations and changes in udder health were assessed. Of the recommendations given, 44.3% were completely implemented, 23.1% partially, and 32.6% were not implemented. No differences in implementation of recommendations were noted between the 3 study arms. At study enrollment, farmers were asked for the study arm of their preference but were subsequently randomly assigned to 1 of the 4 study arms. Farmers that were assigned to the study arm of their preference implemented more recommendations than farmers assigned to a study arm not of their preference. No decrease in the within-herd prevalence of cows that had a high (≥200,000 cells/mL) composite somatic cell count was observed in herds that had a YCBMSCC ≥200,000 cells/mL at the start of intervention. However, the 3 study arms with intervention (positive control, the veterinarian, and the study groups) prevented an increase in the within-herd prevalence of cows that had a high somatic cell count in herds with a low YCBMSCC at the start of the intervention compared with the negative control study arm. In the year after sending the report, herds assigned to the study group study arm had a reduced incidence rate of treated mastitis cases in comparison with the year before sending the report.

[Diagnosis and therapy of retained fetal membranes, puerperal metritis and clinical endometritis in cattle: Results of the Online-survey among Swiss practitioners. I Retained fetal membranes]. / Diagnose und Therapie von Nachgeburtsverhalten, puerperaler Metritis und klinischer Endometritis beim Rind: Ergebnisse einer Online-Umfrage bei Schweizer Tierärzten. I Nachgeburtsverhalten.

The aim of this study was to obtain the diagnostic and therapeutic approach among Swiss practitioners in cows with retained fetal membranes (RFM) (part 1). All members of the Association for ruminant health were contacted per email via the newsletter. The survey was completed by 128 veterinarians, partially responded by 140 veterinarians. The manual removal of the fetal membranes is practiced by 129 of the responding veterinarians. Cows with/without fever are treated usually with intrauterine antibiotics. Cows with RFM with/without fever are most commonly treated parenterally with tetracycline or penicillin. The use of cephalosporins and quinolones in cows with fever is more common than in cows without fever. With the present results of the survey veterinarians should critically question the supposed benefits of the manual removal of the placenta and the use of antibiotics in cows with RFM.

[Diagnosis and therapy of retained fetal membranes, puerperal metritis and clinical endometritis in cattle: Results of the Online-survey among Swiss practitioners. II. Puerperal metritis and clinical endometritis]. / Diagnose und Therapie von Nachgeburtsverhalten, puerperaler Metritis und klinischer Endometritis beim Rind: Ergebnisse einer Online-Umfrage bei Schweizer Tierärzten. II. Puerperale Metritis und klinische Endometritis.

The aim of the study was to obtain the diagnostic and therapeutic approach among Swiss practitioners in cows with puerperal metritis and clinical endometritis (part 2). All members of the Association for ruminant health were contacted per email via the newsletter. The survey was completed by 128 veterinarians, partially responded by 140 veterinarians. The following main symptoms of puerperal metritis were stated by the practitioners: purulent vaginal discharge, fever and reduced appetite. A vaginal and rectal examination was performed to diagnose the disease. Usually, an intrauterine treatment with tetracycline or cefapirin was done. Parenteral administration of tetracycline or penicillin was often combined with PGF(2α), NSAIDS or cortisone. Clinical endometritis was also diagnosed by vaginal and rectal examination and the main symptom indicated was purulent vaginal discharge. The therapy consisted of the administration of PGF(2α), uterine infusions predominantly with cefapirin, and rarely with parenteral administration of antibiotics. Further diagnostic tools were not used and normally cows were not rechecked. The success of the therapy of puerperal metritis and clinical endometritis was judged to be satisfactory to excellent.

Metformin and the risk of head and neck cancer: a case-control analysis.

AIMS: Metformin use has been associated with a decreased risk of some cancers, although data on head and neck cancer (HNC) are scarce. We explored the relation between the use of antidiabetic drugs and the risk of HNC. METHODS: We conducted a case-control analysis in the UK-based Clinical Practice Research Datalink (CPRD) of people with incident HNC between 1995 and 2013 below the age of 90 years. Six controls per case were matched on age, sex, calendar time, general practice and number of years of active history in the CPRD prior to the index date. Other potential confounders including body mass index (BMI), smoking, alcohol consumption and comorbidities were also evaluated. The final analyses were adjusted for BMI, smoking and diabetes mellitus (or diabetes duration in a sensitivity analysis). Results are presented as odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: Use of metformin was neither associated with a statistically significant altered risk of HNC overall (1-29 prescriptions: adjusted OR 0.87, 95% CI 0.61-1.24 and ≥ 30 prescriptions adjusted OR 0.80, 95% CI 0.53-1.22), nor was long-term use of sulphonylureas (adjusted OR 0.87, 95% CI 0.59-1.30), or any insulin use (adjusted OR 0.92, 95% CI 0.63-1.35). However, we found a (statistically non-significant) decreased risk of laryngeal cancer associated with long-term metformin use (adjusted OR 0.41, 95% CI 0.17-1.03). CONCLUSIONS: In this population-based study, the use of antidiabetic drugs was not associated with a materially altered risk of HNC. Our data suggest a protective effect of long-term metformin use for laryngeal cancer.

Incidence of and risk factors for severe hypoglycaemia in treated type 2 diabetes mellitus patients in the UK--a nested case-control analysis.

AIMS: To assess incidence rates (IRs) of and identify risk factors for incident severe hypoglycaemia in patients with type 2 diabetes newly treated with antidiabetic drugs. METHODS: Using the UK-based General Practice Research Database, we performed a retrospective cohort study between 1994 and 2011 and a nested case-control analysis. Ten controls from the population at risk were matched to each case with a recorded severe hypoglycaemia during follow-up on general practice, years of history in the database and calendar time. Using multivariate conditional logistic regression analyses, we adjusted for potential confounders. RESULTS: Of 130,761 patients with newly treated type 2 diabetes (mean age 61.7 ± 13.0 years), 690 (0.5%) had an incident episode of severe hypoglycaemia recorded [estimated IR 11.97 (95% confidence interval, CI, 11.11-12.90) per 10,000 person-years (PYs)]. The IR was markedly higher in insulin users [49.64 (95% CI, 44.08-55.89) per 10,000 PYs] than in patients not using insulin [8.03 (95% CI, 7.30-8.84) per 10,000 PYs]. Based on results of the nested case-control analysis increasing age [≥ 75 vs. 20-59 years; adjusted odds ratio (OR), 2.27; 95% CI, 1.65-3.12], cognitive impairment/dementia (adjusted OR, 2.00; 95% CI, 1.37-2.91), renal failure (adjusted OR, 1.34; 95% CI, 1.04-1.71), current use of sulphonylureas (adjusted OR, 4.45; 95% CI, 3.53-5.60) and current insulin use (adjusted OR, 11.83; 95% CI, 9.00-15.54) were all associated with an increased risk of severe hypoglycaemia. CONCLUSIONS: Severe hypoglycaemia was recorded in 12 cases per 10,000 PYs. Risk factors for severe hypoglycaemia included increasing age, renal failure, cognitive impairment/dementia, and current use of insulin or sulphonylureas.

Genotype-specific risk factors for Staphylococcus aureus in Swiss dairy herds with an elevated yield-corrected herd somatic cell count.

Bovine mastitis is a frequent problem in Swiss dairy herds. One of the main pathogens causing significant economic loss is Staphylococcus aureus. Various Staph. aureus genotypes with different biological properties have been described. Genotype B (GTB) of Staph. aureus was identified as the most contagious and one of the most prevalent strains in Switzerland. The aim of this study was to identify risk factors associated with the herd-level presence of Staph. aureus GTB and Staph. aureus non-GTB in Swiss dairy herds with an elevated yield-corrected herd somatic cell count (YCHSCC). One hundred dairy herds with a mean YCHSCC between 200,000 and 300,000cells/mL in 2010 were recruited and each farm was visited once during milking. A standardized protocol investigating demography, mastitis management, cow husbandry, milking system, and milking routine was completed during the visit. A bulk tank milk (BTM) sample was analyzed by real-time PCR for the presence of Staph. aureus GTB to classify the herds into 2 groups: Staph. aureus GTB-positive and Staph. aureus GTB-negative. Moreover, quarter milk samples were aseptically collected for bacteriological culture from cows with a somatic cell count ≥150,000cells/mL on the last test-day before the visit. The culture results allowed us to allocate the Staph. aureus GTB-negative farms to Staph. aureus non-GTB and Staph. aureus-free groups. Multivariable multinomial logistic regression models were built to identify risk factors associated with the herd-level presence of Staph. aureus GTB and Staph. aureus non-GTB. The prevalence of Staph. aureus GTB herds was 16% (n=16), whereas that of Staph. aureus non-GTB herds was 38% (n=38). Herds that sent lactating cows to seasonal communal pastures had significantly higher odds of being infected with Staph. aureus GTB (odds ratio: 10.2, 95% CI: 1.9-56.6), compared with herds without communal pasturing. Herds that purchased heifers had significantly higher odds of being infected with Staph. aureus GTB (rather than Staph. aureus non-GTB) compared with herds without purchase of heifers. Furthermore, herds that did not use udder ointment as supportive therapy for acute mastitis had significantly higher odds of being infected with Staph. aureus GTB (odds ratio: 8.5, 95% CI: 1.6-58.4) or Staph. aureus non-GTB (odds ratio: 6.1, 95% CI: 1.3-27.8) than herds that used udder ointment occasionally or regularly. Herds in which the milker performed unrelated activities during milking had significantly higher odds of being infected with Staph. aureus GTB (rather than Staph. aureus non-GTB) compared with herds in which the milker did not perform unrelated activities at milking. Awareness of 4 potential risk factors identified in this study guides implementation of intervention strategies to improve udder health in both Staph. aureus GTB and Staph. aureus non-GTB herds.

A longitudinal study investigating the prevalence of Staphylococcus aureus genotype B in seasonally communal dairy herds.

Staphylococcus aureus is a major mastitis-causing pathogen. Various genotypes have been recently identified in Switzerland but Staph. aureus genotype B (GTB) was the only genotype associated with high within-herd prevalence. The risk of introducing this Staph. aureus genotype into a herd may be increased by frequent animal movements. This may also be the case when cows from different herds of origin are commingled and share their milking equipment for a limited period of time. The aim of the present study was to determine the prevalence of Staph. aureus GTB in seasonally communal dairy herds before and after a summer period when dairy farming is characterized by mixing cows from different herds of origin in 1 communal operation. In addition, the environment was investigated to identify potential Staph. aureus GTB reservoirs relevant for transmission of the disease. A total of 829 cows from 110 herds of origin in 9 communal operations were included in the study. Composite milk samples were collected from all cows during the first or second milking after arrival at the communal operation and again shortly before the end of the season. Swab samples from the environment, involved personnel, and herding dogs present were collected before the cows arrived. At the end of the season, sampling of personnel was repeated. All samples were analyzed for the presence of Staph. aureus GTB using an established quantitative PCR. At the beginning of the season, Staph. aureus GTB-positive cows were identified in 7 out of 9 communal operations and the within-communal operation prevalence ranged from 2.2 to 38.9%. At the second sampling, all communal operations were Staph. aureus GTB positive, showing within-communal operation prevalence from 1 to 72.1%. The between-herd of origin prevalence increased from 27.3 to 56.6% and the cow-level prevalence increased from 11.2% at the beginning of the season to 29.6% at the end of the season. On 3 different communal operations, Staph. aureus GTB-positive swabs from seasonally employed personnel were identified at the end of the season. The results indicate that Staph. aureus GTB can easily spread in communal operations when cows from different herds of origin are mixed during the summer season. Effective management measures need to be designed to prevent the spread of Staph. aureus GTB in seasonally communal herds.

[Mastitis management in Swiss dairy farms with udder health problems]. / Mastitis-Management in Schweizer Milchviehbetrieben mit Eutergesundheitsproblemen.

The objective of this study was to describe the udder health management in Swiss dairy herds with udder health problems. One hundred dairy herds with a yield-corrected somatic cell count of 200'000 to 300'000 cells/ml during 2010 were selected. Data concerning farm structure, housing system, milking technique, milking procedures, dry-cow and mastitis management were collected during farm visits between September and December 2011. In addition, quarter milk samples were collected for bacteriological culturing from cows with a composite somatic cell count ≥ 150'000 cells/ml. The highest quarter level prevalence was 12.3 % for C. bovis. Eighty-two percent of the pipeline milking machines in tie-stalls and 88 % of the milking parlours fulfilled the criteria for the vacuum drop, and only 74 % of the pipeline milking machines met the criteria of the 10-l-water test. Eighty-five percent of the farms changed their milk liners too late. The correct order of teat preparation before cluster attachment was carried out by 37 % of the farmers only. With these results, Swiss dairy farmers and herd health veterinarians can be directed to common mistakes in mastitis management. The data will be used for future information campaigns to improve udder health in Swiss dairy farms.

Le but de cette étude était de décrire la gestion de la santé de la mamelle dans des exploitations connaissant des problèmes en la matière. Pour cela, on a sélectionné 100 exploitations laitières présentant, en 2010, un nombre théorique de cellules dans le lait de mélange compris entre 200'000 et 300'000 cellules/ml. Les données relatives à la structure de l'exploitation, à la détention des animaux, à la technique et au travail de traite ainsi qu' à la gestion du tarissement et des mammites ont été relevées lors d'une visite d'exploitation dans la période allant de septembre à décembre 2011.En outre des échantillons de lait des vaches avec un nombre de cellules ≥ 150'000/ml ont été analysés bactériologiquement. La prévalence la plus élevée au niveau des quartiers était celle de C. bovis avec 12.3 %. Lors du test des machines à traire relatif à la chute du vacuum, 82 % des installations des stabulations entravées et 88 % des salles de traites étaient dans la zone normale et seules 74 % des installations réussissaient le «test des 10 l d'eau¼. Le changement des manchons était effectué trop tard dans 85 % des exploitations. Seul 37 % des exploitants se tenaient, lors de la préparation des mamelles, à un ordre correct des phases de travail. Ces résultats doivent rendre les producteurs de lait et les vétérinaires d'exploitations attentifs aux erreur de gestion les plus fréquentes. Ces données serviront de bases pour les campagnes d'information futures, dans le but d'améliorer la santé de la mamelle dans les exploitations laitières suisses.

Bovine mastitis: the diagnostic properties of a PCR-based assay to monitor the Staphylococcus aureus genotype B status of a herd, using bulk tank milk.

Staphylococcus aureus genotype B (GTB) is a contagious mastitis pathogen in cattle, occurring in up to 87% of individuals. Because treatment is generally insufficient, culling is often required, leading to large economic loss in the Swiss dairy industry. As the detection of this pathogen in bulk tank milk (BTM) would greatly facilitate its control, a novel real-time quantitative PCR-based assay for BTM has previously been developed and is now being evaluated for its diagnostic properties at the herd level. Herds were initially classified as to their Staph. aureus GTB status by a reference method. Using BTM and herd pools of single-quarter and 4-quarter milk, the herds were then grouped by the novel assay, and the resulting classifications were compared. A total of 54 dairy herds were evaluated. Using the reference method, 21 herds were found to be GTB positive, whereas 33 were found to be negative. Considering the novel assay using both herd pools, all herds were grouped correctly, resulting in maximal diagnostic sensitivities (100%) and specificities (100%). For BTM samples, diagnostic sensitivities and specificities were 90 and 100%, respectively. Two herds were false negative in BTM, because cows with clinical signs of mastitis were not milked into the tank. Besides its excellent diagnostic properties, the assay is characterized by its low detection level, high efficiency, and its suitability for automation. Using the novel knowledge and assay, eradication of Staph. aureus GTB from a dairy herd may be considered as a realistic goal.

First Report of Asian Brown Rot Caused by Monilia polystroma on Apricot in Switzerland.

A survey for Monilia fructicola (G. Winter) Honey on apricots (Prunus armeniaca L.) was conducted in July and August 2009 and 2010 in Canton Wallis, Switzerland. Mummies of fruits showing brown rot were collected and isolations were conducted. Nearly 200 fungal isolates, tentatively identified as M. fructigena, were retested with a multiplex PCR (1). With the Agilent 2100 Bioanalyzer (Agilent Technologies, Basel, Switzerland) instead of 1.5% agarose gels, the 23 bp difference between the diagnostic fragments of M. fructigena and M. polystroma van Leeuwen (1) could clearly be scored. M. polystroma was diagnosed in 3 of 65 and 1 of 132 isolates collected in 2009 (13 orchards) and 2010 (10 orchards), respectively. The internal transcribed spacer (ITS) regions of four isolates (09-G4, 09-P16, 09-S5, and 10-C6) were amplified and sequenced (4). The four sequences (GenBank No. JN128835) as well as those of the Hungarian isolate UFT (AM937114 [3]) were identical and highly similar to the type sequence for M. polystroma (Y17876 [2]). The type sequence had a "T" at position 414, which was lacking in the other five sequences. The genomic region of unknown function used by Côté et al. (1) to develop their PCR diagnostic tool was sequenced for isolate 09-G4 with primers MFG.for (3) and M Poly rev 5'-CCACTTACATTTTTGGCTATTG-3'. The Swiss isolate (GenBank No. JN128836) and the Hungarian isolate UFT (AM937120) sequences were identical. The pathogenicity of isolate 09-G4 was tested on Golden delicious apples. Six apples were surface sterilized (70% ethanol), halved, and placed in sterile plastic boxes cut-side down. Further, six half apples were wounded in the center with a round scalpel with a diameter of 1 cm and inoculated with a round, potato dextrose agar (PDA) plug (1-cm diameter) of actively growing mycelium (5- to 7-day-old culture). Control apples (six halves) were treated with a PDA plug without mycelium. All fruits were incubated at 20°C with a 12-h light 12-h dark cycle. Seven days after inoculation, typical brown rot symptoms were visible on all inoculated fruits. Mock inoculated fruits remained healthy. Three inoculated halves, in addition to the brown rot symptoms, also produced sporodochia and around the inoculation point the tissue become black. With the multiplex PCR (1), M. polystroma was identified as the pathogen causing brown rot symptoms on the inoculated apples. The ellipsoid single-cell hyaline conidia of isolate 09-G4 grown on the Golden delicious apples averaged 15.2 ± 4.0 × 8.97 ± 1.1 µm and were the expected size for M. polystroma conidia (14.9 to 9.1 µm [4]). The first evidence of a new Monilia species was reported by Fulton et al. (2). They found that M. fructigena isolates from Japan were distinguishable from European isolates by five base substitutions in the ITS region (four in ITS1 and one in ITS2). Later, van Leeuwen et al. (4) found that the two groups of isolates could also be distinguished by morphological differences and described the new species as M. polystroma. According to the Centre for Agricultural Bioscience International, the impact of M. polystroma in a new area is presumed to be the same or very similar to that of M. fructigena. To our knowledge, this is the first report of M. polystroma in Swiss orchards. References: (1) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (2) C. E. Fulton et al. Eur. J. Plant Pathol. 105:495, 1999. (3) M. Petróczy and L. Palkovics. Eur. J. Plant Pathol. 125:343, 2009. (4) G. C. M. van Leeuwen et al. Mycol. Res. 106:444, 2002.

[Intramammary use of antibiotics in dairy farms in the canton of Ticino before, during and after Staphylococcus aureus genotype B elimination]. / Intramammärer Antibiotikaeinsatz vor, während und nach einer Staphylococcus aureus Genotyp B Sanierung in Milchkuhbetrieben im Kanton Tessin.

INTRODUCTION: The aim of the study was to evaluate the use of intramammary antibiotics before, during and after the elimination of Staphylococcus aureus genotype B (SAGTB). Data on intramammary antibiotic use was collected in 65 dairy farms as part of a pilot project for SAGTB elimination in the canton of Ticino from 2017 to 2019. The investigated farms were divided into 46 affected farms (with at least one SAGTB-positive animal) and 19 control farms (SAGTB-free farms). Data on antibiotic use were requested from veterinarians and treatment incidence, as a measure of antibiotic use, was calculated based on medical records and veterinary prescriptions. In addition, the treatment incidence was calculated for 47 farms during alpine farming period. In 2018 (elimination year), the mean incidence of treatment during lactation in the SAGTB-positive farms was significantly higher than in the control farms (p=0,003). In 2019 no significantly lower antibiotic use during lactation or dry period was detected between 2017 (before elimination) and 2019 (after elimination). Alpine farming places where only S. aureus genotype B-negative animals had access to had a significantly lower antibiotic use during lactation (p=0,004). The new federal database (Antibiotics Information System in Veterinary Medicine, IS ABV) should allow continuous monitoring and to confirm the reduction of antibiotic use in the coming years.

INTRODUCTION: L'objectif de l'étude était d'évaluer la consommation d'antibiotiques intramammaires avant, pendant et après l'assainissement de Staphylococcus aureus génotype B (SAGTB). Dans le cadre d'un projet pilote d'assainissement SAGTB dans le canton du Tessin, l'étude a recueilli des données sur la consommation d'antibiotiques intramammaires de 2017 à 2019 dans 65 exploitations laitières réparties en 46 exploitations test (avec au moins un animal positif au SAGTB) et 19 exploitations témoins (exploitations exemptes de SAGTB). Les données relatives à la consommation d'antibiotiques ont été demandées aux vétérinaires et, grâce aux enregistrements et aux prescriptions des vétérinaires, il a été possible de calculer l'incidence du traitement en tant que mesure de la consommation d'antibiotiques. En outre, l'incidence des traitements pendant l'alpage a également pu être calculée pour 47 exploitations. En 2018 (année d'assainissement), l'incidence des traitements pendant la lactation était en moyenne significativement plus élevée dans les exploitations SAGTB-positives que pour les exploitations de contrôle (p=0,003). En comparant 2017 (avant l'assainissement) et 2019 (après l'assainissement), il n'y a pas eu de baisse significative de la consommation d'antibiotiques pendant la lactation et le tarissement en 2019. Dans les exploitations d'estivage, on a constaté l'année suivant l'assainissement (2019) une consommation d'antibiotiques significativement plus faible pendant la lactation pour les alpages qui n'accueillaient que des animaux négatifs à S. aureus génotype B (p=0,004). Il faut espérer qu'avec l'aide de la nouvelle base de données fédérale (Système d'information sur les antibiotiques en médecine vétérinaire, IS ABV), le monitoring pourra être poursuivi dans les années à venir et que la réduction de la consommation d'antibiotiques sera confirmée dans les années suivantes.

First Report of the ß-Tubulin E198A Mutation Conferring Resistance to Methyl Benzimidazole Carbamates in European Isolates of Monilinia fructicola.

The causal agent of brown rot on stone and pome fruits, Monilinia fructicola (G. Wint.), is a quarantine pathogen in Europe. It has been detected in Austria (later eradicated), Spain, the Czech Republic, Italy, Germany, and Switzerland (1). In the United States and other countries, M. fructicola isolates were reported to show resistance to different classes of fungicides, including methyl benzimidazole carbamates (MBC) (2). Lichou et al. (2) reported the presence of isolates resistant to the MBC carbendazim in France, but the mechanisms inducing MBC resistance in these isolates were not studied. Ma et al. (3) in California, and more recently, Zhu et al. (4) in South Carolina, demonstrated that the molecular mechanisms accounting for low and high levels of resistance to MBC fungicides in M. fructicola isolates were the mutations H6Y and E198A, respectively, in the ß-tubulin gene. Four M. fructicola isolates each from Italy, France, Spain, and Switzerland (16 isolates total), all having an unknown level of MBC resistance, were selected. In each isolate, the section of the ß-tubulin gene containing the two potentially mutant codons was PCR-amplified with the primers TubA and TubR1 (3) and the amplicons were sequenced directly. Sequence analysis revealed the amino acid histidine (H) at codon 6 in all the isolates, which would not predict MBC resistance, while alanine (A) at codon 198 (the mutation predictive of a high level of MBC resistance) was found in all isolates from Spain and Switzerland and in three isolates each from France and Italy. A representative sequence of the four identical partial ß-tubulin gene sequences from the Swiss isolates was submitted to GenBank under the Accession No. HQ709265. All isolates were tested in a potato dextrose agar (PDA) petri dish assay for resistance to the MBC fungicide thiophanate-methyl (Nippon Soda Co., Ltd., Tokyo, Japan) at the discriminatory dose of 50 µg/ml (4). All isolates with the E198A mutation were able to grow on the media, while the two isolates without the E198A mutation were not able to grow. The result indicated that most isolates had a high level of resistance to the MBC fungicide. To our knowledge, this is the first report of the presence of the E198A mutation conferring resistance to MBC fungicides in European isolates of M. fructicola. As the mutation appears to be widely distributed, we anticipate that MBC fungicides may be ineffective at controlling brown rot in countries with occurrence of M. fructicola. References: (1) M. Hilber-Bodmer et al. Plant Dis. 94:643, 2010. (2) J. Lichou et al. Phytoma 547:22, 2002. (3) Z. H. Ma et al. Appl. Environ. Microbiol. 69:7145, 2003. (4) F. X. Zhu et al. Plant Dis. 94:1511, 2010.

[Systematic sanitation of dairy herds with mastitis caused by Staphylococcus aureus]. / Sanierung von Milchviehbeständen mit Staphylococcus aureus-Mastitiden.

Several strategies are known for sanitizing dairy herd problems caused by Staphylococcus (S.) aureus. They mostly consist of general management measures but specific decision-making at an individual animal level has not been described. A sanitation program in the form of a process chart developed by the Bern Clinic for Ruminants was undertaken in 10 dairy herds with this problem. In an affected herd the cows were divided into 3 groups: healthy, suspect, infected. Three milk samples (MS), taken at two-week intervals were cultivated. The cows were grouped according to the culture results. To measure the success of the sanitation program, the key figures «theoretical tank somatic cell count¼ (target < 150,000 SCC/ml) and «percentage of cows over limit¼ (limit: 150'00 SCC/ml, target < 20 %) were used. These were compared with the corresponding key figures from dairy herds, which were followed-up by the Bern Clinic for Ruminants (control herds). The problem herd sanitation program lasted between 2 and 21 months. A total of 1598 MS were analyzed, of which 241 (15 %) were S. aureus positive (15 %). At the end of the sanitation the key figures between problem herds and control herds were similar. The sanitation program has proved to be practical. The detection of S. aureus positive cows proved to be reliable and the udder health of the herd could be significantly improved.

The ecological importance of habitat complexity to the Caribbean coral reef herbivore Diadema antillarum: three lines of evidence.

When Caribbean long-spined sea urchins, Diadema antillarum, are stable at high population densities, their grazing facilitates scleractinian coral dominance. Today, populations remain suppressed after a mass mortality in 1983-1984 caused a loss of their ecosystem functions, and led to widespread declines in ecosystem health. This study provides three lines of evidence to support the assertion that a lack of habitat complexity on Caribbean coral reefs contributes to their recovery failure. Firstly, we extracted fractal dimension (D) measurements, used as a proxy for habitat complexity, from 3D models to demonstrate that urchins preferentially inhabit areas of above average complexity at ecologically relevant spatial scales. Secondly, controlled behaviour experiments showed that an energetically expensive predator avoidance behaviour is reduced by 52% in complex habitats, potentially enabling increased resource allocation to reproduction. Thirdly, we deployed a network of simple and cost-effective artificial structures on a heavily degraded reef system in Honduras. Over a 24-month period the adult D. antillarum population around the artificial reefs increased by 320% from 0.05 ± 0.01 to 0.21 ± 0.04 m-2 and the juvenile D. antillarum population increased by 750% from 0.08 ± 0.02 to 0.68 ± 0.07 m-2. This study emphasises the important role of habitat structure in the ecology of D. antillarum and as a barrier to its widespread recovery.

First Report of Brown Rot Caused by Monilinia fructicola on Apricot in a Swiss Orchard.

Monilinia fructicola (G. Wint.), causal agent of brown rot on stone and pome fruits, is a quarantine pathogen in Europe (EPPO A2 quarantine pest). Since it was first discovered in French orchards in 2001, this pathogen has been officially identified from orchards in Austria (eradicated), Spain, Czech Republic, Italy, and Germany. M. fructicola has also been reported on imported fruit in Hungary and Switzerland (2). Orchard surveys in Switzerland in 2003 and 2005 found no evidence of natural infections (2). From July to August 2008, a large-scale survey of orchards was conducted in the primary apricot- (Prunus armeniaca Linn.) production region of Switzerland (Canton Valais). Apricots showing brown rot symptoms were collected from 57 different orchards at packinghouses (152 samples). In addition, mummies and fresh fruits showing brown rot symptoms were directly collected from three orchards (70 samples). All samples were tested using the PCR-based assay of Côté et al. (3). Ten apricots, originating from an orchard where the samples were directly collected from the trees, tested positive for M. fructicola. These apricots showed brown, sunken lesions covered with grayish pustules. The remaining brown rot samples were identified as M. laxa and M. fructigena. The positive samples were confirmed by the M. fructicola PCR protocols of Hughes et al. (4), following the EPPO diagnostic protocol (1). Eight amplicons obtained with the PCR protocol of Hughes et al. (4) were sequenced, compared with each other, and blasted to the NCBI database. These amplicons were identical to each other and had a 100% match to 16 M. fructicola isolates originating from several countries including the United States, New Zealand, Japan, and China. The unicellular, hyaline, lemon-shaped conidia of three isolates grown at 22°C on PDA averaged 14.4 ± 1.3 µm long and 8.8 ± 0.77 µm wide, therefore fitting the description for M. fructicola (1). Koch's postulates were fulfilled by reproducing brown rot symptoms on mature apricots inoculated with conidia. Six days after inoculation, typical brown rot symptoms appeared on inoculated fruits while control fruits remained healthy. Molecular tests performed with the protocol of Côté et al. (3) and Hughes et al. (4) confirmed the presence of M. fructicola on the inoculated fruits. In 2009, the presence M. fructicola in the orchard where the pathogen was detected in 2008 was verified. One hundred and thirty-seven apricots showing brown rot symptoms were collected and tested (3). M. fructicola was recovered from two samples, indicating the persistence of the pathogen in the orchard. To our knowledge, this is the first report of natural infection of M. fructicola in a Swiss orchard. References: (1) Anonymous. OEPP/EPPO Bull. 33:281, 2003. (2) E. Bosshard et al. Plant Dis. 90:1554, 2006. (3) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (4) K. J. D. Hughes et al. OEPP/EPPO Bull. 30:507, 2000.