Improvement of the Welfare Quality scoring model for dairy cows to fit experts' opinion.

After several years of implementation, the original Welfare Quality scoring model for dairy cows appears to be highly sensitive to the number and cleanliness of drinkers and not enough to the prevalence of diseases, and as a consequence may not fit the opinion of some animal welfare experts. The present paper aims to improve the Welfare Quality calculations for the criteria 'Absence of prolonged thirst' and 'Absence of disease' in dairy cows, so that the results are more sensitive to input data and better fit experts' opinion. First, we modified the calculation of 'Absence of prolonged thirst' by linearising the calculation for drinkers' availability to avoid threshold effects. Second, we modified the calculation of 'Absence of disease' by applying a Choquet integral on the three lowest spline-based scores for each health disorder to limit compensation between health disorders. Third, we performed a global sensitivity analysis of the original and the alternative scoring models. Fourth, we compared the results obtained with the original and the alternative models with eight experts' opinions on two subsets composed of 44 and 60 farms, respectively, inspected using the Welfare Quality protocol and on which experts gave their opinion on the overall level of animal welfare. Results show that the alternative model significantly reduced the 'threshold effects' related to the number of drinkers and the compensation between health disorders. On the first subset, the alternative model fits the experts' opinion slightly better than the original model (P = 0.061). On the second subset, the models performed equally. In conclusion, the proposed refinements for calculating scores are validated since they significantly reduced 'threshold effects' and the influence of measures related to drinkers. It also reduced the compensation between health disorders by considering only the three lowest scores and thus increasing the influence of measures related to health disorders, and slightly improve at overall score level the accordance with experts' opinion.

Comparing gene expression in deep infiltrating endometriosis with adenomyosis uteri: evidence for dysregulation of oncogene pathways.

BACKGROUND: The pathogenesis of deep infiltrating endometriosis (DIE) is poorly understood. It is considered a benign disease but has histologic features of malignancy, such as local invasion or gene mutations. Moreover, it is not clear whether its invasive potential is comparable to that of adenomyosis uteri (FA), or whether it has a different biological background. Therefore, the aim of this study was to molecularly characterize the gene expression signatures of both diseases in order to gain insight into the common or different underlying pathomechanisms and to provide clues to pathomechanisms of tumor development based on these diseases. METHODS: In this study, we analyzed formalin-fixed and paraffin-embedded tissue samples from two independent cohorts. One cohort involved 7 female patients with histologically confirmed FA, the other cohort 19 female patients with histologically confirmed DIE. The epithelium of both entities was microdissected in a laser-guided fashion and RNA was extracted. We analyzed the expression of 770 genes using the nCounter expression assay human PanCancer (Nanostring Technology). RESULTS: In total, 162 genes were identified to be significantly down-regulated (n = 46) or up-regulated (n = 116) in DIE (for log2-fold changes of < 0.66 or > 1.5 and an adjusted p-value of < 0.05) compared to FA. Gene ontology and KEGG pathway analysis of increased gene expression in DIE compared to FA revealed significant overlap with genes upregulated in the PI3K pathway and focal adhesion signaling pathway as well as other solid cancer pathways. In FA, on the other hand, genes of the RAS pathway showed significant expression compared to DIE. CONCLUSION: DIE and FA differ significantly at the RNA expression level: in DIE the most expressed genes were those belonging to the PI3K pathway, and in FA those belonging to the RAS pathway.

Pan-cancer analysis of genomic scar patterns caused by homologous repair deficiency (HRD).

Homologous repair deficiency (HRD) is present in many cancer types at variable prevalence and can indicate response to platinum-based chemotherapy and PARP inhibition. We developed a tumor classification system based on the loss of function of genes in the homologous recombination repair (HRR) pathway. To this end, somatic and germline alterations in BRCA1/2 and 140 other HRR genes were included and assessed for the impact on gene function. Additionally, information on the allelic hit type and on BRCA1 promoter hypermethylation was included. The HRDsum score including LOH, LST, and TAI was calculated for 8847 tumors of the TCGA cohort starting from genotyping data and for the subcohort of ovarian cancer also starting from WES data. Pan-cancer, deleterious BRCA1/2 alterations were detected in 4% of the tumors, while 18% of the tumors were HRD-positive (HRDsum ≥ 42). Across 33 cancer types, both BRCA1/2 alterations and HRD-positivity were most prevalent in ovarian cancer (20% and 69%). Pan-cancer, tumors with biallelic deleterious alterations in BRCA1/2 were separated strongly from tumors without relevant alterations (AUC = 0.89), while separation for tumors with monoallelic deleterious BRCA1/2 alterations was weak (AUC = 0.53). Tumors with biallelic deleterious alterations in other HHR genes were separated moderately from tumors without relevant alterations (AUC = 0.63), while separation for tumors with such monoallelic alterations was weaker (AUC = 0.57). In ovarian cancer, HRDsum scores calculated from WES data correlated strongly with HRDsum scores calculated from genotyping data (R = 0.87) and were slightly (4%) higher. We comprehensively analyzed HRD scores and their association with mutations in HRR genes in common cancer types. Our study identifies important parameters influencing HRD measurement and argues for an integration of HRDsum score with specific mutational profiles.

Tranexamic acid reduced blood transfusions in acute burn surgery: A retrospective case-controlled trial.

OBJECTIVE: Early excision and grafting of burn wounds are key for reducing prevalence of infection and sepsis. However, it is associated with massive blood loss and patients frequently require large numbers of blood transfusions. This study aimed to evaluate the effect of tranexamic acid (TXA) on blood transfusion requirements in acute burn surgery. METHODS: Patients admitted to the burn intensive care unit between January 2018 and May 2021 and received TXA before first surgery for wound excision and grafting were matched in a 1:2 ratio to patients that did not receive TXA (confounders age,sex,total body surface area (TBSA) burned). Primary endpoint of the analysis was the total number of transfused units of red blood cells (RBC) intra- and postoperatively up to 48 h. Fresh frozen plasma (FFP) and platelets were evaluated. Endpoints were compared between groups using van Elteren tests adjusting for strata variable age, gender, TBSA. RESULTS: Twenty-six TXA patients were matched with 52 control patients resulting in similar distributions of gender (77.9%(TXA)vs. 82.7%(control) males, p=0.542), age (51.7±21.3vs.48.3 ±17.4years,p = 0.459) and %TBSA burned (33.5%(IQR34)vs. 38.5% (IQR 30.5),p = 1.000). TXA group received significantly less RBC units intraoperatively (2.5(IQR 2.0)vs.4.0 units (IQR4.0), p = 0.038) and in total (4.0(IQR3.0)vs.6.0(IQR4.0),p = 0.017). TXA patients also received less blood products in general (RBC, FFP, platelets) in each period and in total. We found no significant difference in length of stay (24.0(IQR26.0)vs.33.0 days (IQR 0.5),p = 0.367) or mortality (15.4%vs.21.2%, p = 0.542). DISCUSSION: This study shows that necessity for blood transfusions in acute burn surgery may be reduced significantly by administration of TXA perioperatively. Randomized-controlled trials are needed to prove these findings.

The immune microenvironment in EGFR- and ERBB2-mutated lung adenocarcinoma.

BACKGROUND: Targeted therapies have improved survival and quality of life for patients with non-small-cell lung cancer with actionable driver mutations. However, epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 gene (HER2, also known as ERBB2) exon 20 insertions (Ex20mut) are characterized by a poor response to currently approved tyrosine kinase inhibitors and immunotherapies. The underlying immune biology is not well understood. MATERIALS AND METHODS: We carried out messenger RNA expression profiling of lung adenocarcinomas (ADCs) with ERBB2 (n = 19) and EGFR exon 20-insertion mutations (n = 13) and compared these to tumors with classical EGFR mutations (n = 40, affecting EGFR exons 18, 19 or 21) and EGFR/ERBB2 mutation-negative lung ADC (EGFR/ERBB2wt, n = 26) focusing on immunologically relevant transcripts. Tumor-infiltrating immune cells were estimated from gene expression profiles. RESULTS: Cytotoxic cells were significantly lower in EGFR-mutated tumors regardless of the affected exon, while Th1 cells were significantly lower in EGFR-Ex20mut compared to EGFR/ERBB2wt tumors. We assessed the differentially expressed genes of ERBB2-Ex20mut and EGFR-Ex20mut tumors compared to EGFR-Ex18/19/21mut and EGFR/ERBB2wt tumors. Of these, the genes GUSB, HDAC11, IFNGR2, PUM1, RASGRF1 and RBL2 were up-regulated, while a lower expression of CBLC, GBP1, GBP2, GBP4 and MYC was observed in all three comparison groups. The omnibus test revealed 185 significantly (FDR = 5%) differentially expressed genes and we found these four most significant gene expression changes in the study cohort: VHL and JAK1 were overexpressed in ERBB2-Ex20mut and EGFR-Ex20mut tumors compared to both EGFR-Ex18/19/21mut and EGFR/ERBB2wt tumors. RIPK1 and STK11IP showed the highest expression in ERBB2-Ex20mut tumors. CONCLUSIONS: Targeted gene expression profiling is a promising tool to read out the characteristics of the tumor microenvironment from routine diagnostic lung cancer biopsies. Significant immune reactivity and specific immunosuppressive characteristics in ERBB2-Ex20mut and EGFR-Ex20mut lung ADC with at least some degree of immune infiltration support further clinical evaluation of immune-modulators as partners of immune checkpoint inhibitors in such tumors.

Risk stratification of EGFR+ lung cancer diagnosed with panel-based next-generation sequencing.

OBJECTIVE: Panel-based next-generation sequencing (NGS) is increasingly used for the diagnosis of EGFR-mutated non-small-cell lung cancer (NSCLC) and could improve risk assessment in combination with clinical parameters. MATERIALS AND METHODS: To this end, we retrospectively analyzed the outcome of 400 tyrosine kinase inhibitor (TKI)-treated EGFR+ NSCLC patients with validation of results in an independent cohort (n = 130). RESULTS: EGFR alterations other than exon 19 deletions (non-del19), TP53 co-mutations, and brain metastases at baseline showed independent associations of similar strengths with progression-free (PFS hazard ratios [HR] 2.1-2.3) and overall survival (OS HR 1.7-2.2), in combination defining patient subgroups with distinct outcome (EGFR+NSCLC risk Score, "ENS", p < 0.001). Co-mutations beyond TP53 were rarely detected by our multigene panel (<5%) and not associated with clinical endpoints. Smoking did not affect outcome independently, but was associated with non-del19 EGFR mutations (p < 0.05) and comorbidities (p < 0.001). Laboratory parameters, like the blood lymphocyte-to-neutrophil ratio and serum LDH, correlated with the metastatic pattern (p < 0.01), but had no independent prognostic value. Reduced ECOG performance status (PS) was associated with comorbidities (p < 0.05) and shorter OS (p < 0.05), but preserved TKI efficacy. Non-adenocarcinoma histology was also associated with shorter OS (p < 0.05), but rare (2-3 %). The ECOG PS and non-adenocarcinoma histology could not be validated in our independent cohort, and did not increase the range of prognostication alongside the ENS. CONCLUSIONS: EGFR variant, TP53 status and brain metastases predict TKI efficacy and survival in EGFR+ NSCLC irrespective of other currently available parameters ("ENS"). Together, they constitute a practical and reproducible approach for risk stratification of newly diagnosed metastatic EGFR+ NSCLC.

A sensitive method for the recovery of Escherichia coli serogroup O55 including Shiga toxin-producing variants for potential use in outbreaks.

AIM: Shiga toxin-producing Escherichia coli (STEC) cause bloody diarrhoea, kidney failure and occasionally death. However, identifying the source of infection caused by STEC other than serogroup O157 is hampered by the availability of sensitive methods for detecting these pathogens. In this study, we developed novel tools for detecting E. coli O55 that is potentially associated with human outbreaks. METHODS AND RESULTS: Overall specificity of immuno-magnetic separation (IMS) beads coated with anti-O55 serum was good with exception of cross-reactivity with E. coli O22 and O23, which was eliminated using an O55-specific PCR. Limit of detection for E. coli O55 using O55-IMS beads in spiked cattle faeces was on average 50 CFU per ml (range 1-90), and improved to <10 CFU per ml using the O55-specific PCR, following IMS on samples enriched for 2 h with E. coli O55. Application of these tools to test cattle faeces collected on-farm allowed the isolation of O55:H19, which through whole genome sequencing was compared to STEC O55:H7 human outbreak strains. CONCLUSION: These tools provide a sensitive method which could be used to screen samples for STEC O55, whether environmental or human clinical. SIGNIFICANCE AND IMPACT OF THE STUDY: Several human outbreaks reported in England were caused by STEC O55:H7. Tools developed here could assist in identification of the environmental source for these isolates, which has not yet been established.

[From panel diagnostics to comprehensive genomic analysis : Infobesity or empowerment?] / Von der Paneldiagnostik zu umfassenden genomischen Analysen : Informationsüberfluss oder Zugewinn?

Precision oncology is obtaining a central role in the therapy of malignant diseases. The indication for targeted therapy is based on the identification of molecular targets for which next-generation sequencing (NGS) is commonly used nowadays. All approved predictive biomarkers and molecular targets, including gene fusions and copy number alterations, can be identified depending on panel design and method applied. Some clinical scenarios, however, may require more holistic genomic approaches, such as whole-genome/whole-exome and transcriptome analysis, which must be embedded in a clinical trial. Here, key aspects and applications of each method are summarized and discussed.

Identification of a highly lethal V3+ TP53+ subset in ALK+ lung adenocarcinoma.

Tyrosine kinase inhibitors (TKI) have improved prognosis in metastatic anaplastic lymphoma kinase (ALK)-driven lung adenocarcinoma, but patient outcomes vary widely. We retrospectively analyzed the clinical course of all cases with assessable baseline TP53 status and/or ALK fusion variant treated at our institutions (n = 102). TP53 mutations were present in 17/87 (20%) and the echinoderm microtubule-associated protein-like 4 (EML4)-ALK variant 3 (V3) in 41/92 (45%) patients. The number of metastatic sites at diagnosis was affected more by the presence of V3 than by TP53 mutations, and highest with both factors (mean 5.3, p < 0.001). Under treatment with ALK TKI, progression-free survival (PFS) was shorter with either TP53 mutations or V3, while double positive cases appeared to have an even higher risk (hazard ratio [HR] = 2.9, p = 0.015). The negative effect of V3 on PFS of TKI-treated patients was strong already in the first line (HR = 2.5, p = 0.037) and decreased subsequently, whereas a trend for PFS impairment under first-line TKI by TP53 mutations became stronger and statistically significant only when considering all treatment lines together. Overall survival was impaired more by TP53 mutations (HR = 4.9, p = 0.003) than by V3 (HR = 2.4, p = 0.018), while patients with TP53 mutated V3-driven tumors carried the highest risk of death (HR = 9.1, p = 0.02). Thus, TP53 mutations and V3 are independently associated with enhanced metastatic spread, shorter TKI responses and inferior overall survival in ALK+ lung adenocarcinoma. Both markers could assist selection of cases for more aggressive management and guide development of novel therapeutic strategies. In combination, they define a patient subset with very poor outcome.

Risk factors associated with beef cattle losses on intensive fattening farms in Austria, Germany and Italy.

The aim of this study was to investigate management and feeding practices associated with on-farm loss rate (mortality) on 63 beef cattle farms in Austria, Germany and Italy with housing systems other than fully slatted pens. Information on mortality and 56 categorised factors relating to the cleanliness of animal facilities, health and feeding management, animal-human interaction, cattle transport and origin were gathered during on-farm visits. Samples of total mixed rations (TMRs) were collected and analysed for chemical composition and particle size distribution. Twenty-eight categorised factors were removed from the initial 56 due to exclusion criteria (missing data≥20% and/or monolevel factors with≥80% answers in one category). Mortality was the response variable in the risk factor analysis and the remaining 10 continuous covariates from TMR analyses and 28 categorised factors were independent predictors. Mean (±standard deviation) mortality, representing the proportion of dead, euthanased and early culled animals over the total number of animals bought in or reared in the previous year, was 2.8±3.5%. Fourteen factors were significantly associated with mortality in the bivariable analyses; seven factors were not considered further in the multivariable analysis due to collinearity. None of the factors related to TMR were associated with mortality. Four categorical factors, referring to biosecurity measures and management, were retained in the final multivariable model, with country effect. Buying cattle from only one farm, no mixing of animals during transport, presence of a dedicated sick pen and keeping production records were associated with lower percentage mortality.

Longitudinal study on the occurrence in pigs of colistin-resistant Escherichia coli carrying mcr-1 following the cessation of use of colistin.

AIMS: In 2015, colistin-resistant Escherichia coli and Salmonella with the mcr-1 gene were isolated from a pig farm in Great Britain. Pigs were subsequently monitored over a ~20-month period for the occurrence of mcr-1-mediated colistin resistance and the risk of mcr-1 E. coli entering the food chain was assessed. METHODS AND RESULTS: Pig faeces and slurry were cultured for colistin-resistant E. coli and Salmonella, tested for the mcr-1 gene by PCR and selected isolates were further analysed. Seventy-eight per cent of faecal samples (n = 275) from pigs yielded mcr-1 E. coli after selective culture, but in positive samples only 0·2-1·3% of the total E. coli carried mcr-1. Twenty months after the initial sampling, faecal samples (n = 59) were negative for E. coli carrying mcr-1. CONCLUSIONS: The risk to public health from porcine E. coli carrying mcr-1 was assessed as very low. Twenty months after cessation of colistin use, E. coli carrying mcr-1 was not detected in pig faeces on a farm where it was previously present. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that cessation of colistin use may help over time to reduce or possibly eliminate mcr-1 E. coli on pig farms where it occurs.

Mortality, diarrhea and respiratory disease in Danish dairy heifer calves: Effect of production system and season.

Diarrhea and respiratory disease are major health problems for dairy calves, often causing calf mortality. Previous studies have found calf mortality to be higher in organic dairy herds compared to conventional herds. The aim of this study was to investigate the association between production system (conventional/organic), season (summer/winter) and calf mortality risk, diarrhea, signs of respiratory disease and ocular discharge, respectively, for dairy heifer calves aged 0-180 days. Sixty Danish dairy herds, 30 conventional and 30 organic, were visited once during summer and once during winter. During the herd visits, calves were clinically examined for signs of diarrhea, hampered respiration, nasal discharge, coughing and ocular discharge. Data on mortality were obtained from the Danish Cattle Database. Data were analyzed using logistic regression models, with mortality risk and disease measures as outcome variables for each of three calf age groups: 0-28, 29-90 and 91-180 days. In organic herds, odds of mortality among calves aged 0-28 days were 2.09 (95% confidence interval (CI): 1.38-3.15) times higher during winter compared to summer. Odds of nasal discharge for calves 0-28 days in organic herds were 10.3 (95% CI: 2.27-46.6), 10.7 (95% CI: 2.40-40.0) and 5.97 (95% CI: 1.29-27.6) times higher for organic and conventional herds during winter (OW and CW) and conventional herds during summer (CS) respectively, compared to organic herds during summer (OS). For calves aged 29-90 days, odds of nasal discharge were 8.22 (95% CI: 3.88-17.4), 8.06 (95% CI: 3.18-20.4) and 2.86 (95% CI: 1.08-7.55) times higher for OW, CW and CS respectively, compared to OS. Odds of nasal discharge for calved aged 91-180 days were 7.03 (95% CI: 3.95-12.5) and 4.27 (95% CI: 1.81-10.1) times higher for OW and CW respectively compared to OS. For calves aged 29-90 days, odds of coughing were 2.23 (95% CI: 1.06-4.71) and 3.82 (95% CI: 1.76-8.21) times higher for OW and CW compared to OS, while odds of coughing for calves aged 91-180 days were 2.09 (95% CI: 1.19-3.67) and 2.55 (95% CI: 1.39.4.67) times higher for OW and CW compared to OS. Odds of ocular discharge for calves aged 29-90 days were 0.22 (95% CI: 0.10-0.52), 0.27 (95% CI: 0.11-0.66) and 0.42 (95% CI: 0.18-0.99) times higher for OW, CW and CS compared to OS. In conclusion, mortality and morbidity of Danish dairy heifer calves are, for some variables and in certain age groups, dependent on production system and season.

Trained-user opinion about Welfare Quality measures and integrated scoring of dairy cattle welfare.

The Welfare Quality (WQ) protocol for on-farm dairy cattle welfare assessment describes 27 measures and a stepwise method for integrating values for these measures into 11 criteria scores, grouped further into 4 principle scores and finally into an overall welfare categorization with 4 levels. We conducted an online survey to examine whether trained users' opinions of the WQ protocol for dairy cattle correspond with the integrated scores (criteria, principles, and overall categorization) calculated according to the WQ protocol. First, the trained users' scores (n = 8-15) for reliability and validity and their ranking of the importance of all measures for herd welfare were compared with the degree of actual effect of these measures on the WQ integrated scores. Logistic regression was applied to identify the measures that affected the WQ overall welfare categorization into the "not classified" or "enhanced" categories for a database of 491 European herds. The smallest multivariate model maintaining the highest percentage of both sensitivity and specificity for the "enhanced" category contained 6 measures, whereas the model for "not classified" contained 4 measures. Some of the measures that were ranked as least important by trained users (e.g., measures relating to drinkers) had the highest influence on the WQ overall welfare categorization. Conversely, measures rated as most important by the trained users (e.g., lameness and mortality) had a lower effect on the WQ overall category. In addition, trained users were asked to allocate criterion and overall welfare scores to 7 focal herds selected from the database (n = 491 herds). Data on all WQ measures for these focal herds relative to all other herds in the database were provided. The degree to which expert scores corresponded to each other, the systematic difference, and the correspondence between median trained-user opinion and the WQ criterion scores were then tested. The level of correspondence between expert scoring and WQ scoring for 6 of the 12 criteria and for the overall welfare score was low. The WQ scores of the protocol for dairy cattle thus lacked correspondence with trained users on the importance of several welfare measures.

Prevalence and Spatial Distribution of Salmonella Infections in the Pennsylvania Raccoon (Procyon lotor).

A study was conducted to determine the prevalence and spatial distribution of Salmonella infection in Pennsylvania raccoons (Procyon lotor), common wildlife mammals known to occupy overlapping habitats with humans and domestic food animals. The Pennsylvania Game Commission provided a total of 371 raccoon intestinal samples from trapped and road-killed raccoons collected between May and November 2011. Salmonella was isolated from the faeces of 56 (15.1%) of 371 raccoons in 35 (54%) of 65 counties across Pennsylvania. The five most frequently isolated serotypes were Newport (28.6%), Enteritidis (19.6%), Typhimurium (10.7%), Braenderup (8.9%) and Bareilly (7.1%). Pulsed-field gel electrophoresis (PFGE) analysis of the Salmonella isolates and subsequent comparison to the Pennsylvania Department of Health human Salmonella PFGE database revealed 16 different pulsetypes in Salmonella isolates recovered from raccoons that were indistinguishable from pulsetypes of Salmonella collected from clinically ill humans during the study period. The pulsetypes of seven raccoon Salmonella isolates matched those of 56 human Salmonella isolates by month and geographical region of sample collection. Results from Clustered Regularly Interspaced Short Palindromic Repeats and Multi-Virulence Locus Sequence Typing (CRISPR-MVLST) analysis corroborated the PFGE and serotyping data. The findings of this study show that several PFGE pulsetypes of Salmonella were shared between humans and raccoons in Pennsylvania, indicating that raccoons and humans might share the same source of Salmonella.

Application of the Welfare Quality® assessment system on European beef bull farms.

Welfare concerns for intensive beef production have often been raised, but on-farm welfare assessment studies are rare. The aim of this study was to apply the Welfare Quality® (WQ) welfare assessment system for fattening cattle on beef bull farms to evaluate the state of welfare at the level of WQ measures and of aggregated scores, as well as overall classification. In addition, the purpose was to evaluate two ways of providing feedback information to the farmers with regard to possible welfare improvements on the farms. The study was conducted in Austria, Germany and Italy on a total of 63 beef bull farms with deep litter or cubicle-housing systems. Assessments were carried out 3 times (1 month and 7 months apart from the initial visit). In every country, farmers were assigned to two treatment groups (feedback from initial visit as written report, F, written feedback plus oral advice, FA) and a control group (C), which did not receive any feedback. At the criterion level, the highest average welfare scores were obtained from 'Absence of prolonged hunger' (94/100 points) followed by 'Absence of pain induced by management procedures' (88/100) and 'Comfort around resting' (77/100). Most welfare concerns related to the criteria 'Absence of disease' (40/100), 'Expression of social behaviour' (44/100) and 'Positive emotional state' (48/100), thus indicating room for improvements. Two-thirds of the farms achieved the 'Enhanced' level, about one-third was judged 'Acceptable' and only one farm 'Excellent'. After 6 months of monitoring period, there was no significant welfare improvement in both the treatment groups as compared with the control group. Reasons for the lack of effect may mainly be seen in the short monitoring period and a lack of external incentives. In conclusion, the WQ assessment system revealed areas for improvement, but longer term studies and investigations on alternative ways of transferring outcomes from on-farm welfare assessments to farmers should be carried out in future.

Transmission of shiga toxin-producing Escherichia coli O104:H4 at a family party possibly due to contamination by a food handler, Germany 2011.

We investigated a cluster of shiga toxin-producing Escherichia coli (STEC) O104:H4 infections after a family party during a large STEC O104:H4 outbreak in Germany. To identify the vehicle we conducted a retrospective cohort study. Stool samples of party guests, and food and environmental samples from the catering company were tested for STEC. We defined cases as party guests with gastrointestinal symptoms and laboratory-confirmed STEC infection. We found 23 cases among 71 guests. By multivariable analysis consumption of salmon [odds ratio (OR) 15, 95% confidence interval (CI) 2.3-97], herb cream (OR 6.5, 95% CI 1.3-33) and bean salad (OR 6.1, 95% CI 1.4-26) were associated with STEC infection. STEC O104:H4 was detected in samples of bell pepper and salmon. The food handler developed STEC infection. Our results point towards transmission via several food items contaminated by a food handler. We recommend regular education of food handlers emphasizing their role in transmitting infectious diseases.

On-farm animal welfare assessment in beef bulls: consistency over time of single measures and aggregated Welfare Quality(®) scores.

Consistency over time of (on-farm) animal welfare assessment systems forms part of reliability, meaning that results of the assessment should be representative of the longer-term welfare state of the farm as long as the housing and management conditions have not changed considerably. This is especially important if assessments are to be used for certification purposes. It was the aim of the present study to investigate consistency over time of the Welfare Quality(®) (WQ(®)) assessment system for fattening cattle at single measure level, aggregated criterion and principle scores, and overall classification across short-term (1 month) and longer-term periods (6 months). We hypothesized that consistency over time of aggregated criterion and principle scores is higher than that of single measures. Consistency was also expected to be lower with longer intervals between assessments. Data were obtained using the WQ(®) protocol for fattening cattle during three visits (months 0, 1 and 7) on 63 beef farms in Austria, Germany and Italy. Only data from farms where no major changes in housing and management had taken place were considered for analysis. At the single measure level, Spearman rank correlations between visits were >0.7 and variance was lower within farms than between farms for six and two of 19 measures after 1 month and 6 months, respectively. After aggregation of single measures into criterion and principle scores, five and two of 10 criteria and three and one of four principles were found reliable after 1 and 6 months, respectively. At the WQ(®) principle level, this was the case for three and one of four principles. Seventy-nine per cent and 75% of the farms were allocated to the same overall welfare category after 1 month and 6 months. Possible reasons for a lack of consistency are seasonal effects or short-term fluctuations that occur under normal farm conditions, low prevalence of clinical measures and probably insufficient sample size, whereas poor inter-observer agreement leading to inflation of correlation can be ruled out. At the criterion and principle level, aggregation of information into scores appears to partly smoothen undirected variation at the single measure level without losing sensitivity in terms of welfare evaluation. Reliable on-farm animal welfare assessments should therefore be based on repeated assessments. Further long-term studies are recommended to better understand the factors influencing consistency over time.

Why air quality in the Alps remains a matter of concern. The impact of organic pollutants in the alpine area.

In the middle of Europe, the Alps form a geographical and meteorological trap for atmospheric pollutants including volatile and semi-volatile organic compounds emitted in the surrounding lowlands. This is due to their barrier effects, high precipitation rates, and low ambient temperatures. Also the pollutants emitted in the cities inside the Alps spread in the region depending on orographic and meteorological conditions. Although a number of studies on the distribution and effect of pollutants in the Alps has been published, comprehensive information on potential hazards, and ways to improve this sensible environment are lacking. This opinion paper is the result of a discussion during the Winterseminar of the AlpsBioCluster project in Munich. It summarizes the current literature and presents some case studies on local pollution sources in the Alps, and the possibility of using biomonitoring techniques to assess critical pollution loads and distributions.

Dural ectasia in Loeys-Dietz syndrome: comprehensive study of 30 patients with a TGFBR1 or TGFBR2 mutation.

The purpose of this study was to assess the frequency, severity, and clinical associations of dural ectasia (DE) in Loeys-Dietz syndrome (LDS). Database analysis of three German metropolitan regions identified 30 patients with LDS and TGFBR1 mutation in 6 and a TGFBR2 mutation in 24 individuals (17 men; mean age: 31 ± 19 years), as well as 60 age and sex-matched control patients with Marfan syndrome carrying a FBN1 mutation. DE was present in 22 patients with LDS (73%), and it related to skeletal score points (p = 0.008), non-skeletal score points (p < 0.001), and to the presence of ≥7 systemic score points (p = 0.010). Similarly, the severity of DE was related to body height (p = 0.010) and non-skeletal score points (p = 0.004). Frequency (p = 0.131) and severity of DE (p = 0.567) was similar in LDS and Marfan syndrome. DE is a manifestation of LDS that occurs with similar frequency and severity as in Marfan syndrome. Severity of DE may serve as a marker of the overall connective tissue disease severity. LDS may be considered in patients with DE.