Estimation of epidemiological cut-off values for eight antibiotics used for treatment of bovine mastitis caused by Streptococcus uberis and Streptococcus dysgalactiae subsp. dysgalactiae.

Interpretive criteria for antimicrobial susceptibility testing are lacking for most antimicrobials used for bovine streptococcal mastitis. The objectives of this study were to determine (tentative) epidemiological cut-off ((T)ECOFF) values for clinically relevant antibiotics used for treatment of bovine mastitis, and to estimate the proportion of acquired resistance (non-wild-types) in Streptococcus dysgalactiae subsp. dysgalactiae and Streptococcus uberis. A total of 255 S. uberis and 231 S. dysgalactiae subsp. dysgalactiae isolates were obtained in Denmark and Norway from bovine mastitis. The isolates were tested for susceptibility to 10 antibiotics using broth microdilution. In accordance with the European Committee on Antimicrobial Susceptibility Testing (EUCAST) standard operating procedure, additional published MIC distributions were included for the estimation of ECOFFs for cloxacillin, cephapirin, lincomycin and tylosin, and TECOFFs for amoxicillin, benzylpenicillin, cephapirin and oxytetracycline. The proportion of non-wild-type (NWT) isolates for the beta-lactams was significantly higher in the Danish S. uberis (45-55%) compared to the Norwegian isolates (10-13%). For oxytetracycline, the proportion of NWT was significantly higher in the Danish isolates, both for S. uberis (28% vs. 3%) and S. dysgalactiae (22% vs. 0%). A bridging study testing in parallel MICs in a subset of isolates (n = 83) with the CLSI-specified and the EUCAST-specified broths showed excellent correlation between the MICs obtained with the two methods. The new ECOFFs and TECOFFs proposed in this study can be used for surveillance of antimicrobial resistance, and - for antimicrobials licensed for streptococcal bovine mastitis - as surrogate clinical breakpoints for predicting their clinical efficacy for this indication.

Development of the Breast Cancer Survivors Symptom Checklist for Use in Follow-Up Multidisciplinary Appointments.

Background: Breast cancer survivors are primarily followed up to monitor the effectiveness of treatment and complications and to detect recurrences. Many breast cancer survivors may experience prolonged adverse physical and psychological effects, which should also be addressed at follow-ups. The objective of this study was to develop a brief symptom assessment tool for breast cancer survivors to be used as a guideline for the survivors and all health care professionals conducting the routine follow-up. The second objective was to describe the women's individual experiences with follow-ups. Methods: A literature review, a focus group of 6 healthcare professionals using a nominal group technique process, and the experience and feedback via qualitative interviews with 16 breast cancer survivors was used to develop the Breast Cancer Survivors Symptom Assessment Checklist (BCS-SC). Results: The BCS-SC consists of a set of 13 symptoms/burdens and one question. On a scale from 0 (no symptom) to 10 (worst imaginable), survivors indicated the extent to which they experience each symptom. All survivors perceived the annual follow-ups as important, but none prepared for them. Eight of the 16 survivors reported that they had 2 or more of the symptoms/burdens listed in the BCS-SC. However, only one of the survivors had mentioned her symptom to the doctor at follow-up. Conclusion: The BCS-SC is a comprehensive assessment tool for symptoms/burdens that are common among breast cancer survivors and can aid efforts to optimize their follow-up. Furthermore, the BCS-SC allows for a more patient-initiated and focused consultation, leading to more patient-centered quality care.

First Finding of a Methicillin-Resistant Staphylococcus aureus (MRSA) t304/ST6 from Bovine Clinical Mastitis.

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) comprises a global threat to humans and animals. Here, we report and characterize the MRSA t304/ST6 variant which, to our knowledge, represents the first case found in bovine clinical mastitis. In general, the MRSA t304/ST6 variant is rarely described in livestock, contrary to humans where it is widely recognized. Phenotypic and genotypic resistance profiling showed that the bovine-MRSA t304/ST6 isolate expressed low susceptibility toward cefoxitin (MICcefoxitin = 16 µg/mL) and carried the mecA resistance gene in the SCCmec IVa. The bovine-MRSA t304/ST6 isolate carried a plasmid similar to that which has been frequently observed among human-MRSA t304/ST6 isolates in Denmark (GenBank accession no. NZ_CP047022). In addition, a Staphylococcus prophage 3 (ϕSA3) was detected, encoding an immune evasion cluster (IEC) of putative virulence genes associated with human host-specificity (sea, sak, and scn). Taken together, these findings suggest that the MRSA t304/ST6 found in this study represents a recent host-jump event, with human to cow transmission. This study emphasizes the importance of and the need for performance of antimicrobial resistance surveillance among bovine mastitis pathogens, including S. aureus and MRSA.

Association between Antibiotic Consumption and Resistance in Mink Production.

Antibiotic consumption is considered to be a main driver of antibiotic resistant bacteria. Mink breeding follows a distinctive seasonal reproduction cycle, and all of the mink produced in the northern hemisphere are bred, born, and pelted around the same time of year. Some of the diseases are age-related, which is reflected in the seasonal variation of antibiotic consumption. The seasonality makes mink a good model for the investigation of the association between antibiotic consumption and resistance. The objectives of this study were (1) to monitor the farm level of antibiotic resistance during one production cycle and (2) to assess the potential associations between antibiotic consumption and resistance. Twenty-four farms were included in this study (Denmark n = 20, Iceland n = 2, and The Netherlands n = 2), following a cohort of animals born in 2018. Staphylococcus delphini and Escherichia coli were isolated from samples of the carcasses and faeces and were collected randomly. The isolates were susceptibility tested and subsequently divided into the sensitive wildtype (WT) and the resistant non-wildtype (NWT) populations. The antibiotic consumption relative to the sampling periods was assessed as having a short-term or a long-term impact, i.e., in two explanatory factors. For both S. delphini and E. coli, a large between-farm variation of NWT profiles was detected. In the final multivariable, generalized linear mixed models, significant associations between NWT isolates and the consumption of specific antibiotics were found: the short-term use of tetracyclines in the growth period was associated with the occurrence of tetracycline NWT E. coli in the growth period (OR: 11.94 [1.78; 89.28]), and the long-term use of macrolide and tetracyclines was associated with the occurrence of erythromycin NWT S. delphini in the weaning period (OR: 18.2 [2.26; 321.36]) and tetracycline NWT S. delphini in the growth period (OR: 8.2 [1.27; 63.31]), respectively. Farms with zero consumption in the study years prior to sampling also had a substantial proportion of NWT isolates, indicating that NWT isolates are persistent and/or widely spread in the environment. Generally, a high occurrence of tetracycline NWTs was observed. NWT isolates with resistance against the most commonly used antibiotics were found on all the farms, stressing the need for routine surveillance and the prudent use of antibiotics. The results offer a preview of the complex relationship between consumption and resistance, demonstrating some significant associations between use and resistance. Moreover, antibiotic-resistant bacteria are present even on farms with no antibiotic consumption over extended periods, and theoretical explanations supported by the data are offered.

Development and evaluation of a bead-based Multiplexed Fluorescent ImmunoAssay (MFIA) for detection of antibodies to Salmonella enterica serogroup B and C1 in pigs.

BACKGROUND: Since 1995, a surveillance program for Salmonella has been applied in the Danish pig industry in order to reduce cases of human salmonellosis. The objective of this study was to develop a bead-based Multiplexed Fluorometric ImmunoAssay (MFIA) as an improved serological surveillance method compared to the Salmonella mix ELISA, which has been the national reference immunoassay in the Danish Salmonella surveillance program for about 20 years. RESULTS: An MFIA for detection of antibodies to Salmonella serogroup B and C1 was developed and optimized with regard to coupling of beads with Salmonella lipopolysaccharide antigens and establishing suitable assay conditions. The Salmonella MFIA was validated by testing sera from experimentally infected pigs as well as field sera from non-infected and infected pig herds, and by comparing to results from the Salmonella mix ELISA, which was run in parallel. Sensitivity and specificity was evaluated using receiver operating curve analysis showing an area under curve for the serogroup B and C1 MFIA of 0.984 and 0.998, respectively. The Salmonella MFIA was shown to detect more antibody-positive samples in seropositive herds compared to the Salmonella mix ELISA, and Bayesian statistics confirmed that the MFIA had a considerably higher sensitivity (94.5%) compared to the mix ELISA (75.1%). The assay specificity was slightly lower for the Salmonella MFIA (96.8%) compared to Salmonella mix ELISA (99.5%). Coupled beads were stable for at least 1 year at 4˚C, and MFIA reproducibility and repeatability of the Salmonella MFIA were acceptable. Results from proficiency tests also indicated that the Salmonella MFIA was more sensitive than the Salmonella mix ELISA and that they had similar specificity. CONCLUSIONS: A bead-based MFIA for simultaneous detection of porcine serum antibodies to Salmonella enterica serogroup B and C1 was developed and implemented in the Danish porcine serological Salmonella surveillance program in 2018. The Salmonella MFIA can distinguish, as opposed to the Salmonella mix ELISA, between antibodies to serogroup B and C1 and the MFIA shows considerably better sensitivity.

Effects of health related farm-level factors on skin size and quality in commercial mink (Neovison vison) production.

This study investigated the potential effects of management and health related factors on the productivity in the commercial mink production, during 2015-2018. Data were available from the database at Kopenhagen Fur, the national veterinary prescription database, VetStat, and the laboratory database at the Center for Diagnostics, Technological University of Denmark. A cross-sectional study, including 1.464 min. farms grouped into 1.187 epidemiological units, was applied. Data were analyzed in two models with different outcomes representing productivity on the mink farms, namely skin size and economical value (value sum) of the produced skin. The studied risk factors included use of vaccines and antibacterials, herd size, associated feed producer, purchases and sales of live animals, breeding results (litter size after weaning), Aleutian mink disease virus antibody (AMDV status) and stamping out, and laboratory test results. Vaccination against mink enteritis parvovirus and high breeding results were found to have a positive association with both outcomes, skin size and value sum. Both outcomes also varied significantly between farm clusters associated with different feed producers. Significant effects of antibacterial treatment were found, but the results were complex with both positive and negative associations with the outcome variables, depending on season and interactions with feed producer. Positive effects on antibacterial prescription on skin size were observed, except for farms associated with two small feed producers, known to have a variable microbiological feed quality. In farms receiving feed of very high quality, the positive effect of antibacterial prescription was marginal. CONCLUSIONS: The use of mink data has allowed us to assess the impact of feed quality as well as antibacterial prescription on productivity. The results showed a positive quantitative effect of vaccination against mink enteritis parvovirus on skin size and value, with an optimal effect by vaccination of the whole litter. Antibacterial prescription in the growth period, particularly around weaning, was found to have a positive quantitative effect on productivity in some farms, and the results suggest that the effect was associated with the feed quality. Use of antibacterials to counteract negative effects of low feed quality is not in accordance with principles for prudent use of antibacterials.

Corrigendum: Close to a Decade of Decrease in Antimicrobial Usage in Danish Pig Production-Evaluating the Effect of the Yellow Card Scheme.

[This corrects the article DOI: 10.3389/fvets.2020.00109.].

Close to a Decade of Decrease in Antimicrobial Usage in Danish Pig Production-Evaluating the Effect of the Yellow Card Scheme.

The emergence and dissemination of antimicrobial resistant pathogens have led to rising concern about the widespread and excessive use of antimicrobials in veterinary medicine. Denmark has implemented several initiatives to reduce antimicrobial use in animals since the 90s, with the Yellow Card scheme implemented in 2010. The aim of the present study is to investigate the effects on antimicrobial use in Danish pig farms of changes in the legislation regarding the Yellow Card, based on analysis of temporal trends in monthly antimicrobial consumption calculated at herd-level from 2010 to 2017. A dynamic linear model with a linear growth component was used to model the data. The percentage of herds with a negative growth component and a significant decline below zero (based on 95% credible intervals) was assessed for the entire study period. The analysis was run separately for the different age groups within each farm: weaners (up to 30 kg), sows and finishers herds, and categorized according to antimicrobial use before the Yellow Card initiative (2008-2009), as groups of herds with "low," "moderate," or "high" consumption. The results evidence a decline of the antimicrobial consumption in Danish pig herds, more pronounced during two periods: with the introduction of the Yellow Card and in relation to the announcements and changes in the legislation in 2014. Weaner herds in the high consumption category had the highest percentage of herds with a negative growth component, reaching 82% of herds in January 2011, followed by weaner herds with moderate antimicrobial consumption reaching 71% of herds in October 2012. For finisher herds in the moderate consumption category, the proportion with a negative growth increased from 38% in January 2010 to 71% in July 2011. A decline in antimicrobial consumption was also evident after announcements and changes in the legislation in 2014, particularly for sows and finishers. Our findings suggest that changes in the Yellow Card scheme promoted a continuous reduction in antimicrobial use in Danish pig herds, particularly in herds with high antimicrobial consumption.

Male breast cancer: clinicopathological characterization of a National Danish cohort 1980-2009.

BACKGROUND: To describe relevant pathological parameters of Danish male breast cancer patients (MBCP) diagnosed from 1980 to 2009, and to relate these data to treatment, overall survival (OS) and standardized mortality rate (SMR). MATERIALS AND METHODS: The MBCP cohort was defined from national Danish registers. A total of 643 MBCP were identified with tissue available in 457. Among these, 384 were primary operable. Where tissue blocks were available, tumor type, grade, estrogen receptor (ER), progesteron receptor (PgR) and androgen-receptor (AR) status as well as HER 2 and Ki67 were performed. OS was quantified by Kaplan-Meier estimates and SMR was calculated based on mortality rate among patients relative to the mortality rate in the general population. RESULTS: Male breast cancer was more often of ductal type, grade II and a very high proportion were ER and AR positive and HER2 negative. Intrinsic subtypes based on immunohistochemical evaluation showed luminal subtype. Ki67 ratio increased over period of study. OS declined by increased age, bigger tumor size, positive lymph node status, higher grade and Luminal B subtype. Hazard ratio and relative risk of SMR were highest for patients aged < 60 years. CONCLUSION: Male breast cancer is of luminal subtype, but more often Luminal B. Ki67 is crucial in evaluation of subtypes by immunohistochemistry, but have limitations. Subtyping seems to be of major importance. AR also can have a role in future treatment.

Unweaving tangled mortality and antibiotic consumption data to detect disease outbreaks - Peaks, growths, and foresight in swine production.

As our capacity to collect and store health data is increasing, a new challenge of transforming data into meaningful information for disease monitoring and surveillance has arisen. The aim of this study was to explore the potential of using livestock mortality and antibiotic consumption data as a proxy for detecting disease outbreaks at herd level. Changes in the monthly records of mortality and antibiotic consumption were monitored in Danish swine herds that became positive for porcine reproductive and respiratory syndrome (PRRS) and porcine pleuropneumonia. Laboratory serological results were used to identify herds that changed from a negative to a positive status for the diseases. A dynamic linear model with a linear growth component was used to model the data. Alarms about state changes were raised based on forecast errors, changes in the growth component, and the values of the retrospectively smoothed values of the growth component. In all cases, the alarms were defined based on credible intervals and assessed prior and after herds got a positive disease status. The number of herds with alarms based on mortality increased by 3% in the 3 months prior to laboratory confirmation of PRRS-positive herds (Se = 0.47). A 22% rise in the number of weaner herds with alarms based on the consumption of antibiotics for respiratory diseases was found 1 month prior to these herds becoming PRRS-positive (Se = 0.22). For porcine pleuropneumonia-positive herds, a 10% increase in antibiotic consumption for respiratory diseases in sow herds was seen 1 month prior to a positive result (Se = 0.5). Monitoring changes in mortality data and antibiotic consumption showed changes at herd level prior to and in the same month as confirmation from diagnostic tests. These results also show a potential value for using these data streams as part of surveillance strategies.

Healthcare use before and after changing disability pension policy: a regional Danish cohort study.

BACKGROUND: The social security policy for disability pension (DP) was changed in Denmark in 2013 and eligibility requirements were tightened. We describe and compare the use of healthcare among individuals with incident DP before and after the policy change. METHODS: This was a follow-up study based on data from nationwide databases. The study included individuals with incident DP aged 18-64 years and living in The North Denmark Region. We included individuals with incident DP before (2010-12, n = 6286) and after (2014-15, n = 1042) the 2013 policy change. Poisson regression was used to examine group differences in (i) contact to healthcare and (ii) hospitalization. For this purpose, we used incidence rate ratios stratified on type of contact before being awarded DP. RESULTS: We found a change of diagnoses for healthcare use towards higher proportions of cardiovascular, pulmonary, neurological and cancer diseases and lower proportion with musculoskeletal disorder in the populations being granted DP after policy changes. For individuals with psychiatric contact before being granted DP, we found no significant differences between periods in psychiatric healthcare after DP was awarded. For individuals with somatic contact before being granted DP, we found an increased risk of contact to somatic healthcare and hospitalization after DP requirements were tightened. CONCLUSION: The study demonstrated that individuals who were granted DP after the eligibility requirements had been tightened suffered from more medical conditions and had an ongoing need for healthcare. In contrast, no significant difference in risk of psychiatric contact or hospitalization after DP was demonstrated.

Within-farm prevalence and environmental distribution of livestock-associated methicillin-resistant Staphylococcus aureus in farmed mink (Neovison vison).

The aim of the present study was to identify the animal prevalence and environmental reservoir of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in mink farms. LA-MRSA on mink constitutes a human health hazard to farmers and farm workers, who handle the animals and are at risk of bites and scratches from colonized sites. The primary route of LA-MRSA colonization of mink is suspected to be by ingestion of contaminated pig by-products. We performed a cross-sectional study with repeated measurements during May-July 2017. A total of 644 mink carcasses (542 mink kits and 102 breeding animals) from five Danish farms were sampled. From each carcass, pharynx was swabbed and the right forepaw dissected. In addition, environmental samples covering feed, air, glove, cages (top and between) and nest boxes were collected on the farms. MRSA was selectively cultured from each sample and suspect colonies were assessed using matrix-assisted laser desorption ionisation (MALDI-TOF) for species confirmation. Further, from each farm, three isolates from mink and one isolate per positive environmental site were sent for whole genome sequencing. We isolated LA-MRSA from mink in four out of the five farms, but LA-MRSA bacterium was detected on all farms. On farms with LA-MRSA positive animals, the overall apparent animal prevalence ranged from 20% [13;29] CI95% to 29% [22;38]CI95%. LA-MRSA was isolated from kits before weaning, most likely due to a contaminated environment or transfer from the dam. Further, the apparent prevalence of LA-MRSA in kits tended to increase during the first months of age. On farms where LA-MRSA was isolated from mink, LA-MRSA was also isolated from the environment. LA-MRSA was isolated from all environmental sites tested (i.e. glove, on top of and between the cages and in the nest boxes), apart from air. The negative air samples contrast with the high concentrations of LA-MRSA in air found in the pig production. Hence, the risk of human exposure to LA-MRSA on mink-farms tends to be associated mainly with direct contact with contaminated environmental sites and the handling of colonized mink. All sequenced isolates were confirmed as LA-MRSA CC398 and genetically similar to clones previously isolated from the Danish pig production, supporting the hypothesis of LA-MRSA being transmitted by contaminated pig by-products.

Outcomes From Using Mortality, Antimicrobial Consumption, and Vaccine Use Data for Monitoring Endemic Diseases in Danish Swine Herds.

The aim of this study was to assess the potential of using multiple data sources currently available in Denmark for monitoring swine diseases. The study included farms that, based on serology, changed from "negative" to "positive" status for Porcine Reproductive and Respiratory Syndrome (PRRS), enzootic pneumonia (Mycoplasma hyopneumonia), and porcine pleuropneumonia (Actinobacillus pluropneumoniae) between January 2014 and September 2017. These corresponded to 45 swine farms working as individual operation units (i.e., their disease status is independent from other farms) and 81 farms that were part of joint operation units (i.e., 2 or more farms considered to be an epidemiological unit, having swine and personnel are transferred among them, that have the same disease status). Additionally, a total of 95 farms with a negative status for these three diseases during the study period were also included in the study. Changes in mortality data, antimicrobial consumption, and vaccine use at herd level were monitored using Shewhart control charts prior to, during, and after these farms were found positive for the three diseases. The analysis was run separately for the different age groups-weaners (up to 30 kg), sows and finishers herds-within each farm. Briefly, the highest percentage of herds generating alarms was generated up to 3 months before they changed their disease status based on mortality (30%) and 1 month after based on antimicrobial use for respiratory diseases (100%). Porcine pleuropneumonia showed to be the disease with the highest impact on these data at herd level; alarms based on the three data streams were generated in the same month that herds changed their status to porcine pleuropneumonia-positive, as well as the following months. Alarms based on vaccine use generally occurred within the same month or after changes in disease status. False alarms were found in 2% (median value) of the herds for the different age groups based on mortality and antimicrobial use for respiratory diseases in healthy farms. Monitoring changes in mortality data, antimicrobial consumption, and vaccine use showed changes (i.e., warnings) at herd level prior to confirmation from diagnostic tests.

Risk factors for the occurrence of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in Danish pig herds.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is widespread in many European countries including Denmark, where 88% of randomly selected production herds tested positive in 2016. In the present study, we investigated herd-level risk factors for farms being classified as LA-MRSA positive (study 1), in addition to herd-level risk factors for farms changing status from LA-MRSA negative to LA-MRSA positive during a 2-year period (study 2). Risk factors previously identified in other studies were confirmed in study 1: large herd size, herd type (lower risk in herds with sows) and number of pig suppliers. Due to the effect of herd type, data from sow herds (N = 41) and herds without sows (N = 166) were analysed separately. A univariable analysis found that the variables significantly associated with LA-MRSA status for sow herds were: use of wet feed in the sow units; higher weights of piglets at weaning; availability of a delivery room on the farm; cleaning of aisles after pigs were moved; number of pigs per weaner section; number of pigs purchased in the past year, and factors related to rodent control and human traffic in the herd. In herds without sows, the univariable analysis showed that the presence of other species of animal on the farm; negative pressure ventilation; full sectioning; frequent visits from the veterinarian; peroral use of tetracyclines for weaners; number of pigs purchased in the past year, and factors related to rodent control and human traffic in the herd were significantly associated with LA-MRSA status. For herds that changed from LA-MRSA negative to positive (study 2), having a company contract for mouse control, having more than one pig supplier and using group medication in the drinking water were the variables associated with LA-MRSA status in the univariable analysis. We did not succeed in building a biologically meaningful multivariable model based on any of the datasets and, as observed in similar studies, many of the risk factors identified in the univariable analysis were related to herd size. It was therefore not possible to determine whether it was the size of the herd or related factors that were the causal risk factors for being LA-MRSA positive.

Building the foundation for veterinary register-based epidemiology: A systematic approach to data quality assessment and validation.

Epidemiological studies often use data from registers. Data quality is of vital importance for the quality of the research. The aim of this study was to suggest a structured workflow to assess the quality of veterinary national registers. As an example of how to use the workflow, the quality of the following three registers was assessed: the Central Husbandry Register (CHR), the database for movement of pigs (DMP) and the national Danish register of drugs for veterinary use (VetStat). A systematic quantitative assessment was performed, with calculation the proportion of farms and observations with "poor quality" of data. "Poor" quality was defined for each measure (variable) either as a mismatch between and/or within registers, registrations of numbers outside the expected range, or unbalanced in- and outgoing movements. Interviews were conducted to make a complementary qualitative assessment. The proportion of farms and observations within each quality measure varied. This study highlights the importance of systematic quality assessment of register data and suggests a systematic approach for such assessments and validations without the use of primary data.

The association of diabetes mellitus and insulin treatment with expression of insulin-related proteins in breast tumors.

BACKGROUND: The insulin receptor (INSR) and the insulin growth factor 1 receptor (IGF1R) play important roles in the etiology of both diabetes mellitus and breast cancer. We aimed to evaluate the expression of hormone and insulin-related proteins within or related to the PI3K and MAPK pathway in breast tumors of women with or without diabetes mellitus, treated with or without insulin (analogues). METHODS: Immunohistochemistry was performed on tumor tissue of 312 women with invasive breast cancer, with or without pre-existing diabetes mellitus, diagnosed in 2000-2010, who were randomly selected from a Danish breast cancer cohort. Women with diabetes were 2:1 frequency matched by year of birth and age at breast cancer diagnosis to those without diabetes. Tumor Microarrays were successfully stained for p-ER, EGFR, p-ERK1/2, p-mTOR, and IGF1R, and scored by a breast pathologist. Associations of expression of these proteins with diabetes, insulin treatment (human insulin and insulin analogues) and other diabetes medication were evaluated by multivariable logistic regression adjusting for menopause and BMI; effect modification by menopausal status, BMI, and ER status was assessed using interactions terms. RESULTS: We found no significant differences in expression of any of the proteins in breast tumors of women with (n = 211) and without diabetes (n = 101). Among women with diabetes, insulin use (n = 53) was significantly associated with higher tumor protein expression of IGF1R (OR = 2.36; 95%CI:1.02-5.52; p = 0.04) and p-mTOR (OR = 2.35; 95%CI:1.13-4.88; p = 0.02), especially among women treated with insulin analogues. Menopause seemed to modified the association between insulin and IGF1R expression (p = 0.07); the difference in IGF1R expression was only observed in tumors of premenopausal women (OR = 5.10; 95%CI:1.36-19.14; p = 0.02). We found no associations between other types of diabetes medication, such as metformin, and protein expression of the five proteins evaluated. CONCLUSIONS: In our study, breast tumors of women with pre-existing diabetes did not show an altered expression of selected PI3K/MAPK pathway-related proteins. We observed an association between insulin treatment and increased p-mTOR and IGF1R expression of breast tumors, especially in premenopausal women. This observation, if confirmed, might be clinically relevant since the use of IGF1R and mTOR inhibitors are currently investigated in clinical trials.

Male breast cancer: a nation-wide population-based comparison with female breast cancer.

OBJECTIVE: Describe prognostic parameters of Danish male breast cancer patients (MBCP) diagnosed from 1980-2009. Determine all-cause mortality compared to the general male population and analyze survival/mortality compared with Danish female breast cancer patients (FBCP) in the same period. MATERIAL AND METHODS: The MBCP cohort was defined from three national registers. Data was extracted from medical journals. Data for FBCP is from the DBCG database. Overall survival (OS) was quantified by Kaplan-Meier estimates. Standardized mortality ratios (SMRs) were calculated based on mortality rate among patients relative to the mortality rate in the general population. The association between SMR and risk factors were analyzed in univariate and multivariable Poisson regression models. Separate models for each gender were used for the analyses. RESULTS: We found a marked difference in OS for the two genders. For the total population of MBCP, 5- and 10-year survivals were 55.1% and 31.7%, respectively. For FBCP, the corresponding figures were 76.8% and 59.3%. Median age at diagnosis for FBCP was 61 years and 70 years for MBCP. By applying SMR, the difference in mortality between genders equalized and showed pronounced age-dependency. For males <40 years, SMR was 9.43 and for females 19.56 compared to SMR for males 80 + years (0.95) and females 80 + years (0.89). During the period 1980-2009, the risk of dying gradually decreased for FBCP (p < .0001). The risk 1980-1984 was 35% higher than 2005-2009 (RR 1.35). Although the risk of dying for MBCP was also lowest in 2005-2009, there was no clear tendency (p = .1439). The risk was highest in 1990-1994 (RR =2.48). CONCLUSION: We found better OS for FBCP than for MBCP. But SMR showed similar mortality rate for the two genders, except for very young FBCP, who had higher SMR. Furthermore, significantly improved survival over time for FBCP was observed, with no clear tendency for MBCP.

Computional algorithm for lifetime exposure to antimicrobials in pigs using register data-The LEA algorithm.

Accurate and detailed data on antimicrobial exposure in pig production are essential when studying the association between antimicrobial exposure and antimicrobial resistance. Due to difficulties in obtaining primary data on antimicrobial exposure in a large number of farms, there is a need for a robust and valid method to estimate the exposure using register data. An approach that estimates the antimicrobial exposure in every rearing period during the lifetime of a pig using register data was developed into a computational algorithm. In this approach data from national registers on antimicrobial purchases, movements of pigs and farm demographics registered at farm level are used. The algorithm traces batches of pigs retrospectively from slaughter to the farm(s) that housed the pigs during their finisher, weaner, and piglet period. Subsequently, the algorithm estimates the antimicrobial exposure as the number of Animal Defined Daily Doses for treatment of one kg pig in each of the rearing periods. Thus, the antimicrobial purchase data at farm level are translated into antimicrobial exposure estimates at batch level. A batch of pigs is defined here as pigs sent to slaughter at the same day from the same farm. In this study we present, validate, and optimise a computational algorithm that calculate the lifetime exposure of antimicrobials for slaughter pigs. The algorithm was evaluated by comparing the computed estimates to data on antimicrobial usage from farm records in 15 farm units. We found a good positive correlation between the two estimates. The algorithm was run for Danish slaughter pigs sent to slaughter in January to March 2015 from farms with more than 200 finishers to estimate the proportion of farms that it was applicable for. In the final process, the algorithm was successfully run for batches of pigs originating from 3026 farms with finisher units (77% of the initial population). This number can be increased if more accurate register data can be obtained. The algorithm provides a systematic and repeatable approach to estimating the antimicrobial exposure throughout the rearing period, independent of rearing site for finisher batches, as a lifetime exposure measurement.

Antimicrobial resistance among pathogenic bacteria from mink (Neovison vison) in Denmark.

BACKGROUND: For proper treatment of bacterial infections in mink, knowledge of the causative agents and their antimicrobial susceptibility patterns is crucial. The used antimicrobials are in general not registered for mink, i.e. most usage is "off-label". In this study, we report the patterns of antimicrobial resistance among pathogenic bacteria isolated from Danish mink during the period 2014-2016. The aim of this investigation was to provide data on antimicrobial resistance and consumption, to serve as background knowledge for new veterinary guidelines for prudent and optimal antimicrobial usage in mink. RESULTS: A total number of 308 Escherichia coli isolates, 41 Pseudomonas aeruginosa, 36 Streptococcus canis, 30 Streptococcus dysgalactiae, 55 Staphylococcus delphini, 9 Staphylococcus aureus, and 20 Staphylococcus schleiferi were included in this study. Among E. coli, resistance was observed more frequently among the hemolytic isolates than among the non-hemolytic ones. The highest frequency of resistance was found to ampicillin, 82.3% and 48.0% of the hemolytic of the non-hemolytic isolates, respectively. The majority of the P. aeruginosa isolates were only sensitive to ciprofloxacin and gentamicin. Among the Staphylococcus spp., the highest occurrence of resistance was found for tetracycline. Regarding the nine S. aureus, one isolate was resistant to cefoxitin indicating it was a methicillin-resistant Staphylococcus aureus. Both ß-hemolytic Streptococcus species showed high levels of resistance to tetracycline and erythromycin. The antimicrobial consumption increased significantly during 2007-2012, and fluctuated at a high level during 2012-2016, except for a temporary drop in 2013-2014. The majority of the prescribed antimicrobials were aminopenicillins followed by tetracyclines and macrolides. CONCLUSIONS: The study showed that antimicrobial resistance was common in most pathogenic bacteria from mink, in particular hemolytic E. coli. There is a need of guidelines for prudent use of antimicrobials for mink.

Risk factors associated with diarrhea in Danish commercial mink (Neovison vison) during the pre-weaning period.

BACKGROUND: Pre-weaning diarrhea in mink, also known as "sticky kits", is a syndrome and outbreaks occur every year on commercial mink farms in all mink producing countries. Morbidity and mortality can be considerable on a farm with huge economic consequences for the farmer as well as compromised welfare for the mink kits. Although efforts have been taken to identify etiologic agents involved in outbreaks, the syndrome is still regarded as multifactorial and recurring problems on the same farms draw attention to management and environmental risk factors. In the pre-weaning period from May to June 2015, a case control study was carried out on 30 Danish mink farms. Data concerning management, biosecurity, hygiene, feed consumption, antibacterial prescription and production efficiency were analyzed. RESULTS: The proportion of 1-year old females, farm size (total number of females), energy supply per female in the late gestation period, and dogs accessing the farm area were significantly associated with being a case farm. Case farms were prescribed almost twice the amount of antibacterials per gestational unit (female and litter) as in control farms. Farmers on case farms spent significantly more time nursing and treating the animals and experienced more females with mastitis compared to farmers on control farms. No significant differences in cleaning practices or hygienic measures between case and control farms were found and there were no differences in drinking water quality, bedding material, composition neither of color types nor in management regarding litter equalization. CONCLUSIONS: Results from this study showed an association between the occurrence of pre-weaning diarrhea on mink farms and parity profile, farm size and feeding intensity in the gestational period. The access of dogs to the farm area was a significant risk factor, but needs further clarification.