Palliativ care in nursing home - Health professionals experience

Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages

Absence of Actinobacillus pleuropneumoniae in semen from serologically positive tested AI-boars

IntroductionTransmission of Actinobacillus pleuropneumoniae (Ap) between animals occurs mainly due to direct contact between pigs, due to nose-nose contact or uptake of nasal/oral fluids. To assess the risk of transmission of Ap by semen, first it isneeded to know whether Ap can be detected in semen. The aim of this study was to evaluate the performance of a qPCR for the ApxIVA gene, on semen and test semen of seropositive but healthy boars for the presence of Ap.Materials and MethodsTo enable detection of the ApxIVA gene by qPCR in semen, the validated protocol for tonsil brush samples was adjusted, according to a validated protocol for detection of Brucellosis in semen. In short, DNA isolation was performed using the DNEasy kit. To assess the qPCR efficiency and minimal detection limit for Ap in fresh and undiluted semen a pooled semen sample was spiked with a 10-fold serial dilution of Ap in triplicate. Thereafter DNA isolation was performed and the qPCR was performed as described earlier (Tobias, 2012). Finally, 19 fresh and undiluted semen samples of serologically positive boars (by ApxIV ELISA and/or LC-LPS ELISA) were processed and tested by ApxIVA qPCR.ResultsThe minimal detection limit of the ApxIVA qPCR in semen was >34 - ”340 Ap DNA copies per reaction, when testing spiked semen. None of the semen samples of serologically positive tested boars (0/19) returned a positive qPCR result.ConclusionThis study shows that ApxIVA qPCR testing of semen for Ap is feasible, but that detection limit increased from 5 copies / reaction in tonsil brush material to between 34 and 340 Ap DNA copies /reaction, which equals ~103 – 104 CFU /mL semen. As seminal transmission is already considered unlikely and in addition targeted screening of semen samples of serologically positive boars resulted in negative results, the risk of Ap seminal transmission by serological positive boars seems low

Absence of Actinobacillus pleuropneumoniae in semen from serologically positive tested AI-boars

IntroductionTransmission of Actinobacillus pleuropneumoniae (Ap) between animals occurs mainly due to direct contact between pigs, due to nose-nose contact or uptake of nasal/oral fluids. To assess the risk of transmission of Ap by semen, first it isneeded to know whether Ap can be detected in semen. The aim of this study was to evaluate the performance of a qPCR for the ApxIVA gene, on semen and test semen of seropositive but healthy boars for the presence of Ap.Materials and MethodsTo enable detection of the ApxIVA gene by qPCR in semen, the validated protocol for tonsil brush samples was adjusted, according to a validated protocol for detection of Brucellosis in semen. In short, DNA isolation was performed using the DNEasy kit. To assess the qPCR efficiency and minimal detection limit for Ap in fresh and undiluted semen a pooled semen sample was spiked with a 10-fold serial dilution of Ap in triplicate. Thereafter DNA isolation was performed and the qPCR was performed as described earlier (Tobias, 2012). Finally, 19 fresh and undiluted semen samples of serologically positive boars (by ApxIV ELISA and/or LC-LPS ELISA) were processed and tested by ApxIVA qPCR.ResultsThe minimal detection limit of the ApxIVA qPCR in semen was >34 - ”340 Ap DNA copies per reaction, when testing spiked semen. None of the semen samples of serologically positive tested boars (0/19) returned a positive qPCR result.ConclusionThis study shows that ApxIVA qPCR testing of semen for Ap is feasible, but that detection limit increased from 5 copies / reaction in tonsil brush material to between 34 and 340 Ap DNA copies /reaction, which equals ~103 – 104 CFU /mL semen. As seminal transmission is already considered unlikely and in addition targeted screening of semen samples of serologically positive boars resulted in negative results, the risk of Ap seminal transmission by serological positive boars seems low

Faecal carriage, risk factors, acquisition and persistence of ESBL-producing Enterobacteriaceae in dogs and cats and co-carriage with humans belonging to the same household

BACKGROUND: ESBL-producing Enterobacteriaceae (ESBL-E) are observed in many reservoirs. Pets might play an important role in the dissemination of ESBL-E to humans since they live closely together. OBJECTIVES: To identify prevalence, risk factors, molecular characteristics, persistence and acquisition of ESBL-E in dogs and cats, and co-carriage in human-pet pairs belonging to the same household. METHODS: In a nationwide study, one person per household was randomly invited to complete a questionnaire and to submit a faecal sample. Dog and cat owners were invited to also submit a faecal sample from their pet. Repeated sampling after 1 and 6 months was performed in a subset. ESBL-E were obtained through selective culture and characterized by WGS. Logistic regression analyses and random forest models were performed to identify risk factors. RESULTS: The prevalence of ESBL-E carriage in these cohorts was 3.8% (95% CI: 2.7%-5.4%) for human participants (n=550), 10.7% (95% CI: 8.3%-13.7%) for dogs (n=555) and 1.4% (95% CI: 0.5%-3.8%) for cats (n=285). Among animals, blaCTX-M-1 was most abundant, followed by blaCTX-M-15. In dogs, persistence of carriage was 57.1% at 1 month and 42.9% at 6 months. Eating raw meat [OR: 8.8, 95% CI: 4.7-16.4; population attributable risk (PAR): 46.5%, 95% CI: 41.3%-49.3%] and dry food (OR: 0.2, 95% CI: 0.1-0.5; PAR: 56.5%, 95% CI: 33.2%-66.6%) were predictors for ESBL-E carriage in dogs. Human-dog co-carriage was demonstrated in five households. Human-cat co-carriage was not observed. CONCLUSIONS: ESBL-E prevalence was higher in dogs than in humans and lowest in cats. The main risk factor for ESBL-E carriage was eating raw meat. Co-carriage in dogs and household members was uncommon.</p

Genomic analysis of European bovine Staphylococcus aureus from clinical versus subclinical mastitis

Intramammary infections (IMI) with Staphylococcus aureus are a common cause of bovine mastitis and can result in both clinical (CM) or subclinical mastitis (SCM). Although bacterial isolates of S. aureus differ in their virulence potential it is largely unclear which bacterial virulence factors are responsible for increased clinical severity. We performed a genome wide association study and used a generalized linear mixed model to investigate the correlation between gene carriage, lineage and clinical outcome of IMI in a collection of S. aureus isolates from cattle with CM (n = 125) and SCM (n = 151) from 11 European countries. An additional aim was to describe the genetic variation of bovine S. aureus in Europa. The dominant lineages in our collection were clonal complex (CC) 151 (81/276, 29.3%), CC97 (54/276, 19.6%), CC479 (32/276, 11.6%) and CC398 (19/276, 6.9%). Virulence and antimicrobial resistance (AMR) gene carriage was highly associated with CC. Among a selection of nine virulence and AMR genes, CC151, CC479 and CC133 carried more virulence genes than other CCs, and CC398 was associated with AMR gene carriage. Whereas CC151, CC97 were widespread in Europe, CC479, CC398 and CC8 were only found in specific countries. Compared to CC151, CC479 was associated with CM rather than SCM (OR 3.62; 95% CI 1.38-9.50) and the other CCs were not. Multiple genes were associated with CM, but due to the clustering within CC of carriage of these genes, it was not possible to differentiate between the effect of gene carriage and CC on clinical outcome of IMI. Nevertheless, this study demonstrates that characterization of S. aureus CC and virulence genes helps to predict the likelihood of the occurrence of CM following S. aureus IMI and highlights the potential benefit of diagnostics tools to identify S. aureus CC during bovine mastitis