Mycobacterium avium paratuberculosis infection of calves – The impact of dam infection status

Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic condition of dairy cattle, and is endemic in the UK. Lack of understanding of the relative importance of different transmission routes reduces the impact of control scheme recommendations. The long incubation period for Johne’s disease makes evaluation of control schemes difficult, and so this long-term cohort study offers a rare and valuable insight into the disease epidemiology. A longitudinal study was carried out following a cohort of 440 UK dairy cows in 6 herds recruited in 2012-2013. Individuals entering the milking herd were routinely monitored for the presence of MAP using quarterly milk ELISA testing. Using a Cox proportional-hazards regression model the relationship between time until first detection of infection and dam MAP status was investigated. We then compared the magnitude of the effect of dam status with that of other risk factors in order to understand its relative importance. Dam status was found to be the only observed factor that was significantly associated with time to an individual testing MAP-positive (p = 0.012). When compared to negative dams, we found a marginally significant effect of having a positive dam at time of calving, that increased the hazard of an individual testing positive by a factor of 2.6 (95% confidence interval: 0.89–7.79, p = 0.081). Further positive associations were found with dams becoming positive after the birth of the subject; a dam seroconverting within 12 months post parturition being associated with a 3.6 fold increase in hazard (95% confidence interval: 1.32–9.77, p = 0.013), and dams seroconverting more than a year after calving increased the hazard by a factor of 2.8 (95% confidence interval: 1.39–5.76, p = 0.004). These results suggest that cows may be transmitting MAP to their offspring at an earlier stage than had previously been thought, and so raise important questions about how this transmission may be occurring. The results of the study may have important practical implications for the management on-farm of the offspring of MAP-positive animals, with the potential to vastly reduce the time required to eliminate this chronic disease

Replication of fifteen loci involved in human plasma protein N-glycosylation in 4,802 samples from four cohorts

Human protein glycosylation is a complex process, and its in vivo regulation is poorly understood. Changes in glycosylation patterns are associated with many human diseases and conditions. Understanding the biological determinants of protein glycome provides a basis for future diagnostic and therapeutic applications. Genome-wide association studies (GWAS) allow to study biology via a hypothesis-free search of loci and genetic variants associated with a trait of interest. Sixteen loci were identified by three previous GWAS of human plasma proteome N-glycosylation. However, the possibility that some of these loci are false positives needs to be eliminated by replication studies, which have been limited so far. Here, we use the largest set of samples so far (4,802 individuals) to replicate the previously identified loci. For all but one locus, the expected replication power exceeded 95%. Of the sixteen loci reported previously, fifteen were replicated in our study. For the remaining locus (near the KREMEN1 gene) the replication power was low, and hence replication results were inconclusive. The very high replication rate highlights the general robustness of the GWAS findings as well as the high standards adopted by the community that studies genetic regulation of protein glycosylation. The fifteen replicated loci present a good target for further functional studies. Among these, eight genes encode glycosyltransferases: MGAT5, B3GAT1, FUT8, FUT6, ST6GAL1, B4GALT1, ST3GAL4, and MGAT3. The remaining seven loci offer starting points for further functional follow-up investigation into molecules and mechanisms that regulate human protein N-glycosylation in vivo

Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water

Introduction and objective Mycobacteria have been isolated from almost all types of natural waters, as well as from man-made water distribution systems. Detection of mycobacteria using PCR has been described in different types of water; however, currently, there is no standardised protocol for the processing of large volumes of water. Material and Methods In the present study, different filtering methods are tested and optimised for tap or river water filtration up to 10 L, as well as filter processing and DNA isolation using four commercially available kits. Results The PowerWater DNA isolation kit (MoBio, USA), together with a kit used for soil and other environmental samples (PowerSoil DNA isolation kit, MoBio), had the highest efficiency. Filtration of 10 L of water and elution of the filter in PBS with the addition of 0.05% of Tween 80 is suggested. Conclusions The described protocol for filter elution is recommended, and the use of the PowerWater DNA isolation kit for the highest mycobacterial DNA yield from water samples. The described protocol is suitable for parallel detection of mycobacteria using cultivation

Management Accountants’ Empathy and Their Violation of Fiduciary Duties: a Replication and Extension Study Using fMRI

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Detection of viable Mycobacterium avium subspecies paratuberculosis in powdered infant formula by phage-PCR and confirmed by culture

Surveys from different parts of the world have reported that viable Mycobacterium avium subsp. paratuberculosis (MAP) can be cultured from approximately 2% of samples of retail pasteurised milk samples. Pasteurised milk is used for the production of powdered infant formula (PIF) and therefore there is a concern that MAP may also be present in these products. Several studies have previously reported the detection of MAP in PIF using PCR-based assays. However, culture-based surveys of PIF have not detected viable MAP. Here we describe a phage amplification assay coupled with PCR (page-PCR) that can rapidly detect viable MAP in PIF. The results of a small survey showed that the phage-PCR assay detected viable MAP in 13% (4/32) of PIF samples. Culture detected viable MAP in 9% (3/32) PIF samples, all of which were also phage-PCR positive. Direct IS900 PCR detected MAP DNA in 22% (7/32) of PIF samples. The presence of viable MAP in PIF indicates that MAP either survived PIF manufacturing or that post-production contamination occurred. Irrespective of the route of MAP contamination, the presence of viable MAP in PIF is a potential public health concern