RESUMO
This study aimed to evaluate the state of welfare of a group of dogs during the first month after entering the shelter by using different stress parameters. Blood and fecal samples were collected from a group of 71 dogs at the time of admission to the shelter. In 46 of these dogs, sampling was repeated after four weeks. Well-recognized welfare biomarkers, such as fecal cortisol and leukocytes, as well as some innovative parameters (ß-endorphin and lysozyme) were determined. Uni- and multivariate statistical analyses were used to evaluate their interactions and changes over time. Neutrophils (p < 0.01), lysozyme (p < 0.05), and fecal cortisol (p < 0.05) decreased, while lymphocytes (p < 0.05) increased after four weeks compared to the first days of being in the shelter, suggesting an improvement in the dogs' welfare over time. A principal component analysis extracted three bipolar components (PCs), explaining 75% of the variance and indicating negative associations between neutrophil and lymphocyte (PC1), lysozyme and ß-endorphin (PC2), cortisol and lysozyme (PC3). The associations between these variables within each PC also confirmed the intricate relationships between the hypothalamic-pituitary-adrenal (HPA) axis and the immune system as well as the importance of a multiparametric approach in evaluating welfare.
RESUMO
To date, a plenty of techniques for the detection of JAK2V617F is used over different laboratories, with substantial differences in specificity and sensitivity. Therefore, to provide reliable and comparable results, the standardization of molecular techniques is mandatory.A network of 19 centers was established to 1) evaluate the inter- and intra-laboratory variability in JAK2V617F quantification, 2) identify the most robust assay for the standardization of the molecular test and 3) allow consistent interpretation of individual patient analysis results. The study was conceived in 3 different rounds, in which all centers had to blindly test DNA samples with different JAK2V617F allele burden (AB) using both quantitative and qualitative assays.The positivity of samples with an AB < 1% was not detected by qualitative assays. Conversely, laboratories performing the quantitative approach were able to determine the expected JAK2V617F AB. Quantitative results were reliable across all mutation loads with moderate variability at low AB (0.1 and 1%; CV = 0.46 and 0.77, respectively). Remarkably, all laboratories clearly distinguished between the 0.1 and 1% mutated samples.In conclusion, a qualitative approach is not sensitive enough to detect the JAK2V617F mutation, especially at low AB. On the contrary, the ipsogen JAK2 MutaQuant CE-IVD kit resulted in a high, efficient and sensitive quantification detection of all mutation loads. This study sets the basis for the standardization of molecular techniques for JAK2V617F determination, which will require the employment of approved operating procedures and the use of certificated standards, such as the recent WHO 1st International Reference Panel for Genomic JAK2V617F.