Riables were derived and used to select dogs to the study groups. The variables that

Riables were derived and used to select dogs to the study groups. The variables that we were interested the most were fear towards unfamiliar people (human fear_frequency, human fear_intensity), fearfulness total and noise sensitivity. Human fear_frequency was simply the owner reported frequency of dog showing fearful reaction when meeting a stranger (frequency scoring 0 = never; 1 = 0?0 of the occasions; 2 = 40?0 of the occasions; 3 = 60?00 of the occasions: 4 = alwayswhen meeting unfamiliar people). Human fear_intensity was calculated as follows: the frequency of showing fearful reaction when meeting unfamiliar people was multiplied with the sum of owner recorded fearful behavioral reactions. Each type of behavior equaled 1, except the avoidance-reaction which was weighted by multiplying it with 5. Fearfulness variable was calculated as a sum of frequencies of showing fearful behavioural reactions towards unfamiliar people (see scoring above 0?), unfamiliar dogs (0?) and in new situations (0?), and thus the score varied between 0 and 12. In addition, we calculated a variable describing the dog’s fear of loud noises (noise sensitivity), by calculating a sum of frequencies of showing a fearful reaction towards thunder (see scoring above 0?), fireworks (0?) and gunshot (0?). The behaviour of seven of the dogs was verified by a short 5-min test conducted by same person for all the dogs–not all dogs were tested as some had already died between the blood sampling and behavioral testing, or lived too far. Shortly, test consisted of three parts; meeting an unfamiliar person, exploration in the novel space, and novel object test. More details of the test can be found from Tiira and Lohi, 2014. We selected 10 fearful and 10 non-fearful Great Danes for the study, and detailed information about all the individual dogs is presented in Table 1. Our criteria for non-fearful dogs was that all the variables (human fear_ frequency, human fear_intensity, fearfulness total and noise sensitivity) had to have score 0. In the case group, our main inclusion criterion was that the dog had to show fear towards unfamiliar people at 40?00 of all situations (human fear_frequency score 2?). In addition, dog’s needed to have fearfulness score >2. Additionally to these criteria, we used matched pairs with approximately same age for blood samples between case and control groups. We aimed, at first, to get only males, however, in order to keep the age of blood sampling approximately same in both control and case groups we also had to include two females for both groups. EDTA-blood samples were collected from each dog and stored in -20 degrees. The blood samples were collected from the privately owned Finnish dogs with owners consent under a valid ethical license (Finnish National Animal Experiment Board, ELLA, license number ESAVI/6054/04.10.03/2012).Dietary informationThe owners were retrospectively asked to report the diet of the dog at the time of blood sampling to help us consider possible nutritional effects on order EPZ004777 26266977″ title=View Abstract(s)”>PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26266977 metabolite profiles. Dietary information was collected from 17 out of 20 dogs (two cases and one control missing). Comparison of the diet profiles indicated only minor differences betweenPuurunen et al. Behav Brain Funct (2016) 12:Page 3 ofTable 1 Demographics of the dogsAge (years) Mean age (SD) Sex 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 1.1 1.5 2.4 2.8 4.2 5.4 4.4 9.3 1.8 1.6 3.2 4.5 3.3 1.1 8.5 4.8 3.3 1.6 2 3.4 (2.2) 3.5 (2.5) Male.