Racehorse welfare is gaining increasing public attention, however scientific evidence in this area is lacking. In order to develop a better understanding of racehorse welfare, it must be measured and monitored. This is the first study to assess racehorse welfare using scientific objective methods across a training season. The aim of this study was threefold, firstly to investigate welfare measures which could be used in the first welfare assessment protocol for racehorses. Secondly, to understand the effect that a racing and training season had on individual racehorses and thirdly to identify risk factors for both good and poor welfare. Thirteen racehorse training yards were visited at the beginning and the peak of the racing season in England. Behavioral observations along with individual environmental and animal-based welfare measures were carried out on 353 horses in 13 training yards selected for variability. In our sample the horses were generally in good physical health: 94% of horses recorded as an ideal body condition score, no horses had signs of hoof neglect and 77.7% had no nasal discharge. The overall prevalence of external Mouth Corner Lesions was 12.9% and was significantly higher for Flat racing than Jump racing horses. The majority of horses (67.5%) showed positive horse human interactions. When stabled 54.1% horses had physical social contact and nasal discharge was not associated with increased physical contact. The training season significantly affected Human Reactivity Tests, Horse Grimace Scale scores and time spent resting and feeding. A total of 14.5% of horses displayed stereotypic behavior on at least two occasions. Horses with windows in their stables spent more time surveying their surroundings. Overall, in this population of racehorses, horses spent around a third of their daytime feeding (33.7%) followed by time spent standing resting (22.6%). The welfare assessment protocol used in this study is suitable for use in industry to collect welfare data on racehorses.