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Dogs can be trained for various odour detection tasks, and have been used to search for explosives, drugs or cancer biomarkers. However, there are only a few studies that have attempted to train dogs for oestrus detection using vaginal fluid, milk or urine of cows. In this study, seven dogs were trained utilizing special training equipment to discriminate oestrus against dioestrus scent in vaginal fluid, urine and milk of dairy cows. We set out (1) to determine the length of the learning phase of dogs using vaginal discharge to identify cows in oestrus, (2) to prove that dogs can generalize oestrus specific odour of cows and are not limited to an individual cow scent in oestrus, and (3) to study if dogs can identify oestrus specific odour in urine and milk, after they have been trained with vaginal fluid. Our results demonstrate that dogs can be trained for oestrus detection with vaginal swabs or urine of cows using special laboratory equipment within 52 positive reinforced contacts to the target scent. The dogs were correctly identifying the oestrus sample with a maximum accuracy of 80.3%. They could verify this accuracy in a blinded test situation testing novel samples. The dogs in this study were able to detect oestrus in urine and milk samples spontaneously, after being trained with vaginal fluid samples.