In social insects, colony fission is a particular mode of dispersal by which an adult colony splits into two or more independent nests. In the monogynous ant Aphaenogaster senilis, field data suggest that new queens may be produced in queenless daughter nests after nest relocation. Because workers do not fly, colony fission limits dispersal distance, leading young sister colonies to compete together and with the mother queen. In the present study we analysed the effects of queen loss and diet change on nestmate recognition. Queenright colonies were separated into two queenless and one queenright fragments. One queenless group received the same food as the queenright group, while the other queenless group received a different diet for 150 days. Recognition bioassays revealed that aggression between queenright and queenless former nestmates increased progressively until day 20, when they could no longer be reunited. Different diets also induced aggression between orphaned groups. Chemical analyses indicated that cuticular hydrocarbon profiles were already different between groups after 5 days. Overall, our results are in accordance with the graded model of nestmate recognition and suggest that the loss of the mother queen progressively leads to the independence of the related nests after fission. This may also allow queenless ants to merge again with their mother colony during a short time window after fission.