Summary1. It has been proposed that growth and reproduction of animals is frequently limited by multiple nutrients simultaneously. To improve our understanding of the consequences of multiple nutrient limitations (i.e. co-limitation) for the performance of animals, we conducted standardized population growth experiments using an important aquatic consumer, the rotifer Brachionus calyciflorus. 2. We compared nutrient profiles (sterols, fatty acids and amino acids) of rotifers and their diets to reveal consumer-diet imbalances and thus potentially limiting nutrients. In concomitant growth experiments, we directly supplemented potentially limiting substances (sterols, fatty acids, amino acids) to a nutrient-deficient diet, the cyanobacterium Synechococcus elongatus, and recorded population growth rates. 3. The results from the supplementation experiments corroborated the nutrient limitations predicted by assessing consumer-diet imbalances, but provided more detailed information on co-limiting nutrients. While the fatty acid deficiency of the cyanobacterium appeared to be of minor importance, the addition of both cholesterol and certain amino acids (leucine and isoleucine) improved population growth rates of rotifers, indicating a simultaneous limitation by sterols and amino acids. 4. Our results add to growing evidence that consumers frequently face multiple nutrient limitations and suggest that the concept of co-limitation has to be considered in studies assessing nutrient-limited growth responses of consumers.