Animal vocalizations contain information about individual identity that could potentially be used for the monitoring of individuals. However, the performance of individual discrimination is subjected to many biases depending on factors such as the amount of identity information, or methods used. These factors need to be taken into account when comparing results of different studies or selecting the most cost-effective solution for a particular species. In this study, we evaluate several biases associated with the discrimination of individuals. On a large sample of little owl male individuals, we assess how discrimination performance changes with methods of call description, an increasing number of individuals, and number of calls per male. Also, we test whether the discrimination performance within the whole population can be reliably estimated from a subsample of individuals in a pre-screening study. Assessment of discrimination performance at the level of the individual and at the level of call led to different conclusions. Hence, studies interested in individual discrimination should optimize methods at the level of individuals. The description of calls by their frequency modulation leads to the best discrimination performance. In agreement with our expectations, discrimination performance decreased with population size. Increasing the number of calls per individual linearly increased the discrimination of individuals (but not the discrimination of calls), likely because it allows distinction between individuals with very similar calls. The available pre-screening index does not allow precise estimation of the population size that could be reliably monitored. Overall, projects applying acoustic monitoring at the individual level in population need to consider limitations regarding the population size that can be reliably monitored and fine-tune their methods according to their needs and limitations.