Abstract Animals modify their behaviours and interactions in response to changing environments. In bats, environmental adaptations are reflected in echolocation signalling that is used for navigation, foraging and communication. However, the extent and drivers of echolocation plasticity are not fully understood, hindering our identification of bat species with ultrasonic detectors, particularly for cryptic species with similar echolocation calls. We used a combination of DNA barcoding, intensive trapping, roost and emergence surveys and acoustic recording to study a widespread European cryptic species complex (Pipistrellus pipistrellus and Pipistrellus pygmaeus) to investigate whether sibling bat species could exhibit extreme echolocation plasticity in response to certain environmental conditions or behaviours. We found that P. pygmaeus occupied the acoustic niche of their absent congeneric species, producing calls with P. pipistrellus’ characteristic structure and peak frequencies and resulting in false positive acoustic records of that species. Echolocation frequency was significantly affected by the density of bats and by maternity rearing stage, with lower frequency calls emitted when there was a high density of flying bats, and by mothers while juveniles were non-volant. During roost emergence, 29% of calls had peak frequencies typical of P. pipistrellus, with calls as low as 44 kHz, lower than ever documented. We show that automatic and manual call classifiers fail to account for echolocation plasticity, misidentifying P. pygmaeus as P. pipistrellus. Our study raises a vital limitation of using only acoustic sampling in areas with high densities of a single species of a cryptic species pair, with important implications for bat monitoring. Significance statement Ultrasonic acoustic detectors are widely used in bat research to establish species inventories and monitor species activity through identification of echolocation calls, enabling new methods to study and understand this elusive understudied group of nocturnal mammals. However, echolocation call signalling in bats is intrinsically different to that of other taxa, serving a main function of navigation and foraging. This study demonstrates an extreme level of plasticity, showing large variation in call frequency and structure in different situations. We showcase the difficulty and limitation in using acoustic sampling alone for bat monitoring and the complications of setting parameters for species identification for manual and automatic call classifiers. Our observations of call frequency variation correlated with density and absence of congenerics provide novel insights of behavioural echolocation plasticity in bats.