Since the spread of white-nose syndrome in North America, several bat species have shown precipitous declines in abundance and distribution. With lower netting detection probabilities for the currently threatened but proposed endangered northern long-eared bat (Myotis septentrionalis) and endangered Indiana bats (Myotis sodalis), determination of presence or absence for regulatory clearance often has shifted to the use of acoustic sur- veys. However, acoustic surveys are unable to differentiate between non-reproductive individuals versus a maternity colony. We used recorded nightly echolocation pass counts of bat species-specific probabilities with maximum likelihood estimator (MLE) scores to determine thresholds by cover type and reproductive period whereby the potential for northern long-eared bat or Indiana bat maternity colonies occurs. Where nightly MLE P-values were

<0.05, mean predicted nightly pass counts were significantly higher in areas of known northern long-eared bat and Indiana bat maternity colonies versus sites that were in either species’ distribution but with no maternity activity known. Nightly pass counts (MLE P < 0.05) were higher for sites with observed maternity activity for both bat species across forest, forest-field edge, and riparian areas versus sites where no maternity activity was known. For northern long-eared bats, nightly pass counts were highest in the juvenile volancy period (after 15 July) whereas, for Indiana bats, nightly pass counts were highest in the lactation period (16 June to 15 July). Except for edge conditions for northern long-eared bats, a MLE P < 0.05 combined with nightly pass counts above thresholds developed from surveys at known maternity colony sites for both species may indicate potential presence of a maternity colony locally and provide a tool to more efficiently use targeted mist-netting for further determination.