Research investigating the behavioral and physiological responses of bats to echoes typically includes analysis of acoustic signals from microphones and/or microphone arrays, using time difference of arrival (TDOA) between array elements or the microphones to locate flying bats (azimuth and elevation). This has provided insight into transmission adaptations with respect to target distance, clutter, and interference. Microphones recording transmitted signals and echoes near a stationary bat provide sound pressure as a function of time but no directional information. This dissertation introduces spatial audio techniques to bat biosonar studies as a complementary method to the current TDOA based acoustical study methods. This work proposes a couple of feasible methods based on spatial audio techniques, that both track bats in flight and pinpoint the directions of echoes received by a bat. A spatial audio/soundfield microphone array is introduced to measure sounds in the sonar frequency range (20-80 kHz) of the big brown bat (Eptesicus fuscus). The custom-built ultrasonic tetrahedral soundfield microphone consists of four capacitive microphones that were calibrated to match magnitude and phase responses using a transfer function approach. Ambisonics, a signal processing technique used in three-dimensional (3D) audio applications, is used for the basic processing and reproduction of the signals measured by the soundfield microphone. Ambisonics provides syntheses and decompositions of a signal containing its directional properties, using the relationship between the spherical harmonics and the directional properties. As the first proposed method, a spatial audio decoding technique called HARPEx (High Angular Resolution Planewave Expansion) was used to build a system providing angle and elevation estimates. HARPEx can estimate the direction of arrivals (DOA) for up to two simultaneous sources since it decomposes a signal into two dominant planewaves. Experiments proved that the estimation system based on HARPEx provides accurate DOA estimates of static or moving sources. It also reconstructed a smooth flight-path of a bat by accurately estimating its direction at each snapshot of pulse measurements in time. The performance of the system was also assessed using statistical analyses of simulations. A signal model was built to generate microphone capsule responses to a virtual source emitting an LFM signal (3 ms, two harmonics: 40-22 kHz and 80-44 kHz) at an angle of 30° in the simulations. Medians and RMSEs (root-mean-square error) of 10,000 simulations for each case represent the accuracy and precision of the estimations, respectively. Results show lower d (distance between a capsule and the soundfield microphone center) or/and higher SNR (signal-to-noise ratio) are required to achieve higher estimator performance. The Cramer-Rao lower bounds (CRLB) of the estimator are also computed with various d and SNR conditions. The CRLB which is for TDOA based methods does not cover the effects of different incident angles to the capsules and signal delays between the capsules due to a non-zero d, on the estimation system. This shows the CRLB is not a proper tool to assess the estimator performance. For the second proposed method, the matched-filter technique is used instead of HARPEx to build another estimation system. The signal processing algorithm based on Ambisonics and the matched-filter approach reproduces a measured signal in various directions, and computes matched-filter responses of the reproduced signals in time-series. The matched-filter result points a target(s) by the highest filter response. This is a sonar-like estimation system that provides information of the target (range, direction, and velocity) using sonar fundamentals. Experiments using a loudspeaker (emitter) and an artificial or natural target (either stationary or moving) show the system provides accurate estimates of the target's direction and range. Simulations of imitating a situation where a bat emits a pulse and receives an echo from a target (30°) were also performed. The echo sound level is determined using the sonar equation. The system processed the virtual bat pulse and echo, and accurately estimated the direction, range, and velocity of the target. The simulation results also appear to recommend an echo level over -3 dB for accurate and precise estimations (below 15% RMSE for all parameters). This work proposes two methods to track bats in flight or/and pinpoint the directions of targets using spatial audio techniques. The suggested methods provide accurate estimates of the direction, range, or/and velocity of a bat based on its pulses or of a target based on echoes. This demonstrates these methods can be used as key tools to reconstruct bat biosonar. They would be also an independent tool or a complementary option to TDOA based methods, for bat echolocation studies. The developed methods are believed to be also useful in improving man-made sonar technology. / Doctor of Philosophy / While bats are one of the most intriguing creatures to the general population, they are also a popular subject of study in various disciplines. Their extraordinary ability to navigate and forage irrespective of clutter using echolocation has gotten attention from many scientists and engineers. Research investigating bats typically includes analysis of acoustic signals from microphones and/or microphone arrays. Using time difference of arrival (TDOA) between the array elements or the microphones is probably the most popular method to locate flying bats (azimuth and elevation). Microphone responses to transmitted signals and echoes near a bat provide sound pressure but no directional information. This dissertation proposes a complementary way to the current TDOA methods, that delivers directional information by introducing spatial audio techniques. This work shows a couple of feasible methods based on spatial audio techniques, that can both track bats in flight and pinpoint the directions of echoes received by a bat. An ultrasonic tetrahedral soundfield microphone is introduced as a measurement tool for sounds in the sonar frequency range (20-80 kHz) of the big brown bat (Eptesicus fuscus). Ambisonics, a signal processing technique used in three-dimensional (3D) audio applications, is used for the basic processing of the signals measured by the soundfield microphone. Ambisonics also reproduces a measured signal containing its directional properties. As the first method, a spatial audio decoding technique called HARPEx (High Angular Resolution Planewave Expansion) was used to build a system providing angle and elevation estimates. HARPEx can estimate the direction of arrivals (DOA) for up to two simultaneous sound sources. Experiments proved that the estimation system based on HARPEx provides accurate DOA estimates of static or moving sources. The performance of the system was also assessed using statistical analyses of simulations. Medians and RMSEs (root-mean-square error) of 10,000 simulations for each simulation case represent the accuracy and precision of the estimations, respectively. Results show shorter distance between a capsule and the soundfield microphone center, or/and higher SNR (signal-to-noise ratio) are required to achieve higher performance. For the second method, the matched-filter technique is used to build another estimation system. This is a sonar-like estimation system that provides information of the target (range, direction, and velocity) using matched-filter responses and sonar fundamentals. Experiments using a loudspeaker (emitter) and an artificial or natural target (either stationary or moving) show the system provides accurate estimates of the target's direction and range. Simulations imitating a situation where a bat emits a pulse and receives an echo from a target (30°) were also performed. The system processed the virtual bat pulse and echo, and accurately estimated the direction, range, and velocity of the target. The suggested methods provide accurate estimates of the direction, range, or/and velocity of a bat based on its pulses or of a target based on echoes. This demonstrates these methods can be used as key tools to reconstruct bat biosonar. They would be also an independent tool or a complementary option to TDOA based methods, for bat echolocation studies. The developed methods are also believed to be useful in improving sonar technology.