Canines are one of the best biological detectors of energetic materials available; however, canine detection of explosives is impacted by a number of factors, including environmental conditions. The objectives of this study were: 1) determine how canine detection limits vary when both the canine and odorant are tested in varying temperature and humidity conditions (canine and odor interactive effects); and 2) determine if an acclimatization plan can improve detection limits in an adverse environmental condition. Eight working line canines were trained to detect four energetics: prill ammonium nitrate (AN), Composition 4 (C4), trinitrotoluene (TNT) and double base smokeless powder (SP). In Experiment 1, canines completed a 3-alternative forced choice 3-down-1-up staircase threshold assessment in five environmental conditions: 40°C and 70% relative humidity (RH), 40°C and 40% RH, 0°C and 90% RH, 0°C and 50% RH and 21°C and 50% RH. Canines showed a 3.5-fold detection limit increase (poorer detection) for C4 in 40°C and 70% RH compared to their detection limit at 21°C and 50% RH. In Experiment 2, the eight canines were split into two groups (n = 4), control and acclimation groups. The control group completed the threshold assessment for C4 at 21°C and 50% RH each day for 20 days, with 5 minutes of petting prior to testing. The acclimation group completed the same assessment daily starting at 21°C and 50% RH but temperature and RH were incremented daily over the course of 6 days to the 40°C and 70% RH condition. After the initial six days, the acclimation group completed daily assessments at 40°C and 70% RH condition for the remainder of the experiment. All acclimatization group canines started their session with 5 minutes of toy or food retrieves. Detection limits for C4 for all dogs were tested in 40°C and 70% RH on day 11 and day 22. The acclimatization plan improved detection limits in the 40°C and 70% RH condition for C4 compared to the non-acclimated group. In this set of experiments, canine detection limits for four explosive odorants were found to vary based on environmental condition and were mostly driven by impacts on the canine rather than odor availability. The acclimatization plan did result in lower detection limits (i.e., increased performance). Future work should determine what factor (exercise or environmental exposure) is more effective in acclimatization for odor detection work.