As the opioid epidemic continues across the United States, law enforcement K9s (LEK9s) are at increased risk of accidental exposure and overdose. This study evaluated a novel training program teaching handlers to administer naloxone to their LEK9 in the event of an overdose. Seventy-five LEK9 handlers from a governmental agency attended a naloxone training session. A presurvey given to the handlers evaluated their knowledge of opioid overdose in LEK9s and their confidence administering naloxone. Officers were educated via a PowerPoint presentation about naloxone and how to administer it to their LEK9. A postsurvey evaluated changes in their knowledge and confidence as a result of the presentation. Sixty-two presurveys and 47 postsurveys were completed. Nearly all handlers had never given their LEK9 an intramuscular or intranasal injection. Most handlers were not comfortable monitoring their LEK9’s vital signs for an opioid overdose. After the training, handlers demonstrated a mild increase in comfort level administering intramuscular and intranasal naloxone (15 and 14% increase, respectively). Comfort level monitoring vital signs and symptoms of an opioid overdose increased 38 and 32%, respectively. Handlers may not be fully prepared to assess and treat their LEK9 and may benefit from a targeted training program teaching them to administer naloxone.