Introduction French Guiana is a French territory in South America covered by 90% of tropical forest. Despite regular first aid standards training, some of the workers in isolated areas remain untrained in common problems they may encounter. We present here the knowledge assessment of these workers concerning first aid and a new training program. Methods Thirty-two workers operating in remote areas attended a 1-d training course. The program covered infectious diseases, wildlife envenomation, and first aid and basic life support with a wilderness medicine focus. Questionnaires were issued to assess their knowledge and satisfaction with the course. Results The learners were 78% male and of intermediate age (26–55 y). The most common profession was forester (53%). Professionals had been working in remote areas for 12 (4–19) y, and 91% had already received first aid training. The common life-threatening emergencies were not well known; only 6 learners (19%) knew how to recognize a cardiac arrest, 7 (22%) recognized symptoms of severe anaphylaxis, and 23 (72%) performed maneuvers not recommended in cases of upper airway partial obstruction. A total of 26 learners (81%) completed the satisfaction questionnaire. The training met the learners’ expectations. However, 92% would like further training, and 96% requested regular renewal. Conclusions This training was the first step toward the creation of a specialized wilderness first aid course. This work supports the need to reinforce training for nonmedical personnel by integrating regionally specific topics. This experience permitted the rise of a working group that proposed a medical kit as well as first aid protocols for workers in isolated areas.