Ecotourism is considered an effective agent to conserve environmentally fragile areas while bringing economic opportunities to local communities at the same time. In the past decades, empirical studies about ecotourism's conservation effects on wetland ecosystems attracted relatively less academic attention than rainforests and coral reefs. Florida, listed as one of the states with the greatest share of wetland loss in the U.S. due to rapid growth in agriculture, tourism, and urban development, has a small number of existing wetlands under the protection of the park and reserve system. To generate long-term positive environmental impacts, ecotourism stakeholders' contributions to planning and management activities could be an alternative beyond land use controls for conserving Florida wetlands. The major objective of this study is to explore the relationship between ecotourism and wetland conservation contributions which lead to long-term environmental sustainability. The study surveyed 97 nature-based tour operators in Florida on their activeness in a set of wetland planning and management behavior, the proportion of tourism revenue from ecotourism, and the perceived tourism benefits regarding economic, socio-cultural, and ecological aspects. Factor analysis was employed to identify indicators for the composite factors, such as the conservation contributions and incentives (i.e., economic benefits, socio-cultural benefits, and ecological benefits). The effects of ecotourism involvement and conservation incentives on tour operators' participation in wetland conservation practices were statistically modeled. The results demonstrated the business characteristics, degrees of the perceived tourism benefits, and frequency of participation in wetland conservation activities of responding tour operators. On average, tour operators were not as active in wetland management and planning approaches leading to the long-term environmental health as theories suggest they should be. Generally, the regression analysis results illustrated the significant association between ecotourism involvement and tour operators' conservation contribution. It is noteworthy that the effects of ecotourism involvement on tour operators' participation in environmental planning and water management processes were relatively important. In addition, the incentive of the perceived socio-cultural benefits was identified as the leading factor by regression analysis. The findings lent to the policy suggestions in expanding the incentives to drive major stakeholders' active engagement in wetland conservation planning and management, which is critical for collaborative and adaptive management.