Conducting a periodical systematic review on a given field of study enables to comprehend the conceptual development of a discipline. A decade has passed since the last in-depth systematic review on contemporary ecotourism research has been published. This study is, therefore, executed to systematically analyze ecotourism research articles published in the previous twenty-five years and thereby to reassess the past, understand the present and envisage the future of ecotourism development. 470 articles published in nine top-tier tourism journals are screened and analyzed. Thematic and summative content analyses are employed to examine the contents of each publication. The findings highlight that over the years, researchers have examined myriads of ecotourism research themes. Despite fluctuations over the years, the overall publication trend shows growth. Most ecotourism studies are conducted in developing countries, with Asia taking the lead followed by Africa and Latin America. However, in terms of researchers’ domicile, the USA becomes the first followed by Australia, the UK, and Canada. Despite modest successes, overall, the contribution of ecotourism towards sustainable destination development remains indiscernible. The current study provides up-to-date insight into the ecotourism literature by highlighting the main themes and trends of ecotourism research over the last twenty-five years. Research findings also enhance our understanding of the evolution of ecotourism research and ignite further study in ecotourism by suggesting issues that merit rigorous investigation.