Considering the interrelationship between the landscape, ways, and walking, and the significance of walking trails with the theoretical lens of landscape studies, this study analyzes connections with and characteristics of human and non-human elements in the landscape of the Shin-etsu Trail. It is a 110 km track passing along the border between Nagano and Niigata Prefectures and is a well-known nature trail in Japan; many other trails have been created or re-established based on it. Addressing the issues that research on recreational walkways is currently insufficient especially in the non-Western context, and that natural components of landscapes are not sufficiently studied in tourism research, this inquiry explores the ecological characteristics of the trail landscape and how local stakeholders perceive them. This case study adopted a qualitative method with the Actor-Network Theory, involving walking, observation, and interviews. In the trailscape embracing many actors, this paper focuses on networks between the natural environment and Japan’s traditional hunter-gatherers known as the matagi, whose practice has been conducted in the Akiyama-go, the settlement area through which the trail passes, since the 18th century at the latest. This hunting practice can be currently the marginal relativity interconnecting with natural actants in the uncertain environmental and social changes such as global warming, technological advances, and aging and depopulation. However, some stakeholders of the trail do not fully perceive such characteristics and they are poorly reflected in trail management. This study contributes to suggesting the prospect and importance of ecological anthropology in tourism and landscape research in terms of the non-anthropocentric argument on nature-human connectedness and concludes that raising their awareness of the traits, applying them to the management, and inducing hikers to be more aware of them are crucial for sustainable trail-based tourism leading to enlivening and envisioning the matagi’s traditional ecological knowledge and practices.