Nowadays, Laos remains one of the scientifically least known countries of Asia in terms of herpetological knowledge. Here, we evaluate composition of species in freshwater ecosystems (main river courses) and terrestrial ecosystems (forests) in two distinct regions using Visual Encounter Surveys along designed transects, examination of fishers’ catches and standardized interviews. In Northern Laos, we recorded only 18 reptile individuals (2 turtle and 1 snake species). Interview surveys demonstrated that in Nam Xam River, fishers are more likely to hunt turtles and we identified one potential site where the world’s rarest turtle, Rafetus swinhoei, could be still present in the wild. In Nam Et Phou Louey National Park, we found 19 reptile species (8 lizard, 10 snake, 1 turtle species) in the different study sites, demonstrating a low species density in all different surveyed habitats. In Central/Southern Laos, we observed 30 species of reptiles (14 snake, 16 lizard, 35 amphibian species). Our study offers preliminary insights into the composition of amphibians and reptiles in Laos. The great majority of the species were non-threatened or not evaluated, and a few were threatened, suggesting that more research is needed to proper understand the conservation status of Laos’ herpetofauna. We highlighted, indirectly by interviews with local fishers, the possible presence of the turtle Rafetus swinhoei, thus providing a new hope for avoiding the extinction of this species. Finally, we observed a relatively low number of species in each habitat type, which is remarkably lower than in tropical forests of other continents or of nearby south-east Asian countries, indicating that the herpetofauna communities in Laos are depleted, reflecting an ‘empty forest syndrome’.