Light pollution has become established as a global environmental issue, driven by urbanisation and the growing use of artificial light at night (ALAN), which disrupts the natural rhythms of ecosystems. This phenomenon negatively affects biodiversity; particularly nocturnal species whose ecological interactions depend on darkness. This study assesses the impact of light pollution on protected forests in Guayaquil, Ecuador, specifically Cerro Blanco, Papagayo, Estero Salado, and Prosperina. Using VIIRS satellite imagery from 2014, 2019, and 2024, variations in nighttime radiance and their impact on local species such as <em>Engystomops guayaco</em> and <em>Sylvilagus dauleensis</em> were analysed. The results show an increase in light pollution near urban boundaries, particularly in Cerro Blanco, disrupting biological rhythms and threatening the survival of these nocturnal species. This study highlights the urgent need for public policies that promote sustainable lighting and the creation of “dark zones” to mitigate these effects and preserve biodiversity in these critical ecosystems.