Nitrate nitrogen (N-NO3-), the most common pollutant in groundwater, is a result of the effect of diffuse sources of pollution like agriculture and animal husbandry intensive. The land use for these economic activities is very common in the Los R?os sub-region in the state of Tabasco, Mexico, where the Los R?os and Boca del Cerro aquifers are located. The aim of this research was to assess the concentrations of nitrate-nitrogen (N-NO3-) in groundwater, determine the quality in agreement with the maximum permissible limits established by national and international regulations, and the risks to the public health and aquatic life. The spatial distribution of N-NO3- was determined using the inverse distance weighted (IDW) interpolation technique. The average nitrate-nitrogen concentration was 0.76 mg.L-1, while the maximum concentration observed was 3.98 mg.L1. This does not exceed the maximum permissible limit (MPL) established in the national and international normativity for drinking water. However, in 50% of the sampling sites, the concentrations of N-NO3- exceed the MPL established in Mexico for the protection of the life of seawater. Relatively low concentrations of N-NO3- have shown to be toxic to certain aquatic organisms, and the aquifers studied discharge a third of the water to the rivers in the area, which flow into the Laguna de T?rminos Campeche and the Gulf of Mexico. Laguna de T?rminos Campeche is one of the most diverse and rich environmental systems on earth, where numerous ecosystems converge such as coastal lagoons, wetlands, mangroves, seagrasses, and coral reefs.