© 2018 Universidad Autonoma de Tlaxcala. All rights reserved. Free-living amoebae (FLA) are widely distributed protozoa in the environment which has been isolated from a variety of sources including, water, soil and, dust, although diseases caused by them are considered neglected waterborne pathogens due to the lack of research concerning them. Among these organisms, Acanthamoeba spp. and Naegleria fowleri, the ones of interest for the present study, are reported to be pathogenic and opportunistic agents of severe pathologies. Furthermore, these organisms act as “Trojan horses” for pathogenic bacteria, fungi and viruses resistant to its’ phagocytosis, which represents a threat to public health due to the resistance of these protozoans to inactivation methods. Currently, there is no available data about their distribution in environmental sources in the state of Tamaulipas, thus the aim of this study was to evaluate the presence of these organisms in the City of Reynosa, Tamaulipas using microbiological culture methods. Their identification is based on morphological features observed with light microscopy and the sampling places are evaluated to determine the biological diversity and presence of other organisms due to the observed elevated levels of pollution. The results revealed that 50% of the assessed sampled places were positive for the presence of Acanthamoeba spp.