Information on Influenza A virus (IAV) prevalence in shorebird populations in Guatemala is scarce, and no live captures of shorebirds have been made for this purpose. The only reported case of Influenza A in shorebirds in Guatemala corresponded to a harvested individual. To update and obtain information of IAV in shorebirds, a transversal study was made on the South Coast of Guatemala. The main objective of this study was to evaluate presence of Influenza A among migratory shorebirds in stopover areas in the country. Captures and sampling took place in three sites during migration (October–April) over three migratory periods (2016–2017, 2017–2018, and 2018–2019). In total, 127 shorebirds belonging to 14 species of genera Calidris, Charadrius, Tringa, and Actitis were captured using mist nets. Tracheal and cloacal swabs were collected from each bird using a cotton swab, then tested with RT-PCR techniques targeting Influenza A. Blood samples were taken from the jugular or basilic vein to obtain serum that later was tested using a commercial ELISA Kit for detection of antibodies against Influenza A. Birds were released after sampling. All samples were PCR negative (0/127).One sandpiper specimen (Calidris minutilla) tested sero-positive to Influenza A through the ELISA test (1/122). These results suggest that IAV infection in this region may be minimal, as other studies have reported; nevertheless, it is important to consider that sample size was small in this study, and that the species included may not be the best indicators.