This investigation aimed at examining physicochemical properties and characteristics of house and cave edible bird's nest (EBN) collected from locations in Southern Thailand. The house EBN and the cave EBN samples were gathered from eight provinces in Southern Thailand. Color, proximate composition, mineral, nitrate, nitrite sialic acid (Sia-a) content, amino acid composition, and microbial load of all the collected samples were determined. Protein was the major component found in both the house EBN (52.68-54.73%) and the cave EBN (52.65-55.25%). Nonessential amino acids, i.e., aspartic acid/asparagine and serine; and essential amino acids, i.e., leucine, threonine, and phenylalanine, were dominant amino acids for both EBN. The average Sia-a content of the house EBN (3570 mg/100 g) was higher than that of the cave EBN (3128.7 mg/100 g). On the other hand, higher calcium, nitrate, and nitrite contents were found in the cave EBN. However, the microbiological quality of all the samples was within the limits issued by the Thai National Bureau of Agricultural Commodity and Food Standards. The quality of the EBN samples is varied depending on the production origin or geographical location. EBN is recommended as a high-quality animal product rich in Sia-a and amino acids.