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Edible bird’s nest (EBN) is well known as a natural food product rich in glycoproteins such as sialic acid, minerals, and essential amino acids. Evidence from epidemiological studies suggests that EBN dietary supplementation improved brain cognitive functions. In mammals, the highest absorption of sialic acid from EBN occurs in the brain where it participates as an integral part of ganglioside structure in synaptogenesis and neural transmission. Sialic acid in EBN is vital during rapid brain growth particularly for preterm infants. While EBN dietary supplementation has been associated to enhance brain functions in infants, the effects of multiple generations of dietary EBN on cognitive function remain unclear. Thus, this study aimed to determine the effects of dietary EBN supplementation from different locations (e.g south (S) and north (N) of Peninsular Malaysia, Sabah (B) and commercial (C) on the cognitive function of transgenerational mice. To address these issues, C57BL/6 breeder mice (F0) were fed with different sources of EBN for six weeks (10 mg/kg) using oral gavage. Then, all animals were bred to obtain first generation (F1) until the second generation (F2) animals. At six weeks of age, F1 and F2 animals were tested for brain cognitive function by Y-maze test. Histological study for neuron density and distribution were analyzed using the hematoxylin and eosin procedure. The active compounds of EBN were determined using HPLC and the brain genes expression associated with cognitive function (e.g GNE, ST8SiaIV, SLC17A5, and BDNF mRNA) were analyzed using real-time PCR. Results showed that dietary EBN supplementation improved cognitive performance of F0, F1 and F2 mice by significantly increased the number of entries (9.04 ± 0.15; P < 0.05) and the time spent (2.40 ± 0.4 min; P < 0.05) in the novel arm of Y-maze test compared to control. This could indicate that the breeder mice (F0) absorbed sialic acid from EBN which crossed the placenta to contribute fetal