Aim: To isolate and identify Shigella species from faecal samples based on cultural and biochemical tests. Material and Methods: For the isolation of Shigella spp., faecal samples from cattle, poultry and humans were collected from various locations of Pantnagar. Fecal specimens were processed according to standard protocols. Results: Out of 511 fecal samples (311 human, 100 cattle and 100 poultry faecal samples) analyzed, 8 isolates of Shigella species were confirmed on the basis of Gram stain, morphology, cultural identification and biochemical characters. All the 8 Shigella isolates were obtained from human stool samples giving a prevalence rate of 2.57%. Conclusion: Under the conditions of the current study, Shigella species were prevalent in human population although in small numbers, whereas it was not isolated from cattle and poultry samples.