The present study aimed to rule out the potential of multiple pathogens strains coexisting in a single sample and whether single colony could represent all microorganism present in the sample. A total of 20 samples (cloacal and tracheal) were collected from 10 apparently healthy quails in EL-Beheira governorate, Egypt. The samples were individually cultured on EMB agar. Eight out of 10 tracheal samples and 9 out of 10 cloacal samples were positive with green metallic sheen colonies. Fifteen colonies were individually collected from each plate of 8 tracheal samples and 20 colonies from each plate of 9 cloacal samples. As a result, 300 colonies were collected, with each colony being treated as a single sample. All 300 colonies were tested by Gram staining, motility test, biochemical tests, sugar fermentation and enzymatic reaction. The findings demonstrated that mixed infections were more common than single infections. Escherichia coli was the most frequently isolated organism (54.6%) followed by Enterobacter spp (22.3%), Klebsiella spp (11.6%), Citrobacter spp (9%), and Serratia liquefaciens (2.3%). Serological identification of E. coli (n=164) showed diversity of the serotypes among the samples derived from the same plate. Sixteen different serogroups in all the isolates were identified. Moreover, antimicrobial susceptibility of E. coli serotypes showed variable patterns with high sensitivity to gentamycin, doxycycline, ciprofloxacin, amikacin, tetracycline, and ampicillin indicating that these antimicrobials could be of great promise in combating E. coli infection in quails. These findings imply that diverse bacterial populations could be present in a single sample, making analysis of single colony from a bacterial culture plate insufficient to represent the actual population of pathogens. Further, quails could possibly play a significant role in harboring and spread of a variety of pathogens from the same or different strains.