Milk comprises proteins, fats, minerals, carbohydrates, multiple vitamins, and various special substances dispersed in water. Different types of pathogenic microbes present in unpasteurized raw milk, cause gastrointestinal tract infections and food-borne infections. The current study was based on evaluating pathogenic microbe’s frequencies among the mammal (buffalo, cow, camel, goat and sheep) milk and assessing their nutritional parameters along with the screening of antibiogram of these pathogens in the district Peshawar, Pakistan. A total of 25 milk samples (05 from each mammals) were collected. Among the analyzed milk samples, S. aureus and Shigella spp. were observed n= 04 (16%), while E. coli, Salmonella spp., and P. aeruginosa were n= 03 (12%). The S. aureus was found highly resistant (100%) to Meropenem, Gentamicin, Amoxicillin, Aztreonam, and Doxycycline, while found highly sensitive (50%) to Amikacin, and Levofloxacin. E. coli showed high resistant (100%) to Meropenem, Ciprofloxacin, Amoxicillin, Ampicillin, and Doxycycline, while found to be highly sensitive (50%) to Levofloxacin, and Cefotaxime. The Salmonella spp. was found highly resistant (100%) to Amikacin, Meropenem, Amoxicillin, Cefotaxime, Ampicillin, Aztreonam, and Doxycycline, while found highly sensitive (50%) to Ciprofloxacin, and Gentamicin. The Shigella spp. was found highly resistant (100%) to Meropenem, Amoxicillin, Ampicillin, and Doxycycline, while found highly sensitive (25%) to Amikacin, Levofloxacin, Ciprofloxacin, Gentamicin, Cefotaxime, and Aztreonam. The P. aeruginosa was highly resistant (100%) to Amikacin, Levofloxacin, Ciprofloxacin, Gentamicin, Cefotaxime, Amoxicillin, Aztreonam, Doxycycline, while found highly sensitive (25%) to Meropenem, and Ampicillin. The high ESBL-producing bacterial pathogen was found E. coli (66.66%), followed by Shigella spp. (50%), Salmonella spp., and P. aeruginosa (33.33%), respectively. The nutritional analysis of mammal milk samples significantly (p < 0.05) with different parameters. Among these parameters, the fat Mean ± SD (3.36 ± 0.11), protein content Mean ± SD (2.91 ± 0.04), lactose content Mean ± SD (4.31 ± 0.10), and total solid content Mean ± SD (11.61 ± 0.27) were observed. In conclusion, of the current study, pathogenic microbes are present in unpasteurized milk, which shows high resistance to various antibiotics, and the nutritional values of milk were also low.