Sheep plays a significant economic role in the breeding and production of raw materials, such as meat, milk, wool, and hair in Qena City, Egypt. K. pneumoniae is an opportunistic bacterium able to cause diseases in humans and animals. K. pneumoniae is mostly resistant to antibiotics due to the formation of biofilm that causes huge losses in sheep breeding farms in Qana City. This study aimed to find the best antibiotic to treat bacterial infections caused by K. penumoniae and the relationship between antimicrobial resistance and biofilm formation. 150 nasal swabs were collected from various locations in Qena City and classified into three groups (Fifty nasal swabs from sheep suffering from respiratory infestations, 50 samples from apparently clinical health sheep and 50 from sheep dead or slaughtered accidentally). The samples were plated onto MacConkey agar and then confirmed by PCR assay. Antibiotic sensitivity test of identified K. pneumoniae isolates were done. Diagnosis of biofilm formation by tissue culture plates among isolates showed multi-drug resistances and detection of virulence genes (Fim A and Mrk A) responsible for biofilm formation by PCR. It was found that out of 150 samples 104 isolates were identified biochemically as K. pneumoniae . Using PCR technique for 11 K. pneumoniae isolates out of 104 biochemically identified K. pneumoniae isolates were K. pneumoniae 16s-23SITS coding gene (species–specific gene). It was also found that Fim A and Mrk A were the virulence genes that were responsible for biofilm formation in this study.