The antimicrobial resistance currently impedes and threatens the future of effective prevention and treatment of the continually expanding range of infections caused by bacteria. This study aimed to identify the bacterial causes the wound infection among animals and using the antibiotic/nanoparticles mixture as a new attempt for the treatment the wound infection induced in rats. For this purpose, 112 swabs wound infection cases in the different animal types (36 sheep, 21 goats, 12 cows, 4 horses, 8 dogs, 9 rabbits, 7 genies pigs and 15 rats) were studied in the for bacterial isolation. The Pseudomonas aeruginosa was tested for its sensitivity to the antibiotics and the nanoparticles (CoFe2O4 and NiFe2O4) in vitro by using the MIC method. Also the wound infection was induced in the rats and the effect of nanoparticles/antibiotics mixture were tested in vivo. The results showed that P. aeruginosa was the predominant bacterial type that the caused wound infection. The minimum inhibitor concentration of NiFe2O4 and CoFe2O4 nanoparticles were 32 µg /ml and 16 µg /ml respectively. A clear synergistic effect of antibiotic/ nanoparticles as antibacterial were noticed which appear as a decrease in MIC and increase of the inhibitory diameter zone. According to the result of Random Amplification of Polymorphic DNA test, the nanoparticles effects on genetic material of P. aeruginosa observed as an appearance/disappearance of bands, increase in thickness and clarity of the bands.