Pseudomonas aeruginosa and Acinetobacter spp. are the microorganisms most frequently associated with serious healthcare-associated infections and death. The objective of the research consisted of characterizing the isolates of P. aeruginosa and A. spp. in respiratory secretions of patients hospitalized in the Clinical Surgical Hospital of Santa Clara, Villa Clara, Cuba between January 2014 and December 2018. A descriptive, longitudinal and retrospective study was carried out with the isolates of P. aeruginosa and A. spp. from the Clinical Surgical Hospital "Arnaldo Milián Castro" in the city of Santa Clara, Villa Clara province, Cuba between the months of January 2014 to December 2018. For the realization of this work, a discretionary and intentional sampling by criterion was selected. The population consisted of 2 207 isolates, and the sample consisted of 249 P. aeruginosa and 705 A. spp. The data were obtained from the record books of the respiratory samples section of the Microbiology Laboratory and were arranged in a documentary observation guide. A. spp., was the most frequently isolated microorganism in the intensive care units, with a uniform behavior during the entire series studied, while P. aeruginosa predominated in the first and fourth trimesters. A. spp., showed almost absolute levels of resistance throughout the study, while P. aeruginosa showed low percentages of resistance with a propensity to increase resistance. It is concluded that both microorganisms were the most frequently isolated in respiratory secretions in ICUs, surpassing in recent years’ other microorganisms, such as Gram-positive cocci and enterobacteria in general isolates and in HAIs, with marked prevalence rates for both genera, as well as an evident resistance, characterized by differences in relation to percentages, but with a propensity to increase it.