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This study aimed to identify <jats:italic>P. aeruginosa<\/jats:italic> efflux pumps and examine the effect of Zinc sulfate on these pumps. This study was conducted on 104 <jats:italic>P. aeruginosa<\/jats:italic> clinical isolates collected from different types of specimens. The antimicrobial susceptibility pattern of all isolates was examined by disc diffusion and microdilution methods. A Cartwheel test was done for phenotypic detection of <jats:italic>P. aeruginosa<\/jats:italic> efflux pumps in the presence and absence of zinc sulfate. The expression of efflux pumps encoding genes and their regulators in the presence and absence of zinc sulfate was measured using real-time reverse transcriptase PCR. A high rate of multidrug-resistant (MDR) pattern was observed among the tested isolates (55.7%). Combining different antibiotics with Zinc sulfate exhibited synergistic effects against most of the studied isolates. The addition of sub-inhibitory concentrations of Zinc sulfate to the cartwheel test was found to inhibit efflux pump activity in all tested isolates. The expression of the positive regulator gene, <jats:italic>mexT<\/jats:italic>, was significantly decreased. Additionally, Zinc sulfate significantly upregulated the expression of the <jats:italic>mexR<\/jats:italic> gene (P-value\u2009&lt;\u20090.05), a negative regulator of the MexAB-OprM efflux pump. Zinc sulfate demonstrated a strong antimicrobial effect against <jats:italic>P. aeruginosa<\/jats:italic> clinical isolates. We recommend the use of Zinc sulfate as an antibiotic adjuvant and efflux pump inhibitor in the treatment of <jats:italic>P. aeruginosa<\/jats:italic> infections to enhance the bacterium\u2019s susceptibility to antimicrobial agents.<\/jats:p>","DOI":"10.1007\/s11274-025-04352-4","type":"journal-article","created":{"date-parts":[[2025,4,28]],"date-time":"2025-04-28T01:00:42Z","timestamp":1745802042000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Zinc sulfate acts as an efflux pump inhibitor on Pseudomonas aeruginosa clinical isolates"],"prefix":"10.1007","volume":"41","author":[{"given":"Wedad M.","family":"Abdelraheem","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hadeer Ibrahim","family":"Yassin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaimaa","family":"Zaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mona Abdel Monem","family":"Esmail","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,28]]},"reference":[{"key":"4352_CR1","doi-asserted-by":"publisher","DOI":"10.1016\/j.genrep.2021.101301","volume":"24","author":"M Abavisani","year":"2021","unstructured":"Abavisani M, Goudarzi M, Ghalavand Z, Hajikhani B, Rad ZR, Rad ZR, Hashemi A (2021) Evaluation of efflux pumps overexpression and \u03b2-lactamase genes among colistin resistant Pseudomonas aeruginosa. 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