Objective: Multi and extensively drug-resistant (MDR and XDR), Pseudomonas aeruginosa (P. aeruginosa) and Acinetobacter baumannii (A. baumannii) are two main causative agents of nosocomial infections leading to increased morbidity and mortality. We aim to study the prevalence of MDR and XDR-A. baumannii and P. aeruginosa phenotypes in clinical specimens. We conducted this for 1 year (2017-2018) and isolated bacteria from the clinical samples. Then, XDR and MDR strains were determined by susceptibility testing (disc diffusion). Results: Out of 3248 clinical samples, A. baumannii and P. aeruginosa strains were detected in 309(9.51%) of them. Susceptibility testing indicated that (16.50%) and (15.53%) of the P. aeruginosa and (74.75%) and (73.13%) of the A. baumannii isolates were screened as the MDR and XDR strains. The frequency of MDR isolates was higher in wound samples 222 (71.8%). This rate in behavioral intensive care unit (BICU) and restoration ward, were 187 (60.5%) and 63 (20.4%). The frequency of XDR isolates in BICU 187 (59.54%), restoration 58(18.77%), and burns 30 (9.70%) were assessed as well. Considering high isolation rates of MDR and XDR of mentioned strains, it is necessary to apply prevention criteria for eradication of the mentioned bacteria from hospital wards.

The integrons, as the mobile exogenous elements, play a prominent role in the spreading of antimicrobial resistance genes from Pseudomonas aeruginosa clinical isolates to other bacteria. This study aimed to investigate the frequency of class 1 integrons andresistance gene cassettes carrying by them in clinical isolates as well as multidrug resistant P. aeruginosa. Materials and Methods: A total of 100 clinical isolates of P. aeruginosa were collected from 5 hospitals in Mazandaran province, north Iran. The antibiotic susceptibility pattern of the isolates was evaluated using the disk agar diffusion method. Genomic DNAs were extracted and then the presence of class 1 integrons was detected by the PCR test. All PCR products of the positive isolates were sequenced for the detection of resistance gene cassettes by the Sanger method. Results: Forty-one percent of the clinical isolates were multi-drug resistant. Also, 42% of the isolates were contained class 1 integron, and 61.9% of the integron positive isolates were detected as MDR. We detected 10 different gene cassettes sizing from 0.6 to 3.5 kb in the present study. The sequencing analysis of the internal variable regions of the class 1 integrons showed that the 0.75 kb gene cassette (aadB) was the most frequent resistance gene (54.76%) among all clinical isolates, as well as the MDR isolates. Other resistance genes detected in this study were included: aadA6-orfD (35.71%), aacA4-bla OXA-10 (21.42%), aadB-aacA4-bla OXA-10 (19.04%), bla OXA-10-aacA4-VIM1 (11.9%), aacA4-cat B10 (7.14%), aacA5-aadA1-cmlA5 (7.14%), bla OXA31-aadA2 (4.76%), and aac(3)-Ic-aac A5-cmlA5 (4.76%). To the best of our knowledge, bla OXA-10-aacA4-VIM1 cassette array is detected for the first time in this study. Conclusion: The treatment of infections caused by P. aeruginosa in this region of Iran is a major problem due to the high prevalence of class 1 integrons. It seems that the high prescription of beta-lactams and aminoglycosides for the treatment of these infections may be replaced by other combination therapy stewardships.

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In recent years, the prevalence of resistance to aminoglycosides among clinical isolates of Pseudomonas aeruginosa is increasing. The aim of this study was to investigate the role of aminoglycoside-modifying enzymes (AMEs) in resistance to aminoglycosides in clinical isolates of P. aeruginosa. The clinical isolates were collected from different hospitals. Disk agar diffusion test was used to determine the antimicrobial resistance pattern of the clinical isolates, and the minimum inhibitory concentration of aminoglycosides was detected by microbroth dilution method. The PCR was performed for discovery of aminoglycoside-modifying enzyme-encoding genes. Among 100 screened isolates, 43 (43%) isolates were resistant to at least one tested aminoglycosides. However, 13 (13%) isolates were resistant to all tested aminoglycosides and 37 isolates were detected as multidrug resistant (MDR). The resistance rates of P. aeruginosa isolates against tested antibiotics were as follows: ciprofloxacin (41%), piperacillin-tazobactam (12%), cefepime (32%), piperacillin (26%), and imipenem (31%). However, according to the MIC method, 13%, 32%, 33%, and 37% of the isolates were resistant to amikacin, gentamicin, tobramycin, and netilmicin, respectively. The PCR results showed that AAC(6 ′ )-Ib was the most commonly (26/43, 60.4%) identified AME-encoding gene followed by AAC(6 ′ )-IIa (41.86%), APH(3 ′ )-IIb (34.8%), ANT(3 ″ )-Ia (18.6), ANT(2 ″ )-Ia (13.95%), and APH(3 ″ )-Ib (2.32%). However, APH(3 ′ )-Ib was not found in any of the studied isolates. The high prevalence of AME-encoding genes among aminoglycoside-resistant P. aeruginosa isolates in this area indicated the important role of AMEs in resistance to these antibiotics similar to most studies worldwide. Due to the transmission possibility of these genes between the Gram-negative bacteria, we need to control the prescription of aminoglycosides in hospitals.

Background & Objective: Efflux pump inhibitors (EPIs) can block efflux pumps and are helpful in potentiating the activity of aminoglycosides against Pseudomonas aeruginosa. The present study compared the effects of phenylalanine-arginine beta naphthylamide (PAβN) and curcumin on aminoglycoside minimum inhibitory concentration (MIC) on Pseudomonas aeruginosa clinical isolates. Materials & Methods: For this descriptive-analytical study, 100 clinical isolates of Pseudomonas aeruginosa were collected and identified by differential diagnostic tests. The MICs of amikacin, gentamicin, and tobramycin were evaluated before and after adding EPIs using a micro-broth dilution test. Results: The bacteria were isolated from different types of samples, including urine (26 isolates), sputum (37 isolates), ulcers (20 isolates), catheters (eight isolates), blood (five isolates), feces (two isolates), and eyes (two isolates). Overall, 60% of the isolates were obtained from males (mean age = 47.85), and 40% from females (mean age = 44.76). In the MIC test, 11 (25.5%), 15 (34.8%), and 18 (41.8%) isolates were resistant to amikacin, gentamicin, and tobramycin, respectively. Significant reductions in the MICs of amikacin, gentamicin, and tobramycin were observed after adding curcumin in 54-100% of aminoglycoside-resistant isolates, while fewer changes in the MICs of aminoglycosides were seen against these clinical isolates after adding PAβN (36-55%). Conclusion: Curcumin and PAβN can potentiate the effect of aminoglycosides on clinical isolates of Pseudomonas aeruginosa and change their susceptibility pattern due to efflux pump inhibition. However, our outcomes detected that curcumin was more effective than the PAβN against the aminoglycoside-resistant isolates of P. aeruginosa.