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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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.

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: According to the reports from World Health Organization, increased use of antibiotics and bacterial resistance has become a worldwide issue. Resistance to antimicrobial agents in Escherichia coli clinical strains is increasing. Objectives: The objective of this study was to determine the antibiotic resistance patterns and frequency of multidrug resistance (MDR) phenotype in Escherichia coli isolated from patients in two major hospitals of Sari, north Iran. Methods: In this descriptive-analytical study, a total of 13322 clinical specimens were collected from patients. All the specimens were evaluated to determine the presence of E. coli strains using conventional biochemical tests and API kit. Susceptibility testing against twelve antibiotics was determined using the disk-diffusion method. Results were interpreted in accordance to the Clinical and Laboratory Standard Institute (CLSI) protocol. Results: Out of 13322 studied samples, 964 (7.23%) E. coli strains were identified. In two hospitals, high resistance to ampicillin and cefalexin was presented in 621 (64.4 %) and 402 (41.7 %) isolates, respectively. The highest antibiotic resistance was observed in the burn unit, the burn intensive care unit (ICU) and the burn restoration section, while all the strains (eight) that were isolated from the neonatal-ICU, were sensitive to all the tested antibiotics except cefalexin, nitrofurantoin, nalidixic acid, and ampicillin. Also, strains isolated from urine, wound, stool, and blood samples were resistant to all tested antibiotics. Conclusion: Increased resistance to different antibiotics in burn hospitals has created increasing concern. Very high resistance to some antibiotics indicates that these drugs are misused in therapeutic centers and highlights that infection control measures should be arranged in the ICUs of our hospitals.

Background: Multi-drug resistant (MDR) Citrobacter freundii (C. freundii) as a causative agent of nosocomial infections is a health threat, especially in hospitals. This study was conducted to determine the prevalence of MDR C. freundii, considering isolation sites and a variety of utilized antibiotics. Materials and Methods: In this cross-sectional study, the clinical samples of C. freundii strains were collected and screened using traditional bacteriological tests in Zareh Hospital, Sari City, Iran, during 2016-2017. We used disk diffusion methods to assess the susceptibility patterns of isolates according to the Clinical Laboratory Standard Institute (CLSI) guidelines. Results: Out of 3248 clinical samples, C. freundii strains were detected in 109 samples (32.1% females and 67.9% males). Susceptibility tests indicated that 89 isolates (81.65%) were MDR strains. Frequencies of MDR C. freundii strains were higher in the Behavioral Intensive Care Unit (BICU) (37.61%) and restoration ward (29.35%) compared with other hospital wards. Conclusion: Considering the MDR C. freundii strains detected from burn hospital wards, it is necessary to implement prevention criteria for their eradication from burn hospitals. The results indicate the urgent need to design more practical methods for controlling infection in hospital wards.

Objectives: Metallo-beta-lactamases play a major role in the resistance of Pseudomonas aeruginosa to carbapenems. The aim of this study was the phenotypic and molecular detection of IMP and SPM carbapenemase genes in 100 carbapenem-resistant clinical isolates of P. aeruginosa. The isolates identified using standard microbiological tests, and their antibiotic susceptibility pattern determined by disk agar diffusion (Kirby Bauer) method. Phenotypic identification of Metallo-beta-lactamase-producing strains assessed by the combined disk test (CDT). Then, PCR was used to detect the presence of IMP and SPM genes.Results: The highest and lowest levels of antibiotic resistance were observed against gentamicin (40%) and piperacillin-tazobactam (13%), respectively. Besides, 40 isolates (40%) had the Multi-drug Resistant (MDR) phenotype, while 5 (12.5%) MDR isolates were resistant to all antibiotics tested. The results of the CDT showed that among 43 carbapenem non-susceptible clinical isolates of P. aeruginosa, 33 (76.74%) isolates were Metallo-beta-lactamase-producing strains. Also, the frequency of the IMP gene was determined to be 9%, while none of these isolates carried the SPM gene. Due to the high prevalence of carbapenem-resistant and MDR P. aeruginosa in this study, routine antibiotic susceptibility testing and phenotypic identification of carbapenemase production by this bacterium are necessary for proper selection of antibiotics.