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Urinary tract infection (UTI) is a common type of infectious disease globally. The aim of this study was to detect the frequency of fosA3 and fosC2 genes in extended-spectrum β-lactamases (ESBL) and bla DHA, bla CMY-2, and bla CMY-42 genes in AmpC β-lactamases-producing isolates of Escherichia coli. In total, 120 isolates of E. coli were collected from three teaching hospitals between March 2014 and February 2015. Antibiotic susceptibility tests were carried out by disk diffusion method. The presence of bla CMY-2, bla CMY-42, bla DHA, fosA3, and fosC2 genes was detected by polymerase chain reaction (PCR) and sequencing. Of the 120 strains, 92 (76.6%) were identified as ESBL producers, 30 (25%) were determined as AmpC β-lactamase producers, and 24 (20%) had both ESBL and AmpC β-lactamase enzymes. Imipenem, fosfomycin, and nitrofurantoin had the best effect against isolates of E. coli. PCR assay demonstrated that the frequency of bla CMY-2, bla CMY-42, and bla DHA genes among AmpC β-lactamases-producing strains were 39%, 1%, and 17.5%, respectively. This study reports the first detection of fosfomycin resistance in Iran. This study indicated the increasing prevalence of UTI isolates of E. coli-harboring ESBL and AmpC β-lactamases genes in Iran. Therefore, due to the high rate of bla DHA and bla CMY genes and emergence of fosfomycin-resistant E. coli isolates, we recommend continuous monitoring of antibiotic resistance as well as attention to guidelines of infection controls.

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Acta Microbiologica et Immunologica Hungarica
Authors: Sara Davoudabadi, Hossein Goudarzi, Mehdi Goudarzi, Abdollah Ardebili, Ebrahim Faghihloo, Javad Yasbolaghi Sharahi, and Ali Hashemi

Abstract

In this study, we focused on the emergence of extensively drug-resistant (XDR), pandrug-resistant (PDR), and hypervirulent Klebsiella pneumoniae (hvKP) in Iran. During 2018 to 2020 a total of 52 K. pneumoniae isolates were collected from different clinical specimens. The hvKP isolates were identified by PCR amplification of virulence and capsular serotype-specific genes. Hypermucoviscous K. pneumoniae (hmKP) were identified by string test. Carbapenem-resistant hvKP (CR-hvKP), multidrug-resistant hvKP (MDR-hvKP), extensively drug-resistant hvKP (XDR-hvKP), and pandrug-resistant hvKP (PDR-hvKP) were determined by disc diffusion method, Carba-NP test and PCR method. XDR-hvKP isolates were typed by multilocus sequence typing (MLST). Among all K. pneumoniae isolates 14 (26.9%) were identified as hvKP and 78.6% (11/14) of them were hmKP however, none of the classic K. pneumoniae (cKP) isolates were hmKP. The predominant capsular serotype of hvKP was K2 (42.85%) followed by K1 (35.71%). The prevalence of MDR-hvKP, XDR-hvKP and PDR-hvKP isolates were 6 (42.9%), 5 (35.7%) and 1 (7.1%), respectively. ESBL production was found in 85.7% of hvKP isolates and most of them carried bla TEM gene (78.6%) and 6 isolates (42.9%) were CR-hvKP. Among hvKP isolates, 1 (7.1%), 2 (14.3%), 3 (21.4%), 8 (28.6%), and 11 (78.6%) carried bla NDM-6, bla OXA-48, bla CTX-M, bla SHV, and bla TEM genes, respectively. According to MLST analysis, 2, 1, 1, and 1 XDR-hvKP isolates belonged to ST15, ST377, ST442, and ST147, respectively. The occurrence of such isolates is deeply concerning due to the combination of hypervirulence and extensively drug-resistance or pandrug-resistance.

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Acta Microbiologica et Immunologica Hungarica
Authors: Seyedeh Marzieh Jabbari Shiadeh, Leila Azimi, Taher Azimi, Ali Pourmohammad, Mehdi Goudarzi, Bahare Gholami Chaboki, and Ali Hashemi

Abstract

Antibiotic resistance and especially multiresistance in Enterococci, is a serious public health issue especially in infections of immunocompromised patients. EfrAB is a heterodimeric multidrug ATP-binding cassette (ABC) transporter that causes endogenous resistance to antimicrobials including fluoroquinolones in Enterococcus spp. The aim of this study was to seek the gene expression rate and role of efrAB efflux pump in ciprofloxacin resistant Enterococcus faecalis and Multilocus Sequence Typing (MLST) of multiresistant isolates. Phenotypic and genotyping identification of 80 E. faecalis isolates were performed. Minimum inhibitory concentrations (MICs) to ciprofloxacin (CIP) were measured with and without carbonylcyanide 3-chlorophenylhydrazone (CCCP) by broth microdilution. After DNA extraction and sequencing for detection of efrA and efrB genes, the efrAB efflux positive isolates that were resistant to ciprofloxacin and showed decrease of ciprofloxacin MIC range were identified. Isolates that exhibited decrease in ciprofloxacin MIC range from two to ten folds were assessed for biofilm formation and finally, the expression levels of efrB, efrA genes were measured by quantitative Real-Time PCR (qRT-PCR). High rates of resistance to tetracycline and minocycline and low rates of resistance to the most antibiotics used in this study were detected. The results in this study indicated that the incidence of Multiple drug resistance (MDR) was 23.7% and all isolates that were resistant to ciprofloxacin revealed several degrees of overexpression in efrA and efrB genes. Our study found two ST480 and one ST847 in E. faecalis isolates. In conclusion, despite of low frequency of resistance to the most antibiotics and MDRs in our region, we found one ST480 isolate with resistance to eight antibiotics that also exists in other parts of the world.

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Acta Microbiologica et Immunologica Hungarica
Authors: Elham Abbasi, Hossein Goudarzi, Ali Hashemi, Alireza Salimi Chirani, Abdollah Ardebili, Mehdi Goudarzi, Javad Yasbolaghi Sharahi, Sara Davoudabadi, Ghazaleh Talebi, and Narjes Bostanghadiri

Abstract

A major challenge in the treatment of infections has been the rise of extensively drug resistance (XDR) and multidrug resistance (MDR) in Acinetobacter baumannii. The goals of this study were to determine the pattern of antimicrobial susceptibility, bla OXA and carO genes among burn-isolated A. baumannii strains. In this study, 100 A. baumannii strains were isolated from burn patients and their susceptibilities to different antibiotics were determined using disc diffusion testing and broth microdilution. Presence of carO gene and OXA-type carbapenemase genes was tested by PCR and sequencing. SDS-PAGE was done to survey CarO porin and the expression level of carO gene was evaluated by Real-Time PCR. A high rate of resistance to meropenem (98%), imipenem (98%) and doripenem (98%) was detected. All tested A. baumannii strains were susceptible to colistin. The results indicated that 84.9% were XDR and 97.9% of strains were MDR. In addition, all strains bore bla OXA-51 like and bla OXA-23 like and carO genes. Nonetheless, bla OXA-58 like and bla OXA-24 like genes were harbored by 0 percent and 76 percent of strains, respectively. The relative expression levels of the carO gene ranged from 0.06 to 35.01 fold lower than that of carbapenem-susceptible A. baumannii ATCC19606 and SDS – PAGE analysis of the outer membrane protein showed that all 100 isolates produced CarO. The results of current study revealed prevalence of bla OXA genes and changes in carO gene expression in carbapenem resistant A.baumannii.

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Acta Microbiologica et Immunologica Hungarica
Authors: Zohreh Riahi Rad, Zahra Riahi Rad, Hossein Goudarzi, Mehdi Goudarzi, Hesam Alizade, Fariba Naeimi Mazraeh, Javad Yasbolaghi Sharahi, Abdollah Ardebili, and Ali Hashemi

Abstract

Carbapenems are employed to treat infections caused by Gram-negative bacteria including Klebsiella pneumoniae. This research is aimed to perform phenotypic detection of β-lactamases and molecular characterization of NDM-1 positive K. pneumoniae isolates. Another objective is to investigate NDM-1 producing K. pneumoniae among children in Iran. From 2019 to 2020, altogether 60 K. pneumoniae isolates were acquired from various patients in certain Iranian hospitals. Antimicrobial susceptibility testing was performed by disk diffusion and broth microdilution methods. In addition, mCIM and eCIM were used to confirm the production of carbapenemases and metallo-beta-lactamases (MBLs), respectively. Detection of resistance genes namely, bla NDM-1, bla IMP, bla VIM, bla KPC, bla OXA-48-like, bla CTX-M, bla SHV, bla TEM, and mcr-1 was performed by PCR and confirmed by DNA sequencing. Multilocus sequence typing (MLST) was employed to determine the molecular typing of the strains. According to the findings, the highest rate of carbapenem resistance was detected against doripenem 83.3% (50). Moreover, 31.7% (19) were resistant to colistin. Further to the above, altogether 80% (48) were carbapenemase-producing isolates and among them 46.7% (28) of the isolates were MBL and 33.3% (20) isolates were serine β-lactamase producer. According to the PCR results, 14 isolates produced bla NDM-1. Remarkably, four bla NDM-1 positive isolates were detected in children. In addition, these isolates were clonally related as determined by MLST (ST147, ST15). Altogether ten bla NDM-1 positive isolates were ST147 and four bla NDM-1 positive isolates were ST15. Based on the results, the emergence of NDM-producing K. pneumoniae among children is worrying and hence, it is necessary to develop a comprehensive program to control antibiotic resistance in the country.

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Acta Microbiologica et Immunologica Hungarica
Authors: Seyedeh Marzieh Jabbari Shiadeh, Ali Hashemi, Fatemeh Fallah, Parnian Lak, Leila Azimi, and Marjan Rashidan

Enterococcus faecalis is one of the most significant pathogen in both nosocomial and community-acquired infections. Reduced susceptibility to antibiotics is in part due to efflux pumps. This study was conducted on 80 isolates of E. faecalis isolated from outpatients with urinary tract infection during a period of 1 year from April 2014 to April 2015. The antibiotic susceptibility patterns of isolates were determined by the disk diffusion method and presence of efrA and efrB genes was detected by PCR and sequencing. Minimum inhibitory concentrations (MICs) to ciprofloxacin (CIP) were measured with and without carbonyl cyanide 3-chlorophenylhydrazone (CCCP) by broth microdilution. The highest resistance rate was observed to erythromycin (83.3%) and the prevalence of efrA and efrB genes in all E. faecalis isolates was 100%. This study showed that 9 out of 13 (69.2%) ciprofloxacin-resistant isolates became less resistant at least fourfolds to CIP in the presence of efflux pump inhibitor. Our result showed that CCCP as an efflux inhibitor can increase effect of CIP as an efficient antibiotic and it is suggested that efrAB efflux pumps are involved in resistance to fluoroquinolone.

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Background:

The distribution of drug resistance among clinical isolates of Escherichia coli and Klebsiella pneumoniae has limited the therapeutic options. The aim of this study was to report the prevalence of quinolone resistance genes among E. coli and K. pneumoniae clinical strains isolated from three educational hospitals of Tehran, Iran.

Materials and methods:

A total of 100 strains of E. coli from Labbafinejad and Taleghani Hospitals and 100 strains of K. pneumoniae from Mofid Children and Taleghani Hospitals were collected between January 2013 and May 2014. Antimicrobial susceptibility tests were done by disk diffusion method based on Clinical and Laboratory Standards Institute guidelines. Detection of qepA, aac(6′)-Ib-cr, acrA, and acrB genes was done by polymerase chain reaction (PCR).

Results:

In this study, fosfomycin and imipenem against E. coli and fosfomycin and tigecycline against K. pneumoniae had the best effect in antimicrobial susceptibility tests. PCR assay using specific primers demonstrated that the prevalence of qepA, aac(6′)-Ib-cr, acrA, and acrB genes among the 100 E. coli isolates was 0 (0%), 87 (87%), 92 (92%), and 84 (84%), respectively. The prevalence of qepA, aac(6′)-Ib-cr, acrA, and acrB genes among the 100 K. pneumoniae isolates was 4 (4%), 85 (85%), 94 (94%), and 87 (87%), respectively.

Conclusion:

The distribution of qepA, aac(6′)-Ib-cr, acrA, and acrB resistance determinants in E. coli and K. pneumoniae is a great concern. Therefore, infection control and prevention of spread of drug-resistant bacteria need careful management of medication and identification of resistant isolates.

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Acta Microbiologica et Immunologica Hungarica
Authors: Mohsen Heidary, Alireza Salimi Chirani, Saeed Khoshnood, Gita Eslami, Seyyed Mohammad Atyabi, Habibollah Nazem, Mohammad Fazilati, Ali Hashemi, and Saleh Soleimani

Acinetobacter baumannii is a major opportunistic pathogen in healthcare settings worldwide. In Iran, there are only few reports on the prevalence of aminoglycoside resistance genes among A. baumannii isolates. The aim of this study was to investigate the existence of aminoglycoside-modifying enzyme (AME) genes from A. baumannii strains collected at a university teaching hospital in Iran. One hundred A. baumannii strains were collected between 2014 and 2015 from hospitalized patients at Loghman Hakim Hospital, Tehran, Iran. Antimicrobial susceptibility was determined by disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. The DNA was extracted using a kit obtained from Bioneer Co. (Korea) and was used as a template for polymerase chain reaction. The most active antimicrobial agent against these strains was colistin. The rate of extended-spectrum cephalosporin resistance was 97%. The aadA1, aadB, aac(6′)-Ib, and aac(3)-IIa genes were found in 85%, 77%, 72%, and 68% of A. baumannii isolates, respectively. This study showed a high prevalence rate of AME genes in A. baumannii. This prevalence rate has explained that further aminoglycoside resistance genes may have role in the resistance of clinical isolates of A. baumannii. Therefore, control and treatment of serious infections caused by this opportunistic pathogen should be given more consideration.

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Abstract

Staphylococcus aureus as an opportunistic bacterial pathogen with intrinsic and acquired resistance to many antibiotics is a worldwide problem. The current study was undertaken to evaluate the resistance pattern, and determine the genetic types of multidrug-resistant S. aureus isolated from wound.

This cross-sectional study was conducted over the period of two years (from December 2018 to November 2020) at the hospitals affiliated to Shahid Beheshti University of Medical Sciences, Tehran, Iran. In present study, 75 multidrug-resistant S. aureus isolates collected from wound infections were investigated. Phenotypic resistance was assessed by Kirby–Bauer disk diffusion method. Conventional PCR was performed for the detection of virulence encoding genes. Genotyping of strains was performed based on coa gene polymorphism using multiplex-PCR assay. SCCmec typing, spa typing and MLST were also used to characterize the genotype of the mupirocin, tigecycline and vancomycin resistant multidrug-resistant S. aureus isolates.

All 75 multidrug-resistant S. aureus isolates in the study were confirmed as MRSA. Coagulase typing distinguished isolates into five genotypic patterns including III (40%), I (24%), IVb (16%), V (10.7%) and type X (9.3%). Resistance to tigecycline was detected in 4% of MDR-MRSA isolates and all belonged to CC8/ST239- SCCmec III/t421 lineage. According to our analysis, one VRSA strain was identified that belonged to coa type V and CC/ST22-SCCmec IV/t790 lineage. Resistance to mupirocin was detected in 9.3% of strains. All 7 mupirocin resistant MDR-MRSA isolates exhibited resistance to mupirocin in high level. Of these, 4 isolates belonged to CC/ST8-SCCmec IV/t008 (57.1%), 2 isolates belonged to CC/ST8-SCCmec IV/t064 (28.6%) and one isolate to CC/ST22-SCCmec IV/t790 (14.3%).

Altogether, current survey provides a snapshot of the characteristics of S. aureus strains isolated from patients. Our observations highlighted type III as predominant coa type among multidrug-resistant MDR strains indicating low heterogeneity of these isolates. Our study also indicates the importance of continuous monitoring of the genotypes of MDR-MRSA isolates to prevent nosocomial outbreaks and the spread of MDR isolates.

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Journal of Radioanalytical and Nuclear Chemistry
Authors: Mehdi Ardestani, Ali Arabzadeh, Zahra Heidari, Amirreza Hosseinzadeh, Hamidreza Ebrahimi, Elham Hashemi, Mona Mosayebnia, Mohammad Shafiee-Alavidjeh, Abbas Alavi, Mohammad Babaei, Arman Rahmim, Seyed Ebrahimi, and Massoud Amanlou

Abstract  

DTPA is a very strong metal chelator widely utilized in radiopharmaceutical chemistry for conjugation of chemicals which do not have enough potency for direct metalo-labeling and also to manage toxic radioactive materials such as plutonium, americium, and curium. It is difficult to conjugate DTPA to an amine group in a singular direction and such reactions usually also coincidently produce a mixture of DTPA-bis-amides and DTPA-mono-amide resulting in considerable insufficiencies/difficulties in synthesis and especially yield/separation procedures. In this paper, novel methods for the exclusive synthesis of DTPA-mono-amide have been established which extensively reduce the difficulties otherwise encountered and increase the reaction’s yield considering the green chemistry approaches. This is expected to be of interest to radiopharmaceutical researchers interested in the DTPA (Radio)-metallic-conjugate. Overall, this paper provides a framework to achieve a higher degree of propriety from DTPA as a chelator to conjugate to the chemical compounds.

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