Farhad B. Hashemi

Frequency of Chlamydia trachomatis in Women with Cervicitis in Tehran, Iran

Chlamydia trachomatis (CT) is the most common cause of bacterial sexually transmitted infection (STI) worldwide, but current data concerning the prevalence of CT among women in Iran is scarce. Data regarding the frequency of CT infection among Iranian women can help to justify the implementation of a national CT screening program that can reduce the high morbidity associated with sequelae of CT infections by treating infected women. Endocervical secretions from 123 married women (20–55 years) with cervicitis were tested by a PCR-EIA method using primers to amplify a CT-specific plasmid. The digoxigenin-labeled amplicon was measured by hybridization to a biotin-labeled probe and a strepavidin-coated plate, followed by an enzyme-linked colorimetric analysis. Overall frequency of CT infection among women was 17% (21/123). The range of CT frequency among various age groups was 12–25%. The 31–40-year-age group comprised the majority (49%) of CT positive samples, followed by 20–30 year group (33%). Although the 20-to-30-year-old women reported the highest frequency of STI history, they had the lowest relative frequency of CT infection (12%). There is a high frequency of CT infection among women with cervicitis in Tehran, Iran, thus indicating a necessity to implement a routine CT screening program in the major cities of Iran and possibly nationwide. Identification of CT-infected women may prevent its spread, and thereby reduce the high morbidity associated with CT infections among women in Iran.

Antibiotic Resistance of Acinetobacter baumannii in Iran: A Systemic Review of the Published Literature

Objectives Acinetobacter baumannii is a bacterium responsible for health care-associated infections, and it frequently develops multiple drug resistance (MDR). The prevalence of antibiotic-resistant A. baumannii in Iran has increased, and this may cause significant clinical problems. Therefore, in order to elucidate the development of antibiotic resistance, we performed a systematic review of the literature published on antibiotic-resistant A. baumannii reported in Iran. Methods Thirty-six publications that met the criteria for inclusion were reviewed from an initial 87 papers. Selected papers published between 2008 and September 2014, were categorized on the basis of the sample collecting year been between 2001 and 2013. Results Analysis of data revealed that, in general, there was an increase in antimicrobial resistance. During the initial time point of these studies (2001–2007) there was a high rate of resistance to all antibiotics, with the exception of carbapenems, lipopeptides, and aminoglycosides that had a low resistance rate in comparison with the others. Also, the resistance rate was increased in one group of these three antimicrobial groups from 2010 to 2013. In particular, there was an increase in resistance to carbapenems (imipenem and meropenem) from 2010–2011 and 2012–2013, whereas no significant change in the resistance rate of the other two antimicrobial groups (lipopeptides and aminoglycosides) during the study time was observed, although we did observe certain trends in amikacin (aminoglycoside group antibiotic) between 2011–2012 and 2012–2013. Conclusion These findings indicate that antimicrobial resistance of A. baumannii in Iran has increased, which may very well affect the antimicrobial resistance of this organism worldwide. Based on these results, novel prevention and treatment strategies against A. baumannii infections are warranted. Furthermore, these data may assist in revising treatment guidelines and regional policies in care units to slow the emergence of antimicrobial resistance.

Wide distribution of carbapenem resistant Acinetobacter baumannii in burns patients in Iran

Antimicrobial resistance in carbapenem non-susceptible Acinetobacter baumannii (CNSAb) is a major public health concern globally. This study determined the antibiotic resistance and molecular epidemiology of CNSAb isolates from a referral burn center in Tehran, Iran. Sixty-nine CNSAb isolates were tested for susceptibility to antimicrobial agents using the E test methodology. Multiple locus variable number tandem repeat analysis (MLVA), Multilocus sequence typing (MLST) and multiplex PCR were performed. PCR assays tested for ambler classes A, B, and D β-lactamases. Detection of ISAba1, characterization of integrons, and biofilm formation were investigated. Fifty-three (77%) isolates revealed XDR phenotypes. High prevalence of blaOXA-23-like (88%) and blaPER-1 (54%) were detected. ISAba1 was detected upstream of blaADC, blaOXA-23-like and blaOXA51-like genes in, 97, 42, and 26% of isolates, respectively. Thirty-one (45%) isolates were assigned to international clone (IC) variants. MLVA identified 56 distinct types with six clusters and 53 singleton genotypes. Forty previously known MLST sequence types forming 5 clonal complexes were identified. The Class 1 integron (class 1 integrons) gene was identified in 84% of the isolates. The most prevalent (33%) cassette combination was aacA4-catB8-aadA1. The IC variants were predominant in the A. baumannii lineage with the ability to form strong biofilms. The XDR-CNSAb from burned patients in Iran is resistant to various antimicrobials, including tigecycline. This study shows wide genetic diversity in CNSAb. Integrating the new Iranian A. baumannii IC variants into the epidemiologic clonal and susceptibility profile databases can help effective global control measures against the XDR-CNSAb pandemic.

In Vitro Evaluation of the Antimicrobial Activity of Nanosilver‑Mineral Trioxide Aggregate against Frequent Anaerobic Oral Pathogens by A Membrane‑enclosed Immersion Test

Background: Nanoparticles of silver (NanoAg) have been shown to control the growth of bacteria, but application of NanoAg in endodontics has not been evaluated. This in vitro study evaluates the antimicrobial activity of NanoAg to enhance the inhibitory effects of mineral trioxide aggregate (MTA). Methods: The antibacterial activities of NanoAg and NanoAg-MTA against four types of anaerobic pathogens were tested in vitro using (1) agar diffusion test (ADT) and (2) a newly devised membrane-enclosed immersion test (MEIT). Results: Both NanoAg and NanoAg-MTA inhibited the growth of all four test bacteria at 25 ppm concentration. MEIT analysis consistently showed that NanoAg enhanced the antimicrobial activity of MTA significantly, and the bacterial susceptibility to lower concentrations of NanoAg varied depending on the type of bacteria. Overall, NanoAg-MTA showed significant inhibitory effect which was time and dose dependent. Conclusions: Our data support that NanoAg can serve as an excellent MTA additive against anaerobic endodontic-periodontal pathogens with clinical applications for infection control in endodontics.

Antimicrobial Resistance of Acinetobacter baumannii to Imipenem in Iran: A Systematic Review and Meta-Analysis

Imipenem-resistant multi-drug resistant (IR-MDR) Acinetobacter baumannii has been emerged as a morbidity successful nosocomial pathogen throughout the world.To address imipenem being yet the most effective antimicrobial agent against A. baumannii to control outbreaks and treat patients, a systematic review and meta-analysis was performed to evaluate the prevalence of IR-MDR A. baumannii. We systematically searched Web of Science, PubMed, MEDLINE, Science Direct, EMBASE, Scopus, Cochrane Library, Google Scholar, and Iranian databases to identify studies addressing the antibiotic resistance of A. baumannii to imipenem and the frequency of MDR strains in Iran. Out of 58 articles and after a secondary screening using inclusion and exclusion criteria and on the basis of title and abstract evaluation, 51 studies were selected for analysis. The meta-analysis revealed that 55% [95% confidence interval (CI), 53.0–56.5] of A. baumannii were resistant to imipenem and 74% (95% CI, 61.3–83.9) were MDR. The MDR A. baumannii population in Iran is rapidly changing toward a growing resistance to imipenem. Our findings highlight the critical need for a comprehensive monitoring and infection control policy as well as a national susceptibility review program that evaluates IR-MDR A. baumannii isolates from various parts of Iran.

Frequency ofChlamydia trachomatisin Women with Cervicitis in Tehran, Iran

Chlamydia trachomatis (CT) is the most common cause of bacterial sexually transmitted infection (STI) worldwide, but current data concerning the prevalence of CT among women in Iran is scarce. Data regarding the frequency of CT infection among Iranian women can help to justify the implementation of a national CT screening program that can reduce the high morbidity associated with sequelae of CT infections by treating infected women. Endocervical secretions from 123 married women (20–55 years) with cervicitis were tested by a PCR-EIA method using primers to amplify a CT-specific plasmid. The digoxigenin-labeled amplicon was measured by hybridization to a biotin-labeled probe and a strepavidin-coated plate, followed by an enzyme-linked colorimetric analysis. Overall frequency of CT infection among women was 17% (21/123). The range of CT frequency among various age groups was 12–25%. The 31–40-year-age group comprised the majority (49%) of CT positive samples, followed by 20–30 year group (33%). Although the 20-to-30-year-old women reported the highest frequency of STI history, they had the lowest relative frequency of CT infection (12%). There is a high frequency of CT infection among women with cervicitis in Tehran, Iran, thus indicating a necessity to implement a routine CT screening program in the major cities of Iran and possibly nationwide. Identification of CT-infected women may prevent its spread, and thereby reduce the high morbidity associated with CT infections among women in Iran.

Immunogenicity of Pseudomonas aeruginosa recombinant b-type fagellin as a vaccine candidate: Protective efficacy in a murine burn wound sepsis model

Pseudomonas aeruginosa (PA) is a formidable opportunistic pathogen among patients with burn wound infections. Antimicrobial therapy is often unsuccessful because PA can develop multi-drug resistance; thus, immunotherapy can be a rational alternative. The goal of this study was to evaluate the immunogenicity recombinant type b flagellin (r-b-flagellin) as a potential vaccine against P. aeruginosa in a mouse model for burn wound sepsis. Primary immunization with r-b-flagellin (10μg) followed by two booster shots was sufficient to generate a robust humoral response, which was predominantly a T helper 2 (Th2) type response consisting mainly of subtype IgG1 and low levels of IgG2a. Analysis of the Th1-Th2 response among immunized mice showed an increased production of IL-4, INF-γ and IL-17 by splenocytes upon stimulation by r-b-flagellin. Opsono-phagocytosis assays confirmed the enhanced killing of bacteria by anti r-b-flagellin immune sera. These antibodies were also able to inhibit motility of P. aeruginosa and afforded protection to immunized mice by reducing bacterial load in the site of original infection into the liver of challenged mice. The reduction of systemic bacterial spread resulted in an increase in the survival rate of challenged immunized mice. In conclusion, immunization of mice with r-b-flagellin protein increased the level of humoral and cellular immune response and led to an efficacious protection against P. aeruginosa infection in the burn mouse model.

Genotypic and Antimicrobial Susceptibility of Carbapenem-resistant Acinetobacter baumannii: Analysis of ISAba Elements and blaOXA-23-like Genes Including a New Variant

Carbapenem-resistant Acinetobacter baumannii (CR-AB) causes serious nosocomial infections, especially in ICU wards of hospitals, worldwide. Expression of blaOXA genes is the chief mechanism of conferring carbapenem resistance among CR-AB. Although some blaOXA genes have been studied among CR-AB isolates from Iran, their blaOXA-23-like genes have not been investigated. We used a multiplex-PCR to detect Ambler class A, B, and D carbapenemases of 85 isolates, and determined that 34 harbored blaOXA-23-like genes. Amplified fragment length polymorphism (AFLP) genotyping, followed by DNA sequencing of blaOXA-23-like amplicons of CR-AB from each AFLP group was used to characterize their blaOXA-23-like genes. We also assessed the antimicrobial susceptibility pattern of CR-AB isolates, and tested whether they harbored insertion sequences ISAba1 and ISAba4. Sequence comparison with reference strain A. baumannii (NCTC12156) revealed five types of mutations in blaOXA-23-like genes; including one novel variant and four mutants that were already reported from China and the USA. All of the blaOXA-23-like genes mutations were associated with increased minimum inhibitory concentrations (MICs) against imipenem. ISAba1 and ISAba4 sequences were detected upstream of blaOXA-23 genes in 19 and 7% of isolates, respectively. The isolation of CR-AB with new blaOXA-23 mutations including some that have been reported from the USA and China highlights CR-AB pervasive distribution, which underscores the importance of concerted national and global efforts to control the spread of CR-AB isolates worldwide.

Immunogenicity and protective efficacy of Pseudomonas aeruginosa type a and b flagellin vaccines in a burned mouse model

Immunogenicity and efficacy of Pseudomonas aeruginosa type a and b flagellins (hereafter, flagellins) as candidate vaccines were evaluated using an experimental burned mouse model. The protection afforded and the reduction in bacterial burden achieved by these vaccine candidates were determined. Primary immunization with flagellins followed by two booster shots generated a robust immune response. Cytokine analysis demonstrated the secretion of interleukin-4 more than interferon-γ from immunized T-cells in response to in vitro antigen stimulation. IgG response was of Th2 type, predominantly with IgG1 and lower IgG2a levels before and after challenge. In vitro opsonophagocytosis assays confirmed protective potential of immune sera via enhanced bacterial cell killing. Immune sera also inhibited P. aeruginosa motility. Serum cytokine analysis demonstrated high IL-12 and low IL-10 levels in flagellin-immunized mouse sera. Reduced systemic bacterial spread from original infection site into liver and spleen was associated with increased survival. Immunization of mice with flagellins increased the humoral immune response and protection against P. aeruginosa infection in our mouse model.

Immunization with Bivalent Flagellin Protects Mice against Fatal Pseudomonas aeruginosa Pneumonia

Pseudomonas aeruginosa lung infections present a major challenge to healthcare systems worldwide because they are commonly associated with high morbidity and mortality. Here, we demonstrate the protective efficacy of type a and b flagellins (bivalent flagellin) against acute fatal pneumonia in mice. Mice immunized intranasally with a bivalent flagellin vaccine were challenged by different flagellated strains of P. aeruginosa in an acute pneumonia model. Besides the protective effect of the vaccine, we further measured the host innate and cellular immunity responses. The immunized mice in our study were protected against both strains. Remarkably, active immunization with type a or b flagellin significantly improved survival of mice against heterologous strain compared to flagellin a or b antisera. We also showed that after an intranasal challenge by P. aeruginosa strain, neutrophils are recruited to the airways of vaccinated mice, and that the bivalent flagellin vaccine was proved to be protective by the generated CD4+IL-17+ Th17 cells. In conclusion, bivalent flagellin vaccine can confer protection against different strains of P. aeruginosa in an acute pneumonia mouse model by eliciting effective cellular and humoral immune responses, including increased IL-17 production and improved opsonophagocytic killing.