Detection and distribution of zoonotic pathogens in wild Norway rats (Rattus norvegicus) from Tehran, Iran.

This is the first study on the prevalence of vector-borne zoonotic pathogens found in Rattus norvegicus (R. norvegicus) in urban areas of Tehran, Iran. Serological tests were used to detect IgG antibodies against Coxiella burnetii (C. burnetii) and Rickettsia spp. using a commercial qualitative rat ELISA kit. The frequency of Streptobacillus moniliformis (S. moniliformis) and Bartonella spp. was determined using a conventional PCR method. Molecular detection and characterization of Leptospira spp. were conducted using TaqMan real-time PCR based on lipL32 gene and SecY typing methods. A total of 100 R. norvegicus rats were collected from five regions in Tehran, Iran, and investigated to determine their zoonotic pathogens. S. moniliformis and Bartonella spp. were detected in 23 of 100 (23%) and 17 of 100 (17%) R. norvegicus populations, respectively. The highest prevalence of S. moniliformis and Bartonella spp. with similar frequency rates (n = 6/20; 30%) was seen among the R. norvegicus rats captured from the northern and southern parts of Tehran, respectively. Seroreactivity against C. burnetii and Rickettsia spp. was detected in 4% and 1% of R. norvegicus, respectively. C. burnetii. was identified only in one rat captured from the eastern part of Tehran. Results showed that Leptospira spp. was detected only in two rats, collected from the southern part (n = 2/20; 10%) of Tehran. The secY typing method identified two different Leptospira species including L. interrogans and L. kirschneri. The results showed that urban rats might play an important role in transmission of zoonotic pathogens to humans.

Multicentre study of the main carbapenem resistance mechanisms in important members of the Enterobacteriaceae family in Iran.

Resistance to carbapenems has been increasingly reported from the Enterobacteriaceae family, with different mechanisms in different geographic parts of the world. This study investigated the mechanisms of carbapenem resistance in Escherichia coli, Klebsiella pneumoniae and Enterobacter spp. carried out as a multicentre study (n = 10). All third-generation cephalosporin-resistant E. coli, K. pneumoniae and Enterobacter spp. that had been recovered from the selected provinces were included. Modified Hodge test and Carba NP test were done as a phenotypical method for detection of carbapenemase; the most common carbapenemase was detected by PCR. We evaluated the presence of an active efflux pump by using cyanide 3-chlorophenylhydrazone. Overexpression of AcrA/B and presence of OqxAB was detected by real-time PCR and conventional PCR respectively. Microorganisms in this study included 58 E. coli, 95 K. pneumoniae and 60 Enterobacter spp. Modified Hodge test showed a sensitivity of 41% and a specificity of 83%, and the Carba NP test showed a sensitivity of 26% and a specificity of 92% for detection of carbapenemase. OXA-48 was the most frequently detected carbapenemase, followed by NDM-1. Thirty-nine percent and 27% of positive cyanide 3-chlorophenylhydrazone test organisms included active AcrA/B and OqxAB efflux pumps respectively. The result showed the Carba NP test was more specific than MHT. Data confirmed the involvement of AcrA/B and OqxAB efflux pump as a carbapenem resistance mechanism in selected bacteria. Similar to other reports from the Middle East, we found OXA-48 and NDM-1 to be the most frequent carbapenemase.

Coagulase-negative staphylococci (CoNS) meningitis: a narrative review of the literature from 2000 to 2020.

Coagulase-negative staphylococci (CoNS) are recognized as comprising the main part of human normal microbiota and are rarely associated with severe and intensive infections. However, these organisms can cause a number of infections in humans, especially immunocompromised patients and neonates. Bacterial meningitis, as an important and acute infection in the central nervous system, is still a major global health challenge and a serious infectious disease, causing a high rate of mortality and morbidity. CoNS as causative agents of meningitis are generally related to trauma or direct implantation of foreign bodies and the presence of a cerebrospinal fluid shunt. Numerous epidemiologic and clinical studies have shown that different CoNS isolates such as Staphylococcus capitis, Staphylococcus lugdunensis, Staphylococcus hominis, Staphylococcus epidermidis, Staphylococcus schleiferi, Staphylococcus saprophyticus, Staphylococcus warneri and Staphylococcus haemolyticus are more frequently associated with meningitis. This study attempts to determine the role of CoNS in meningitis and reviews the reported cases of meningitis induced by CoNS from the year 2000 to 2020 in the literature.

Evaluating the antimicrobial resistance patterns and molecular frequency of bla oxa-48 and bla GES-2 genes in Pseudomonas aeruginosa and Acinetobacter baumannii strains isolated from burn wound infection in Tehran, Iran.

The aim of this study is to evaluate the antimicrobial resistance patterns and molecular frequency of bla GES-2 and bla oxa-48 genes in Pseudomonas aeruginosa and Acinetobacter baumannii strains isolated from burn wound infection in Tehran, Iran. In this study, 50 isolates of A. baumannii and 48 isolates of P. aeruginosa were collected from the Burn Unit of Shahid Motahari Hospital at Tehran, Iran. Antibiotic susceptibility tests of all isolates were carried out using the disc diffusion method, and the production of extended-spectrum ß-lactamases (ESBLs) in isolates was surveyed by the double disc synergy method and based on CLSI (2019 AST M100) criteria. Finally, the frequency of bla GES-2 and bla oxa-48 genes was surveyed by PCR. Antibiotic susceptibility tests showed that 48/48 (100%) of P. aeruginosa isolates and 49/50 (98%) of A. baumannii isolates were resistant to ceftriaxone and cefotaxime, respectively. Ceftazidime exhibited the lowest (26/48; 54.1%) resistance rates against P. aeruginosa isolates. The production of ESBLs was seen in 8/48 (16.6%) and 3/50 (6%) of P. aeruginosa and A. baumannii isolates, respectively. On the basis of conventional PCR and sequencing, the frequencies of the bla GES-2 gene among P. aeruginosa and A. baumannii was 87.5% and 58%, respectively. Moreover, bla oxa-48 gene was detected in 70.83% and 92% of P. aeruginosa and A. baumannii isolates, respectively. Results suggest that antibiotic-resistant A. baumannii and P. aeruginosa strains isolated from burn patients are frequently found; therefore, it is absolutely necessary to implement continuous screening and follow-up programmes for detecting antimicrobial resistance.

High-resolution melt curve analysis for rapid detection of rifampicin resistance in Mycobacterium tuberculosis: a single-centre study in Iran.

This study aimed to compare the diagnostic accuracy of high-resolution melting (HRM) analysis in comparison with Xpert MTB/RIF as well as conventional drug susceptibility testing (DST) for the detection of rifampicin (RIF) resistance in Mycobacterium tuberculosis in Iran. A comparative cross-sectional study was carried out from April 2017 to September 2018. A total of 80 culture-positive clinical samples selected during the study period were analysed for detection of RIF-resistant TB by conventional DST, Xpert MTB/RIF, and sequencing. Sensitivity and specificity of the HRM calculated according to DST was our reference standard test in this study. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of HRM assay were found to be 100%, 89.33%, 38.46%, and 100% respectively. The analysis demonstrated that the diagnostic accuracy of HRM tests is insufficient to replace Xpert MTB/RIF and conventional DST. HRM tests have the advantage of time to result and may be used in combination with culture. Further work to improve molecular tests would benefit from standardized reference standards and the methodology.

Clinical characteristics and molecular epidemiology of children with meningitis in Tehran, Iran: a prospective study.

The molecular epidemiology of meningitis in children is unclear in Iran, and data are scarce. We aimed to characterize its clinical and paraclinical features as well as to determine the distribution of genotype/capsular types of common bacterial meningitis agents in children in Iran. All children suspected to have meningitis aged 4 days to 15 years were enrolled onto a prospective cross-sectional study from January 2015 to September 2017. Diagnostic values of clinical features, cerebrospinal fluid and serum parameters were evaluated independently and in combination with each other by multivariate logistic regression to develop a diagnostic rule. Genotype/capsular types of all the isolates were determined by targeting serotype-specific genes with uniplex or multiplex PCR. Among 119 patients suspected of having meningitis, 43 had bacterial meningitis, 19 aseptic and one tuberculous; and there were 56 nonmeningitis cases (NMC). Presentation of four features at the same time-cerebrospinal fluid white blood cell count, protein, polymorphonuclear leukocytes and serum C-reactive protein-revealed 100% sensitivity and 86.4% specificity for diagnosis of bacterial meningitis. Haemophilus influenzae type b (60%), Streptococcus pneumoniae serotype 3 (28.5%) and Neisseria meningitidis B (63.5%) were the most prevalent serotypes. This study demonstrated that a well-designed combination of clinical and paraclinical features is useful, but these combinations are not good enough to be relied on as stand-alone exclusionary tests for the diagnosis of bacterial meningitis. In addition, public immunization of infants with the most prevalent bacterial meningitis serotypes is recommended.

Role of efflux pumps in reduced susceptibility to tigecycline in Acinetobacter baumannii.

Acinetobacter baumannii is an important human pathogen responsible for a various type of infections. These bacterial strains are generally resistant to numerous antibiotics. Therefore, eradication of such strains is problematic and related to high mortality. We investigated the effect of cyanide 3-chlorophenylhydrazone (CCCP) efflux pump inhibitor in tigecycline-resistant strains of Acinetobacter baumannii. In a cross-sectional study, from July until the end of February 2017, eighty isolates of A. baumannii were recovered. Antimicrobial susceptibility testing against tigecycline was performed by the disc diffusion method and determination of minimum inhibitory concentration by broth microdilution method, according to Clinical and Laboratory Standards Institute guidelines. Active efflux pumps were detected by CCCP as an efflux pumps inhibitor, and the gene expression of some of the resistance/nodulation/division (RND)-type efflux pumps was measured by semiquantitative RT-PCR (qRT-PCR). Antibiotic susceptibility tests in this study showed that 78 of 80 A. baumannii isolates were resistant to tigecycline. The results of phenotypic detection of efflux pumps revealed that 23.07% of tigecycline-resistant A. baumannii isolates can contain active efflux pumps. On the basis of conventional PCR, genes coding for adeF and adeJ were detected in 76 (98%) A. baumannii isolates. The results of qRT-PCR showed that the transcript level of the adeJ gene increased in 66.6% A. baumannii isolates with CCCP-positive tests and was correlated with tigecycline resistance. The results of this study indicate that RND-type efflux pumps appear to play a significant role in the tigecycline resistance of A. baumannii.

Mycobacterium simiae pulmonary disease in Iran: systematic review and meta-analysis.

Mycobacterium simiae is one of the most common nontuberculous mycobacteria (NTM) microorganisms causing lung disease in many countries in the world. A reliable estimate of the extent of M. simiae pulmonary disease has not been well investigated in Iran. We systematically searched multiple databases to identify relative studies. Studies were excluded if they did not use the American Thoracic Society (ATS) and Infectious Diseases Society of America (IDSA) diagnostic criteria for NTM diseases. Data were extracted independently and in duplicate. We assessed pooled estimate by using a random model effect, and sources of heterogeneity were assessed by using Cochran's Q and the I 2 statistic. The potential for publication bias was explored by using Begg's and Egger's tests. All analyses were conducted with Stata 14.0 (StataCorp, College Station, TX, USA). Of 172 articles identified, seven met the inclusion criteria. Of 355 patients who were culture positive for NTM, 82 had M. simiae pulmonary disease according to the ATS/IDSA diagnostic criteria. The pooled frequency of M. simiae pulmonary disease among patients with NTM was 25.0% (95% confidence interval, 16.8-33.2). No evidence of publication bias was observed among the included studies (p >0.05 for Begg's and Egger's tests). Clinical isolates of M. simiae are increasingly being recognized as a cause of pulmonary disease in Iran and need further attention by health authorities.

Insertional inactivation of oprD in carbapenem-resistant Pseudomonas aeruginosa strains isolated from burn patients in Tehran, Iran.

In this study, we report the insertion sequence ISPpu21 in the oprD porin gene of carbapenem-resistant Pseudomonas aeruginosa isolates from burn patients in Tehran, Iran. Antibiotic susceptibility tests for P. aeruginosa isolates were determined. Production of metallo-ß-lactamases (MBLs) and carbapenemase was evaluated and the ß-lactamase-encoding and aminoglycoside-modifying enzyme genes were investigated by PCR and sequencing methods. The mRNA transcription level of oprD and mex efflux pump genes were evaluated by real-time PCR. The outer membrane protein profile was determined by SDS-PAGE. The genetic relationship between the P. aeruginosa isolates was assessed by random amplified polymorphic DNA PCR. In all, 10.52% (10/95) of clinical isolates of P. aeruginosa harboured the ISPpu21 insertion element in the oprD gene. The extended-spectrum ß-lactamase-encoding gene in ISPpu21-carrying isolates was blaTEM. PCR assays targeting MBL and carbapenemase-encoding genes were also negative in all ten isolates. The rmtA, aadA, aadB and armA genes were positive in all ISPpu21 harbouring isolates. The relative expression levels of the mexX, mexB, mexT and mexD genes in ten isolates ranged from 0.1- to 1.4-fold, 1.1- to 3.68-fold, 0.3- to 8.22-fold and 1.7- to 35.17-fold, respectively. The relative expression levels of the oprD in ten isolates ranged from 0.57- to 35.01-fold, which was much higher than those in the control strain P. aeruginosa PAO1. Evaluation of the outer membrane protein by SDS-PAGE suggested that oprD was produced at very low levels by all isolates. Using random amplified polymorphic DNA PCR genotyping, eight of the ten isolates containing ISPpu21 were shown to be clonally related. The present study describes a novel molecular mechanism, ISPpu21 insertion of the oprD gene, associated with carbapenem resistance in clinical P. aeruginosa isolates.