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Mycobacterium fortuitum is a clinically important species among nontuberculous mycobacteria (NTM). Treatment of diseases caused by NTM is challenging. The aim of this study was identification of drug susceptibility and detection of mutations in erm(39) related to clarithromycin resistance and in rrl related to linezolid resistance in clinical isolates of M. fortuitum in Iran. In the study, 328 clinical NTM isolates were subjected to identification based on rpoB and 15% of isolates were assigned to M. fortuitum. Minimum inhibitory concentration for clarithromycin and linezolid was determined by E-test. Altogether 64% of M. fortuitum isolates showed resistanc to clarithromycin and 18% of M. fortuitum isolates showed resistance to linezolid. PCR and DNA sequencing were performed in erm(39) and in rrl genes for detection of mutations related to clarithromycin and linezolid resistance, respectively. Sequencing analysis revealed (84.37%) single nucleotide polymorphisms in the erm(39). A total 55.55% of M. fortuitum isolates harbored an A→G, 14.81% harbored an C→A, 29.62% harbored an G→T mutation in erm(39) at position 124, 135, 275. Seven strains harbored point mutation in the rrl gene either at T2131C or at A2358G. Our findings showed M. fortuitum isolates have become a serious problem with high-level antibiotic resistance. The existence of drug resistance to clarithromycin and linezolid indicates more attention to the study of drug resistance in M. fortuitum.

Differentiating Mycobacterium tuberculosis (MTB) from nontuberculous mycobacteria (NTM) is very important in the treatment process of patients. According to the American Thoracic Society guideline (ATS), NTM clinical isolates should be identified at the species level proper treatment and patient management. This study aimed to identify NTM clinical isolates by evaluationg rpoB, ssrA, tuf, atpE, ku, and dnaK genes, and use multilocus sequence analysis (MLSA) to concatenate the six genes.Ninety-six Mycobacterium isolates, including 86 NTM and 10 MTB isolates, from all the patients referred to the certain TB Reference Centres were included. All isolates were evaluated by PCR amplification of rpoB, ssrA, tuf, ku, atpE, and dnaK genes and MLSA.Out of 96 isolates, 91 (94.8%), 87 (90.6%), 72 (75%), 84 (87.5%) and 79 (82.3%) were differentiated to the species level by rpoB, tuf, ssrA, dnaK and atpE genes, respectively. The ku gene was able to identify 69 (80.2%) isolates of the 86 NTM isolates to the species level. We could identify 100% of the isolates to the species level by MLSA.None of the PCR targets used in this study were able to completely differentiate all species. The MLSA technique used to concatenate the six genes could increase the identification of clinical Mycobacterium isolates and all 16 species were well-differentiated.

The global rise in drug-resistant Mycobacterium tuberculosis (M.tb), and especially the significant prevalence of isoniazid (INH)-resistance constitute a significant challenge to global health. Therefore, the present study aimed to investigate mutations in prevalent gene loci-involved in INH-resistance phenotype-among M.tb clinical isolates from southwestern Iran.Drug susceptibility testing (DST) was performed using the conventional proportional method on confirmed 6620 M.tb clinical isolates, and in total, 15 INH-resistant and 18 INH-susceptible isolates were included in the study. Fragments of six genetic loci most related to INH-resistance (katG, inhA promoter, furA, kasA, ndh, oxyR-ahpC intergenic region) were PCR-amplified and sequenced. Mutations were explored by pairwise alignment with the M.tb H37Rv genome.The analysis of gene loci revealed 13 distinct mutations in INH-resistant isolates. 60% (n = 9) of the INH-resistant isolates had mutations in katG, with codon 315 predominately (53.3%, n = 8). Mutation at InhA - 15 was found in 20% (n = 3) of resistant isolates. 26.7% (n = 4) of the INH-resistant isolates had kasA mutations, of which G269S substitution was the most common (20%, n = 3). The percentage of mutations in furA, oxyR-ahpC and ndh was 6.7% (n = 1), 46.7% (n = 7), and 20% (n = 3), respectively. Of the mutations detected in ndh and oxyR-ahpC, 5 were also observed in INH-susceptible isolates. This study revealed seven novel mutations, four of which were exclusively in resistant isolates.This study supports the usefulness of katG and inhA mutations as a predictive molecular marker for INH resistance. Co-detection of katG S315 and inhA-15 mutations identified 73.3% (11 out of 15 isolates) of INH-resistant isolates.

There are limited studies on the antibiotic resistance patterns of slowly growing mycobacteria (SGM) species and their related gene mutations in Iran. This study aimed to elucidate the antibiotic susceptibility profiles and the mutations in some genes that are associated with the antibiotic resistance among SGM isolates from Iran. The SGM strains were isolated from sputum samples of suspected tuberculosis (TB) patients. SGM species were identified by standard phenotypic tests and were assigned to species level by amplification and sequencing of the dnaK gene. The minimum inhibitory concentration (MIC) of eight antibiotics was determined using broth microdilution method. The mutations in rrl, rpoB, gyrA, and gyrB genes were investigated in clarithromycin, rifampin, and moxifloxacin resistant isolates using sequencing method. A total of 77 SGM isolates including 46 (59.7%) Mycobacterium kansasii, 21 (27.3%) Mycbacterium simiae, and 10 (13%) Mycobacterium avium complex (MAC) were detected. The amikacin and linezolid with the susceptibility rates of 97.4% and 1.3% were the most and the least effective antibiotics, respectively. All MAC and M. simiae isolates, and 32 (69.6%) M. kansasii strains had multiple-drug resistance (MDR) profiles. The rrl, rpoB, gyrA, and gyrB genes showed various mutations in resistant isolates. Although the current study showed an association among resistance to the clarithromycin, rifampin, and moxifloxacin with mutations in the relevant genes, further research using the whole-genome sequencing is needed to provide a clearer insight into the molecular origins of drug resistance in SGM isolates.

This study was aimed to evaluate the antibiotic resistance, biofilm formation, and genetic diversity of carbapenem-resistant Pseudomonas aeruginosa (CRPA) strains isolated from four types of nosocomial infections (NIs) including urinary tract infection (UTI), ventilator-associated pneumonia (VAP), surgical site infection (SSI), and bloodstream infection (BSI).In total, 115 isolates of NIs-causing P. aeruginosa were collected from NIs. Antibiotic susceptibility testing (AST) was performed using disk diffusion method and minimum inhibitory concentrations. Biofilm formation was tested on 96-well polystyrene microtiter plates (MTP). CRPA isolates were genotyped using multiple-locus variable number of tandem repeat analysis (MLVA). The most resistance and susceptibility rates were observed to amikacin (70.6%) and colistin (96.1%), respectively. Colistin and meropenem were the most active antimicrobial agents in VAP, SSI, and BSI. While, colistin and cefepime were the most active in UTIs. In total, 52.2% (n = 60/115) of P. aeruginosa isolates were carbapenem resistant, of which 95.0%, 55.0%, and 5.0% were multidrug-resistant, extensively drug-resistant, and pandrug-resistant, respectively. There was a significant association between resistance to carbapenem and resistance to other antibiotics except for piperacillin/tazobactam. The biofilm production of CRPA isolates was 95.0%, of which 23.3% were strong biofilm producers. Based on MLVA, there were 34 different types of CRPA isolates classified into three main clusters and 5 sub clusters.The association of CRPA with other antibiotic resistance, the high rates of biofilm production, and the high genetic diversity of the isolates may be a warning of the need for a careful surveillance program.

Oral candidiasis (OC) is the most frequent opportunistic fungal infection, which is a predictive indicator of immunosuppression and disease progression among people living with HIV/AIDS (PLWHA). In the present study, 109 Candida isolates were collected from 94 PLWHA afflicted with oral Candida infection (OCI) following highly active antiretroviral therapy (HAART). The susceptibility profiles of Candidaspp. to six antifungal agents were evaluated using CLSI broth microdilution. The prevalence of OCI was 34.06%. The susceptibility profile of Candidaspp. revealed 100% sensitivity to caspofungin, while 6.4%, 5.4%, 24.5%, and 2.8% of Candida isolates showed resistance or nonwild-type MICs to fluconazole, itraconazole, posaconazole, and amphotericin B, respectively. Notably, 15.9% of patients and 3.7% of isolates showed mixed Candida infections and multidrug resistance, respectively. The low-level resistance to antifungal agents observed in the present study may be explained by the fact that none of the participants had prior and prolonged exposure to these antifungals. However, more focus should be placed on the mechanisms of reduced susceptibility and low-level resistance in Candida species since they can serve as stepping stones to developing clinical resistance. Alongside this, it seems a must to understand the local epidemiology of Candida spp. and their susceptibility pattern.

Background This study aimed to investigate the occurrence of Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran by culture and the real-time PCR of 23S-5S rRNA gene spacer region. Methods and results A total of 123 clinical respiratory samples including 63 pleural aspirates, 57 bronchoalveolar lavage (BAL), and 3 sputum were collected from 65 males and 58 females with pneumonia symptoms. All samples were cultured on the Modified Wadowsky-Yee (MWY) agar. The Legionella species was identified by routine bacteriological tests. The presence of the 16S-23S rRNA spacer region gene was investigated by real-time PCR. The Legionella species were differentiated by sequencing of 16S-23S rRNA gene. A total of 2 (1.6%) BAL specimens were positive for Legionella species by culture method. No Legionella spp. were identified in pleural aspirates and sputum samples by the culture method. Using real-time PCR, 9 (7.3%) samples including 6 BAL, 1 sputum, and 2 pleural aspirates were positive for legionella species. These species were detected in 3 (5.2%) females and 6 males (9.2%). The results of sequencing showed that eight species were L. pneumophila while one was L. cherrii. Also, the 2 isolates that were identified by culture method, were confirmed as L. pneumophila by sequencing. Conclusions The results showed that using the real-time PCR has a more efficacy for detecting of Legionella species in respiratory samples. Also, L. pneumophila was the most prevalent species circulating in the southwest region of Iran. So, periodic monitoring programs is recommended to prevent epidemics due to this bacterium.

Burn infection continues to be a major issue of concern globally and causes more harm to developing countries. This study aimed to identify the aerobic bacteriological profiles and antimicrobial resistance patterns of burn infections in three hospitals in Abadan, southwest Iran. The cultures of various clinical samples obtained from 325 burn patients were investigated from January to December 2019. All bacterial isolates were identified based on the standard microbiological procedures. Antibiotic susceptibility tests were performed according to the CLSI. A total of 287 bacterial species were isolated from burn patients. Pseudomonas aeruginosa was the most frequent bacterial isolate in Gram-negative bacteria and S. epidermidis was the most frequent species isolated in Gram-positive bacteria. The maximum resistance was found to ampicillin, gentamicin, ciprofloxacin, while in Gram-negative bacteria, the maximum resistance was found to imipenem, gentamicin, ciprofloxacin, ceftazidime, and amikacin. The occurrence of multidrug resistance phenotype was as follows: P. aeruginosa (30.3%), Enterobacter spp (11.1%), Escherichia coli (10.5%), Citrobacter spp (2.1%), S. epidermidis (2.8%), S. aureus, and S. saprophyticus (0.7%). Owing to the diverse range of bacteria that cause burn wound infection, regular investigation, and diagnosis of common bacteria and their resistance patterns is recommended to determine the proper antibiotic regimen for appropriate therapy.

Escherichia coli has been most widely used for production of the recombinant proteins. Over-expression of the recombinant proteins is the mainspring of the inclusion bodies formation. The refolding of these proteins into bioactive forms is cumbersome and partly time-consuming. In the present study, we reviewed and discussed most issues regarding the recovery of “classical inclusion bodies” by focusing on our previous experiences. Performing proper methods of expression, solubilization, refolding and final purification of these proteins, would make it possible to recover higher amounts of proteins into the native form with appropriate conformation. Generally, providing mild conditions and proper refolding buffers, would lead to recover more than 40% of inclusion bodies into bioactive and native conformation.

Mycobacterium tuberculosis resistant to effective first-line drugs (FLDs) has challenged national and global tuberculosis control programs. This study aimed to identify mutations in 4 genes related to rifampin, pyrazinamide, and ethambutol resistance among clinical isolates of M. tuberculosis from southwestern Iran. After drug susceptibility testing of 6620 M. tuberculosis clinical isolates by proportional method, a total of 24 FLD-resistant strains were included in the study. Fragments of rpoB, pncA, embB, and ubiA genes were amplified and sequenced to mine the mutations by pairwise alignment with the corresponding M. tuberculosis H37Rv genes. Phenotypic resistance to rifampin, isoniazid, and ethambutol was detected in 67, 54, and 33% (n = 16, 13, and 8) of the isolates, respectively. Of rifampin-resistant isolates, 31% (5/16) were mono-resistant, and 56% (9/16) were multidrug-resistant (MDR). In 100% of rifampin-resistant isolates, mutations were found in the rifampin resistance-determining region (RRDR) of the rpoB, with S450L substitution being the most common, especially in MDRs (77.8%, 7/9). Resistance-conferring mutations in pncA were present in 12.5% (3/24) of FLD-resistant isolates. The embB and ubiA mutations were found in 62.5 and 12.5% (5/8 and 1/8) of ethambutol-resistant isolates, respectively, of which the embB D354A was the most common substitution (37.5%, 3/8). Sixteen distinct mutations were identified, one of which was novel. The sequence analysis of the RRDR segment was the best way to detect rifampin resistance. The rpoB S450L substitution could be a helpful molecular marker to predict MDR. In other genes, no mutation was identified as a reliable marker.

Background Non-tuberculous mycobacterial (NTM) infections are growing concern in many countries around the globe including Iran. Among them, Mycobacterium kansasii (M. kansasii) causes both pulmonary and extra-pulmonary infections. Despite the high prevalence of M. kansasii isolates in Iran, unfortunately little is known about the epidemiological aspects of M. kansasii infection. Hence, the aim of the present study was to investigate the molecular identification, determination of subtypes variation and geographic distribution of clinical isolates of M. kansasii isolates. Methods In the present study, 108 clinical pulmonary isolates suspected to NTM were collected from four Tuberculosis Regional Reference Laboratories in Iran during 2016-2018. The isolates were confirmed as NTM using conventional and molecular methods. Among them, M. kansasii isolates were subjected to rpoB gene sequencing. For determination of subtyping of M. kansasii isolates, polymerase chain reaction-restriction enzyme analysis (PCR-REA) based on the hsp65 gene was performed. Results Based on the rpoB gene sequence analysis, 33 (30.5%) isolates were identified as M. kansasii species, compared to 31 (28.7%) isolates using phenotypic methods. The subtype I was the most frequent subtype (n=24; 72.7%), followed by subtype II (n=8; 24.2%). Conclusion We indicated that the rate of M. kansasii isolation with clinical significance appears to be increasing in Iran, especially in highly industrialized cities. The high rate of M. kansasii subtype I may suggest that this genotype has a particular potency for colonization, and a higher epidemiological potential for causing infection in humans. More studies are needed to provide a better understanding of the biology and pathogenicity of M. kansasii subtype I.

Background Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most pathogens associated with health care. Molecular typing methods are vital for outbreak investigations of MRSA. The aim of this study was characterization of SCCmec, spa types and multidrug resistant of methicillin-resistant Staphylococcus aureus isolates in Ahvaz, Iran. Methods A total of 50 MRSA isolates were determined by using the phenotypic method and mecA gene. Antibiotic resistance profile and SCCmec types were screened using disc diffusion method and PCR, respectively. For spa typing of MRSA isolates, two molecular typing methods including the PCR-sequencing and high-resolution melting (HRM) analysis were used. Results In the present study, the highest sensitivity of MRSA was to vancomycin and linezolid and the lowest to clindamycin. In the MRSA isolates, 22% were XDR and 78% were MDR. SCCmec type III was found commonly among MRSA. Based on PCR-sequencing and HRM results, 10 different spa types were identified. The spa types t037 and t030 were the most common in this study. Conclusion This study emphasizes the spa variation among MRSA isolates, which may be considered as an important criterion when treating staphylococcal infections. Accurate and early detection of MDR, XDR, or even PDR MRSA isolates strains must be commenced by all clinical microbiology laboratories to reduce the menace of antimicrobial resistance.

Background Among Non-tuberculous mycobacteria (NTM) which generally cause opportunistic infections, especially in immunocompromised hosts, Mycobacterium simiae (M. simiae) is one of the most important NTM, associated with pulmonary disease. The main concern about M. simiae infections is the extreme resistance of this NTM to antibiotics. There are limited studies about drug susceptibility testing (DST) and the causes of drug resistance in M. simiae. Hence, the current study aimed to identify the M. simiae isolates and to assess the drug resistance of the isolates using phenotypic and molecular methods. Materials and methods In this study, 50 clinical pulmonary isolates suspected of NTM were collected from regional tuberculosis reference laboratories in Iran. The isolates were identified as M. simiae by using standard biochemical tests and molecular methods. DST was performed for identified M. simiae isolates and additional 35 M. simiae isolates from the department archive, against eight drugs. The mutations in gyrA, gyrB, and rrl genes in clarithromycin and moxifloxacin resistant isolates were investigated by polymerase chain reaction (PCR) followed by sequencing. Results Out of 50 suspected NTM isolates, 25 isolates were detected as M. simiae species based on the biochemical tests, and 18 isolates were verified based on the rpoB gene sequence analysis to achieve a total of 53 isolates when the archive isolates were included. DST results showed that all 53 isolates were resistant to isoniazid, rifampin, and clofazimine. The rate of resistance to ethambutol and linezolid were 34 (64%), and 40 (76%) respectively. The highest susceptibility rate was demonstrated for amikacin 53 (100%) and clarithromycin 45(85%), followed by moxifloxacin 35(66%). Sequence analysis showed mutations in positions 2058 and 2059 of the rrl gene, as well non-synonymous mutation at codons 389, 444, and 571 of the gyrB gene. Sequence analysis showed no mutation in the gyrA gene. drug-resistant isolates with mutations showed higher MICs compared to non-mutant resistant isolates. Conclusions This study revealed amikacin, clarithromycin, and moxifloxacin as the most effective antibiotics. However, since M. simiae exhibited a high level of antibiotic resistance in vitro, therefore, species identification and determining the antibiotic susceptibility pattern of the isolates are essential before treatment.

α-l-Guluronic acid (ALG; G2013) has been previously introduced as a new anti-inflammatory agent with promising therapeutic effects. Thus, in the present study, we aimed to evaluate the acute and sub-acute toxicity of ALG through intravenous (i.v.) administration in Balb/C mice. ALG was administrated i.v. to the mice with doses of 300, 600, and 1000 mg/kg of body weight to investigate acute toxicity (single dose) and with doses of 25, 50, and 100 mg/kg of body weight to sub-acute toxicity study (daily injections for a period of 28 days). The mortality rate, food and water intake, behavior, body weight, gross necropsy, hematological and biochemical parameters as well as histopathological presentations of the vital organs (kidneys, liver, lungs, spleen, and heart) were examined in treated groups and compared to the healthy controls. The results of both acute and sub-acute studies showed that i.v. administrations of ALG did not affect the investigated parameters in both sexes, indicating that the LD

Purpose: Dental unit's environment and relevant instruments are a major source of infectious diseases caused by a variety of microorganisms. The application of various disinfectants is one of the most effective methods for reducing or eliminating microbial contamination. The objective of this study was to evaluate the antibacterial effects of deconex and sodium hypochlorite against bacterial taxa isolated from dental unit's environment of Ahvaz Jundishapur University of Medical Sciences, southwest of Iran. Methods: In order to evaluate the quality of disinfection, sampling was performed from different parts of 100 clinical units. For bacterial recovery and isolation, samples were enriched and cultured onto different microbiological culture media. Species identification was carried out using phenotypic and molecular methods (16S rDNA sequence analysis). In vitro activity of sodium hypochlorite and deconex were determined by the broth micro-dilution method. Results: According to conventional techniques, Bacillus spp (48%) was the most frequently encountered isolates, followed by staphylococcus spp (26%). By using both techniques, Bacillus subtilis was the most frequently encountered species (n=23, 21%), followed by Bacillus licheniformis (n=8, 7.4%), Streptococcus pneumonia (n=8, 7.4%), Staphylococcus epidermidis (n=8, 7.4%), Staphylococcus saprophyticus (n=8, 7.4%) and Staphylococcus warneri. The highest levels of contamination were observed in oral medications. The deconex had lower minimum inhibitory concentration (MIC) concentration in comparasion to sodium hypochlorite, which showed that deconex was a much more potent disinfectant. Conclusion: In conclusion, the results of the present in vitro study showed that deconex had promising results for decontamination of the tested microorganism, and it is recommended for disinfecting of dental units and environment. In this study, the high percentage of dental unit's contamination showed the need to improve disinfection procedures, sterilization systems, and the use of an appropriate concentration of deconex and sodium hypochlorite for dental units decontamination .

BackgroundThe drug resistance is expected to be the most important challenge in infection control in Iran, where there is no local report or standard drug resistance monitoring system. Therefore, this study aimed to investigate the aerobic and anaerobic bacterial profile of nosocomial infections and their antibiotic resistance in Ahvaz, southwest Iran.MethodologyThe gram-positive and gram-negative bacteria were identified on the basis of conventional culture and biochemical tests. The antibiotic resistance of the bacterial isolates against antibiotics was determined by the disk diffusion method.ResultsAmong total 1156 collected positive samples,E.coliand coagulase-negative staphylococci (CoNS) were the most frequent pathogenic gram negative bacteria (GNB) and gram positive bacteria (GPB) respectively. Drug susceptibility testing revealed that among GNB,P.aeruginosawas 100% resistant to amikacin, cefepime, ciprofloxacin and tetracycline. In the case ofE.coli, the resistance rate was (98%) for trimethoprim sulfamethoxazole and cefepime. For GPB,S.aureusshowed the highest resistance rates to amikacin (100%) and clindamycin (100%). In addition, CoNS strains showed a high level of resistance to doxycycline (100%), erythromycin (100%) and cefoxitin (97%). InBacteroeides fragilisisolates, the highest resistance rate belonged to clindamycin (72%), andClostridium perfringensstrains showed high level of resistance to penicillin (46%).ConclusionThe results highlighted that there are distinct factors leading to antimicrobial resistance in Ahvaz, southwest Iran. The primary contributors to the resistance development, include poor surveillance of drug-resistant infections, poor quality of available antibiotics, clinical misuse, and the ease of access to antibiotics. Moreover, similar factors such as self-medication and the lack of regulation on medication imports play a role in antibiotic resistance in the region.

Background & objectives: West Nile virus (WNV) is a positive-sense, single-stranded RNA virion, that belongs to the Flaviviridae family. This virus is preserved in a bird-mosquito cycle that is capable of inducing diseases as a dead-end or endpoint host in humans as well as horses. In 2016, a suspicious case of crow population death was reported by the Department of Environment, Ministry of Health, Iran. Considering the mass migration of birds together with the WNV-related symptoms, including uncoordinated walking, ataxia, inability to fly, lack of awareness, and abnormal body posture, it was necessary to further investigate the possible causes of this incident. The objective of this study was molecular detection of WNV in crows utilizing the real-time PCR method in the northern provinces of Iran. Methods: A total of 12 crows (8 dead, 4 alive) with a possible WNV infection, were collected from the northern provinces of Iran (Golestan, Mazandaran, and Guilan). A tissue sample of the liver, kidney, or lung was collected from all the crows, and RNA was isolated using an RNA extraction kit. A one-step real-time PCR method using a TaqMan probe was used for virus detection. Results: All the infected crows were positive for WNV. The 132-bp real-time PCR amplicon of the genome was detected in all the samples. Comparative phylogenetic analysis revealed that WNV isolated from Iran clustered with strains from the USA, Hungary, and Culex pipiens. Interpretation & conclusion: The WNV genome sequence was detected in all the infected crows. The results confirmed the connection of this isolation with clade1a strains. Hence, determining the epidemiologic and prevalence characteristics of the WNV for transmission control is of critical importance in Iran.

Background Nocardia species belong to the aerobic actinomycetes group of bacteria which are gram-positive and partially acid-fast Bacilli. These bacteria may sometimes be associated with nosocomial infections. Nocardia diseases are not required to be reported to public health authorities in Iran. Hence, the present study was designed to determine the prevalence of human Nocardia spp. in Iran by using a systematic review and meta-analysis according to the preferred reporting items for systematic reviews and meta-Analyses statement. Methods The data of the prevalence of Nocardia species were collected from databases such as Embase, PubMed/MEDLINE via Ovid, Web of Science, Scopus and Google Scholar as well as national Iranian databases, including SID, Magiran. Analyses were conducted by STATA 14.0. Results The meta-analyses showed that the proportion of Nocardia spp. in Iranian studies varied from 1.71(1.17, 2.24) to 0.46(0.09, 0.83). N. asteroides (21% [95% CI 1.17, 2.24]), N. cyriacigeorgica (17% [95% CI 0.99, 1.77]), N. facanica (10% [95% CI 0.75, 1.00]) were considered to be common causative agents. Conclusions Our study presents that despite the fact that Nocardia spp. are normally are saprophytic organisms, are currently accounts as emerging pathogens due to an increase in immunocompromised patients among Iranian populations. Considering our results, the establishment of advanced diagnostic facilities for the rapid detection of Nocardia infections are required for optimal therapeutic strategies of Nocardia spp. in Iran. Our findings could help the programmatic management of the disease within the context of Nocardia control programs.

Background: Enteropathogenic Escherichia coli (EPEC) is one of the most important pathogens among young children worldwide. Both eae and bfp genes have been used to identify EPEC strains and categorize them into typical and atypical strains. They may be an emerging pathogen in both developing and developed countries. Objectives: This study was primarily conducted to assess the epidemiology, drug resistance, and β-lactamase distribution of EPEC, as well as the detection of efa1/lifA in atypical strains. Methods: A total of 251 E. coli strains isolated from children with diarrhea were evaluated for their EPEC pathotype by PCR for the presence of eae, stx1, stx2, and bfp genes. Serogrouping with polyvalent antisera was performed to confirm EPEC strains. Atypical EPEC-containing samples were evaluated for the efa1/lifA gene. EPEC isolates were assessed to recognize the antibiotic resistance and screened to detect extended-spectrum β-lactamases (ESBLs). Results: Enteropathogenic E. coli strains were detected in 17 (6.78%) of E. coli isolates by PCR. The prevalence of typical and atypical strains was determined at 35.3% and 64.7%. All strains were completely susceptible to colistin, imipenem, and meropenem. The prevalence of blaCTX-M and blaTEM genes was calculated at 70.58% and 58.82%, respectively. Conclusions: Enteropathogenic E. coli isolates are completely sensitive to carbapenems, and precise therapeutic strategies are required to prevent the spread of these beta-lactamase genes among diarrheagenic E. coli.

Background Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most pathogens associated with health care. Molecular typing methods are vital for outbreak investigations of MRSA. The aim of this study was characterization of SCCmec, spa types and multidrug resistant of methicillin-resistant Staphylococcus aureus isolates in Ahvaz, Iran. Methods A total of 50 MRSA isolates were determined by using the phenotypic method and mecA gene. Antibiotic resistance profile and SCCmec types were screened using disc diffusion method and PCR, respectively. For spa typing of MRSA isolates, two molecular typing methods including the PCR-sequencing and high-resolution melting (HRM) analysis were used. Results In the present study, the highest sensitivity of MRSA was to vancomycin and linezolid and the lowest to clindamycin. In the MRSA isolates, 22% were XDR and 78% were MDR. SCCmec type III was found commonly among MRSA. Based on PCR-sequencing and HRM results, 10 different spa types were identified. The spa types t037 and t030 were the most common in this study. Conclusion This study emphasizes the spa variation among MRSA isolates, which may be considered as an important criterion when treating staphylococcal infections. Accurate and early detection of MDR, XDR, or even PDR MRSA isolates strains must be commenced by all clinical microbiology laboratories to reduce the menace of antimicrobial resistance.

Tuberculosis (TB) is considered as one of the most important infectious diseases in the world, and recent rise and spread of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) strains, have made the matter worsened. Due to the importance of TB prevalence in Iran, this study was designed to investigate the genetic diversity among MDR strains of MTB by MIRU-VNTR typing scheme. A total of 88 drug resistant M. tuberculosis isolates belong to pulmonary TB cases were collected from several TB reference centers of Iran. Drug susceptibility testing for Isoniazid and Rifampin was performed using the agar proportion method and MDR isolates were underwent genotyping by using 12-locus- based MIRU-VNTR typing. On performing proportion method, 22 isolates were identified as MDR. By typing of MDR isolates using 12-loci MIRU-VNTR technique, high diversity were demonstrated in MDR strains and these were classified into 20 distinct MIRU-VNTR genotypes. MIRU loci 10 and 26 were the most discriminatory loci with 8 and 7 alleles respectively; while MIRU loci 2, 20, 24 and 39 were found to be the least discriminatory with 1–2 alleles each. We noticed a mixed infection in isolate 53, as this isolate comprised simultaneous two alleles in MIRU loci 40, 10, 16 and 39. In conclusion, this result represents MIRU-VNTR typing as a useful tool for studying genetic diversity of MDR-MTB in regional settings, and will help the health sectors to construct a preventive program for MDR-TB. Additionally, it can detect mixed infection which can facilitate management of treatment.

In the present study, zinc oxide (ZnO) and iron oxide (FeO) nanoparticles were examined for their possible, in vitro anthelmintic effects against Toxocara vitulorum. The worms were incubated for 24 h with different concentrations (0.004, 0.008 and 0.012% w/v) of the nanoparticles. The parasite mobility, mortality, superoxide dismutase (SOD) activity, malondialdehyde (MDA) and nitric oxide (NO) level were recorded at different time intervals. The results showed that both of the nanoparticles could significantly decrease worm’s mobility, increase mortality rate as well as elevate MDA and NO content as compared to control group in a time- and concentration-dependent manner. SOD activity was elevated with the low concentration of the nanoparticles but it was decreased in higher ones. It can be concluded that ZnO and FeO nanoparticles exert their anthelmintic effects via induction of oxidative/nitrosative stress.

Staphylococcus saprophyticus is the second most frequent community-acquired causative agent of acute urinary tract infection (UTI). Some strains of S. saprophyticus can create biofilms, increasing their virulence. Once biofilms have been produced, antibiotic resistance is exacerbated. Hence, the aims of the present study were the study of biofilm formation, structure and antibiotic resistance in S. saprophyticus strains causing UTIs in women in Ahvaz, Iran. Overall, 43 S. saprophyticus isolates were recovered from UTIs. Antibiotic resistance pattern and the biofilm production and structure were determined using phenotypic methods. Most S. saprophyticus isolates were resistant to erythromycin, but all isolates were sensitive to linezolid and vancomycin. Fifty-eight per cent of S. saprophyticus were multidrug resistant. Twenty-one per cent of S. saprophyticus isolates harbored the mecA gene. Biofilm formation was observed in 65% of S. saprophyticus isolates and most had polysaccharide matrix. Our data indicate high rates of antibiotic resistance and the capability of biofilm formation among S. saprophyticus isolates. The emergence of antibiotic resistance in the management of UTIs is a serious public health issue. The findings of this study could be used to improve treatment plans to control UTIs. Consequently, increased awareness of the mechanisms underlying biofilm formation and the development of drug resistance will allow UTIs to be more efficiently controlled and treated.

Background Immunotherapy, during which the immune system of the patient is manipulated to act against tumors has been among the most successful methods in the treatment of breast cancer, a leading cause of mortality among women worldwide. Objectives To investigate the immunotherapeutic efficacy of Lactobacillus casei lysate as an adjuvant in combination with a heated-4T1 mammary carcinoma cell lysate in a model of breast cancer. Methods After ethics committee approval of all animal procedures, a murine model of breast cancer was induced in BALB/c mice using 4T1 cells. These mice were immunized with a combination of lysates of heated 4T1 cells and L. casei. Subsequent changes in tumor size and weight, and the production of TNF-α, IL-2, IL-12, IL-17, and IL13 were measured. Lung weights were measured as an indicator of metastasis to other organs. Results The tumor size and weight in mice immunized with the combined vaccine were significantly reduced compared with controls. The combined immunotherapy altered the pattern of cytokine production to the advantage of antitumor immunity, and was significantly more potent than immunization with heated-4T1-cell lysate or L. casei lysate alone. Conclusions Coadministration of L. casei lysate enhanced the immunotherapeutic efficacy of the heated-4T1-cell lysate as a source of tumor-associated antigens. L. casei can potentially be used as an adjuvant combined with sources of tumor antigens in the treatment of cancers, and as a safe alternative to the current adjuvants that cause greater irritation to hosts. Further studies are required to clarify the mechanisms underlying these effects.

Lentiviral gene delivery is now considered as a major candidate in the future of cancer therapy. To avoid common side-effects associated with methods of cancer therapy, the survivin promoter shows great promise due to its high expression level in multiple cancers. In this research, using a 2A peptide, Noxa was coexpressed with hemagglutinin neuraminidase (HN) driven by the survivin promoter (Surp). Coding genes of Noxa and HN were connected to the upstream and downstream of a 2A peptide using SOE PCR. After cloning of Noxa-2A-HN and Surp into the lentiviral vector, the final construct was transfected to SK-BR3 tumor cells and human embryonic kidney 293T cells. Quantitative real-time PCR implied efficient transfection and integration of the lentiviral vector in target cells. Western blotting analysis was performed to verify the expression of Noxa and HN and we determined cell viability by MTT calorimetric assay after transfection. Our results showed that the expression of Noxa and HN, though not utterly specific, was relatively higher in tumor cells and led to the selective suppression of tumor cells, which eventually resulted in a cell survival reduction of 76%. In conclusion, the novel pLN2AH-Surp lentiviral vector proved the potential for further in vivo and clinical experiments.

There are accumulating reports of the emergence of drug-resistant strains of HSV-1 that have become a barrier to successful treatment of HSV-1 infection. Therefore, there is a pressing need to identify and evaluate alternative antiherpetic agents. The aim of the present study was to investigate the effect of copper oxide nanoparticles (CuO-NPs) on HSV-1 infection. The MTT assay was applied to examine the cytotoxic effects of CuO-NPs on Vero cells. Antiherpetic potency was determined using the TCID50 and quantitative Real-Time PCR assays. To evaluate the inhibitory impact of CuO-NPs on the expression of viral antigens, an indirect immunofluorescence assay (IFA) was performed. Acyclovir was used as a reference drug in all experiments. Exposure of HSV-1 with CuO-NPs at the highest non-toxic concentration (100 μg/mL) resulted in 2.8 log10 TCID50 reduction in infectious virus titer as compared with virus control (P

Influenza type A is considered as a severe public health concern. The mechanism of drugs applied for the control of this virus depends on two surface glycoproteins with antigenic properties, namely hemagglutinin (HA) and neuraminidase (NA). HA aids the virus to penetrate cells in the early stage of infection and NA is an enzyme with the ability to break glycoside bonds, which enables virion spread through the host cell membrane. Since NA contains a relatively preserved active site, it has been an important target in drug design. Oseltamivir is a common drug used for the treatment of influenza infections, for which cases of resistance have recently been reported, giving rise to health concerns. Flavonoids are natural polyphenolic compounds with potential blocking effects in the neuraminidase active site. Based on their antiviral effect, the flavonoids quercetin, catechin, naringenin, luteolin, hispidulin, vitexin, chrysin and kaempferol were selected in the present study and compared alongside oseltamivir on molecular docking, binding energy and active site structure, in order to provide insight on the potential of these compounds as targeted drugs for the control and treatment of influenza type A. The molecular characterization of flavonoids with binding affinity was performed using AutoDock Vina software. The results indicated that these compounds may effectively block the NA active site. Therefore, these natural compounds derived from fruits have the potential for development into drugs for controlling influenza, which may aid overcome the clinical challenge of the H1N1 strain epidemic.

Introduction Enteroaggregative Escherichia coli (EAEC) has been implicated as an emerging cause of traveler’s diarrhea, persistent diarrhea among children, and immunocompromised patients. The present study aimed to investigate the prevalence of antibiotic resistance, extendedspectrum β-lactamase (ESBL) production, and virulence factors of EAEC isolates obtained from Iranian children suffered from diarrhea. Materials and methods In this cross-sectional study, from March 2015 to February 2016, 32 EAEC isolates were collected from fecal samples of children aged

Azar Dokht Khosravi,1,2 Mitra Roointan,2 Effat Abbasi Montazeri,2 Sajad Aslani,3,4 Mohammad Hashemzadeh,1,2 Moslem Taheri Soodejani5 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Department of Microbiology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; 4Student Research Committee, Kerman University of Medical Sciences, Kerman, Iran; 5Department of Biostatistics and Epidemiology, School of Health, Kerman University of Medical Sciences, Kerman, Iran Introduction: Coagulase-negative staphylococci (CoNS) are normal inhabitants of human skin and mucous membranes. However, CoNS represent one of the major nosocomial pathogens, especially in immunocompromised patients. The increasing incidence of CoNS and mainly methicillin-resistant strains underlines the need for an accurate identification of Staphylococcus isolates at the species level. Analysis of the tuf gene proved to be an accurate tool for the species identification of CoNS. The aims of this study were to identify the CoNS species by tuf gene-based polymerase chain reaction method and sequencing, and to determine the frequency of CoNS clinical isolates resistant to methicillin (MRCoNS) and other antibiotics.Methods: A total of 200 staphylococci isolates were collected from various clinical samples. Phenotyping methods were used for initial identification followed by polymerase chain reaction amplification of tuf gene with subsequent sequencing. The phylogenetic relationships among species were analyzed using the neighbor-joining method based on the partial gene sequence of tuf. Microbroth dilution test was used for screening methicillin resistance, and disk diffusion susceptibility testing was performed for evaluation of antibiotic resistance among the isolates.Results: In the present study, 125 isolates were identified as CoNS; among them, Staphylococcus epidermidis 54(43.2%) and Staphylococcus haemolyticus 50 (40.0%) were demonstrated as the most prevalent species. Resistance to methicillin was detected in 54.4% of the CoNS based on microbroth dilution method. In disk diffusion susceptibility testing, the greatest resistance of CoNS was demonstrated for cefoxitin (65.4%), cotrimethoxazole (54.4%), and clindamycin (49.6%), while daptomycin (87.2%) and linezolid (83.2%) showed the greatest effectiveness for CoNS isolates.Conclusion: Our results confirmed the predominance of S. epidermidis and S. haemolyticus among CoNS isolates. The high prevalence of MRCoNS strains is a serious concern and strongly suggests the need for control program measures in our hospitals in order to reduce MRCoNS infections, especially in immunocompromised patients. Keywords: coagulase-negative staphylococci, tuf gene, antimicrobial resistance, PCR, susceptibility testing

Mansour Amin,1,2,* Mehrandokht Sirous,2,* Hazhir Javaherizadeh,3–5 Mohammad Motamedifar,6,7 Morteza Saki,2,8 Hojat Veisi,2 Saeedeh Ebrahimi,1,9 Sakineh Seyed-Mohammadi,2,8,* Mohammad Hashemzadeh1,2 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Abuzar Children’s Hospital, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Alimentary Tract Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 5Nursing Care Research Center in Chronic Diseases, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 6Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; 7Shiraz HIV/AIDS Research Center, Institute of Health, Shiraz University of Medical Sciences, Shiraz, Iran; 8Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 9Department of Virology, Faculty of Mdicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran *These authors contributed equally to this work Introduction: Enteroaggregative Escherichia coli (EAEC) has been implicated as an emerging cause of traveler’s diarrhea, persistent diarrhea among children, and immunocompromised patients. The present study aimed to investigate the prevalence of antibiotic resistance, extended-spectrum β-lactamase (ESBL) production, and virulence factors of EAEC isolates obtained from Iranian children suffered from diarrhea. Materials and methods: In this cross-sectional study, from March 2015 to February 2016, 32 EAEC isolates were collected from fecal samples of children aged

Background: Puma is a highly robust pro-apoptotic protein. The protein becomes activated by p53 ensuing beyond-repair DNA damage. Downregulation of SIRT 1, by miR-128, elevates activated p53 that foment Puma indirectly. Objectives: In the present study, we used two-expression Adeno-Associated Virus (AAV) system for co-expression of miR-128 and Puma in order to evaluate apoptotic response; both in the tumor and normal cells, respectively. Materials and Methods: Three recombinant AAVs constructs were generated. The First rAAV bearing Puma under the control of hTERT (p-AAV), the second construct designed such that to carry miR-128 downstream of CMV (mi-AAV), and the last construct comprises of the both CMV-miR-128 and hTERT- Puma. Real-Time PCR and western blotting were used to evaluate expression levels of the transduced genes. Results: MTT assay and DAPI staining shown suicidal effect of each recombinant AAV vectors. p-AAV cytotoxicity was recorded for 62% of the tumor cells, while for normal cells it was only 20% cytotoxic. The second construct, mi-AAV, was not as potent and selective as p-AAV. This construct was shown to be 27% and 16% cytotoxic for BT-474 and HEK-293 cells, respectively. Co-expression of Puma and miR-128 (p-mi-AAV) was accomplished with a selective cytotoxicity toward BT-474. This construct was 85% toxic for tumor cells, although it was only 25% toxic for the normal cell line (HEK-293). Conclusions: In this study, we have shown that not only Puma is able to instigate apoptotic response but also its co-expression along with miR-128 could significantly enhance apoptosis in a synergistic manner.