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Background: Enterococcus faecium is one of the members of ESKAPE pathogens. Due to its resistance to antimicrobial agents, treating this bacterium has become challenging. The development of innovative approaches to combat antibiotic resistance is necessary. Phage therapy has emerged as a promising method for curing antibiotic-resistant bacteria. Methods: In this study, E. faecium phages were isolated from wastewater. Phage properties were characterized through in vitro assays (e.g. morphological studies, and physicochemical properties). In addition, whole genome sequencing was performed. A hydrogel-based encapsulated phage was obtained and its structure characteristics were evaluated. Wound healing activity of the hydrogel-based phage was assessed in a wound mice model. Results: The purified phage showed remarkable properties including broad host range, tolerance to high temperature and pH and biofilm degradation feature as a stable and reliable therapeutic agent. Whole genome sequencing revealed that the genome of the EF-M80 phage had a length of 40,434 bp and harbored 65 open reading frames (ORFs) with a GC content of 34.9% (GenBank accession number is OR767211). Hydrogel-based encapsulated phage represented an optimized structure. Phage-loaded hydrogel-treated mice showed that the counting of neutrophils, fibroblasts, blood vessels, hair follicles and percentage of collagen growth were in favor of the wound healing process in the mice model. Conclusion: These findings collectively suggest the promising capability of this phage-based therapeutic strategy for the treatment of infections associated with the antibiotic-resistant E. faecium. In the near future, we hope to expect the presence of bacteriophages in the list of antibacterial compounds used in the clinical settings. [ABSTRACT FROM AUTHOR]

Introduction: One ofthe most hazardous infections for humans is still Mycobacterium tuberculosis. Finding the connections between various clinical strains has been a top focus in epidemiological research. Methods: In this work, we created a very strong phylogeny of Mycobacterium TB using multilocus sequence analysis, or MLSA. To identify polymorphic nucleotide sites, five gene fragments from the Rpsl (302 bp), MprA (559 bp), LipR (322 bp), KatG (488 bp), and Fgd1 (266 bp) regions were subjected to MLSA, a single nucleotide polymorphism (SNP)-based technique. The Hunter-Gaston Index (HGI) was used to measure the discriminatory power of each locus for all genes. Results: Each distinct allelic profile in this investigation was given a sequence type (ST) number; nine sequence types from a total of twenty strains were found, suggesting a significant degree of strain diversity in this region. Conclusion: Our results showed that the presence of high genetic diversity among clinical isolates of M. tuberculosis in Northeast of Iran. There is no evidence for recent transmission. [ABSTRACT FROM AUTHOR]

Background: The global spread of plasmids carrying carbapenemase genes within carbapenem resistant Acinetobacter baumannii (CRAB) strains poses a worldwide public health issue. In this study, we conducted a comprehensive genetic analysis of plasmids and chromosomes harboring the major carbapenemase genes (blaNDM, blaKPC, blaVIM, blaIMP, blaGES, blaOXA-58-like, blaOXA-24/40-like, blaOXA-143-like, and blaOXA-23-like) in CRAB strains using bioinformatic tools. Methods: We retrieved plasmids and chromosomes carrying the major carbapenemase genes from GenBank. The size, replicon type, and conjugal apparatus of the plasmids were also determined. Furthermore, allele types, coexistence of other antimicrobial resistance genes alongside carbapenemases in plasmids or chromosomes, co-occurrence of carbapenemase genes, gene repetition, and sequence types (ST) of whole genomes were characterized. Results: The database contained 113 plasmids and 38 chromosomes harboring carbapenemase genes. This investigation revealed that blaNDM and blaOXA-58-like were the predominant allele types in both the plasmids and chromosomes. Nine (7.96%) plasmids with blaNDM-1 were potentially conjugative. The most common replicon types of the plasmids were R3-T1, R3-T8, R3-T2, R3-T23, and RP-T1. The analysis revealed that blaNDM-1 and blaOXA-58-like genes possessed the highest variety of co-existence with other antibiotic resistance genes. The cooccurrence of dual carbapenemases was identified in 12 plasmids and 19 chromosomes. Carbapenemase gene repetitions were identified in 10 plasmids and one chromosome. Circular alignment revealed that the plasmids carrying the co-occurrence of blaNDM-1 and blaOXA-58 were more homogeneous. However, there was heterogeneity in certain regions of these plasmids. According to the minimum spanning tree (MST) results, the majority of the plasmids belonged to the genomes of ST2Pas, ST1Pas, ST422Pas, ST622Pas, and ST85Pas. Conclusion: A. baumannii appears to have a strong ability for genome plasticity to incorporate carbapenemase genes on its plasmids and chromosomes to develop resistance against carbapenems. Mobilizable plasmids harboring carbapenemases significantly contribute to the dissemination of these genes. The genetic structure of the plasmids revealed a strong associations of class I integrons, ISAba-like structures, Tn4401 elements, and aac (6')-Ib with carbapenemases. Furthermore, gene repetition may also be associated with carbapenem heteroresistance. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Acinetobacter baumannii strains are increasing worldwide. In this study, samples were collected from hospital environments, extra hospital environments, and fecal carriages. 76% (89/117) of bacterial isolates were detected as A. baumannii strains. The imipenem resistance in the hospital environment, fecal carriages, extra hospital environments, and clinical isolates was 37.7% (17/45), 100% (9/9), 0% (0/45), and 92.9% (92/99), respectively. The blaVIM and blaOXA-23 were detected in 6.6% (3/45) and 2.2% (1/45) of strains isolated from hospital environments. Interestingly, strains isolated from fecal carriages had blaVIM, blaOXA-23, and blaIMP genes which resembled carbapenem resistance genes in clinical strains. The structure of clonal relatedness among all non-clinical isolates was as follows: CC2, 37% (33/89); CC1, 22.4% (20/89); CC3, 12.3% (11/89); CC25, 7.8% (7/89); CC10, 4.4% (4/89) and CC15, 2.2% (2/89). Comparison of clonal relatedness among clinical and non-clinical isolates indicated that widespread clones including CC2, CC3, and CC10 were common clonal complexes between two categories. [ABSTRACT FROM AUTHOR]

The emergence of carbapenem resistance among Escherichia coli is a serious threat to public health. The objective of this study was to investigate resistance genes and clonality of carbapenem resistant E. coli in Iran. Between February 2015 and July 2016, a total of 32 non-duplicate E. coli isolates that were ertapenem resistant or intermediate (R/I-ETP) were collected from patient clinical or surveillance cultures (rectal swabs) at two university hospitals. Resistance genes were identified by PCR and sequencing. Conjugation experiments, PCR-based replicon typing, PFGE and multilocus sequence typing (MLST) were performed. PCR assays showed, among the 32 isolates, twenty-nine strains produced carbapenemase genes. The predominant carbapenemase was bla OXA-48 (82.8%), followed by bla NDM-1 (31%), bla NDM-7 (6.9%) and bla OXA-181 (3.4%). Seven of the bla NDM positive isolates co-harbored bla OXA-48 carbapenemases. The bla NDM and bla OXA-48 were found in IncA/C and IncL/M conjugative plasmids, respectively. The bla CTX-M-15 , qnrA and intI1 genes were also present in most isolates. The PFGE revealed genetic diversity among the 28 E. coli isolates, which belonged to six minor PFGE clusters and 14 isolates were singletons. The 26 isolates were distributed into 18 STs, of which two were dominant (ST648 and ST167). We identified one bla NDM-1 -positive ST131 E. coli isolates that harbor the bla CTX-M-15 and bla TEM genes. Horizontal transfer of IncA/C and IncL/M plasmids has likely facilitated the spread of the bla OXA-48 and bla NDM genes among E. coli . Their clonal diversity and the presence of faecal carriers in isolates suggest an endemic spread of OXA-48 and NDM. Therefore, it emphasizes the critical importance of monitoring and controlling the spread of carbapenem resistant E. coli . [ABSTRACT FROM AUTHOR]

Background: Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. The main mechanism of resistance of this bacterium to fluoroquinolones and carbapenems are the modification of type II topoisomerases (DNA gyrase and topoisomerase IV) and alterations in the OprD porin, respectively. The aim of this study was to examine for the occurrence of mutations related to fluoroquinolone resistance of gyrA and parC genes and mutational inactivation of oprD gene of clinical isolates using DNA sequencing technique. Methods: A total of 60 P. aeruginosa isolates were collected from the hospitalized patients in the Intensive Care Units (ICUs) of Al-Zahra hospital located in Isfahan, Iran. The pattern of sensitivity to antibiotics was determined using CLSI disk diffusion and MIC methods. The assay was based on a DNA sequencing method using polymerase chain reaction (PCR) for amplification and sequencing of t h e selected genes. Results: The results show that replacement of Ile for Thr-83 in gyrA was the only replacement, while other substitutions not observed. No mutations were found in parC. The most frequent amino acid alterations were E185Q, P186G, and V189T, found in five resistance isolates, However, nucleotide insertions and deletions mutations not observed. Conclusion: Our study suggested that mutation of gyrA and oprD genes may play a minor role in fluoroquinolone and carbapenem resistance and other mechanisms may contribute to the fluoroquinolone and carbapenem resistance of P. aeruginosa. [ABSTRACT FROM AUTHOR]

Background: Since December 2019, the world is struggling with an outbreak of coronavirus disease-2019 (COVID-19) infection mostly represented as an acute respiratory distress syndrome and has turned into the most critical health issue worldwide. Limited information is available about the association between dynamic changes in the naso/oropharyngeal viral shedding in infected patients and biomarkers, aiming to be assessed in the current study. Materials and Methods: This quasi-cohort study was conducted on 31 patients with moderate severity of COVID-19 manifestations, whose real-time polymerase chain reaction (RT-PCR) test was positive for severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) RNA at baseline. RT-PCR was rechecked for patients every 3–4 days until achieving two negative ones. In parallel, biomarkers, including lymphocyte count, lactate dehydrogenase (LDH), and C-reactive protein (CRP), were assessed every other day, as well. Viral shedding also was assessed. Results: Spearman's correlation test revealed a significant direct correlation between the viral shedding from the symptom onset and the time, in which CRP (P = 0.0015, r = 0.54) and LDH (P = 0.001, r = 0.6207) return to normal levels after symptom onset, but not for lymphocyte count (P = 0.068, r = 0.34). Conclusion: Based on the current study's findings, the duration of SARS-CoV-2 RNA shedding was directly correlated with the required time for LDH and CRP return to normal levels. Therefore, these factors can be considered the determinants for patients' discharge, isolation, and return to social activities; however, further investigations are required to generalize the outcomes. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter species (spp.) have unique ability to survive in the hospital environments for long periods. This feature increases the rate of acquisition of infection of hospitalized patients by this high-resistant bacterium. The purpose of this study was determination of prevalence and antibiotic resistance pattern of Acinetobacter spp. isolated from Alzahra hospital, Isfahan, Iran. Methods: During a three-month period, using the swab method, 40 samples were taken from different parts of the intensive care unit (ICU) and surgery wards of the hospital. All samples were examined for the presence of Acinetobacter spp. After isolation, Acinetobacter spp. were identified using several phenotypic and biochemical tests. Antibiotic susceptibility test was done by disk diffusion method according to CLSI-2011 (Clinical and Laboratory Standards Institute) guidelines. Findings: From 40 swab samples, 202 isolates were obtained that 109 (53.96%) were belonging to gram positive bacilli and coccoci and 93 (46.03%) were gram negative. In the culture of 21 samples (52.5%) Acinetobacter spp. was grown. Most of the isolates were obtained from sink and bed of patients' room, respectively. All of isolates were multidrug resistant (MDR) and 18 isolates (85.71%) were extensively drug resistant (XDR) because of their resistance to carbapenems. Conclusion: Regard increasing rate of health-care associated infections (HAI) caused by Acinetobacter spp. and spreading of distribution of the high-resistance strains in the hospital environments, infection control procedures seems to be necessary to eliminate potential sources of infection. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas aeroginosa (P. aeroginosa) is one of the most important causes of hospital infections, especially in the intensive care unit (ICU). Increasing antibiotic-resistance of P. aeroginosa cause a lot of problems for patients and in some cases, lead to septicemia and death. The most important antibiotics used in the treatment of P. aeruginosa infection are ceftazidime, ciprofloxacin, imipenem and piperacillin mainly used in hospitals. Unsuitable and prolonged use of antibiotics has led to the emergence of multidrug (MDR) and pandrug resistant (PDR) strains. In fact, infections with MDR and PDR strains often result in increased cost of treatment, lengthy stay, and overall morbidity and mortality. This study aimed to determine the pattern of drug resistance in P. aeruginosa infection in ICU of Al-Zahra hospital (Isfahan, Iran). Methods: 66 isolates of P. aeruginosa from different clinical specimens from ICU wards of Al-Zahra hospital were isolated. Antibacterial susceptibility test for imipenem, meropenem, amikacin, gentamicin, ciprofloxacin, levofloxacin, aztreonam, cefepime, piperacillin, piperacillin/tazobactam and ceftazidime was performed using disk diffusion (Kirby-Bauer) method. Findings: Of 66 separated isolates, 51 (77.2%) were of MDR and 33 (50%) were of PDR strain. 75.8% were resistant to ceftazidime and 72.7% to piperacillin. In fact, most of isolates were resistant to both antibiotics. Conclusion: This study shows that overuse and misuse of antibiotics in hospitals has increased drug resistance and creation of the PDR and MDR strains. The results suggest that antibiotic resistance can be determined by choosing the appropriate drug to treat patients. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is an important nosocomial pathogen which causes a wide range of infections. In this study, we addressed the role of class 1 integron, ISAba1 and ISAba125 associated with antimicrobial resistance in 72 clinical isolates of A. baumannii collected from clinical settings in Tehran, Iran. Moreover, to study the clonal relatedness of strains, repetitive extragenic palindromic-PCR (rep-PCR) assay was carried out. PCR revealed that blaOXA-51-like, blaOXA-23-like, blaOXA-24/40-like, blaOXA-58-like, blaNDM, integrase gene (intI1), ISAba1, and ISAba125 were present in 86.11% (62/72), 84.72% (61/72), 30.55% (22/72), 0% (0/72), 0% (0/72), 58.33% (42/72), 97.22% (70/72), and 65.27% (47/72) of the strains, respectively. Sequencing of 39 internal variable regions of class 1 integrons showed seven gene cassette arrays, including aadA4-catB8-aadA1 (2.77%), aadA1-aadA4 (1.38%), aacC4-aadA1 (2.77%), aacC4 (22.22%), aadA1 (13.88%), aadA4 (5.55%), and catB8 (5.55%). We detected ISAba1 in the upstream of blaOXA-23-like, blaOXA-51-like, and blaADC in 54.16% (39/72), 9.72% (7/72), and 56.94% (41/72) of the strains, respectively. Whereas, there was a low frequency of disruptions in carO and dacD genes: 5.55% (4/72) and 4.16% (3/72). Rep-PCR analysis revealed that the isolates were genetically diverse. However, Cl-12 and Cl-15 were the largest clusters and they were recovered from various hospitals. Our analysis showed the high rates of class 1 integrons as a repertoire of aminoglycoside-modifying enzymes. It seems that linkages of ISAba1-blaOXA-23-like and ISAba1-blaOXA-69, and disruptions in carO or dacD can develop resistance to carbapenems among clinical isolates of A. baumannii. [ABSTRACT FROM AUTHOR]

Background: Enterococcus faecium is one of the members of ESKAPE pathogens. Due to its resistance to antimicrobial agents, treating this bacterium has become challenging. The development of innovative approaches to combat antibiotic resistance is necessary. Phage therapy has emerged as a promising method for curing antibiotic-resistant bacteria., Methods: In this study, E. faecium phages were isolated from wastewater. Phage properties were characterized through in vitro assays ( e.g. morphological studies, and physicochemical properties). In addition, whole genome sequencing was performed. A hydrogel-based encapsulated phage was obtained and its structure characteristics were evaluated. Wound healing activity of the hydrogel-based phage was assessed in a wound mice model., Results: The purified phage showed remarkable properties including broad host range, tolerance to high temperature and pH and biofilm degradation feature as a stable and reliable therapeutic agent. Whole genome sequencing revealed that the genome of the EF-M80 phage had a length of 40,434 bp and harbored 65 open reading frames (ORFs) with a GC content of 34.9% (GenBank accession number is OR767211). Hydrogel-based encapsulated phage represented an optimized structure. Phage-loaded hydrogel-treated mice showed that the counting of neutrophils, fibroblasts, blood vessels, hair follicles and percentage of collagen growth were in favor of the wound healing process in the mice model., Conclusion: These findings collectively suggest the promising capability of this phage-based therapeutic strategy for the treatment of infections associated with the antibiotic-resistant E. faecium . In the near future, we hope to expect the presence of bacteriophages in the list of antibacterial compounds used in the clinical settings., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Khazani Asforooshani, Elikaei, Abed, Shafiei, Barzi, Solgi, Badmasti and Sohrabi.)