Virulence-associated genes and drug susceptibility patterns of uropathogenic Escherichia coli isolated from patients with urinary tract infection.

Background: Different Escherichia coli phylogenetic groups, such as A, B1, B2, and D, have four functional groups - adhesins, microcins, toxins, and capsules - which can cause urinary tract infections (UTIs). A phylogenetic group with a high virulence content becomes a worldwide health concern. Resistance to antimicrobial agents increasingly complicates the management of E. coli extraintestinal infections, as a major source of illness, death, and increased health care costs. The aim of this study was to determine the virulence content and the antimicrobial susceptibility pattern of different uropathogenic E. coli (UPEC) phylogenetic groups in Ahvaz, Iran. Methods: Phylogenetic groups, virulence-associated genes (VAGs), and antimicrobial susceptibility tests were detected by molecular and phenotypic methods in a total of 232 clinically well-characterized E. coli strains, isolated from two collections of patients with hospital-acquired (HA) and community-acquired (CA) UTIs. Results: Our results revealed that among 232 UPEC strains, the most frequent phylogenetic group was phylogroup D (58%) with the greatest content in virulence factors, including kpsM (23%), neuA (76.3%, capsule), cnf (29.6%, toxin), and Pap (54.8%, adhesin). Phylogroups D and, to a lesser extent, B2 were the most drug-resistant phylogroups. In addition, phylogroup D was responsible for the majority of HA (64.7%) and CA (48.4%) infections. Conclusion: Among UPEC strains causing UTIs, different phylogroups, through different VAGs, could cause severe infection. Knowledge about the distribution of the four functional groups and VAGs belonging to these phylogroups would significantly help to confine and prevent the development of lethal infection caused by these strains.

WITHDRAWN: Phylotyping based virulence associated genes and drug susceptibility of Uropathogenic Escherichia coli in urinary tract infection patients.

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Mycobacterium ahvazicum sp. nov., the nineteenth species of the Mycobacterium simiae complex.

Four slowly growing mycobacteria isolates were isolated from the respiratory tract and soft tissue biopsies collected in four unrelated patients in Iran. Conventional phenotypic tests indicated that these four isolates were identical to Mycobacterium lentiflavum while 16S rRNA gene sequencing yielded a unique sequence separated from that of M. lentiflavum. One representative strain AFP-003T was characterized as comprising a 6,121,237-bp chromosome (66.24% guanosine-cytosine content) encoding for 5,758 protein-coding genes, 50 tRNA and one complete rRNA operon. A total of 2,876 proteins were found to be associated with the mobilome, including 195 phage proteins. A total of 1,235 proteins were found to be associated with virulence and 96 with toxin/antitoxin systems. The genome of AFP-003T has the genetic potential to produce secondary metabolites, with 39 genes found to be associated with polyketide synthases and non-ribosomal peptide syntases and 11 genes encoding for bacteriocins. Two regions encoding putative prophages and three OriC regions separated by the dnaA gene were predicted. Strain AFP-003T genome exhibits 86% average nucleotide identity with Mycobacterium genavense genome. Genetic and genomic data indicate that strain AFP-003T is representative of a novel Mycobacterium species that we named Mycobacterium ahvazicum, the nineteenth species of the expanding Mycobacterium simiae complex.

Mycobacterium aquaticum sp. nov., a rapidly growing species isolated from haemodialysis water.

The characterization of five Iranian isolates, four from hospital haemodialysis water and one from the sputum of a patient, led to the detection of a novel mycobacterium species. The strains were characterized by mucoid colonies developing in 3-5 days at temperatures ranging from 25 to 37 °C. The biochemical test pattern was unremarkable while the HPLC profile of mycolic acids resembled that of Mycobacterium fortuitum. The sequences of three major housekeeping genes (16S rRNA, hsp65 and rpoB) were unique and differed from those of any other mycobacterium. Mycobacterium brisbanense, which is the species that shared the highest 16S rRNA gene sequence similarity (99.03 %), was distinct, as shown by the average nucleotide identity and by the genome to genome distance values (91.05 and 43.10 %, respectively). The strains are thus considered to represent a novel species of the genus Mycobacterium, for which the name Mycobacterium aquaticum sp. nov. is proposed. The type strain is RW6T (=DSM 104277T=CIP111198T).

Mycobacterium persicum sp. nov., a novel species closely related to Mycobacterium kansasii and Mycobacterium gastri.

Four strains isolated in Iran from pulmonary specimens of unrelated patients are proposed as representative of a novel Mycobacterium species. Similarity, at the phenotypic level, with Mycobacterium kansasii is remarkable with the photochromogenic yellow pigmentation of the colonies being the salient feature. They differ, however, genotypically from this species and present unique sequences in 16S rRNA, hsp65 and rpoB genes. The average nucleotide identity and the genome-to-genome distance fully support the status of an independent species. The name proposed for this species is Mycobacterium persicum sp. nov. with AFPC-000227T (=DSM 104278T=CIP 111197T) as the type strain.

In Vitro Antimicrobial Susceptibility of Nontuberculous Mycobacteria in Iran.

Many species of nontuberculous mycobacteria (NTM) have long been identified as important causes of human disease, the incidence of which is rising. Several reports have suggested increasing trend of both in vitro and in vivo resistance to available treatment regimes. The aim of this study was to evaluate antibiotic susceptibility of clinically relevant NTM isolates using standard microbroth dilution test. Antimicrobial susceptibility testing was performed following National Committee for Clinical Laboratory Standards methods for NTM isolates, including 85 Mycobacterium fortuitum, 39 Mycobacterium chelonae, and 30 Mycobacterium abscessus subsp. abscessus as rapidly growing mycobacteria and 48 Mycobacterium simiae and 40 Mycobacterium kansasii as slowly growing mycobacteria. All isolates were recovered from various types of clinical samples and identified by multilocus sequence analysis. Trimethoprim-sulfamethoxazole (TMP-SMZ), amikacin, tobramycin, clarithromycin, moxifloxacin, linezolid, and imipenem showed better activity against M. fortuitum rather than meropenem, ciprofloxacin, cefoxitin, and doxycycline. Amikacin was active against 93% of M. abscessus subsp. abscessus. Linezolid, clarithromycin, cefoxitin, ciprofloxacin, imipenem, moxifloxacin, tobramycin, TMP-SMZ, doxycycline, and meropenem showed some activities on M. abscessus subsp. abscessus as well. The majority of M. abscessus subsp. abscessus and M. chelonae strains were multidrug resistant. Among the 40 isolates of M. kansasii, all were susceptible to ethambutol, isoniazid, clarithromycin, moxifloxacin, and linezolid. These isolates were also resistant to doxycycline and 50% were resistant to rifampicin and ciprofloxacin. M. simiae was resistant to clarithromycin, doxycycline, isoniazid, and TMP-SMZ, and the majority of isolates showed high levels of resistance to linezolid, ethambutol, ciprofloxacin, streptomycin, and rifampicin. The majority of M. simiae isolates were multidrug resistant. Our data confirm the need for performing of standard susceptibility testing of any clinically important NTM isolate.

Genetic diversity and antimicrobial susceptibility of Nocardia species among patients with nocardiosis.

The aim of this multicenter study was to determine the genetic diversity and antibiotic susceptibility of clinically isolated Nocardia species. One hundred twenty-seven patients with nocardiosis were randomly selected from 5 provinces of Iran. Molecular diagnosis of Nocardia species was performed using multilocus sequence analysis of gyrase B of the ß subunit of DNA topoisomerase (gyrB), and 16S rRNA and subunit A of SecA preproteintranslocase (secA1). Antimicrobial susceptibility testing was performed following the Clinical and Laboratory Standards Institute recommendations. Thirty-five N. cyriacigeorgica, 30 N. asteroides, 26 N. farcinica, 12 N. otitidiscaviarum, and 10 N. abscessus cultures were studied. All isolates were susceptible to linezolid. All isolates of N. cyriacigeorgica, N. asteroides, N. abscessus, and N. otitidiscaviarum were susceptible to trimethoprim-sulfamethoxazole, while 8% of N. farcinica isolates were resistant to this drug. All N. otitidiscaviarum isolates were highly resistant to imipenem, but N. cyriacigeorgica, N. asteroides, N. farcinica, and N. abscessus were only moderate resistant. The susceptibility patterns vary with different species of Nocardia. Resistance to trimethoprim-sulfamethoxazole in Iran is low and this drug should be first line therapy, unless drug susceptibility testing shows resistance. Linezolid also covers Nocardia well and could be a second line agent.

Mycobacterium celeriflavum sp. nov., a rapidly growing scotochromogenic bacterium isolated from clinical specimens.

Six strains of a rapidly growing scotochromogenic mycobacterium were isolated from pulmonary specimens of independent patients. Biochemical and cultural tests were not suitable for their identification. The mycolic acid pattern analysed by HPLC was different from that of any other mycobacterium. Genotypic characterization, targeting seven housekeeping genes, revealed the presence of microheterogeneity in all of them. Different species were more closely related to the test strains in various regions: the type strain of Mycobacterium moriokaense showed 99.0 % 16S rRNA gene sequence similarity, and 91.5-96.5 % similarity for the remaining six regions. The whole genome sequences of the proposed type strain and that of M. moriokaense presented an average nucleotide identity (ANI) of 82.9 %. Phylogenetic analysis produced poorly robust trees in most genes with the exception of rpoB and sodA where Mycobacterium flavescens and Mycobacterium novocastrense were the closest species. This phylogenetic relatedness was confirmed by the tree inferred from five concatenated genes, which was very robust. The polyphasic characterization of the test strains, supported by the ANI value, demonstrates that they belong to a previously unreported species, for which the name Mycobacterium celeriflavum sp. nov. is proposed. The type strain is AFPC-000207(T) ( = DSM 46765(T) = JCM 18439(T)).

Heterogeneity of Iranian clinical isolates of Mycobacterium fortuitum.

BACKGROUND AND OBJECTIVES: The increase of infections caused by nontuberculous mycobacteria (NTM) is receiving increasing attention worldwide. Mycobacterium fortuitum is encountered with increasing frequency in clinical laboratories of Iran. MATERIALS AND METHODS: Sequence variation of 48 M. fortuitum clinical isolates, were investigated by sequence analysis of the 16S-23S Internal Transcribed Spacer. RESULTS: Twelve different sequence types (sequevar) were identified by sequence analysis of ITS region. Seven previously described sequevar including MfoA, MfoB, MfoC, MfoD, MfoE, MfoF and MfoG identified. Five novel sequevar namely MfoH, MfoI, MfoJ, MfoK and MfoL that were distinctly different from the previously described sequevar were detected among different clinical strains of M. fortuitum, from Iran. CONCLUSION: This study showed that the ITS region possesses high discriminatory power between the clinical isolates up to the clonal level. The results also suggest the possibility of the existence of predominant clone of M. fortuitum in affected patients in Iran. The data also point to the conclusion that a large variety of M. fortuitum clone can produce disease although certain clones seem to be predominant.