Hasan Shojaei

Mycobacterium novocastrense sp. nov., a rapidly growing photochromogenic mycobacterium.

A strain isolated from a biopsy sample taken from a slowly spreading skin granulation on a childs hand was found to have properties consistent with its classification in the genus Mycobacterium. An almost complete gene sequence of the 16S rRNA of the strain was determined following the cloning and sequencing of the amplified gene. The sequence was aligned with those available for mycobacteria, and phylogenetic trees were inferred with four tree-making algorithms. The organism, which formed a distinct phyletic line within the evolutionary radiation occupied by rapidly growing mycobacteria, was readily distinguished from members of validly described species of rapidly growing mycobacteria on the basis of its mycolic acid pattern and a number of other phenotypic features, notably its ability to form yellow pigmented colonies when incubated in the light. The name proposed for this new species is Mycobacterium novocastrense. The type strain is DSM 44203.

Mycobacterium elephantis sp. nov., a rapidly growing non-chromogenic Mycobacterium isolated from an elephant.

A strain isolated from a lung abscess in an elephant that died from chronic respiratory disease was found to have properties consistent with its classification in the genus Mycobacterium. An almost complete sequence of the 165 rDNA of the strain was determined following the cloning and sequencing of the amplified gene. The sequence was aligned with those available on mycobacteria and phylogenetic trees inferred by using three tree-making algorithms. The organism, which formed a distinct phyletic line within the evolutionary radiation occupied by rapidly growing mycobacteria, was readily distinguished from members of validly described species of rapidly growing mycobacteria on the basis of its mycolic acid pattern and by a number of other phenotypic features, notably its ability to grow at higher temperatures. The type strain is Mycobacterium elephantis DSM 44368T.

Mycobacterium iranicum sp. nov., a rapidly growing scotochromogenic species isolated from clinical specimens on three different continents.

The isolation and characterization of a novel, rapidly growing, scotochromogenic mycobacterial species is reported. Eight independent strains were isolated from clinical specimens from six different countries of the world, two in Iran, two in Italy and one in each of following countries: Greece, The Netherlands, Sweden and the USA. Interestingly, two of the strains were isolated from cerebrospinal fluid. The strains were characterized by rapid growth and presented orange-pigmented scotochromogenic colonies. DNA-based analysis revealed unique sequences in the four regions investigated: the 16S rRNA gene, the rRNA gene internal transcribed spacer 1 and the genes encoding the 65 kDa heat-shock protein and the beta-subunit of RNA polymerase. The phylogenetic analysis placed the strains among the rapidly growing mycobacteria, being most closely related to Mycobacterium gilvum. The genotypic and phenotypic data both strongly supported the inclusion of the strains investigated here as members of a novel species within the genus Mycobacterium; the name Mycobacterium iranicum sp. nov. is proposed to indicate the isolation in Iran of the first recognized strains. The type strain is M05(T) ( = DSM 45541(T) = CCUG 62053(T) = JCM 17461(T)).

Drug resistance pattern of Mycobacterium tuberculosis isolates from patients of five provinces of Iran.

OBJECTIVE To determine the patterns of resistance to first line anti-tuberculosis (TB) drugs among a collection of Mycobacterium tuberculosis (MTB) isolates from 5 provinces of Iran. METHODS A total of the 6 426 clinical specimens from patients suspected of active TB were collected from March 2010 to June 2012. All specimens were subjected for microscopy and culture tests in the TB centers of studies provinces. Drug susceptibility testing to the first line anti-TB drugs for culture positive MTB was performed on Löwenstein-Jensen (LJ) medium using proportion method. RESULTS Of 6 426 clinical specimens, 261 were culture positive for mycobacteria, of which 252 were MTB and 9 were MOTT (mycobacteria other than tuberculosis). Of 252 MTB isolates, 211 (83.7%) were pan-susceptible and 41 (16.3%) were resistant to at least one drug. Resistance was most common to streptomycin, 30 isolates (12.0%), followed by isoniazid, 20 isolates (8.0%), rifampin, 15 isolates (6.0%) and ethambutol, 14 isolates (5.5%). Sixteen (6.3%) MTB isolates were MDR. A clear evidence of heterogeneity amongst the 5 provinces in the proportions with resistance to one or more drugs was observed [χ(2); = 12.209 (4 degrees of freedom), P values = 0.015 9]. CONCLUSIONS The prevalence of drug resistance in this study area underscoring the need for further enforcement of TB control strategies in the Iran. Drug susceptibility testing for all TB cases to provide optimal treatment, establishing advanced diagnostic facilities for rapid detection of MDR-TB and continuous monitoring of drug resistance are recommended for prevention and control of drug-resistant TB.

Mycobacterium arupense infection in HIV-infected patients from Iran

Summary Here we report two cases of infection caused by Mycobacterium arupense in HIV-infected patients who had received Mycobacterium avium complex medication after primary treatment with antituberculous drugs. The causative agents were isolated from the respiratory and blood specimens of the patients. The identification was based on conventional and molecular tests. Our study provides further evidence on the role of this microorganism in clinical cases.

Mycobacterium iranicum infection in HIV-infected patient, Iran.

To the Editor: The species Mycobacterium iranicum was described in 2013 (1) on the basis of 8 clinical strains isolated in various countries (Iran, Italy, Greece, the Netherlands, Sweden, and the United States). Recently, the isolation of M. iranicum from the sputum of a woman also was reported (2). We report the isolation of this newly recognized species from an HIV-positive patient. A scotochromogenic, rapidly growing strain was isolated in 2012 from respiratory specimens of an HIV-positive 44-year-old Iranian man with chronic pulmonary disease. The patient had been found to be HIV seropositive (viral load >1,000 copies/mL, CD4 lymphocyte count 120/µL) in 2004 when he was hospitalized because of fever, weight loss, and oral candidiasis. Treatment with antiretroviral drugs, including stavudine, lamivudine, and nevirapine, was begun. The patient rapidly improved; the fever disappeared, he gained weight, and he was discharged from the hospital. At a 6-month follow-up visit, viral load was 1,000 copies/mL and CD4 lymphocyte count was 420/µL. He continued to receive antiretroviral treatment until 2010 when treatment was discontinued because of its high cost. The man was hospitalized again in 2012 with mild fever, weight loss, chronic chest pain, and nonproductive cough. At that time, the viral load was >100,000 copies/mL, and CD4 count was 5 lymphocytes/µL. Tuberculin skin test results were negative, radiograph of the chest showed no abnormalities, and routine cultures of sputum and blood were negative for common bacteria. Lactate dehydrogenase level (98 U/L [reference <600 U/L]) was within normal limits, whereas liver function was abnormal (alanine aminotransferase level 95 U/L [reference <36 U/L], L-aspartate aminotransferase level 85 U/L [reference <29 U/L], alkaline phosphatase 180 U/L [reference 44–147 U/L], and total bilirubin 1.4 mg/dL [reference 0.3–1mg/dl]). Antiretroviral therapy was resumed, which led to an increase in CD4 cells (205 lymphocytes/µL after 1 month). The examination by microscopy (Ziehl-Neelsen staining) of 3 sputum samples did not reveal acid-fast bacilli; culture for mycobacteria was not done. Oral treatment with tetracycline was started, but the patient’s fever and chest pain remained unchanged. After bronchoscopy, 2 of 3 bronchial lavage (BAL) samples were found to be positive for acid-fast coccobacilli by microscopy, and rapidly growing, deep orange mycobacteria grew in all 3 cultures. Giemsa stain did not show Pneumocystis jirovecii in BAL samples. A standard antituberculosis regimen was undertaken but did not result in substantial improvement. At 1 month follow-up, 1 sputum sample was negative for acid-fast organisms, and 1 BAL specimen was positive by microscopy and in culture. When the isolate was identified as M. iranicum, therapy was replaced with a combination of amikacin and ciprofloxacin for 3 months (standard treatment used in Iran for infections caused by rapidly growing mycobacteria), and the patient improved rapidly. Mycobacteria were neither observed nor grew in culture in a BAL specimen obtained 1 month after the change in therapeutic regimen. Identification of the isolates was initially attempted with biochemical tests, and they were negative for niacin production, nitrate reduction, Tween 80 hydrolysis, and semiquantitative catalase. The tests were positive for urease activity, iron uptake, tellurite reduction, arylsulfatase (3 days after the start of the test), 5% NaCl tolerance, and heat-stable (68°C) catalase. The genetic sequencing of almost-complete (1,450 bp) 16S rRNA gene (3), a 710-bp fragment of the β-subunit of the RNA polymerase gene (4), and the hypervariable region (402 bp) of the 65-kDa heath-shock protein (5) revealed 99.8%, 99.4%, and 100% identity, respectively, with sequences of the type strain found in GenBank and definitively confirmed the identification. The clinical criteria required by the American Thoracic Society and Infectious Disease Society of America (3) to assess the importance of the isolation of a nontuberculous mycobacterium from pulmonary specimens include, in adjunct to a specific symptomatology, the presence of nodular or cavitary lung lesions and the exclusion of any other possible cause of the disease. The normal thoracic radiograph findings for the case-patient described here cannot, however, be considered a definitive exclusion criterion: in highly immunocompromised patients, a chest radiograph may show no abnormalities, even when substantial pathologic features of infection are present (6). The microbiological criteria were clearly fulfilled by isolating the organism from multiple sputum specimens and the BAL specimens. The patient’s response to the treatment and the disappearance of thoracic symptoms further support the assertion. Our report confirms the potential pathogenicity of M. iranicum. In addition to the case described here, 9 isolations of this species have been reported so far. Among them, the clinical relevance has been demonstrated for 2 strains grown from respiratory specimens of patients with pulmonary disease and for 1 strain isolated from a cutaneous lesion (1,2). The role of an accurate identification, in conjunction with symptoms and radiographic findings, is central to understanding the clinical significance of mycobacteria isolated from pathologic specimens.

Chronic Pulmonary Disease Due to Mycobacterium monacense Infection: The First Case from Iran

We herein report a case in which the recently characterized species Mycobacterium monacense was isolated from the sputum of an Iranian patient. This case represents the first isolation of M. monacense from Iran. The isolate was identified by conventional and molecular techniques. Our findings show that M. monacense infection is not restricted to developed countries.

Pulmonary and extrapulmonary infection caused by Mycobacterium conceptionense: the first report from Iran

We report what we believe to be the first clinical isolation of Mycobacterium conceptionense in a developing country.

Chronic pelvic pain due to Mycobacterium parascrofulaceum in an Iranian patient: first report of isolation and molecular characterization from Asia

1Dr, Associate Professor; PhD in Molecular Microbiology, Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran 2Dr., Research Associate, Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran 3Dr, Research Associate, Microbiology Group, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 4Research Assistant; Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

Molecular analysis and species diversity of Nocardia in the hospital environment in a developing country, a potential health hazard

Purpose. Despite hundreds of reports on the isolation of Nocardia from clinical samples, the presence and diversity of Nocardia species that are capable of survival in a harsh and adverse condition, such as a hospital environment, have not been comprehensively studied. The aim of this study was to assess Nocardia species diversity in a hospital environment to provide a better insight into their potential threat as a reservoir for the development of nosocomial infections. Methodology. A total of 90 samples of hospital water, dust and soil, collected from 30 hospitals, were analysed for the presence of Nocardia using standard protocols for isolation and characterization of the isolates. Conventional tests were used for preliminary identification, and PCR amplification of the 596 bp amplicon of the 16S rRNA and sequence analysis of 16S rRNA were performed for genus and species identification. Results. A total of 25 Nocardia isolates (27.7 %) from 10 species were recovered from 90 samples. The three most prevalent species were N. cyriacigeorgica, 24 %, N. asteroides, 16 % and N. kroppenstedtii, 12 %, followed by N. salmonicida‐like, 8 % and single isolates of N. otitidiscaviarum, N. flavorozea‐like, N. neocaledoniensis‐like and N. sungurluensis‐like. Thirteen out of twenty five isolates showed characteristics of six novel species. Conclusion. Our study showed that the hospital environment is a potential reservoir of a diverse range of Nocardia species, due to the remarkable survival capability of these bacteria in an adverse hospital environment, which carries a threat to the health of patients.