Pulmonary Nocardia ignorata Infection in Gardener, Iran, 2017.

Nocardia ignorata, which was first described in 2001, is a rare human pathogen. We report a case of pulmonary nocardiosis caused by this bacterium in a 55-year-old man from Iran. The patient, a gardener, had frequent exposure to soil and may have acquired the infection from that source.

Multiple Myeloma or Brucellosis: A Case Report.

BACKGROUND: Brucellosis, a major health problem in developing countries, is a multisystem infection with a broad spectrum of clinical manifestations. Hematological complications, ranging from an intravascular coagulopathy to mild homeostasis disorders (such as gammopathy), have been reported in brucella infection. These signs and symptoms may lead to misdiagnosis of brucellosis with other hematological diseases. CASE: A 65-year-old male whose occupation was shepherding was referred to our hospital as a known case of multiple myeloma with continuous fever, muscle weakness, and night sweating after taking 2 courses of chemotherapy. The laboratory diagnosis of multiple myeloma had been based on the observation of a high percent of plasma cells in the bone marrow aspiration. At follow- up, the result of patient's fever workup, with 2 sets of blood cultures, was positive for Brucella melitensis. Isolated brucella was confirmed as B. melitensis by 16S rRNA sequencing. Brucellosis serologic test was performed by agglutination test and positive results were obtained. The patient was discharged with the cessation of fever and general improvement after the end of the parental treatment phase of brucella bacteremia. CONCLUSIONS: Brucella infection may cause a severe disease, mimicking a primary hematological disease, which could complicate the correct diagnosis. In brucellosis cases, due to the wide range of symptoms, in addition to cultivation and serological methods, molecular methods should also be used to prevent inappropriate diagnosis and additional costs.

The Chemical Composition and Anti-mycobacterial Activities of Trachyspermum copticum and Pelargonium graveolens Essential Oils.

BACKGROUND: Microbial resistance to antibiotics and their adverse effects related to these antibiotics are a matter of global public health in the 21th century. The emergence of drug-resistant strains, has gained the interest of the scientists to discover new antimicrobial agents from the essential oil of medicinal plants. METHODS: Anti-mycobacterial effects of Trachyspermum copticum and Pelargonium graveolens essential oils were determined against multi-drug resistant clinical strains of Mycobacterium tuberculosis, Mycobacterium kansasii, Mycobacterium fortuitum and standard strain of Mycobacterium tuberculosis H37Rv by a Broth micro-dilution method. Pelargonium graveolens plant named Narmada was discovered by Kulkarni R.N et al. (Patent ID, USPP12425P2) and a formulation comprising thymol obtained from Trachyspermum is useful in the treatment of drug-resistant bacterial infections (Patent ID, US6824795B2). The chemical composition of hydro-distilled essential oils was determined by GC and GC-MS. RESULTS: Minimum Inhibitory Concentration (MIC) values for T. copticum essential oil against tested isolates were ranged from 19.5 µg/mL to 78 µg/mL. The least minimum inhibitory concentration of P. graveolens extract against M. Kansasii and MDR-TB was 78 µg/ml. CONCLUSION: The results of the present research introduced T. copticum and P. graveolens essential oils as a remarkable natural anti-mycobacterial agent, but more pharmacological studies are required to evaluate their efficacy in animal models.

In Vitro Anti-mycobacterial Activity of Three Medicinal Plants of Lamiaceae Family.

BACKGROUND: Mycobacterium tuberculosis as an intracellular pathogen causes Tuberculosis (TB). Due to the long time required for treatment, hepatotoxicity of drugs and also emergence of Multidrug-Resistant (MDR) and Extremely Drug Resistant (XDR) strains, TB is currently a major public health problem. Some medicinal plants possess remarkable activity against Mycobacterium. Among them, Lamiaceae family are of pharmaceutical interest because of their potential antimicrobial properties. The aim of the study was to evaluate the in vitro activities of Satureja rechingeri, Satureja khuzestanica and Zataria multiflora against MDR M. tuberculosis and two Non-Tuberculous Mycobacteria (NTM). METHODS: The essential oils were prepared by the standard method. The confirmed strains were obtained from the microbial collection of Tehran University of Medical Sciences. Minimum Inhibitory Concentrations (MICs) of essential oils of plants against mycobacterial strains were determined using standard broth microdilution method. RESULTS: MDR M. tuberculosis was completely inhibited by Z. multiflora at 78µg/ml concentration. S. rechingeri and S. khuzestanica also showed same anti-mycobacterial activity against MDR M. tuberculosis with MICs of 156 µg/ml. The MICs of the essential oils against M. tuberculosis H37Rv, M. kansasii and M. fortuitum were in the range from 39 to 156 µg/ml. CONCLUSION: The studied medicinal plants showed notable effects against mycobacterial strains. Our results indicated that utilization of Lamiaceae family can be helpful for treatment of mycobacterial infections.

New Insights in to the Intrinsic and Acquired Drug Resistance Mechanisms in Mycobacteria.

Infectious diseases caused by clinically important Mycobacteria continue to be an important public health problem worldwide primarily due to emergence of drug resistance crisis. In recent years, the control of tuberculosis (TB), the disease caused by Mycobacterium tuberculosis (MTB), is hampered by the emergence of multidrug resistance (MDR), defined as resistance to at least isoniazid (INH) and rifampicin (RIF), two key drugs in the treatment of the disease. Despite the availability of curative anti-TB therapy, inappropriate and inadequate treatment has allowed MTB to acquire resistance to the most important anti-TB drugs. Likewise, for most mycobacteria other than MTB, the outcome of drug treatment is poor and is likely related to the high levels of antibiotic resistance. Thus, a better knowledge of the underlying mechanisms of drug resistance in mycobacteria could aid not only to select the best therapeutic options but also to develop novel drugs that can overwhelm the existing resistance mechanisms. In this article, we review the distinctive mechanisms of antibiotic resistance in mycobacteria.

MgrB Alterations Mediate Colistin Resistance in Klebsiella pneumoniae Isolates from Iran.

Colistin is one of the last-resort therapeutic agents to combat multidrug-resistant Gram-negative bacteria (GNB) including Klebsiella pneumoniae. Although it happens rarely, resistance to colistin has been reported for several GNB. A total of 20 colistin resistant (col-R) and three colistin susceptible (col-S) clinical isolates of K. pneumoniae were studied to explore the underlying mechanisms of colistin resistance. The presence of plasmid encoded resistance genes, mcr-1, mcr-2, mcr-3, and mcr-4 genes were examined by PCR. The nucleotide sequences of pmrA, pmrB, phoP, phoQ, and mgrB genes were determined. To evaluate the association between colistin resistance and upregulation of pmrHFIJKLM and pmrCAB operons, transcriptional level of the pmrK and pmrC genes encoding for lipopolysaccharide target modifying enzymes was quantified by RT-qPCR analysis. None of the plasmid encoded resistance genes were detected in the studied isolates. Inactivation of MgrB due to nonsense mutations and insertion of IS elements was observed in 15 col-R isolates (75%). IS elements (IS5-like and IS1-like families) most commonly targeted the coding region and in one case the promoter region of the mgrB. Complementation with wild-type MgrB restored colistin susceptibility in isolates with altered mgrB. All col-R isolates lacked any genetic alterations in the pmrA, phoP, and phoQ genes and substitutions identified in the pmrB were not found to be involved in resistance conferring determined by complementation assay. Colistin resistance linked with upregulation of pmrHFIJKLM and pmrCAB operons with the pmrK and pmrC being overexpressed in 20 and 11 col-R isolates, respectively. Our results demonstrated that MgrB alterations are the major mechanisms contributing to colistin resistance in the tested K. pneumoniae isolates from Iran.

Rapid Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis using TaqMan Allelic Discrimination.

OBJECTIVES: Multidrug-resistant tuberculosis (MDR-TB) is a global problem that many countries are challenged with. Rapid and accurate detection of MDR-TB is critical for appropriate treatment and controlling of TB. The aim of the present study was to evaluate the TaqMan allelic discrimination without minor groove binder (MGB) as a rapid, efficient, and low-cost method for detection of drug resistant strains of Mycobacterium tuberculosis. METHODS: A total of 112 M. tuberculosis isolates from cases with diagnosed TB were subjected to drug susceptibility testing (DST), using the proportion method. Resistant isolates were tested for characterization of mutations in the rpoB and KatG genes by TaqMan genotyping. RESULTS: Of 112 M. tuberculosis isolates for which DST was performed, three, one, and two isolates were MDR, rifampin (RIF) resistant, and isoniazid (INH) resistant, respectively. According to the threshold cycle (Ct) and curve pattern of mutants, TaqMan probes detect all of the mutations in the analyzed genes (katG 315, AGC→ACC, rpoB 531, TCG→TTG, and rpoB 531, TCG→TGG). CONCLUSION: The present study suggests that drug-resistant strains of M. tuberculosis can be detected by pattern's curve or Ct with TaqMan probes without MGB in real-time polymerase chain reaction (PCR).

Molecular characterization of class 1 integrons in MDR Pseudomonas aeruginosa isolated from clinical settings in Iran, Tehran.

Seven hundred and fifty nonreplicated clinical isolates of Pseudomonas aeruginosa were collected from five hospitals during March 2007 to May 2009. Forty-one isolates (5.46%) were characterized as multidrug-resistant P. aeruginosa (MDRPA). PCR assays were used to detect class 1 integrons. Amplifications of internal variable regions (IVRs) of class 1 integrons revealed three different arrays (0.8, 1.3, and 1.7 kb) with different distributions in clinical isolates. The amplified IVRs were sequenced and three gene cassette arrays including aadB (0.8 kb), aadA6-orfD (1.3 kb), and bla(OXA10)-aacA4 (1.7 kb) were identified. In conclusion, we confirmed the high prevalence of class 1 integons with limited diversity of gene cassette arrays in MDRPA clinical isolates found from five hospitals. This is the first report showing gene cassette contents of class 1 integrons in P. aeruginosa isolates in Iran.

Distribution of Helicobacter pylori cagA, cagE, oipA and vacA in different major ethnic groups in Tehran, Iran.

BACKGROUND AND AIM: There are geographical variations in Helicobacter pylori virulence genes; cagA, cagE, vacA and oipA. The present study compared the distribution of these genotypes in major ethnic groups residing in Tehran, Iran and their association with clinical outcomes. METHODS: A total of 124 H. pylori-positive patients living in Tehran were enrolled in this study. The ethnic distribution was 74 Persians, 33 Turks and 17 other ethnics including Kurds, Lurs, Afghanis and Arabs. The presence of the cagA, cagE and oipA genes and vacA alleles (signal [s] and middle [m] region) were determined by polymerase chain reaction (PCR) from H. pylori DNA. RESULTS: The cagA-positive status was predominant in all three ethnic groups (e.g. 65% in Persians and 73% in Turks). In contrast, the cagE-positive status was less than half in Persians (47%) and Turks (30%), whereas it was 77% in other ethnicities (P = 0.008). The predominant vacA genotypes were s1 and m1 in all three ethnic groups (e.g. 68% in Persians and 70% in Turks were s1). There was no significant association between cagA and cagE status or vacA genotypes and clinical outcomes. The oipA-positive strains were more common in non-ulcer dyspepsia (NUD) (63%) than in peptic ulcer patients (15%) (P = 0.001) in Persians, but the association was not observed in other ethnic groups. CONCLUSION: There are some differences in the H. pylori genotypes among the ethnic groups in Iran. However, none of these markers seemed to be clinically helpful in predicting the clinical presentation of a H. pylori infection in Iran.

Molecular characterization of methicillin-resistant Staphylococcus aureus clones from a teaching hospital in Tehran.

A total of 52 methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates were collected from patients attending the teaching hospital of Tehran University of Medical Sciences. Disks containing antibiotics were used to determine the susceptibility of MRSA isolates. Analysis of SmaI macrorestriction profiles of the 52 MRSA isolates were grouped into three PFGE types. The majority of isolates (n=49) were clustered into only one major PFGE type, designated as pulsotype A; these belonged to SCCmec type III or IIIA and showed resistance to ampicillin, ciprofloxacin, cotrimoxazole, erythromycin, gentamicin, and tetracycline. The remaining isolates fell into pulsotypes B and C, both belonging to SCCmec-type IV. All MRSA isolates were susceptible to vancomycin, teicoplanin, quinupristin-dalfopristin, linezolid, and tigecycline. The present study shows that a MRSA clone similar to the Brazilian clone (ST 239) of MRSA, which is a multiresistant MRSA clone with a high level of methicillin resistance, is very common in this teaching hospital in Tehran.

Characterisation of genes encoding aminoglycoside-modifying enzymes among meticillin-resistant Staphylococcus aureus isolated from two hospitals in Tehran, Iran.

Meticillin-resistant Staphylococcus aureus (MRSA) is a major hospital pathogen and typically shows resistance to multiple antimicrobial agents. The susceptibility patterns of 109 MRSA isolates to aminoglycoside antibiotics were determined by the disk diffusion method. Genes encoding the aminoglycoside-modifying enzymes (AMEs) were targeted by polymerase chain reaction (PCR) assays. All isolates were also subjected to multiplex PCR to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) types in the study population. The rates of resistance to various antibiotics were as follows: kanamycin, 97%; tobramycin, 96%; gentamicin, 87%; amikacin, 93%; and netilmicin, 80%. The most prevalent AME genes were aac(6')-Ie-aph(2'') (83%) followed by aph(3')-IIIa (71%). Coexistence of three AME genes was detected in 21% of isolates. The ant(4')-Ia gene was the least frequent AME gene among MRSA isolates (26%). Of the 109 isolates, 106 (97%) were identified as SCCmec type III or IIIA and 3 (3%) as SCCmec type IV. The majority of MRSA isolates belonged to SCCmec III or IIIA and carried the aac(6')-Ie-aph(2'') gene, which is consistent with results of susceptibility testing of these isolates against aminoglycosides.

The effect of dietary oils on cecal microflora in experimental colitis in mice.

OBJECTIVES: In spite of growing evidence indicating the benefits of probiotics, the effects of different dietary oils on intestinal microflora and probiotics have not been elucidated. This study aimed to examine the effects of different dietary oils on intestinal microflora in an experimental model of colitis. METHODS: Eight-week mice were fed isocaloric diets varying only in fat composition for 4 weeks. The oils used were fish oil, canola oil, safflower oil, and chow diet containing beef tallow. Colitis was induced by intracolonic administration of acetic acid on day 21. The inflammation and fecal microflora and serum lipid profiles were evaluated 1 week after induction. RESULTS: Inflammation was highest in the chow diet group followed by safflower, canola, and fish oil fed groups, respectively. The number of fecal bacteroideceae was greater, whereas the number of fecal bifidobacteria was lower in mice fed beef tallow than the other ones. In addition, fish oil reduced the plasma level of triacylglycerole significantly. CONCLUSION: Polyunsaturated fatty acids (PUFAs) can affect intestinal microflora increasing the number of probiotics. PUFAs might be recommended in addition to probiotics for the prevention and/or maintenance treatment of colitis.

Staphylococcal cassette chromosome mec (SCCmec) analysis and antimicrobial susceptibility patterns of methicillin-resistant Staphylococcus aureus (MRSA) isolates in Tehran, Iran.

Oxacillin resistance was present in 99 of 277 (36%) consecutive Staphylococcus aureus isolates collected from hospital patients in Tehran during a 15-month period (January 2004-March 2005). The majority of isolates (77/99 = 78%) had been cultured from wounds or blood. The staphylococcal cassette chromosome mec (SCCmec) types and antimicrobial susceptibility patterns of 99 methicillin-resistant S. aureus (MRSA) strains were determined. Disk diffusion and agar dilution methods were used to determine the susceptibility of isolates to antimicrobial agents as instructed by Clinical and Laboratory Standards Institute. The presence of mecA and SCCmec types was determined by PCR and multiplex PCR. All MRSA isolates were susceptible to vancomycin (MIC90

Use of multilocus enzyme electrophoresis to examine genetic relationships amongst isolates of Mycobacterium intracellulare and related species.

As part of a larger study investigating diversity and distribution of Mycobacterium spp. in Australia, multilocus enzyme electrophoresis was used to assess genetic relationships at 17 enzyme loci amongst a collection of reference strains and isolates initially identified on biochemical and other grounds as M. intracellulare (70), "X' mycobacteria (10), M. scrofulaceum (7), M. avium (8) and M. avium subsp. paratuberculosis (2). Two of the isolates initially identified as M. intracellulare were shown to be quite distinct from the others. Both gave negative results in a species-specific DNA probe test, whilst one was positive by PCR. These results emphasize the uncertainties involved in identifying members of this group. The other M. intracellulare isolates formed a cohesive but diverse group, being divided into 48 electrophoretic types (ETs), with a mean genetic diversity of 0.38. Forty-three of these ETs contained only single isolates. There was no clear relationship between the serovar and ET designation. The index of association calculated for M. intracellulare was significantly different from zero, suggesting that it is a clonal species. PFGE was also applied to selected isolates from the ETs containing multiple isolates, and some of these could be differentiated further. The strains of M. scrofulaceum and "X' mycobacteria were distinct from M. intracellulare, but themselves were highly heterogeneous, with mean genetic diversities of 0.66 and 0.65, respectively. Each of these groups may represent more than one species. M. avium strains were distinct from the two M. avium subsp. paratuberculosis strains, as well as from the other mycobacteria studied.