Prevalence and molecular characterization of non-tuberculous mycobacteria in hospital soil and dust of a developing country, Iran.

The presence and diversity of mycobacteria that are capable of survival in a harsh and adverse condition, such as hospital environments, have not been comprehensively studied. This study aimed to assess the frequency and diversity of mycobacteria in hospital soil and dust of a developing country using a combination of molecular and conventional methods. A total of 318 hospital dust and soil samples collected from 38 hospitals were analysed using standard protocols for characterization of mycobacteria. The conventional tests were used for preliminary identification and Runyon's classification, the PCR amplification of the hsp65 gene and sequence analyses of 16SrRNA were applied for genus and species identification. In total, 28 samples (8.8 %) were positive for mycobacteria. The isolates included 33 mycobacteria species including 19 rapidly growing and 14 slowly growing organisms. The most prevalent species were M. setense and M. lentiflavum, five isolates (15.1 %) each, M. fortuitum, four isolates (12.12 %) and M. kumamotonense and M. massiliense/abscessus complex three isolates (9.1 %) each, M. arupense and M. frederiksbergense, two isolates (6 %) each. The remaining isolates consisted the single strains of eight various mycobacterium species, the results of our study revealed that soil and dust in hospitals can be the reservoir of mycobacteria. This reaffirms the fact that these organisms due to intrinsic resistance can persist in hospitals and create a threat to patient's health, in particular to those who suffer from weakness of immunity.

Clostridium difficile isolated from faecal samples in patients with ulcerative colitis.

BACKGROUND: Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) that is widely identified worldwide. This study aimed to investigate the phenotypic characterization and molecular typing of Clostridium difficile isolates among patients with UC at an inflammatory bowel disease clinic in Iran. METHODS: In this cross-sectional study, conducted from April 2015 to December 2015, 85 UC patients were assessed for C.difficile infection (CDI). C. difficile isolates were characterized based on their toxin profile and antimicrobial resistance pattern. Multi-locus sequence typing analysis (MLST) and PCR ribotyping were performed to define the genetic relationships between different lineages of toxigenic strains. RESULTS: The prevalence of C. difficile isolates was 31.8% (27/85) in patients, of those 15 patients (17.6%) had CDI. Three different sequence types (STs) identified based on MLST among the toxigenic isolates, that is ST54 (33.3%), ST2 (53.3%), and ST37 (13.6%). C. difficile strains were divided into four different PCR-ribotypes (012, 014, 017 and IR1). The most common ribotype was 014 accounting for 48.3% (7/15) of all strains. The strains isolated during the first episode and recurrence of CDI usually belonged to PCR ribotype 014 (ST2). A high rate of CDI recurrence (14.1%, 12/85) experienced in UC patients. Colonization of the gastrointestinal tract with non-toxigenic C. difficile strains was frequent among patients with mild disease. All C. difficile isolates were susceptible to metronidazole, and vancomycin, 86 and 67% of isolates were resistant to clindamycin and erythromycin respectively. There was no correlation between the toxin type and antibiotic resistance (p > 0.05). CONCLUSION: Overall CDI is rather prevalent in UC patients. All patients with CDI experienced moderate to severe disease and exposed to different antimicrobial and anti-inflammatory agents. Close monitoring and appropriate management including early detection and fast treatment of CDI will improve UC outcomes.

Species diversity and molecular characterization of nontuberculous mycobacteria in hospital water system of a developing country, Iran.

BACKGROUND: Hospital environment is of crucial importance in cross-transmission of opportunistic pathogens to the patients. Nontuberculous mycobacteria have the remarkable capability to withstand the adverse condition of hospital environments and pose a potential threat to the health of patients. The current study aimed to assess the frequency and diversity of mycobacteria in hospital water of a developing country using a combination of conventional and molecular methods. METHODS: A total of 148 hospital water samples collected from 38 hospitals were analyzed for the presence of mycobacteria using standard protocols for isolation and characterization of the isolates. The conventional tests were used for preliminary identification and Runyon's classification, the PCR amplification of hsp65 gene and sequence analysis of 16S rRNA were applied for the genus and species identification. RESULTS: A total of 71 [48%] isolates including 30 rapidly growing and 41 slowly growing mycobacteria were recovered. The three most prevalent species were M. lentiflavum, 28.2%, M. paragordonae, 21.1%, and M. fredriksbergense, 9.8%, followed by M. simiae and M. novocastrense, 7%, M. canariasense and M. cookii like, 5.6%, M. setense, 4.2%, M. fortuitum and M. gordonae, 2.8%, and the single isolates of M. austroafricanum, M. massiliense, M. obuense, and M. phocaicum like. CONCLUSION: The results of our study show that the hospital water resources, drinking or non-drinking can be the reservoir of a diverse range of mycobacteria. This reaffirms the fact that these organisms due to intrinsic resistance to common antiseptic and disinfectant solutions persist in hospitals and create a threat to the patient's health and in particular to those that suffer from weakness of immunity.

First report of isolation of Mycobacterium canariasense from hospital water supplies.

Mycobacterium canariasense was first isolated as a novel species in 2004 from clinical specimens in Spain. Since then there have only been a few additional reports from Spain, the USA, and Lebanon on the isolation of this rare species from clinical specimens. We herein present the first report on isolation of this organism from hospital water, which provides evidence for determining the natural habitat of this rare species. The water samples were collected from hospital departments and cultured on Löwenstein-Jensen and Sauton's media. The isolates, i.e. WP5, WP20, and AW2-3, were subjected to identification by conventional and molecular tests including sequencing analysis of 16S rRNA. The water isolates revealed phenotypic and molecular features consistent with M. canariasense including a genus-specific amplicon of the hsp65 gene and 100% similarities with those of M. canariasense CIP: 107998(T) 16S rRNA gene sequences. The current report might be of value in tracing the probable source of infection in patients.

Gut-Lung Microbiota Characterization in Patients with Non-Small Cell Lung Carcinoma and COVID-19 Coinfection.

BACKGROUND: Non-small cell lung cancer (NSCLC) patients with COVID-19 have an excessive chance of morbidity and mortality. The fecal-nasopharyngeal microbiota compositions of NSCLC patients were assessed in this study. METHODS: In total, 234 samples were collected from 17 NSCLC patients infected with COVID-19, 20 NSCLC patients without confirmed COVID-19, 40 non NSCLC patients with COVID-19, and 40 healthy individuals. RESULTS: In lung microbiota, the abundance of Streptococcus spp. in NSCLC patients with confirmed COVID-19 was significantly higher than the two control groups. Pseudomonas aeruginosa and Staphylococcus aureus were listed as the most frequent pulmonary bacterial groups that colonized COVID-19 patients. In fecal specimens, the numbers of Bacteroidetes, Firmicutes, and Actinobacteria phyla were significantly higher amongst NSCLC patients with COVID-19. NSCLC patients infected with COVID-19 showed lower levels of Lactobacillus spp., Akkermansia muciniphila, and Bifidobacterium spp. The counts of Streptococcus spp., in NSCLC patients with COVID-19 were significantly higher than those of healthy individuals (8.49±0.70 log CFU/g wet feces vs 8.49±0.70 log CFU/g wet feces). Prevotella spp. were enriched in the gut and respiratory tracts of COVID-19 patient groups. The unbiased analysis showed an increment in Enterococcus spp., Streptococcus spp., and Prevotella spp. CONCLUSION: Eventually, it was found that compared to control groups, COVID-19 patients with NSCLC showed diminished gut bacteria diversity and increase in Lactobacillus spp., A. muciniphila, and Bifidobacterium spp. The overgrowth of Enterococcus spp., Streptococcus spp., and Prevotella spp. could be potential predictive biomarkers in the gut-lung axis of NSCLC patients with COVID-19.

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)).

The anticancer and antibacterial potential of bioactive secondary metabolites derived From bacterial endophytes in association with Artemisia absinthium.

The continuous search for secondary metabolites in microorganisms isolated from untapped reservoirs is an effective prospective approach to drug discovery. In this study, an in-depth analysis was conducted to investigate the diversity of culturable bacterial endophytes present in the medicinal plant A. absinthium, as well as the antibacterial and anticancer potential of their bioactive secondary metabolites. The endophytic bacteria recovered from A. absinthium, were characterized via the implementation of suitable biochemical and molecular analyses. Agar well diffusion and broth microdilution were used to screen antibacterial activity. SEM was performed to assess the impact of the extracted metabolite on MRSA strain cell morphology. Apoptosis and cytotoxicity assays were used to evaluate anticancer activity against MCF7 and A549. The FTIR, GC-MS were used to detect bioactive compounds in the active solvent fraction. Of the various endophytic bacteria studied, P. aeruginosa SD01 showed discernible activity against both bacterial pathogens and malignancies. The crude ethyl acetate extract of P. aeruginosa SD01 showed MICs of 32 and 128 µg/mL for S. aureus and MRSA, respectively. SEM examination demonstrated MRSA bacterial cell lysis, hole development, and intracellular leaking. This study revealed that the crude bioactive secondary metabolite SD01 has potent anticancer activity. In this study, 2-aminoacetophenone, 1,2-apyrazine-1,4-dione, phenazine and 2-phenyl-4-cyanopyridine were the major bioactive secondary metabolites. In conclusion, our findings indicate that the bacteria recovered from A. absinthium plants and in particular, P. aeruginosa SD01 is a remarkable source of untapped therapeutic, i.e., antimicrobial and anticancer compounds.

Genetic diversity of Iranian clinical isolates of Mycobacterium tuberculosis.

In the current study we aimed to execute a rather less complicated molecular tying method, i.e. the random amplification of polymorphic DNA (RAPD) analysis to find the heterogeneity of Iranian strains of Mycobacterium tuberculosis. The isolates comprised a total of 96 strains of M. tuberculosis collected from clinical specimens of patients in Isfahan and Tehran. The isolates were assigned to the species M. tuberculosis by the key conventional and molecular methods. They were then subjected to RAPD analysis by four arbitrary primers, namely, the primers 27F, 1525R, MS- GF and INS-2. They were then evaluated for the number and intensity of the band patterns. The RAPD profiles of the Iranian isolates showed a degree of heterogeneity which varied based on the primer used. However, analysis of the isolates by primer INS-2 revealed the highest degree of diversity yielding 31 distinguishable RAPD types. RAPD analysis provides a rapid and easy means of identifying heterogeneity among the M. tuberculosis isolates. This typing system might be considered a valuable alternative molecular typing for countries with limited resources provided that the reproducibility and reliability of the method is carefully assured.

The co-administration effects of florfenicol and lasalocid on performance, biochemical and pathological parameters of muscle, heart, liver, kidney and sciatic nerve in broiler chickens.

The study aimed to examine the effect of simultaneous application of florfenicol and lasalocid on the performance and vital organ function of chickens. For this, 300 chicks were divided into four groups. Group one to three received florfenicol, lasalocid and lasalocid plus florfenicol, respectively. Group four as the control group received a basic diet without lasalocid or florfenicol. Lasalocid was used from 7 to 35 days old, continuously. Florfenicol was used at 21 days old for 5 days. The growth indices were measured at the end of each week. The chickens were euthanized at the ages of 28 and 35 days old after collecting blood samples with and without anticoagulants. The liver, heart, muscle, kidney and sciatic nerve were collected in formalin 10% for histopathological examination. The blood and serum samples were used to determine clinical pathologic and hematologic indices. The ratio of internal organs to body weight and ratio of the right ventricle to the total ventricles (RV/TV) of the heart was measured. Results showed, the use of lasalocid decreased feed conversion rate and triglyceride, and increased total protein. Simultaneous administration of lasalocid and florfenicol affected histopathology of the liver and heart and significantly increased creatine phosphokinase, uric acid and the ratio of RV/TV of heart. The eosinophil percentage in the chickens who received florfenicol plus lasalocid was significantly higher than chickens who received florfenicol alone (p < 0.05). In conclusion, it seems that simultaneous administration of the florfenicol and lasalocid induces side-effects especially on cardiac function and it is not recommended.

Gut microbiota in burned patients with Clostridioides difficile infection.

BACKGROUND: The survival rate of patients with severe burn is positively associated with increasing the incidence of the Clostridioides difficile (C. difficile) infection (CDI). The surviving rate of severe burn patients now has an improved but the incidence of Clostridioides difficile (C. difficile) infection (CDI) has been continues increasing during recent two decades. This study assessed the molecular typing and phenotypic characterization isolates of C. difficile in burn patients with diarrhea, as well as environmental and skin infections with C. difficile spores at a referral burn hospital in Isfahan, Iran. It mainly aimed to evaluate the dominant bacterial structure in the gut microbiome of burned subjects with and without CDI. METHODS: In general, 309 samples were collected from 189 burned patients with hospital-acquired diarrhea and 120 swabs were collected from the healthcare workers' dominant hands, different sites of patients' skin, and medical tools. In addition, C. difficile isolates were characterized considering the existence of antibiotic resistance and toxin genes. Clinical cultures with identification of organisms and antibiotic susceptibility were done. C. difficle isolates were then genotyped and compared to clinical outcomes. Finally, the clinical characteristics of the participants were gathered through their records, and the bacterial targets of the gut microbiome were detected using quantitative real-time polymerase chain reaction (PCR). RESULTS: Based on the findings, 51 C. difficile isolates were detected from 189 severe burn patients hospitalized in the hospital. Further, PCR amplification tcdB and tcdA showed 23 isolates (12.2%) as toxigenic. Overall, 18.3% (22/120) of skin and environment samples demonstrated a positive result for C. difficile colonization. A low concentration of metronidazole and vancomycin (MIC90, 0.5, and 1.2 mg/L) inhibited all toxigenic C. difficile strains. Moreover, these isolates represented the highest rates of resistance to moxifloxacin and clindamycin (MIC90, 0.5, and 1.6 mg/L). A significantly reduced abundance of Clostridium spp., Bacteroidetes, and Bifidobacterium and an increase in the quantity of Firmicutes was observed in the gastrointestinal microbiome of burn patients (P < 0.01). Burn patients with CDI showed a significant decrease in Faecalibacterium prausnitzii (F. prausnitzii) while higher Akkermansia muciniphila (A. muciniphila) loads in comparison with healthy controls (P < 0.001 and P < 0.05). Contrarily, burned cases displayed increased levels of opportunistic pathogenic bacteria including the members of Enterococcus spp. and Escherichia coli (P < 0.05). CONCLUSIONS: Despite appropriate infection control strategies in the burn intensive care unit, CDI remains prevalent in severe burn patients. Eventually, the overgrowth of A. muciniphila and the decreased abundance of F. prausnitzii in burn cases with CDI could be potential predictive microbiome biomarkers in burned patients.

Molecular Typing of Community-acquired Methicillin-Resistant Staphylococcus aureus Isolated from 2- to 6-year old Children by Staphylococcal Protein A and Agr Typing in Isfahan, Iran.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has become a considerable public health concern in the entire world due to the rapid spread of this bacterium in human community; also the epidemiology of MRSA has changed, as the isolation of MRSA strains from healthy and non-healthy patients. Therefore, the objective of this study is to determine the genetic diversity and antibiotic resistance profile of community-acquired (CA)-MRSA nasal carriage in the Iranian samples. MATERIALS AND METHODS: A total of 25 CA-MRSA were isolated from the anterior nares of 410 healthy preschool children. All MRSA isolates were characterized by the detection of the toxic shock syndrome toxin-1 (TSST-1) and typed by γ-hemolysin genes, agr groups, and staphylococcal protein A (spa) typing. Kirby-Buyer antibiotic susceptibility testing was performed and interpreted as per the standard guidelines. RESULTS: A total of 25 (6.1%) MRSA isolates were recovered from the anterior nares of 410 preschool children. Sixteen isolates (64%) were positive for the TSST-1 gene. Three agr specificity groups were determined, as follows: eight (32%) isolates belonged to agr Group I, five (20%) isolates belonged to agr Group II, and 12 (48%) isolates belonged to agr Group III. The repeated profiles of these spa types of 25 isolates were organized into eight different lineages groups. Five of lineages contained a single strain, three of lineages contained two strains, and three of lineages consisted of more than three strains. CONCLUSIONS: The results of our study show that the rate of MRSA in our region is significantly high. Additionally, spa type t037 was the predominant type among CA S. aureus.

Biodegradation of polycyclic aromatic hydrocarbons, phenol and sodium sulfate by Nocardia species isolated and characterized from Iranian ecosystems.

Anthropogenic pollutants are known to have adverse effect on ecosystem, biodiversity and human health. Bioremediation is an option that has been widely used to remediate organic contaminants and reduce the risk of these hazardous materials. Microorganisms are readily available to screen and can be rapidly characterized to be applied in many extreme environmental conditions. Actinomycetes have a great potential for the production of bioactive secondary metabolites which have biodegradation activity. This study aimed to screen and characterize Nocardia species with biodegradation potential from diverse Iranian ecosystems. The isolates were screened from 90 collected environmental samples, identified and characterized using conventional and molecular microbiological methods including the PCR amplification and sequencing analysis of 16S rRNA and rpoB genetic markers. Growth rate in presence of pollutants, chromatography, Gibbs and turbidometric methods were used to determine bioremediation ability. A total of 19 Nocardia isolates were recovered from the cultured samples (21.1%) that belonged to 10 various species. The most prevalent Nocardia species was N. farcinica; 4 isolates (21%), followed by N. cyriacigeorgica and N. cashijiensis like; 3 isolates each (15.7%) and N. asteroides and N. kroppenstedtii; 2 isolates each (10.5%). Our results showed that various Nocardia species have great potential for bioremediation purposes, although they have not received much attention of the scholars for such significant usage.

Phenotypic and Genotypic Characteristics of Clostridium difficile Isolates in Patients with Type 2 Diabetes in Iran.

PURPOSE: This study aimed to investigate the phenotypic and genotypic characterization of Clostridium difficile isolates in type 2 diabetes patients with hospital-acquired diarrhea in four teaching hospitals in Isfahan, Iran. PATIENTS AND METHODS: A total of 104 hospitalized patients with type 2 diabetes and nosocomial diarrhea were included in the current study over a 2-year period (2015-2017). C. difficile isolates were characterized by conventional microbiological methods including the presence of toxin genes, antibiotic resistance testing and molecular methods including multilocus sequence typing (MLST) and random amplification of polymorphic DNA (RAPD). RESULTS: All 21 C. difficile isolates (20.2%) were detected from 104 studied patients. All isolates were susceptible to metronidazole and vancomycin. The antimicrobial resistance rates were distinctly higher for clindamycin and for moxifloxacin. Based on PCR amplification of tcdA and tcdB, 13 isolates (12.5%) carried both of these genes and were considered toxigenic. Thirteen toxigenic C. difficile strains were classified into two sequence types (STs), that is, ST54 and ST2 types. The RAPD-PCR amplification patterns of the detected toxigenic C. difficile revealed three distinct but related RAPD clusters. RAPD cluster 1 had the highest similarity with RAPD types 2 and 3. CONCLUSION: A relatively high rate of CDI was observed in patients with type 2 diabetes and was associated with poorer health outcomes. These patients were exposed to multiple antibiotics and other therapeutic agents. We recommend close screening for the coexistence of CDI and type 2 diabetes in patients with diarrhea using a combination of conventional and molecular methods.

Identification and molecular characterization of mycobacteria isolated from animal sources in a developing country.

The essential role of animals in the transmission of infectious diseases has long been recognized. Apart from zoonosis due to Mycobacterium bovis in domestic cattle, acquired mycobacterial zoonosis from animals are vastly under-reported worldwide. This is partly the result of not recognizing that animals can be the source of zoonotic nontuberculous mycobacteria (NTM) infection. The present study intended to be a contribution to the knowledge of somewhat neglected role of animals in harboring, maintenance and dissemination of NTM in the environment. A total of 326 samples from 250 animals were collected and analyzed for the presence of mycobacteria using standard protocols. The preliminary identification and Runyon's classification of isolates were performed by conventional tests. The PCR amplification of a 228 bp fragment of 65-kDa heat shock protein (hsp) gene was applied for the genus identification and the partial sequence analysis of 16S rRNA was applied for the species identification. In total 32 isolates including 26 rapidly growing and 6 slowly growing mycobacteria were recovered from 250 animal samples (12.8%). The isolates recovered from 21 (65.60%) fish, 8 (25%) insects and 3 (9.4%) house cats, dogs and mice. M. fortuitum was the most frequent Mycobacterium spp (13 isolates; 40.6% of all isolates), followed by M. abscessus-chelonae-M. saopaulense group, (5 isolates; 15.6% of all isolates), M. iranicum (3 isolates; 9.4% of all isolates),and M. marinum, M. terrae complex and M. chlorophenolicum (2 isolates each; 18.8% of all isolates), and the single isolates of M. mucogenicum, M. neoaurum, M. conceptionense, M. virginiense, and M. gordonae (5 isolates; 15.6% of all isolates). The current study indicates that a variety of animals can be a permanent or transient source of mycobacterial agents. This ensures the life cycle of the bacteria and the chance of their survival in the environment, which may pose a potential threat to human health.

Molecular characterization and genotyping of methicillin-resistant Staphylococcus aureus in nasal carriage of healthy Iranian children.

PURPOSE: Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has become a considerable public health concern in both developed and developing countries due to the rapid spread of this bacterium around the world, also the epidemiology of MRSA has changed, as the isolation of MRSA strains is not limited to health-care settings or patients with predisposing risk factors. Therefore, the objective of this study is to determine the genetic diversity and antibiotic resistance profile of CA-MRSA nasal carriage in Iranian children. METHODOLOGY: A cross-sectional study was conducted from April 2013 to March 2014. A total of 25 CA-MRSA were isolated from the anterior nares of 410 preschool children with no risk factors. All MRSA isolates were characterized by detection of the Panton-Valentine leukocidin (pvl) and γ-hemolysin genes, staphylococcal cassette chromosome mec (SCCmec) typing and multi-locus sequence typing (MLST). RESULTS: In 25 CA-MRSA isolates, Pvl and γ-hemolysin genes were detected in one (4%) and 18 (72 %) isolates; respectively. Overall, 92% (23/25) of isolates belonged to SCCmec type IV and 8% (2/25) of them had SCCmec type V profile. Using MLST, the 25 isolates were grouped into six clonal complexes (CC) and eight sequence types (ST) (CC5/ST6, CC22/ST22 and ST217, CC30/ST30 and ST1107, CC78/ST859, CC398/ST291 and CC97/ST405). The ST859/SCCmec IV (11/25, 44%) was the predominant clone among the isolates. ST859-MRSA-IV-pvl-negative (resistant to tetracycline) have successfully adapted to the Iranian preschool children population. CONCLUSION: Our results suggest that the genomic diversity was observed among the CA-MRSA. In addition, the current study demonstrates that pvl is not a reliable marker for CA-MRSA in our region.

Molecular epidemiology of Clostridium difficile infection in Iranian hospitals.

Background: Clostridium difficile infection (CDI) is known as one of the most important causes of nosocomial infections. The main objective of this study was to evaluate the presence of Clostridium difficile in the stool of hospitalized patients with diarrhea as well as in their environments. Methods: C. difficile isolates were characterized according to the presence of toxin genes and antibiotic resistance. Multilocus Sequence Typing Analysis (MLST) was applied for finding the genetic polymorphism and relationship among strain lineages. Results: A total of 821 samples (574 stools and 247 swabs) were collected between April 2015 and May 2017. The prevalence of C. difficile isolates was 28.6% (164/574) in patients and 19% (47/247) in swabs taken from medical devices, hands of healthcare workers and skin patient sites. Finally, 11.5% (66/574) toxigenic C. difficile strains isolated from stool samples of inpatients and 4.4% (11/247) from hands of healthcare workers and skin patient sites. All the toxigenic isolates were inhibited by a low concentration of vancomycin (MIC < 0.5 µg/ml). About 43% (33/77) and 39% of isolates were resistant to Clindamycin and moxifloxacin respectively. All isolates were susceptible to metronidazole. Toxigenic C. difficile strains were analyzed by MLST and were divided into 4 different STs. The detected types were ST-54 (57.9%), followed by ST-2 (31.6. %), ST-15 (5.3%) and ST-37 (5.3%), while none of the isolates were identified as ST-1 or ST-11. Significant risk factors for CDI appear to be advanced age, undergoing chemotherapy, previous surgery, and residence in the nursing home. Conclusions: CDI is common in Iran and further studies are recommended to monitor its epidemiological variations. Moreover, greater attempts must be made to encourage antibiotic stewardship by healthcare workers and the public.

Mycobacteriosis and Tuberculosis: Laboratory Diagnosis.

BACKGROUND: Tuberculosis is one of the most important infectious diseases that has claimed its victims throughout much of known human history. With Koch's discovery of the tubercle bacillus as the etiologic agent of the disease, his sanitary and hygienic measures, which were based on his discovery and the development of a vaccine against tuberculosis by Albert Calmette and Camille Guérin in 1921, an attenuated Mycobacterium bovis strain, bacilli Calmette-Guérin (BCG), and the discovery of the first antibiotic against tuberculosis, streptomycin by Selman Waksman in 1943, soon led to the opinion that appropriate control measures had become available for tuberculosis and it had been assumed that the disease could ultimately be eradicated.The emergence of resistant strains of this bacteria and widespread distribution of the disease in the world, and the emergence of the AIDS epidemic destroyed any possibility of global control of tuberculosis in the foreseeable future. OBJECTIVES: The purpose of this review is to highlight the current scientific literature on mycobacterial infections and provide an overview on the laboratory diagnosis of tuberculosis and non-tuberculosis infections based on conventional phenotypic and modern molecular assays. METHOD: In this study, a number of 65 papers comprising 20 reviews, 9 case reports, and 36 original research in association with mycobacteriosis and the laboratory diagnosis of mycobacterial infections, were reviewed. RESULTS: Based on our analysis on the published documents methods applied for the laboratory diagnosis of tuberculosis are continually assessed and developed in order to achieve more rapid, less expensive, and accurate results. Acid-fast staining and culture for mycobacteria remain at the core of any diagnostic algorithm with the sensitivity of 20-70% and specificity of 95-98% for AFB microscopy and the sensitivity of 95% and the specificity of 98% for culture based diagnosis. Following growth in culture, molecular tests such as nucleic acid hybridization probes and DNA sequencing may be used for definitive species identification. Nucleic acid amplification methods provide the means for direct detection of Mycobacterium tuberculosis in respiratory specimens without the prerequisite to isolate or culture the organism, leading to more rapid diagnosis and better patient care. CONCLUSION: As the researchers in a developing country, we strongly believe that despite significant advances in laboratory capacity, in many countries reliable confirmation of suspected mycobacterial diseases is hindered by a lack of knowledge on proper standardized methods, sufficient funds, suitably trained staff and laboratory supplies.

A case report of brain abscess caused by Nocardia cyriacigeorgica in a diabetic patient.

INTRODUCTION: Nocardia are Gram-positive partially acid-fast bacilli capable of inducing a wide range of infections in patients with immunodeficiency, AIDS, cancer, lupus erythematous and diabetes. Nocardia cyriacigeorgica was first isolated in 2001 from a patient with chronic bronchitis. Since then, there have been reports on the clinical significance of this organism in patients with bronchitis, brain abscess and lung diseases. We, herein, report a case of brain abscess in an elderly diabetic patient from Iran. CASE PRESENTATION: The patient was a 73 year-old woman admitted to hospital due to severe headache and shortness of breath. The patient had lived with diabetes for 20 years and suffered from chronic foot ulcer. She was admitted to hospital with fever, weakness, drowsiness and vomiting. Clinical examination and the head CT scan of the left frontal lobe of the brain revealed a metastatic carcinoma involving skull bone in the tumor that resulted in two surgical operations in the following two years. The brain abscess biopsy revealed an infection with Nocardia cyriacigeorgica confirmed by phenotypic and molecular tests including a PCR-based amplification of a target genetic marker, a 596 bp fragment of 16S rRNA gene, followed by almost full 16S rRNA sequencing. CONCLUSION: The rare infections, such as brain abscess with Nocardia, are easily neglected or misdiagnosed due to the fastidious nature of the organism and inadequate microbiological experience of laboratories in the hospitals of developing countries. This case shows that hospitals should consider a better laboratory protocol to deal with the clinical cases in which fastidious organisms, and in particular Nocardia, are involved.