Evaluation of the Effect of Lactobacillus acidophilus ATCC 4356 Bacteriocin against Staphylococcus aureus.

Background: Lactobacillus acidophilus is lactic acid bacteria that produce bacteriocins. Bacteriocins are antimicrobial peptides or proteins that exhibit activity against closely related bacteria. The aim of this study was to determine the effect of L. acidophilus ATCC 4356 bacteriocin against Staphylococcus aureus. Material and Methods. We used four different phenotypic methods for antimicrobial activities against two standard strains: methicillin-resistant S. aureus (MRSA) ATCC 33591 and methicillin-susceptible S. aureus (MSSA) ATCC 25923. The methods were (1) agar well diffusion, (2) overlay soft agar, (3) paper disk, and (4) modification of punch hole. The ammonium sulfate method was used to concentrate crude bacteriocin, and ultrafiltration and dialysis tubes were used to remove ammonium sulfate from the bacteriocins. Each method was repeated in triplicate. Result: L. acidophilus ATCC 4356 showed antimicrobial activity against both MRSA and MSSA standard strains only by the overlay soft agar method and not by the agar well diffusion, punch hole modification, and paper disk methods. No antimicrobial effects were observed in crude bacteriocins concentrated. Conclusion: The growth inhibition of S. aureus in overlay soft agar method may be due to the production of bacteriocin-like substances. The overlay soft agar method is a qualitative test, so there is a need for further study to optimize the conditions for the production of bacteriocin-like substances in the culture supernatant and precise comparison between the inhibitory activity and pheromone secretion of different strains.

Antimicrobial Effects of Mouse Adipose-Derived Mesenchymal Stem Cells Encapsulated in Collagen-Fibrin Hydrogel Scaffolds on Bacteroides fragilis Wound Infection in vivo.

Background: Anaerobes are the causative agents of many wound infections. B. fragilis is the most prevalent endogenous anaerobic bacterium causes a wide range of diseases, including wound infections. This study aimed to assess the antibacterial effect of mouse adipocyte derived-mesenchymal stem cell (AD-MSCs) encapsulated in collagen-fibrin (CF) hydrogel scaffolds on B. fragilis wound infection in an animal model. Methods: Stem cells were extracted from mouse adipose tissue and confirmed by surface markers using flow cytometry analysis. The possibility of differentiation of stem cells into osteoblast and adipocyte cells was also assessed. The extracted stem cells were encapsulated in the CF scaffold. B. fragilis wound infection was induced in rats, and then following 24 h, collagen and fibrin-encapsulated mesenchymal stem cells (MSCs) were applied to dress the wound. One week later, a standard colony count test monitored the bacterial load in the infected rats. Results: MSCs were characterized as positive for CD44, CD90, and CD105 markers and negative for CD34, which were able to differentiate into osteoblast and adipocyte cells. AD-MSCs encapsulated with collagen and fibrin scaffolds showed ameliorating effects on B. fragilis wound infection. Additionally, AD-MSCs with a collagen scaffold (54 CFU/g) indicated a greater effect on wound infection than AD-MSCs with a fibrin scaffold (97 CFU/g). The combined CF scaffold demonstrated the highest reduction in colony count (the bacteria load down to 29 CFU/g) in the wound infection. Conclusion: Our findings reveal that the use of collagen and fibrin scaffold in combination with mouse AD-MSCs is a promising alternative treatment for B. fragilis.

Anti-Biofilm Potential of Lactobacillus casei and Lactobacillus rhamnosus Cell-Free Supernatant Extracts against Staphylococcus aureus.

Background: Biofilm production is an important virulence factor in Staphylococcus aureus. Most of the infections associated with biofilms of this bacterium are very difficult to treat using antibiotics. The present research studied the effects of the two probiotic Lactobacillus species L. casei and L. rhamnosus on S. aureus biofilm. Materials and Methods: Cell-free supernatant (CFS) extracts of L. casei ATCC 39392 and L. rhamnosus ATCC 7469 culture were prepared. The effects of sub-minimum inhibitory concentrations of the CFS extracts on cell surface hydrophobicity (CSH), initial attachment, biofilm formation, and their ability in eradicating S. aureus ATCC 33591 biofilms were assessed. In addition, the effects of CFS extracts on expression of the genes involved in formation of S. aureus biofilms (cidA, hld, sarA, icaA, and icaR) were also evaluated through real-time polymerase chain reaction. Results: CFSs of both Lactobacillus spp. significantly reduced CSH, initial attachment, and biofilm formation and eradicated the biofilms. The above findings were supported by scanning electron microscopy results. These two Lactobacillus CFSs significantly changed the expression of all studied biofilm-related genes. Expression levels of cidA, hld, and icaR genes significantly increased by 4.4, 2.3, and 4.76 fold, respectively, but sarA and icaA genes were significantly downregulated by 3.12 and 2.3 fold. Conclusion: The results indicated that CFS extracts of L. casei and L. rhamnosus had desirable antagonistic and anti-biofilm effects against S. aureus. Consequently, carrying out further research enables us to prepare pharmaceuticals from these CFSs in order to prevent and treat infections caused by S. aureus biofilms.

Quantification of human adenovirus in irrigation water-soil-crop continuum: are consumers of wastewater-irrigated vegetables at risk?

Because of health concerns regarding the presence of enteric viruses in wastewater effluents, this study was designed to investigate the occurrence of human adenovirus (HAdV) in the irrigation water-soil-crop continuum. Viral particles were extracted from wastewater and wastewater- or water-irrigated soil and crop samples and analyzed using real-time PCR. Concentration of fecal indicator bacteria (FIB) were also determined. Quantitative microbial risk assessment was performed to determine the HAdV illness risk associated with the consumption of wastewater-irrigated vegetables. HAdV-F was detected in 74% of wastewater effluent samples with a mean concentration of 38 Genomic Copy (GC)/mL. HAdV was also detected in wastewater-irrigated soil (2 × 102 GC/g) and crop (< 10 GC/g) samples, with no statistically significant difference in concentrations between wastewater- and freshwater-irrigated samples. The results showed no correlation between concentrations of FIB and HAdV in the analyzed samples. Mean probability of illness risk from consumption of wastewater-irrigated vegetables was 4 × 10-1 per person per year (pppy) which was about two orders of magnitude higher than the proposed value by WHO (10-3 pppy) for safe reuse of wastewater. This finding suggests that the wastewater reuse for irrigation of vegetables eaten raw could pose a threat to human health with respect to the risk of viral illness, signifying stricter management of wastewater reuse. However, because of uncertainties in the QMRA model, particularly the ratio of infectious to non-infectious virus particles, more data is required to validate the predicted risk. This information is especially important in arid and semi-arid regions where high temperatures, UV radiation intensity, and desiccation can efficiently inactivate microorganisms in the environment.

Evaluation of the effect of Saccharomyces cerevisiae on the expression of enterotoxin genes in Escherichia coli O157: H7 (EHEC) and Escherichia coli H10407 (ETEC).

Enterotoxigenic (ETEC) and enterohemorrhagic Escherichia coli (EHEC) are the most important intestinal pathogens. Probiotics play an effective role in reducing the expression of virulence factor genes in intestinal pathogenic bacteria. The aim of the present study is to investigate the effect of probiotic Saccharomyces cerevisiae S3 on the expression of enterotoxin genes in both ETEC and EHEC. Supernatant and lysate of S. cerevisiae S3 are prepared. Subminimal inhibitory concentrations (sub-MIC) of supernatant and lysate are individually exerted to O157: H7 and H10407. The genes' expression of enterotoxins (elt, est, stx1, and stx2) are then determined using real-time PCR technique. The results showed, the yeast supernatant could decrease the expression of the elt gene in ETEC and that of stx1 in EHEC. Of note, in other cases, stx1 and est genes' expression increased. The lysate had no inhibitory effect on the expressions of elt, est, and stx2 genes, but it increased the expression of genes in both ETEC and EHEC. Lysate extract only decreased the expression of stx1 in O157: H7. Our study shows some interesting results regarding the effectiveness of the compounds produced by S. cerevisiae S3 in the expressions of toxin genes in both ETEC and EHEC. We recommend more similar studies be performed in this regard.

Evaluation of Antibiotic Resistance Pattern, Alginate and Biofilm Production in Clinical Isolates of Pseudomonas aeruginosa.

BACKGROUND: Pseudomonas aeruginosa is one of the most common opportunistic bacteria causing nosocomial infections, which has significant resistance to antimicrobial agents. This bacterium is a biofilm and alginate producer. Biofilm increases the bacterial resistance to antibiotics and the immune system. Therefore, the present study was conducted to investigate the biofilm formation, alginate production and antimicrobial resistance patterns in the clinical isolates of P. aeruginosa. METHODS: One hundred isolates of P. aeruginosa were collected during the study period (from Dec 2017 to Jul 2018) from different clinical samples of the patients admitted to Milad and Pars Hospitals at Tehran, Iran. Isolates were identified and confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was specified by the disk diffusion method. Biofilm formation and alginate production were measured by microtiter plate and carbazole assay, respectively. RESULTS: Sixteen isolates were resistant to all the 12 studied antibiotics. Moreover, 31 isolates were Multidrug-Resistant (MDR). The highest resistance rate was related to ofloxacin (36 isolates) and the least resistance was related to piperacillin-tazobactam (21 isolates). All the isolates could produce the biofilm and alginate. The number of isolates producing strong, medium and weak biofilms was equal to 34, 52, and 14, respectively. Alginate production was more than 400 µg/ml in 39 isolates, 250-400 µg/ml in 51 isolates and less than 250 µg/ml in 10 isolates. CONCLUSION: High prevalence of MDR, biofilm formation, and alginate production were observed among the clinical isolates of P. aeruginosa. The results also showed a significant relationship between the amount of alginate production and the level of biofilm formation.

Inhibitory effect of probiotic yeast Saccharomyces cerevisiae on biofilm formation and expression of α-hemolysin and enterotoxin A genes of Staphylococcus aureus.

BACKGROUND AND OBJECTIVES: Staphylococcus aureus, as an opportunistic pathogen, is the cause of a variety of diseases from mild skin infections to severe invasive infections and food poisoning. Increasing antibiotic resistance in S. aureus isolates has become a major threat to public health. The use of compounds produced by probiotics can be a solution to this problem. Thus, the purpose of this study was to investigate the effect of Saccharomyces cerevisiae on some virulence factors (biofilm, α-hemolysin, and enterotoxin A) of S. aureus. MATERIALS AND METHODS: Supernatant and lysate extracts were prepared from S. cerevisiae S3 culture. Sub-MIC concentrations of both extracts were separately applied to S. aureus ATCC 29213 (methicillin-sensitive S. aureus; MSSA) and S. aureus ATCC 33591 (methicillin-resistant S. aureus; MRSA) strains. Biofilm formation of these strains was measured by microtiter plate assay and expression level of α-hemolysin and enterotoxin A genes (hla and sea, respectively) using real-time PCR technique. RESULTS: The supernatant extract has reduced both biofilm formation and expression of sea and hla genes, while lysate extract had only anti-biofilm effects. The MRSA strain showed more susceptibility to yeast extracts than MSSA strain in all tests. CONCLUSION: The present study exhibited favorable antagonistic effects of S. cerevisiae S3, as a probiotic yeast, on MSSA and MRSA strains. Based on the findings of this study, the compounds produced by this yeast can be used to control S. aureus infections; however, further similar studies should be conducted to confirm the findings of the present study.

Genetic and Phylogenetic Characterization of the M Gene of Influenza A Virus Isolated from Iranian Patients.

BACKGROUND: A few studies have been done on the molecular analysis of Iranian influenza A isolates M gene. METHODS: In 2014, nasal swabs collected from outpatients with clinical symptoms in the hospital clinics of Tehran, Iran were subjected for influenza detection and subtyping using Real-Time RT-PCR. Sequence and phylogenetic analysis performed on four randomly selected isolates from each subtype (H1N1 and H3N2) using neighbor-joining method. RESULTS: Phylogenetic dendrograms drawn based on M nucleotide sequence of H1N1 isolates showed close relatedness with Omanian isolates while the most isolates of H3N2 have clustered with Kuwait isolates and isolates from outside of geographical location. Amino acid sequence analysis showed S31N substitution in all isolates rendering the virus resistant to adamantanes. CONCLUSION: This study determined the sequence identity and phylogenetic relatedness of M gene sequence got from Iranian influenza A isolates to elucidate the modality of relationship of this gene in comparison with its counterparts from other regions.

Screening and characterization of in-vitro probiotic criteria of Saccharomyces and Kluyveromyces strains.

BACKGROUND AND OBJECTIVES: Probiotics are defined as live micro-organisms conferring a health benefit on the host. Although most probiotics are bacteria, some yeasts such as Saccharomyces and Kluyveromyces, has been found to have effective probiotic properties. The objective of this study was to isolate and identify indigenous Saccharomyces and Kluyveromyces yeast strains and to compare some probiotic characteristics between these two strains in vitro. MATERIALS AND METHODS: Strains were isolated on yeast glucose chloramphenicol agar medium from 205 samples and identified by morphological, physiological and biochemical assays. The effects of different conditions such as pH and temperature on the survival and growth of the isolates were studied. In addition, resistance to acidic pH (1.5, 2, 3 and 5), pepsin and different concentrations of bile salts (1%, 3% and 5%), as well as proteolytic, lipolytic and hemolytic activity of selected isolates were assessed. Finally, the best isolates were selected for investigation of their viability in samples of dairy products. RESULTS: 126 isolates were identified using biochemical and molecular techniques as yeast strains. Five isolates were found to have effective probiotic properties, belonging to Kluyveromyces marxianus (S97, S101 and S106) and Saccharomyces cerevisiae (S28, S34). These isolates were able to grow at 37°C, pH=1.5, withstand to concentration of 5% oxbile and pepsin and exhibit the proteolytic activity. The isolates of K. marxianus showed better viability in dairy (yogurt). CONCLUSION: In the in-vitro comparative experiments, the isolates of K. marxianus showed better probiotic potentials.

Frequency of genes encoding erythromycin ribosomal methylases among Staphylococcus aureus clinical isolates with different D-phenotypes in Tehran, Iran.

BACKGROUND AND OBJECTIVES: Macrolide, lincosamide and streptogramin type B (MLSB) antibiotics are important in the treatment of Staphylococcus aureus infections and existence of isolates with ability to resist against MLSB antibiotics is worrisome. MATERIALS AND METHODS: In this cross sectional study, 101 S. aureus isolates were collected from patients of five selected hospitals in Tehran over a period of five months. Disk diffusion tests and differentiation between constitutive and inducible resistances were carried out by D-test. The presence of mecA, msrA, ermA and ermC genes were detected using PCR or multiplex PCR. RESULTS: Out of 101 S. aureus isolates, 58 (57.4%) were methicillin resistant and 57 (56.4%) expressed resistance to erythromycin. The prevalence of constitutive MLSB (cMLSB), inducible MLSB (iMLSB) and MS (Negative) phenotype in all erythromycin resistant isolates were 71.9, 26.3 and 1.7%, respectively. Out of all the erythromycin resistant isolates, 57.8% harbored both ermA and ermC genes which possessed constitutive resistance. 8.7% of the isolates contained ermA gene alone which possessed inducible resistance with D phenotype and 5.2% of isolates just contained ermC gene which had inducible resistance with D+ phenotype. msrA gene was detected in 3.5% of the erythromycin resistant S. aureus isolates with constitutive resistance. None of the genes were detected among MS phenotypes. CONCLUSION: In this study, most of S. aureus isolates carried both ermA and ermC genes and there was a significant relationship (P value ≤ 0.05) between different resistance phenotypes and erm genes.

Relation between Resistance to Antipseudomonal ß-Lactams and ampC and mexC Genes of Pseudomonas aeruginosa.

BACKGROUND: In order to select a better antibiotic choice for treatment of Pseudomonas aeruginosa infections, this study was conducted to determine the frequency of resistance to some antipseudomonal ß-lactams in P. aeruginosa isolates from patients in Tehran, Iran. In addition, the relation between presence of genes known to be responsible for resistance to ß-lactams (ampC, mexC 1,2, and mexC 3,4 genes) and resistance phenotype among P. aeroginosa isolates was evaluated. METHODS: P. aeruginosa strains were isolated and identified by routine methods and PCR for oprL gene. Disk diffusion method was employed to determine the antimicrobial susceptibility pattern according to CLSI recommendations. PCR was used to detect the resistance genes. RESULTS: Among 100 isolates of P. aeruginosa, 82% had ampC, 86% mexC 1,2 and 89% mexC 3,4 genes and combinations of these genes were seen in most of isolates and only 3% of isolates had none of these genes. Resistance to mezlocillin, cefepime, ceftazidime and piperacillin/ tazobactam was seen in 46%, 41%, 36% and 29% of isolates, respectively. Significant relation (P value ≤0.05 by Chi-square or Fisher Exact test) was observed between the presence of ampC gene and resistance to all the studied ß-lactams in this study. No relation was observed for mexC genes, although many of isolates containing these two genes were phenotypically resistant. DISCUSSION: This study had shown for the first time, the presence of ampC and mexC genes in significant percent of clinical isolates of P. aeruginosa in Tehran, Iran, and relation between presence of ampC gene and resistance to ß-lactams.

Isolation of indigenous Glutathione producing Saccharomyces cerevisiae strains.

BACKGROUND: Glutathione (GSH) is a non-protein thiol compound, which plays an important role in the response to oxidative stress and nutritional stress. The aim of this study was to isolate indigenous S. cerevisiae strains capable of effectively produce GSH. METHODS: One hundred-twenty sweet fruit samples were collected. The strains were isolated on yeast glucose chloramphenicol (YGC) agar medium and identified. The isolates were evaluated for GSH producing on yeast malt (YM) medium. Concentration of glutathione was investigated by recording absorbance of all samples at wavelength 412 nm (Ellman's method). In addition, optimization of glucose and peptone concentration in culture medium and the effects of various environmental conditions such as temperature (20-35 °C), agitation rate (150-250 rpm), and initial pH (4.0-6.0) were assessed on producing of GSH. RESULTS: From 120 samples, 80 isolates were identified by morphological, biochemical and molecular tests as S. cerevisiae. Five isolates were capable to produce effectively GSH. The optimal culture conditions were agitation rate, 200 rpm; temperature, 30 °C; initial pH, 6; glucose, 30 g/l; and peptone concentration, 5 g/l. In optimal conditions, the amount of derived glutathione was improved compared to YM basal medium and highest GSH concentration (296.8 mg/l) was obtained after cultivation with shaking for 72 h. CONCLUSION: The possibility of obtaining S. cerevisiae cells with a high GSH intracellular content can be considered an interesting opportunity of furthering the range of application and utilization of this molecule.

Diarrheagenic Escherichia coli pathotypes frequency in Khuzestan province of Iran.

BACKGROUND AND OBJECTIVES: Diarrheagenic Escherichia coli (DEC) is an emerging agent among pathogens that causes diarrhea. Studies showed that diarrheagenic E. coli such as enterohaemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC), diffusely adhering E. coli (DAEC) and shiga toxin producing E. coli (STEC) strains are among the most frequent causative agents in acute diarrhea. The aim of this study was to determine the frequency of DEC pathotypes in Khuzestan province. MATERIALS AND METHODS: Stool samples were collected from patients with diarrhea in Khuzestan province of Iran. E. coli strains were isolated using conventional culture and standard biochemical tests. The polymerase chain reaction (PCR) technique was used to detect presence of virulence genes, i.e; eae, stx1 and stx2 for EHEC, bfp and eae for EPEC, LT and ST for ETEC, AA for EAEC, invE for EIEC, stx1 and stx2 for STEC. RESULTS: Altogether, 200 stool samples were obtained from patients, of which 158 (79%) were positive for E. coli. DEC was identified in 127 (63%) of stool samples, which frequency of each pathotypes were as follows: atypical EPEC 49 (39%), typical EPEC 1 (0.7%), STEC 50 (39.3%), ETEC 21 (16.3%), EAEC 5 (4.0%) and EIEC 1 (0.7%). Most frequent etiological agents of diarrhea in Khuzestan province of Iran were STEC and EPEC. CONCLUSION: Our findings showed DEC had been agent of diarrhea in Khuzestan. This finding provides evidence that effort should be made to estimate the burden of infection by the etiological agent for better medical approach and should raise notification about antibiotic resistance among bacterial infection.

Detection of Multidrug Resistant (MDR) and Extremely Drug Resistant (XDR) P. Aeruginosa Isolated from Patients in Tehran, Iran.

BACKGROUND: This study was done to detect multidrug resistant (MDR) and extremely drug resistant (XDR) of Pseudomonas aeruginosa among strains isolated from patients in Tehran, Iran, due to importance of these phenotypes in treatment of human infections. METHODS: Eighty eight P. aeruginosa were isolated from patients in Tehran, Iran, and identified by routine methods and PCR for oprL gene. Their antimicrobial susceptibility to 16 antimicrobial agents from 7 antimicrobial categories (aminoglycosides, carbapenems, cephalosporins, fluoroquinolones, penicillins/ß-lactamase inhibitors, monobactams, polymyxins) were determined by disk diffusion method, according to recommendation of Clinical and Laboratory Standards Institute. Characterization of P. aeruginosa isolates as MDR and XDR was done according to standardized international terminology presented by European Centre for Disease Prevention and Control as well as the Centers for Disease Control and Prevention in 2011. MDR was defined as acquired non-susceptibility to at least one agent in ≥3 antimicrobial categories and XDR was defined as non-susceptibility to at least one agent in ≥6 antimicrobial categories. RESULTS: The rates of susceptibility to antimicrobials were as follows: gentamicin 27.3%, tobramycin 54.5%, amikacin 56.8%, netilmicin 36.4%, imipenem 55.7%, meropenem 55.7%, doripenem 60.2%, ceftazidime 63.6%, cefepime 56.8%, ciprofloxacin 59.1%, levofloxacin 60.2%, ticarcillin-clavulanic acid 37.5%, piperacillin-tazobactam 63.6%, aztreonam 43.2%, colistin 90.9%, polymyxin 95.5%. Altogether, 48 (54.5%) and 29 (33%) isolates were characterized as MDR and XDR, respectively. DISCUSSION: The high frequency of antibiotic resistance in clinical isolates of P. aeruginosa in Iran makes epidemiological surveillance of susceptibility of this bacterium more essential for the best selection of empirical antibiotics.

The Prevalence of Resistance to Methicillin in Staphylococcus aureus Strains Isolated from Patients by PCR Method for Detec-tion of mecA and nuc Genes.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is the main cause of hospital infection emerged over the last decades. Rapid detection of MRSA is important for patient care and proper usage of infection control. Detection of mecA genes (encoding resistance to methicillin and other similar antibiotics) and nuc genes (encoding staphylococcal thermostable nuclease) by PCR method is now considered for rapid identification of MRSA strain. The aim of this study was to determine the prevalence of MRSA isolated from patients in Tehran, Iran by PCR method for detection of mecA and nuc genes. METHOD: Phenotypic method such as microscopic and colony morphology and catalase and coagulase tests were used for identification of S. aureus isolates. DNA was extracted from all isolates and the presence of nuc and mecA gene was detected by PCR method. For determination of MRSA by phenotypic methods, oxacillin disk diffusion test were used. Data were analyzed by SPSS software. RESULTS: Out of 126 clinical sample identified by phenotypic method, 101 isolates had nuc gene. In disk diffusion tests by oxacillin disk, 78.2% of isolates were considered to be MRSA, but in PCR method for mecA gene, 69% isolates were positive. CONCLUSIONS: The results showed a high prevalence of methicillin-resistance among S. aureus isolates. Identifying MRSA strains, isolating MRSA-positive patients and carrier's treatment in a hospital to prevent MRSA infection is important in limiting the spread of MRSA. The PCR method for detection of nuc and mecA genes has potential for rapid and accurate diagnosis of MRSA strains.

Phosphate solubilization characteristics of efficient nitrogen fixing soil Azotobacter strains.

BACKGROUND AND OBJECTIVES: Azotobacter is a diazotroph bacterium reported to possess various plant growth-promoting characteristics.The aim of this study was to isolate Azotobacter strains capable of fixing nitrogen and effectively hydrolyzing both organic and inorganic Pi compounds. MATERIALS AND METHODS: In this study, soil samples collected from a diverse range of slightly alkaline soil types were screened for Azotobacter isolates. The inorganic and organic phosphate solubilization potentials of twenty competent phosphate solubilizing Azotobacter isolates were assessed.Variations were noted in the solubilization potentials. RESULT: Three isolates, identified as Azotobacter vinelandii strains O2, O4 and O6, were able to fix atmospheric N2 effectively. The nitrogenase activity of these isolates ranged between 158.6 and 326.4 C2H4h(-1)vial(-1) (ethylene). Bacterial growth rates and phosphate solubilization activities were measured quantitatively under various environmental conditions. A close association was evident between phosphate solubilizing ability and growth rate as an indicator of active metabolism. All three phosphate solubilizing bacteria (PSB) were able to withstand temperature as high as 45°C, high concentration of NaCl (upto 5%) and a wide range of initial pH from 5 to 10 while hydrolyzing phosphate compounds actively. CONCLUSION: Azotobacter vinelandii strains O2, O4 and O6 are superior candidates for biofertilizers that may result in the reduction of chemical nitrogen and phosphate fertilizers leading to increase crop production.

Conjunctival microbial florae in patients with seriously sulfur mustard induced eye injuries.

INTRODUCTION: Ocular surface disorders and infections in sulfur mustard (SM) exposed patients are of particular clinical importance. The aim of the present study is to detect the conjunctival bacterial florae in patients with seriously SM induced eye injuries. MATERIALS AND METHODS: Conjunctival bacterial florae of 143 seriously eye injured subjects as the study group was detected. The results were compared with 26 normal participants. Both groups were matched in age and sex. The samples were taken by sterile swab from interior fornixes of conjunctiva in both groups and were transported to microbiology laboratory by Stuart's Transport Medium. All samples were inoculated onto Blood agar, Mac Conkey agar and Chocolate agar and isolated microorganisms were identified by biochemical tests. The data were analyzed by SPSS and Man Whitney tests. RESULTS: Nineteen cases (13.39%) and none of the controls (0%) had positive culture results (p = .043). Isolated microorganisms from patients included coagulase-negative staphylococci 10 cases (52.6%), Staphylococcus aureus 5 cases (26.3%), non enterobacteriaceae gram negative bacilli 2 cases (10.5%), Penicillium spp. 2 cases (10.5%), Citrobacter sp. 1 case (5.2%), non-spore forming Gram positive bacillus 1 case (5.2%) and α hemolytic streptococcus 1 case (5.2%). Two patients had mixed microorganisms and other patients had just one microorganism. Most of the S. aureus isolates were sensitive to usual antibiotics. CONCLUSIONS: The results of this study showed that the prevalence rate of conjunctival bacterial isolates in patients with seriously SM induced ocular injuries are higher and potentially more dangerous than normal controls.

Phenotypic and genotypic study of macrolide, lincosamide and streptogramin B (MLSB) resistance in clinical isolates of Staphylococcus aureus in Tehran, Iran.

BACKGROUND: Resistance to antimicrobial agents among Staphylococcus aureus is an increasing problem. Two common genes responsible for resistance to macrolide, lincosamide and streptogramin B (MLSB) antibiotics are the ermA and ermC genes. Three resistance phenotypes have been detected to these antibiotics: strains containing cMLSB (constitutive MLSB) and iMLSB (inducible MLSB), which are resistant to macrolide, lincosamide and streptogramin B antibiotics, and MS, which is only resistant to macrolide and streptogramin B antibiotics. The aim of this study was to determine the prevalence of MLSB phenotypes and genotypes in erythromycin-resistant strains of S. aureus isolated from patients in 4 university hospitals in Tehran, Iran. MATERIAL/METHODS: S. aureus strains were isolated from various clinical specimens and identified by routine phenotypic methods and PCR for nuc gene. Erythromycin resistance was determined by disk diffusion testing. Prevalence of MLSB phenotypes was determined by use of the D-test. ermA and ermC genes were detected by PCR. RESULTS: Altogether, 126 erythromycin-resistant strains of S. aureus were detected. Prevalence of cMLSB, iMLSB and MS resistance phenotypes were 92.8%, 6.4%, and 0.8%, respectively; 60.3% of strains had ermA gene and 54.8% ermC gene; 61 strains (48.4%) contained 2 studied erm genes and 42 strains (33.3%) did not have any studied erm genes. CONCLUSIONS: Due to the high prevalence of clindamycin resistance among S. aureus isolated from patients in Iran, we recommend clindamycin therapy only after proper antimicrobial susceptibility testing.

Mupirocin resistance among Iranian isolates of Staphylococcus aureus.

BACKGROUND: The topical agent mupirocin plays a crucial role in strategies designed to control outbreaks of methicillin-resistant Staphylococcus aureus. The rate of high- and low-level mupirocin resistance among S. aureus strains from Iranian hospitals is not known. MATERIAL/METHODS: Two hundred twenty-two nonduplicate S. aureus strains consecutively isolated in four university hospitals in Tehran, Iran, were tested for mupirocin susceptibility by disc diffusion agar method and minimum inhibitory concentration (MIC) determination by the E-test. Susceptibility to 16 other antimicrobial agents was also determined. RESULTS: With the disc diffusion agar method, the majority of strains (97.3%) were susceptible to mupirocin and only 2.7% were resistant. The S. aureus strains showed high resistance (>50%) to most antibiotics, including penicillin G, ampicillin-sulbactam, oxacillin, cefoxitin, ciprofloxacin, erythromycin, tetracycline, clindamycin, gentamicin, and rifampicin, but resistance to linezolid, chloramphenicol, cotrimoxazole, and quinupristin/dalfopristin was low and no isolate was resistant to vancomycin. In the E-test, six strains had MICs of >4 mg/l, i.e. five strains had MICs of 8-256 mg/l (low-level mupirocin resistance) and one strain had 1024 mg/l (high-level mupirocin resistance). One strain was resistant to mupirocin in the disc diffusion agar method but showed sensitivity in the E-test (MIC: 0.94 mg/l). The mupirocin-resistant S. aureus isolates were all methicillin resistant and more resistant to the other antimicrobial agents compared with the mupirocin-susceptible strains. CONCLUSIONS: This study is the first report about mupirocin resistance of S. aureus in Iranian hospitals.

Study of a polymerase chain reaction-based method for detection of herpes simplex virus type 1 DNA among Iranian patients with ocular herpetic keratitis infection.

PURPOSE: To study the presence of the herpes simplex virus type 1 (HSV-1) glycoprotein D gene in tear films of Iranian patients with herpetic keratitis. METHODS: Twenty-five tear film and eye swab specimens from 25 herpetic keratitis patients and 10 specimens from 10 healthy volunteers were collected in the Farabi Eye Hospital, Tehran, Iran. HSV-1 DNA was detected by using the nested polymerase chain reaction (nPCR) method. Viral isolation was done using conventional viral techniques. A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was used for confirmation of positive cytopathic effect cell culture. The results of a diagnosis by an ophthalmologist team were compared with those of nPCR. RESULTS: HSV-1 DNA was identified in tear films of 88% (23/25) of suspected herpetic keratitis patients. All healthy controls (100%) had negative PCR results. HSV-1 was isolated in cell culture and confirmed by ELISA in 12% (3/25) of herpetic keratitis patients who had epithelial keratitis. The kappa value showed a high level of agreement between ophthalmologist team diagnosis and the PCR results (kappa = 0.86, P < 0.0001). CONCLUSIONS: nPCR is a sensitive, rapid, and powerful tool for detection of HSV-1 DNA in tear films of ocular herpetic keratitis patients and can serve as a supplemental method for diagnosis of herpetic keratitis infection.