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Satureja khuzistanica jamzad (SKJ), which is a member of Lamiaceae, has various proven effects such as antispasmodic and anti-inflammatory, anti-diabetic, antioxidant, and antifungal properties. Ho...

Background: Food-borne diseases can cause many problems and harm consumers and societies. Dairy products are an important part of the food pyramid. In addition, dairy products provide many nutritional needs to humans and can also be contaminated with bacteria and cause gastrointestinal diseases and food poisoning. Objective: The purpose of the present research was to evaluate bacterial contamination rate in cheese, raw and pasteurized milk. Methods: For this purpose, 100 samples, including 35 samples of raw milk samples, 35 samples of pasteurized milk and 30samples of cheese, were evaluated. Cultures, biochemical and phenotypic tests were performed. Finally, statistical analysis was performed. Results: Our finding demonstrates that Enterococcus spp was the highest rate in cheese samples, E.coli was the highest rate in raw milk samples, and citrobacter freundii, klebsiella oxytoca and enterococcus spp were of the same rate in pasteurized milk samples. Conclusion: The results obtained in this study demonstrated that some dairy products in Ilam province, were contaminated. So, we suggested that the pasteurized milk and dairy products be done with complete care and raw dairy products should not be used.

Introduction Cancer is a major public health problem in the modern world. Every year, new cases of cancer are diagnosed around the world. Cancer cells are altered cells that have escaped the mechanisms that regulate natural growth. Bacteriocins are cationic peptides synthesized by ribosomes that are secreted by almost all groups of bacteria. Some bacteriocins have shown selective toxicity to cancer cells compared to normal cells. This makes them other candidates for research and clinical trials. Aim Due to the high prevalence of colon cancer and its therapeutic problems, this study was performed on colicin E7 to evaluate its anti-colon cancer properties. Material and methods For this reason, colE7 was cloned in pet32c vector and purified protein was affected on HT-29 cells to evaluate the expression of p53 and bcl2. Results Our in silco analysis demonstrated that colicin E7 has 87.23% confidence as anticancer peptide by ACPred-FL program. First, a PCR reaction was performed using specific primers of the colicin E7 gene, which formed the 1728 bp fragment that belongs to this gene. Conclusions Colicin E7 decreased the expression of bcl2 and increased P53. The results of this study showed the general effect of colicin E7 on cancer cells in vitro, which can be evaluated in the future with further experiments.

Nepeta curvidens is a plant of the mint family and an important species of the genus Nepeta. The main habitat of the N. curvidens in the Garin Mountains in Lorestan Province, Iran. Although locals have used N. curvidens as a food flavoring agent for years, few studies have been conducted on its phytochemical and biological properties. The chemical constituents of N. curvidens essential oil and then its biological properties were studied in detail in this investigation. N. curvidens essential oil was prepared from the aerial parts of the plant using the hydrodistillation method and its chemical composition was analyzed by the GC-MS method. Antimicrobial, antifungal, anti-Leishmania, and cytotoxic activities of the essential oil were investigated in vitro. In addition, antinociceptive and anti-inflammatory effects were evaluated in vivo. Most of N. curvidens essential oil compositions are composed of trans-caryophyllene (17.53%), allo-spathulenol (13.41%), germacrene D (10.12%), bicyclogermacrene (9.36%), α-pinene (9.12%), and β-phellandrene (6.87%). The highest antimicrobial and antifungal effects were observed against L. monocytogenes (MIC=156.25 μg/ml) and K. marxianus (MIC=625 µg/ml), respectively. Concentration-dependent effects were detected in the treatment of L. major, so that the number of macrophages infected with amastigote dropped significantly with higher concentrations. The essential oil showed the highest effect on the lung cancer cell line (A549) with the lowest IC50 (133.2μg/mL). The most resistant cell line to the essential oil was bladder cancer (C450) with an IC50 of 430.69 µg/mL. SI 19.49 index indicated high immunity of the essential oil on macrophage lines. Compared with morphine analgesic, the highest pain control effect of the essential oil was observed in primary and secondary phases at 100 mg/kg concentration in rats. At 100 mg/kg concentration, reduced swelling of the paw and ear was observed in mice compared to the dexamethasone anti-inflammatory drug. The essential oil presents remarkable therapeutic properties such as antifungal, antibacterial, anti-inflammatory properties that probably attributed to the presence of an important trans-caryophyllene compound. Outstandingly, the essential oil has low toxic effects on the normal line and high antitumor properties against cancer cells. Therefore, it is expected to be used in the preparation of less toxic drugs than chemical ones, although more studies are required.

Abstract: Staphylococcus aureus is a human pathogen causing nosocomial infections and increased hospitalization and mortality among human communities. Methicillin-resistant S. aureus strains are considered a severe threat in nosocomial infections and cause complications in the remedy process of bacterial infections. In this study, 137 samples were collected from different departments, staff, and patients in Ilam hospital. Eighty-eight samples of these strains were examined to test antibiotic resistance and diffusion. MIC (minimum inhibitory concentration) and PCR (polymerase chain reaction) were performed on the samples resistant to oxacillin. 36 (40.9%) strains were MRSA, and 52 (59.1%) isolates were MSSA. 44.4% of MRSA strains with IV SCCmec type. Fourteen different spa types were found using spa typing, of which the most abundant types were t037, t030, and t701, and three new types, including t15471, t15474, and t17470, were identified among the strains. The molecular analysis by MLST showed that the strains are classified into 11 different sequence types. Sequence type 239 and clonal complexes of 329 and 22 were dominant. ST239-spat037-SCCmec III was also identified as the most frequent clone of MRSA. The most identified clones were MRSA ST239-spa t037-SCCmec III. The results show the spa-type distribution between samples of patients, personnel, and surfaces, demonstrating MRSA circulation between patients and the environment. The results show the need to control environmental health

To evaluate the anti-bacterial activities of titanium dioxide (TiO) nanoparticles of Origanum (O.) vulgare and Hypericum (H.) perforatum extracts, carvacrol and hypericin against Staphylococcus (S.) aureus.In this study, TiOnanoparticles of O. vulgare and H. perforatum extracts, carvacrol and hypericin, were prepared and their antibacterial effects were evaluated against Staphylococcus (S.) aureus. In this study, scanning electron microscope, fourier transform infrared spectrometer, atomic force microscopy, dynamic light scattering and zeta potential were used to investigate the structure of synthesized drugs.Anti-bacterial activity of synthesized NPs was tested by minimum inhibitory concentration (MIC), minimum bactericidal concentration and disc diffusion method. MICs of TiO-NPs synthesized using O. vulgare, H. perforatum, carvacrol and hypericin and TiO were obtained 250, 62.5, 250, and 250, and 500 μg/mL, respectively. The MBCs for all of these were obtained 1000 μg/mL.Green-synthesized of TiO nanoparticles provides a promising approach to the use of O. vulgare and H. perforatum, carvacrol and hypericin as novel agents and safer antibacterial compounds, especially anti-S. aureus compounds.

Aims: The present study aimed to evaluate Iron uptake protein-related genes in clinical and environmental Klebsiella pneumoniae isolates. Background: Klebsiella pneumoniae as an opportunistic pathogen cause infections in immunocompromised patients. Iron uptake systems play an important role in the pathogenesis of Klebsiella pneumonia. Objectives: This study was designed to investigate the prevalence of iron uptake coding genes among isolates of Klebsiella pneumonia. Materials and Methods: A total of 300 isolates of Klebsiella pneumonia including 150 clinical isolates and 150 environmental isolates were selected. Finally, the frequency of iroN, iucD, kfuA,hmuR, and ybt [yHPI] genes were detected by PCR method. Results: The frequency of kfuA, iucD, iroN, yHPI in clinical isolates were 33.3%, 16.7%, 24.7%, 15.3%, respectively and these genes among environmental isolates were 20.7%, 6%, 49.3% and 0.7%, respectively. Among the clinical isolates, the most frequency genes were kfuA gene [50 isolates] and after that iroN [37 isolates], iucD [25 isolates] and yHPI [23 isolates], the genes with the most frequency among environmental isolates were iroN gene [74 isolates] and following that kfuA [31 isolates], iucD [9 isolates] and yHPI [1 isolate]. No hmuR positive samples among all clinical or environmental isolates were found. Conclusion: The result of this study showed that because of the high frequency of ferric iron system coding gene kfu among clinical isolates, this system might play an important role in the survival of bacteria against its host.

Objective: Staphylococcus aureus, a Gram-positive bacteria, is ranked second among the causes of hospital infections and is one of the three main causes of food poisoning. In recent times, the spread of antibiotic resistance in S. aureus has become very worrisome. Therefore, research for new effective drugs is important. The present study aims to investigate the phytochemical profiles and antibacterial effects of hydroalcoholic extracts of Origanum vulgare (Lamiaceae family) and Hypericum perforatum (Clusiaceae family) and their active compounds on S. aureus (ATCC 12600) in vitro. Methods: The identification of phytochemical compounds in both plants was performed by Highperformance liquid chromatography (HPLC), headspace-solid-phase microextraction (HS-SPME) and Fourier-transform infrared spectroscopy (FTIR). To investigate microbial susceptibility, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and disc diffusion method (DAD) were used. Finally, the results of the study were compared with methicillin. Results: Of the 42 combinations of O. vulgare, carvacrol (48%) and of the 38 combinations of H. perforatum, hypericin (46.2%) were the most abundant. The MIC, MBC and DAD of O. vulgare and H. perforatum, carvacrol, hypericin and methicillin were 625, 625, 312.5, 78.12 and 384 µg/mL, 10000, 10000, 2500, 2500 and 384 µg/mL, and 15.66 ± 4.49, 12.66 ± 0.47 and 22 ± 0.81 mm, respectively. Conclusion: Due to the significant effects of O. vulgare and H. perforatum and their active components against S. aureus, it is expected that in the future, hypericin, carvacrol and their derivatives can be used as effective antibacterial agents against S. aureus.

There is a direct correlation between asymptomatic oropharyngeal colonization by pathogenic bacteria and the prevalence of paediatric respiratory infections. Streptococcus pneumoniae and Haemophilus influenzae are common cause of serious bacterial infections such as meningitis in children. This study aimed to monitor healthy children to detect asymptomatic carriers of S. pneumoniae and H. influenzae. In the present cross-sectional study, real-time PCR assay was developed and evaluated in comparison with culture for direct detection of S. pneumoniae and H. influenzae in 123 oropharynx and nasal cavity specimens from healthy children in Ilam, Iran. In addition, virulence factor (ply and hpd) and iron uptake (tbpA and piuA) genes were evaluated by PCR. Our results demonstrated that among all isolates only 14 S. pneumoniae and eight H. influenzae were identified by phenotypic methods, whereas 37 and 21 S. pneumoniae and H. influenzae were identified, respectively, by real-time PCR assay. All S. pneumoniae and H. influenzae isolates possessed tbpA and piuA genes. Also, 81% (30/37) of S. pneumoniae isolates were positive for ply gene, which encoded pneumolysin, as well as 90% (19/21) of H. influenzae isolates were positive for hpd gene. Simultaneous colonization of S. pneumoniae and H. influenzae could indicate the importance of monitoring of healthy children to identification of carriers. Keywords: Detection, Haemophilus influenzae, iron uptake, real-time PCR assay, Streptococcus pneumoniae, virulence factors

The aim of present investigation was evolution of aminoglycoside resistance in CTX-M-15 producing klebsiella spp. Clinical isolates from general hospitals of Kurdistan Province, Iran. Ninety klebsiella spp. isolates were obtained from different clinical specimens. Antibiotic susceptibility pattern and detection of ESBL producing isolates performed by Kirby-Bauer disc diffusion method according to Clinical and Laboratory Standards Institute’s (CLSI) guidelines. CTX-M-15 gene screened by PCR amplification. Fisher tests was used to analyses by with STATA software program and p-values below 0.05 were considered as significant. Out of 90 clinical isolates the highest and lowest resistance related to cefotaxime and imipenem respectively. The results of detection of ESBL producing isolates showed that 62 isolates (84.4%) were ESBL-positive isolates. Thirty two (42.1%) out of 90 Klebsiella spp. isolates caring CTX-M-15 gene. CTX-M-15 was detected in 56.2% of gentamycin resistance, 34.4% of kanamycin resistance, and 56.2% of amikacin resistance. the present study showed that there was high frequency of CTX-M-15 gene in ESBL producing clinical isolates in Kurdistan Province, Iran. Moreover, there was no statistically significant relationship between CTX-M-15 production and resistance to aminoglycoside antibiotics in Klebsiella spp. isolates.

Aims & Objectives:The aim of this studyisto evaluate genetic relatedness, antibiotic resistance pattern, and virulence characteristics of different types ofS. aureusisolated from air, surfaces, staff, and patients in a Public hospital in Ilam.Methods & Materials:A total of 88 of 140 staphylococci identified asS. aureusby conventional and molecular methods were used in this study. Isolate samples were obtained from surfaces, staff, patients, and hospital indoor air. The sampling from staff and surfaces was done through using swab and air by standard pump. Antimicrobial susceptibility testing and presence different resistant and virulence determinants was assessed. Isolates were then typed by pulsed-field gel electrophoresis (PFGE) and SCCmectyping methods.Results:Out of 88isolates, 36 of them (40.9%) were MRSA. Among MRSA isolates, the range of resistance to antibiotic was 0% in vancomycin to 83.3% in gentamycin. The most prevalent resistant genes among gentamicin resistantS. aureuswereacc (6')/aph (2”)Iaandaph(3”)IIIa. The most common erythromycin resistant gene wasermC. Surprisingly, SCCmectypes I (30.5%), II (25%)were highly distributed. PFGE analysis showed 33 different pulsotypes.Conclusion:This study confirms that different isolates of MSSA and MRSA circulate in Ilam which differ in antimicrobial susceptibility, content of resistance, and virulence determinants.

The existence of infections caused by multidrug resistant (MDR)Acinetobacter baumanniiis a growing problem because of the difficulty to treat them. We examined the published literature and focused our analysis on the investigation of the synergism of colistin and rifampin against MDRA. baumanniiisolatesviasystematic review and meta-analysis. A systematic literature search was performed using the following 4 databases (PubMed, Scopus, EMBASE and ISI Web of Sciences). The related articles were evaluated during the period from December 2014 to January 2015. Information based on resistance and sensitivity to antibiotics, the minimum inhibitory concentration and the effects of two antibiotics on each other including synergism, antagonism, relative synergism and additive antagonism were extracted. A meta-analysis of 17 studies including 448 samples was brought into process and 2% (95% CI 0-4%) and 72% (95% CI 56-89%) resistance to colistin and rifampin were observed, respectively. 42% of all isolates showed MIC = 4 µg/ml (95% CI 14-69%) to rifampin and 30% MIC= 2 µg/ml to colistin (95% CI 3.8-78%). MIC50and MIC90for both rifampin and colistin were 2 µg/ml and 4 µg/ml, respectively. 63% of the strains demonstrated synergy (95% CI 37-90%), 7% were highlighted as relative synergism (95% CI 0.0- 13%), 3% showed an additive effect (95% CI -0.0-7%) and 14% were indifferent (95% CI 6-23%). The antagonistic effect was not observed in this combination. Synergy rates of time-kill assay in rifampin and colistin combinations were generally higher than those of check bored microdilution and E-test method. The results demonstrated that the combination therapy could be more useful when compared to monotherapy and that this strategy might reduce the resistance rate to rifampin in MDRA. baumanniiisolates.

Background: Most cancer studies focus on exploring non-invasive biomarkers for cancer detection. In the present study, we sought to investigate the expression level of microRNA-21 (miR-21), as a potential diagnostic marker, in serum and stool samples from 40 patients with colorectal cancer (CRC) and 40 healthy controls. Methods: Quantitative real-time RT-PCR was applied to determine the relative expression level of miR-21 in serum and stool. At the same time, the sensitivity and specificity of this marker was evaluated by receiver operating characteristic (ROC) curve analysis. Results: miR-21 expression levels of serum and stool were up-regulated 12.1 (P

Carbapenems are the most important therapeutic options that effect against serious infections caused by multidrug resistant Pseudomonas aeruginosa (MDR-PA) isolates. Carbapenems resistant isolates of P. aeruginosa are increasing worldwide. The aim of this study was to determine the carbapenem resistance mechanisms in clinical P. aeruginosa isolates from burn patients, in Tehran, Iran. A total of 53 non-duplicated isolates of carbapenem-resistant P. aeruginosa were collected from burn patients. The presence of carbapenemase genes were determined by PCR. AmpC overproducer isolates were detected by phenotypic method. The mutation and transcription level of oprD were determined by PCR-sequencing and quantitative Real-time PCR (RT-PCR), respectively. Twenty-seven (50.9%) isolates were positive for carbapenemase (blaVIM = 25 and blaIMP = 2) and showed high-level resistance to imipenem and meropenem. Twenty-eight isolates were AmpC overproducers. All isolates had a mutation in the oprD gene and down-regulation of oprD was found in 56.6% of MDR-PA isolates. Although the presence of carbapenemase is the common mechanism of resistant to carbapenem, but carbapenem resistance was found by oprD mutation-driven and the AmpC overproducing isolates in Tehran, Iran.

Aim: This study was designed to evaluate the synergistic activities of hydroalcoholic extracts of medicinal plants Origanum vulgare and Hypericum perforatum and their active components, carvacrol and hypericin against Staphylococcus aureus. Methods: The synergistic effects of the plants, as well as carvacrol and hypericin, were examined using a checkered method against S. aureus (ATCC 12600). Results: A fractional inhibitory concentration of 0.5 was obtained for combination of O. vulgare and H. perforatum and 0.49 for combination of the active ingredients carvacrol and hypericin, both of which indicated a synergistic effect. Conclusion: This preliminary evaluation demonstrated a synergistic property of O. vulgare and H. perforatum extracts in treating S. aureus infection. This study indicates that combination of the plants, as well as combination of carvacrol and hypericin, might be used as a new antibacterial strategy against S. aureus., Lay abstract We studied and evaluated the synergistic activities of hydroalcoholic extracts of oregano and St John's wort and their active components, carvacrol and hypericin, against Staphylococcus aureus. The results suggest that a combination of oregano and St John's wort extracts, as well as a combination of hypericin and carvacrol, have potential for use as natural and effective combinations against S. aureus infections.

The emergence of antibiotic-resistant Staphylococcus aureus in particular methicillin-resistant S. aureus (MRSA) is an important concern in burn medical centers either in Iran or worldwide. A total of 128 S. aureus isolates were collected from wound infection of burn patients during June 2013 to June 2014. Multiplex-polymerase chain reaction (MPCR) assay was performed for the characterization of the staphylococcal cassette chromosome mec (SCCmec). Genes encoding virulence factors and biofilm were targeted by PCR. Of 128 S. aureus isolates, 77 (60.1%) isolates were MRSA. Fifty four (70.1%) isolates were identified as SCCmec type IIIA. The most frequently detected toxin genes among MRSA isolates with SCCmec type IIIA were sea (64.1%) and hla (51.8%). The rate of coexistence of sea with hla and sea with hla and hlb was 37% and12.9%, respectively. The sec, eta, tst, pvl, hla and hlb genes were not detected in any of the MRSA isolates. The most prevalent genes encoding biofilm was eno, found in 61.1% of isolates, followed by fib and icaA found in 48.1% and 38.8% of the isolates, respectively. The rate of coexistence of fib + eno + icaA + icaD and fib + eno was 20.3% and 9.2%, respectively. The ebps gene was not detected in any of the isolates. In conclusion, our study indicated that the sea, hla, fib and icaA were most frequent genes encoding virulence factors among MRSA with SCCmec type IIIA isolated from burn wound infection. Moreover, the results of this study shows that the rate of coexistence of genes encoding different virulence factor were high.

This study investigated the molecular characterizations of 80 methicillin resistant Staphylococcus epidermidis (MRSE) collected during 2012-2013 in Tehran Children's Medical Center, Iran. About 90% of MRSE isolates were multi-drug resistant (MDR) and the highest resistance was observed to cotrimoxazole and they were quite sensitive to quinupristin-dalfopristin and linezolid. Though vanA gene was not detected, the majority of isolates showed intermediate resistance to vancomycin (MIC90 16 μg/ml). Resistance to mupirocin was observed in 18 isolates. Staphylococcal cassette chromosome mec (SCCmec) types V, III, IV and II were detected in 23.75%, 7.5%, 6.25% and 5% of isolates respectively, in some of which the additional parts of mec or ccr complexes were observed. In 57.5% MRSE isolates SCCmec types were not classified. 41.2% of MRSE isolates were carrying intercellular adhesion (ica) operon and 40% had strong or intermediate biofilm. The types of arginine catabolic mobile element (ACME) were limited to type I and II. Nine sequence types (STs) were seen in mupirocin resistant MRSE isolates. The common STs were ST2, ST5 and ST22 with 27.7% (5/18), 22.2% (4/18) and 16.6% (3/18) frequencies, respectively. ST23, ST54 and ST179 plus three novels STs 580, 581,588 were also observed. The majority of STs, 83.3% (15/18) belonged to clonal complex 2 (CC2). The spread of antibiotic resistance and virulence factors among MRSE species is an alarming sign in Children's Hospitals. The combination of these two issues leads to increase the chance of successfully establishing of common STs in hospital environments, and promotes the device-related infections and bacteremia.

Objectives: Out-hospital Cardiac arrest is considered as a global disease, which causes high rate of morbidity and mortality. Although, the return of spontaneous circulation occurs in 10 to 60 percent of cases in OHCA, with variety of treatment, the most patients faced with multiple organ failure and ultimately death. The investigations demonstrated that endogenous vasopressin levels in patients with successful resuscitation is more than died patients. Therefore, it seems the administration of vasopressin during cardiopulmonary resuscitation could be useful. The current study aimed to investigate the administration of vasopressin and epinephrine on neurological surviving of cardiac-respiratory arrest via evaluation of S100b serum factor. Methods: For this reasons, after collecting of sera from two vasopressin-epinephrine and epinephrine receiving patients, sera were subjected for ELISA to evaluate S100b. Results: The findings demonstrated that the great reductions of S100b in sera of patients that receiving vasopressinepinephrine in comparison with those patients that only got the epinephrine. Although no significant difference was observed between two groups, but survival rates after hospital discharge in group that receiving vasopressin-epinephrine was significantly higher than those patients that only got only epinephrine. Conclusion: Today, no advantages of vasopressin over epinephrine have been observed in clinical trials and more studies needed to improve the OHAC patient's surveillance. But, The combination vasopressin-epinephrine in the current study demonstrated that efficacy of this combination should be noted. © 2015 Bentham Science Publishers.

Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.

ObjectivesIn an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase.MethodsEscherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured.ResultsAmylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system.ConclusionThese results indicate that this expression system was appropriate for the production of thermostable α-amylase.

There is a close correlation between asymptomatic oropharyngeal colonization by bacterial pathogens and paediatric respiratory diseases. Evaluation of the frequency of asymptomatic carriers of Neisseria meningitidis and Moraxella catarrhalis in healthy children was the main aim of the current study. In this cross-sectional study, 123 oropharyngeal swabs were collected from children between 2 and 6 years old in kindergartens of Ilam, Iran. Moraxella catarrhalis and N. meningitidis were identified using phenotypic and genotypic assays. In addition, the occurrence of the virulence factors (ctrA and uspA1) and iron uptake (tbpA) genes was evaluated by PCR. Results showed that 21 M. catarrhalis isolates and 17 N. meningitidis isolates were identified by conventional microbiological and biochemical methods, but the RT-PCR assay detected that 18 and 8 isolates were positive for M. catarrhalis and N. meningitidis, respectively. The tbpA gene was positive in all N. meningitidis and M. catarrhalis isolates. Seven isolates were positive for the ctrA gene in N. meningitidis and seven isolates were positive for the uspA1 gene in M. catarrhalis. These pathogenic bacteria often occurred as asymptomatic carriage of N. meningitidis in children from large families with low economic status, which reflects the importance of the environment and socio-economic level of families in the distribution of these potentially pathogenic bacteria in the oropharynx of children. Monitoring for the carriage of potential pathogenic bacteria in the nasopharynx of healthy children is important as this can predispose to infectious diseases; common exposure to human respiratory bacterial pathogens is a further risk factor.

The high incidence of bacterial respiratory infections has led to a focus on evaluating the human respiratory microbiome. Studies based on culture-based and molecular methods have shown an increase in the bacterial community that includes the bacterial phyla Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria in the oropharynx of healthy individuals. Therefore, recognizing this microbial compound and subsequently identifying those carriers of specific pathogens can be of great help in predicting future infections and their control. In this prospective study, we sought to characterize the bacterial communities of the respiratory microbiome in healthy children aged between 3 and 6 years old by combining both cultural techniques and sequencing of the 16S rRNA gene. Seventy-seven oropharynx samples using Dacron swabs were collected from 77 healthy children in the kindergartens of Ilam, Iran. Bacterial identification was performed by phenotypic methods and in house developed PCR-based sequencing (the V1–V9 hypervariable region of the bacterial 16S ribosomal RNA gene). In total, 346 bacterial isolates were characterized based on phenotypic and sequencing-based molecular methods. The 3 most predominant phyla were Firmicutes (74%), Proteobacteria (22%), and Actinobacteria (4%). At the level of the genus, Staphylococci (coagulase-positive and coagulase-negative) and Streptococci were dominant. Also, the most commonly identified potentially pathogenic colonisers were S. aureus (75%), Enterobacteriaceae spp. (40.1%), and A. baumannii (15.6%). The present study identified 3 phyla and 9 family of bacteria in the oropharyngeal microbiome. Remarkably, the presence of potential pathogenic bacteria in the nasopharynx of healthy children can predispose them to infectious diseases, and also frequent exposure to human respiratory bacterial pathogens are further risk factors.

Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.

OBJECTIVES: Nowadays, due to the biofilm formation among of coagulase-negative staphylococci (CoNS), acquisition of antibiotic resistance and virulence genes could be noted. These organisms resembling other staphylococcus have mecA gene which encoded methicillin resistance but the diversity of it among CoNS is higher than other species. Based on increasing of antibiotic resistance in Staphylococcus haemolyticus, analyzing of phenotypic and genotypic biofilm formation, antibiotic resistance and genes involved in this process, SCCmec and ACME typing were placed as aims of current study. Methods. The number of 256 clinical CoNS isolates were collected which 49 isolates were identified as S. haemolyticus. For evaluation antibiotic resistance disk agar diffusion methods was applied, also biofilm assay carried out among methicillin resistance S. haemolyticus and SCCmec and ACME typing was performed. Results. The results of antibiotic susceptibility indicated that the highest resistance belongs to cotrimoxazole and erythromycin (86.6) and the frequency of blaA (96.5) and msrA (79.3) genes were much higher than others. Among MRSH isolates 58.6 have weak biofilm phenotype and 41.4 showed a moderate biofilm density. Also among of biofilm correlated genes, IS 256 (79.31) was the most frequent. The SCCmec typing of MRSH isolates indicated that the type V was dominant. Conclusions. Our finding indicated that correlation is between high rates of existence Is 256 gene and high prevalence of weak biofilm phenotype among MRSH isolates. Current study revealed that the multiple antibiotic resistances among clinical S. haemolyticus isolates are increasing and this is a warning to public health.

The aim of this study was to find different prevalence of genes involved in the biofilm formation process and to assess the phenotypic and genotypic markers of biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections. In this study, 215 S. aureus strains were collected from human and dairy cow's infections. The biofilm forming capacity of the strains was evaluated using a colorimetric microtiter plate assay. The genes encoding microbial surface components, recognizing adhesive matrix molecules (MSCRAMMs) (ebpS, eno, fib, fnbA, fnbB, cna and bap), and the intracellular adhesion (ica) genes (icaA, and icaD) were targeted by polymerase chain reaction (PCR)-based method. Approximately 70% of the isolates produced biofilm. Among these, 59.3% were producers of weakly adherent biofilms while 34.8% and 5.8% produced moderate and strong biofilms, respectively. The most prevalent gene was icaD found in 88.4% of the isolates, followed by icaA, fib and eno found in 87.9%, 75.8% and 75.3% of the isolates, respectively. The bap gene was not detected in any of the isolates. The prevalence of ebpS and fnbA genes among bovine isolates were significantly higher than those in human isolates, whilst the prevalence of cna gene was significantly higher in the human isolates. In this study, a high prevalence of biofilm production was found among S. aureus strains isolated from human and bovine infections. Most biofilm producing isolates were positive for MSCRAMM, icaA, and icaD genes.

Summary: The aim of this study was to analyze antimicrobial resistance patterns and their encoding genes and genotypic diversity of Acinetobacter baumannii isolated from burn patients in Tehran, Iran. The presence of extended-spectrum beta-lactamase- and blaOXA-encoding genes among 37 multidrug resistant (MDR) A. baumannii strains isolated from patients hospitalized in a teaching hospital in Tehran was evaluated. Susceptibility to 7 antibiotics was tested by disk agar diffusion and to polymyxin B and colistin was tested by E-test, according to CLSI guidelines. All isolates were then analyzed by PCR for the presence of blaIMP, blaVIM, blaSIM blaOXA-23, blaOXA-24, and blaOXA-58-like carbapenemase genes, and blaOXA-51-like, blaTEM, blaSHV, blaPER, blaVEB, and blaGIM genes. Genotyping of A. baumannii strains was performed by repetitive sequence-based (REP)-PCR and cluster analysis of REP-PCR profiles. A. baumannii isolates were assigned to international clones by multiplex PCR sequence group analysis. Twenty-five A. baumannii isolates were classified as MDR, and 12 were classified as extensively drug resistant. All isolates were susceptible to colistin and polymyxin B. Eighty-one percent of the isolates was resistant to imipenem or meropenem and harbored at least one or both of the blaOXA-23-like or blaOXA-24-like carbapenemase genes. Co-existence of different resistance genes was found among carbapenem-resistant isolates. Multiplex PCR sequence group analysis most commonly assigned A. baumannii isolates to international clones I (18/37; 48.6%) and II (18/37; 48.6%). An alarming increase in resistance to carbapenems and the spread of blaOXA-23-like and/or blaOXA-24-like carbapenemase genes was observed among A. baumannii strains belonging to clonal lineages I and II, isolated from burn patients in Tehran. Keywords: Acinetobacter baumannii, Lineage, Burn, OXA gene

Background : Extensive use of antibiotics and biocide in treatment of patients and cleaning of surfaces and medical equipment has led to the emergence of resistant microorganisms. The current research goals to determine the antiseptics Minimum Inhibitory Concentration value in Staphylococcus aureus (methicillin -resistant Staphylococcus aureus and methicillin sensitive Staphylococcus aureus) isolates from some major hospitals in Iran and to detect qacA/B, norA , smr and blaZ genes. Methods : Two hundred isolates of S. aureus including 100 MRSA and 100 MSSA clinical isolates were collected from 4 hospitals in the west of Iran during period 2012 to 2013. Detection of disinfectant resistant genes (qac A/B, smr and norA), antimicrobial resistance genes (mecA and blaZ) and SCCmec typing of MRSA isolates was performed by PCR. Results : MIC of chlorhexidine digluconate (CHX) in 70% of MRSA and 30% of MSSA strains was 8-16 µg/ml. High level of MIC of citrimide (>2 µg/ml) in MRSA and MSSA isolates was 20% and 5% ,respectively. MIC of benzalkonium chloride (BC) in 80% of MRSA and 83% of MSSA isolates was less than 2µg/ml; only 9% of MRSA had MIC higher than 2µg/ml. Frequency of antiseptic and antibiotic resistance genes norA, blaZ and qacA/B in MRSA isolates were 83%, 98% and 9%, respectively; while this value for MSSA isolates were 62%, 8% and 0%, respectively. The smr gene was not detected in both MRSA and MSSA isolates. In all biocides high MIC were observed in SCCmec type III and IVc. High frequency of qacA/B gene was found in SCCmec type III,Vc and IVb, which were 66.6% ,22% and 11.1% respectively. Conclusion : We found SCCmec types III, Vc was related to high MIC of biocide in MRSA isolates.