Accessory gene regulator types of Staphylococcus aureus isolated in Gorgan, North of Iran

Background: Staphylococcus aureus is a gram-positive bacterium that has remained a persistent pathogen, causing infections such as endocarditis, meningitis, and toxic shock syndrome in humans. The accessory gene regulator (agr) system of Staphylococcus aureus is responsible for controlling the expression of many genes that code for virulence factors. In this study, we assessed the S.aureus agr Group, based on their source of isolation, in Gorgan, North of Iran. Materials and Methods: DNA of 194 S. aureus isolates was extracted by lysozyme-phenol chloroform method, which included 85 clinical samples, 58 samples which were isolated from noses of health care workers and 51 cases which were obtained from food products in Gorgan, northern Iran. PCR-based assays were used to evaluate agr locus nucleotide polymorphism for the identification of agr specificity Group. Distributions of each agr Group were determined and comparison between different sources was assessed by X2. A p-value of <0.05 was considered as significant. Results: The majority of isolates belonged to agr Group I (43.3%), followed by agr Group III (28.87%), agr Group II (22.68%), and agr Group IV (5.15%). In our study, a majority of S. aureus isolates were recovered from health care workers and food product specimens were of agr Group I and isolates which were recovered from patients were of agr Group III. These differences were statistically significant (P=0.005). There was no statistical difference between the source of isolation of clinical samples of S.aureus and agr type. Conclusion: Agr Group I was predominant among health care workers and food product specimens in Gorgan, North of Iran, but in strains which were isolated from patients, agr Group III was predominant. Investigating the possible role of agr Group III in Staphylococcus aureus infection in future studies is recommended

Staphylococcus aureus typing by digestion of protein A coding gene using Bsp143I

Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with different enzymes has been used for this action. Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene) in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran. Patients and Methods: DNAs of 128 S. aureus subjects (53 from healthy carriers and 75 from patients) were extracted and amplified using specific primers of the spa gene. The product was digested by Bsp143I enzyme and its pattern was assessed by gel electrophoresis. Results: There were seven spa types among the tested S. aureus samples, among which six types differed in the repeated X region of the spa gene, but the seventh type had a deletion on one of BSP143I restriction sites. The frequency of spa types among isolated S. aureus samples as well as healthy carriers was six and five, respectively. S. aureus isolated from wounds showed the most diverse spa types (five) among clinical samples. Types 1, 2 and 4 were observed in all clinical samples, while only one case of type 3 was identified among patients, whereas this type constituted over 32% of the isolates among carriers. We found seven and four spa types among methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates, respectively. Conclusions: Our results showed that typing the spa gene using PCR-RFLP using Bsp143I was an acceptable method for typing S. aureus. Furthermore, this survey showed that the types in healthy carriers and MSSA were more variable than patient and MRSA isolates, respectively. We used the Bsp143I enzyme, which was not used in any previous studies on the spa gene. The results of this study suggested that we can use PCR-RFLP of spa gene by Bsp143I for molecular typing and sequencing of S. aureus, instead of relatively expensive methods. This method is relatively rapid and inexpensive, and can be accomplished in centers with conventional molecular facilities. © 2014, Ahvaz Jundishapur University of Medical Sciences

The Frequency of MRSA carriers in health care workers in Gorgan, North of Iran

Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important pathogen in hospitals. Healthcare personnel are the main source of nosocomial infections and identification and control of MRSA carriers can reduce incidence of infections. The aim of this study was to determine the frequency of methicillin resistant Staphylococcus aureus (MRSA) and their antibiotic susceptibility profile among healthcare workers in Gorgan located in northern Iran. Three hundred and thirty three of healthcare workers were participated in this cross-sectional study in 2010. Samples were taken with sterile cotton swabs from both anterior nares. Swabs were plated onto Mannitol salt agar. S. aureus were identified by Gram stain, Catalase, Coagulase and DNase tests. MIC (micro dilution broth) method was used to determine resistance of strains to methicillin. Antimicrobial susceptibility pattern to other antibiotics was performed by diffusion method. Frequency of S. aureus and MRSA carriers among healthcare workers was 24% (80.33) and 3% (10.33) respectively. MIC of isolates was varied between 0.5 and 65.31 (39%) of cases were showed MIC of intermediate that ranged between 4 and 8. Penicillin and Imipenem resistance were seen in 97.5% and 1.4% of isolated S. aureus strains, respectively. Frequency of S. aureus carriers in healthcare workers in our area was median in compare with other region in Iran but the MRSA carriage in healthy staff was lower than most part of Iran. It would be considering to monitor healthy carrier staff because of high rate intermediate MIC in this group to prevent conversion to MRSA

Antimicrobial effects of folk medicinal plants from the North of Iran against Mycobacterium tuberculosis

Background: Medicinal plants have been used traditionally in Golestan province (north of Iran), against Mycobacterium tuberculosis or the clinical signs of tuberculosis (TB). Objectives: This study aimed to define the inhibitory effects of ethanolic extracts of six of these medicinal plants against Mycobacterium tuberculosis. Materials and Methods: Peganum harmala (seed extract), Punica granatum (peel extract), Digitalis sp. (leaf extract), fruit extract of Citrus lemon, Rosa canina and Berberis vulgaris were extracted in ethanol and their activity against M. tuberculosis isolates were determined by the agar diffusion method. The zone of inhibition (at 200 to 1.6 mg/mL) was measured and the results were compared with isoniazid and rifampin as standard positive controls. Also the concentration of vitamin C of each the extracts was evaluated. Results: The ethanolic extract of Peganum harmala seed and Punica granatum peel exhibited potential activity against all M. tuberculosis isolates with mean inhibitory zone of 18.7 and 18.8 mm, at 200 mg/mL concentration. The mean inhibitory zone around isoniazid and rifampinwere 19.2 and 18.8 mm. Ethanolic extract of Citrus lemon showed moderate inhibitory activity only against sensitive (non MDR; non multi drug resistant) strains of M. tuberculosis, and Digitalis sp. showed inhibitory effects on five isolates. Ascorbic acid content was 43.3 mg/dL in Punica granatum and Digitalis sp. and only 9.1 mg/dL in ethanolic extract of Peganum harmala. Conclusions: The highest content of vitamin C was observed in the extract of Punica granatum, which was observed to be highly active against Mycobacterium tuberculosis, while the P. harmala must have contained other phytochemical constituents that contributed to the anti-tuberculosis effects of this plant. Our findings showed that ethanolic extracts of P. granatum and P. harmala had anti-TB effects comparable to isoniazid and rifampin and can be good candidates for novel and safe natural products against tuberculosis. © 2015, Pediatric Infections Research Center

Comparison of ferment sugars, produce hemolysis and measuring growth in methicillin-resistant and methicillin-sensitive Staphylococcus aureus isolates from inpatients and healthcare workers in Gorgan Hospitals, North of Iran

The mec A gene in Staphylococcus aureus leads to production of new penicillin-binding protein called PBP2a.This change may follow some changes in other phenotypes. The aim of this study was the comparison of Ferment Sugars, Produce Hemolysis and Measuring Growth in MRSA and MSSA isolates. 188 Staphylococcus aureus isolates separated from inpatients and healthcare workers (healthy carriers)were studied.Bacterialcultures in blood agar environment at 37°C during 24h and at 4°C during other 24h were applied for studying hemolysis. Sugar fermentation carried out in phenol red Broth medium, containing glucose, galactose, arabinose, fructose, xylose, ramnose, mannose, sucrose, trehalose, raffinose or maltose. For determining bacterial growth,bacterial concentration of 103was taken each hour during 12 cultured in MHAand colonies were counted after 24h.The mean amount of hemolysis diameter in MRSA isolates was rather more than that of MSSA isolates. The difference between MRSA and MSSA isolates were significant as to fermenting ramnose, trehalose, galactose and xylose. The mean rate of growth in MRSAwere significantly different from that of MSSAisolates (p&lt;0.05).Resistance to methicillin in Staphylococcus aureus isolates accompanies the increase of ability to ferment sugars. This phenomenon may be one of reasons for increased pathogenicity of MRSA isolates; So results shows the logarithmic phase is longer in MRSA isolates, This may implicate that PBP2a production in methicillin-resistant isolates follows slowing down nutrients entrance into the bacterium that in turn may causes slow growth

Prevalence and molecular characterization of Listeria spp. and Listeria monocytogenes isolated from fish, shrimp, and cooked ready-to-eat (RTE) aquatic products in Iran

The prevalence of Listeria spp. and Listeria monocytogenes was investigated by biochemical and molecular methods in a total of 201 fish, shrimp, and ready-to-eat seafood samples collected from Iranian supermarkets. Thirty-six samples were also collected from a seafood processing plant. Twenty-one (8.86) of the total retail and processing plant samples (237) were positive for Listeria spp., confirmed by a simplex PCR assay for the prs gene. Seven (2.95) of the total samples were also positive for L. monocytogenes. The presence of four virulence-associated genes in the seafood isolates (inlA, inlC, inlJ, and hlyA) was examined using PCR and the results were compared with seven clinical L. monocytogenes strains. All virulence genes were detected in six fish isolates. One fish isolate did not show amplification of the inlJ and inlC genes. However, all seven clinical strains were positive for internalin genes. Furthermore, a multiplex PCR assay was employed to evaluate the major L. monocytogenes genoserogroups' distribution. The results revealed that the serotypes of lineage II are most frequently present in clinical and food isolates. In summary, PCR screening for both the major L. monocytogenes serovars and virulence genes revealed the potential public health risk posed by L. monocytogenes in aquatic products. © 2016 Elsevier Ltd

Negative performance of root extract of Onosma dichroanthum Boiss. on the burn wound healing in an animal model

Background: Onosma dichroanthum Boiss. belongs to the Boraginaceae family which locally known as" Hava Chobeh", and it is considered as one of the most important Mountainous medicinal plants in North of Iran, Its red roots have been used by the rural healers as either singular or combination with other plants as antiseptic and anti-inflammatory to treat the burns and wound healing. Methods and Findings: In this study, the acetonic extract of plant root was used for the evaluation of its healing efficiency on burn wound model in rat. Burn induced in Wistar rats, and subsequently they were divided into five groups were treated as follow: Group-I ointment containing of extract 2%, Group-II, traditional ointment containing of goat lipid and root extract, Groups-III ointment alone, Group-IV (positive control) the standard drug (silver sulphadiazine) and Group-V (negative control) untreated rats. The assessment the burn area healing was carried out within 14 days. Our results showed that the treated group in I and II, the intensity of wound increased up to the day 8, even the wound diameter much more expanded than negative control group (group V). The best result was demonstrated by Silver sulphadiazine treated group (group IV). Conclusion: This study shows that Onosma dichroanthum Boiss. root did not have any effect on healing of burn wounds in an animal model. Further studies are requiring for assessment of this plant in combination with other plants on burn wound healing. © Under License of Creative Commons Attribution 3.0 License

Maternal serum copper concentration in premature rupture of membrane: A case-control study

Copper is an integrated parts of metal-protein required far varieties of oxide-reductive metabolic pathways in human. Copper deficiency is considered as risk factors in some pregnancies. Premature rupture of membrane is a pregnancy complication with major adverse effects and is believed maternal Copper deficiency can also be considered as interventional factors. This study was done to evaluate if there is a correlation between maternal serum Copper concentration and premature rupture of membrane in pregnancy. In this case-control study 60 pregnant women with Premature Rupture of Membrane (PROM) were selected as case group including term and pre term the control group consist of 60 pregnant women with normal delivery of term and pre term states. Both group were matched for maternal and pregnancy age. In case and control group the pregnancy at term and pre-term were grouped independently as well. In general the maternal mean serum Copper concentration were 192.4±78.2 and 201.08±82.06 in case and control groups, respectively but this differences statistically was not significant. Data in this study revealed that the absolute value of maternal serum Copper concentration of term or pre term in case groups was slightly lower than related controls. Drop in maternal Copper concentration in some disturbed pregnancies such as premature rupture of membrane is previously demonstrated and based on our data the absolute Copper serum concentration of women with premature rupture of membrane was also slightly lower compared to healthy pregnancy but it was not statistically significant. © Asian Network for Scientific Information

Metallo�β�lactamase�mediated resistance among clinical carbapenem�resistant Pseudomonas aeruginosa isolates in northern Iran: A potential threat to clinical therapeutics

Objective: Carbapenems are effective agents to treat multidrug�resistant (MDR) strains of bacteria, including Pseudomonas aeruginosa. However, there is a potential threat of emergence of carbapenem�resistant P. aeruginosa (CRPA). The aim of this study was to determine antibiotic susceptibility patterns and metallo�beta�lactamase (MBL)�mediated resistance in clinical P. aeruginosa isolates. Materials and Methods: Different clinical specimens were subjected to conventional culture�based identification of P. aeruginosa. Antimicrobial susceptibility patterns and MBL production were evaluated using the Kirby�Bauer and combined double�disk synergy test methods, respectively. Multiplex polymerase chain reaction was performed to investigate the presence of the blaIMP, blaVIM, blaNDM, blaSPM, and blaSIM genes. Results: A total of 71 clinical P. aeruginosa isolates were recovered, of which 28.17 were identified as CRPA. The most active antibiotics were colistin and polymyxin B (92.96 susceptibility to each). A total of 35 and 50 of CRPA isolates were MDR and extensively drug�resistant (XDR), respectively. MBL activity was shown in 20 of CRPA. A total of 90, 40, and 5 of CRPA isolates harbored the blaIMP, blaVIM, and blaNDM genes, respectively. No correlation was found between the MBL�encoding genes of P. aeruginosa and patient characteristics. Conclusion: Although the prevalence of CRPA in our therapeutic centers was relatively low, this rate of carbapenem resistance reflects a threat limiting treatment choices. A high prevalence of MDR/XDR phenotypes among the MBL�producer isolates suggests the need for continuous assessment of antimicrobial susceptibility and surveillance of antibiotic prescription. In addition, infection control measures are needed to prevent further dissemination of these organisms. © 2018 Tzu Chi Medical Journal | Published by Wolters Kluwer - Medknow