Green Synthesis of Silver Nanoparticles by Fusarium oxysporum and its Function Against Aspergillus and Fusarium Fungi.

(NPs) can be produced by various methods such as physical and chemical processes. However, environmentally friendly ways are increasingly requested. In this research, (Ag-NPs) were produced by Fusarium oxysporum, and its antifungal effect on Aspergillus and Fusarium was investigated. Nanoparticles were produced by silver nitrate salt and Fusarium oxysporum native to Isfahan city. In order to optimize the synthesis conditions, optimization of some factors such as volume, concentration, time, temperature, and pH of the extract was performed. The structural and physical properties of NPs were determined by spectrophotometer, XRD, FTIR FESEM, SEM, and TEM microscopy. For the study of the inhibitory effect of NPs on Fusarium and Aspergillus growth, the fungi were cultured in media containing various concentrations of NPs from 50 to 1500 ppm. Then, the colony diameter was measured for over 10 days and the growth inhibition percentage was estimated. For statistical analysis, the 600 Mann-Whitney tests have been applied.The NPs were produced after mixing the powdered fungal mass and silver nitrate salt in optimum conditions which were 2 mM of salt, triple fungal mass volume proportion relative to the salt, pH of 9, and temperature of 28 °C. The existence of a peak at 420 nm in FTIR was due to nanoparticle production. Based on the XRD, the synthesized NPs had suitable properties similar to the standard NPs reported in the studies. Images from TEM, SEM, and FESEM microscopes displayed uniform NPs in variable sizes between 25 and 100 nm. According to the results, the maximum growth inhibition percentage of Ag-NPs on Fusarium was approximately 60% at 1500 ppm, and 88% on Aspergillus at 800 ppm. Biosynthesized Ag-NPs with Fusarium oxysporum have desirable structural traits and can inhibit the growth of Fusarium and Aspergillus at significant levels.

Comparative assessment of the mouse immune responses to colistin-resistant and colistin-sensitive isolates of Acinetobacter baumannii.

Acinetobacter baumannii is known as the most frequent species in clinical samples that is responsible for a large number of nosocomial infection outbreaks. The colistin-resistance of this bacterium has been found to be increasing. This study aimed to evaluate the immune response to colistin-susceptible and colistin-resistant A. baumannii isolates in a mouse model. Samples were prepared from the wounds of patients suspected of A. baumannii admitted to the intensive care unit of Namazi Hospital in Shiraz. Antibiotic susceptibility was studied by disk diffusion and broth microdilution according to CLSI and EUCAST criteria. Among the isolates, one colistin-sensitive isolate and one colistin-resistant isolate were injected intraperitoneally to BALB/C mice. Blood samples were collected after 4 h and cytokines (IL1-ß, IL-12, IFN γ, and IL-10) and surface markers (CD4, CD8, CD3, and CD45) were determined by ELISA and flow cytometry. Then, hematologic and histopathological factors were analyzed, and colony count in lung, liver, kidney, and spleen tissues were also performed. The results showed that levels of cytokines (IL1-ß, IL-12, IFN γ, and IL-10) and markers (CD4, CD8, CD3 and CD45) were higher in mice receiving colistin-resistant A. baumannii isolates than in mice receiving colistin-sensitive A. baumannii isolates, indicating that colistin-resistant isolates 4 h following the intraperitoneal injection stimulated host innate immune system better and produced a stronger immune response. On the other hand, histopathological findings showed inflammatory cell infiltration, hyperemia, and tissue damage. In addition, the bacterial load and tissue damage in the lung was higher than other tissues. The results of this study can have promising potential for the development of a prevention and treatment strategy based on cellular immune response for infection caused by colistin-sensitive and colistin-resistant A. baumannii.

Evaluation of genetic diversity of colistin-resistant Acinetobacter baumannii by BOX-PCR and ERIC-PCR: the first report.

Aim: To control the spread of Acinetobacter baumannii in hospitals, it is necessary to identify the reservoir of organisms and the way they are transmitted. This study analyzed samples by BOX-PCR and enterobacterial repetitive intergenic consensus PCR techniques. Methods: Isolated strains were identified using the Microgen kit and blaOXA-51 gene. The genetic diversity of strains that were sensitive or resistant to colistin was evaluated by BOX-PCR and enterobacterial repetitive intergenic consensus PCR methods. Results: A total of 13% of the isolates were resistant to colistin, whereas 87% of the strains were sensitive to this medication. A. baumannii strains that were resistant or sensitive to colistin were divided into five groups using the BOX-PCR method and six groups using the enterobacterial repetitive intergenic consensus PCR method. Conclusion: Rapid identification and the use of appropriate tools to control colistin-resistant clones are essential to prevent the further spread of A. baumannii.

Evaluation antibiotic resistance and presence of blaOXA-51, blaOXA-58 and blaOXA-23 genes in Acinetobacter baumannii strains via multiplex PCR.

The resistance of Acinetobacter baumannii to most antibiotics is increasing. The presence of metallo-beta-lactamase and carbapenemase enzymes has led to the resistance of these bacteria to carbapenems as one of the major classes of broad-spectrum antibiotics and has raised concerns in human societies. This research evaluated the presence of blaOXA-51, blaOXA-58 and blaOXA-23 genes in A. baumannii strains during a 12 months period. One hundred strains were isolated from the patients hospitalized in ICU of Ali Asghar and Shahid Rajaee trauma hospitals in Shiraz. Bacterial identity was determined by biochemical tests and antibiotic resistance was determined by disk diffusion method. The isolated strains were then evaluated in terms of carrying blaOXA-23, blaOXA-51 and blaOXA-58 genes, using the multiplex PCR method. The results showed that A. baumannii was resistant to carbapenems but most strains were susceptible to tigecyclin and colistin. The majority of strains carried the blaOXA-23 and blaOXA-51 genes, but very few carried the bla OXA-58 gene. The results revealed that the antibiotic resistance of A. baumannii is increasing, which causes a more outbreak of this organism.

Molecular analysis of PmrA and PmrB genes in colistin-resistant Pseudomonas aeruginosa strains via PCR method.

Pseudomonas aeruginosa is one of the most common pathogens in hospitals. Along with the advent of various drug resistance patterns, rising resistance to colistin, the last alternative against this bacterium, is reported as a major clinical concern all over the world. Initially, Pseudomonas aeruginosa strains were identified by diagnostic tests including phenotypic method, growth at 42°C, Gram staining, culture on Blood Agar, EMB Agar, and biochemical oxidase, and catalase tests. The strains were confirmed using Microgen kit. Then, the resistance pattern of the identified strains was evaluated by Antibiogram. The presence of PmrA and PmrB genes were investigated by PCR method.A total of 60 strains of Pseudomonas aeruginosa were isolated and identified using microscopic, macroscopic and microbiological methods. The lowest resistance was observed against chloramphenicol and colistin antibiotics. Most of the strains harbored the PmrA and PmrB genes. The results of this study indicated an increasing trend in the resistance of the bacterium against different antibiotics. Accordingly, it is necessary to establish an infection control and therapeutic strategy in preventing the spread of such as similar resistant organisms.

Molecular evaluation of colistin-resistant gene expression changes in Acinetobacter baumannii with real-time polymerase chain reaction.

BACKGROUND: Acinetobacter baumannii is an important human pathogen which has recently gained increased attention due to the occurrence of drug-resistant nosocomial infections in patients suffering from immune system disorders, and those in hospital intensive care units. The aim of this research was to identify and isolate A. baumannii strains resistant to colistin, determine antibiotic resistance pattern of this bacteria, investigate the presence of colistin-resistant genes, and finally assess the effect of expression changes in pmrA and pmrB genes resistant to A. baumannii against colistin via real-time polymerase chain reaction. METHODS: The samples were initially purified and isolated using biochemical tests and Micro-gen kit. Later, the resistance pattern evaluation of validated samples to different antibiotics and colistin was carried out using two methods viz., disc diffusion and E-test. This was followed by the assessment of genes resistant to colistin via polymerase chain reaction besides gene expression changes via real-time polymerase chain reaction. RESULTS: The results of this study indicated that eleven strains of A. baumannii isolated from Shahid Rajaee Trauma Hospital were resistant to colistin. However, in the resistance pattern evaluation of A. baumannii isolated from Ali Asghar Hospital, all the strains were sensitive to colistin. In the evaluation of genes resistant to pmrA and pmrB, most of the strains resistant to colistin were carriers of these genes. Besides, in the expression assessment of these genes, it was demonstrated that expression of pmrA in the strains resistant to colistin significantly increased in relation to sensitive strains, but the expression of pmrB increased at a lower rate in the strains resistant to colistin as compared to the sensitive strains. CONCLUSION: Thus, it can be safely mentioned that increased expression of pmrA was due to the resistance of A. baumannii to colistin.

Analyzing pmrA and pmrB genes in Acinetobacter baumannii resistant to colistin in Shahid Rajai Shiraz, Iran Hospital by PCR: First report in Iran.

Acinetobacter baumanni is known as a worldwide emerging nosocomial infections and it is classified as one of the six dangerous microorganisms by Diseases Society of America. Multi drug-resistant strains of A. baumannii have been reported in recent decades, which may be a result of the high use of antimicrobial agents. Colistin is the last form of treatment against this organism. The presence of pmrA and pmrB genes in A. baumannii causes the resistance of this organism against Colistin. This cross-sectional study was performed on 100 samples of A. baumannii isolated from ulcer, urinary, respiratory, blood of patients admitted to the intensive care unit of Shahid Rajai Shiraz hospital within a 12-month period. The diagnosis was performed by microscopic and biochemical testing using microgen kits. Determining Colistin resistance was carried out by Diffusion Disc, Colistin antibiotic disc of MAST- England and E-test. The analysis of genes pmrA and pmrB genes was done by PCR. 100 A. baumannii samples were diagnosed out of which using diffusion disk 94 cases were sensitive to Colistin and 6 cases were resistant to it. The E-test results in resistant samples presented an MIC equal to 64 micrograms per milliliter. The PCR results in sensitive and resistant to Colistin samples presented the existence of pmrA and pmrB genes. The results indicated the presence of pmrA and pmrB genes that are the main reason of A. baumannii resistance against the last line of treatment of this organism to Colistin.