International Journal of Molecular and Clinical Microbiology
Volume:9 Issue: 2, Summer Autumn 2019

Silver nanoparticles (AgNPs) are one of the most important nanoparticles which have various biomedical applications. For example as antifungal, antibacterial, anti-cancer and anti-inflammatory agents. Skin infection caused by Trichophyton rubrum and some opportunistic fungi such as Candida albicans and Aspergillus fumigatus are sometimes difficult to be treat. Although silver nanoparticle has long been used as effective inorganic antifungal agent; the antifungal activity of nano-Ag in different size has not been investigated yet. Anti-cancer and antifungal effects of spherical silver nanoparticles (nano-Ag) were investigated in this study and we decided to determination toxicity of Nano-Ag in different diameters (10, 20 and 40 nm) on L929 mouse fibroblasts. TEM microscope has been used to evaluate nanoparticles size and morphology. Nano-Ag’s toxicity were evaluated by MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. This study showed that the toxicity of Nano-Ag in low concentration (16-32µg/ml) are sustainable and Silver nanoparticle effects are size dependent.

Acinetobacter resistant strains have caused medical problems throughout the world. The aim of this study was to evaluate the frequency and pattern of antibiotic resistance of Acinetobacter species isolated from traumatic patients in Shahid Rajaee hospital in Shiraz. In this study, 794 samples were isolated from patients in Shahid Rajaee Hospital. Identification of Acinetobacter was done by biochemical tests and PCR method. Multi-drug resistant (MDR) Acinetobacter were determined using gentamicin, piperacillin, meropenem, colistin, trimethoprim, ciprofloxacin, imipenem, ampicillin, and chloramphenicol antibiotics. In this study, 248 samples of Acinetobacter isolates were identified by molecular and biochemical methods from patients. All of which were isolates of MDR Acinetobacter. The highest percentage of Acinetobacter isolates was reported for upper respiratory tract samples and the lowest for urinary tract. The highest percentage of infection was related to Acinetobacter co-infection with one bacterium in patients aged 45 to 87 years old. As the number of male patients with accident trauma was more than women, the percentage is higher in men. The percentage of patients with Acinetobacter infection in ICUs was higher than in other sections. These results show the evidence of necessity to examine the transmission ways and the increasing incidence of hospital infections.

Ear infection is one of the most common types of infections that can caused by fungal and bacterial agents. The precise diagnosis of the disease and identification of microbial agents is very important to prescribe the right drug and cure patients as soon as possible. The present study was to find the consistency of clinical findings and laboratory tests among patients suspected of ear infections over a one-year period. In this cross-sectional study conducted in 2018 to 2019, 134 ear samples of patients referred to the ENT clinic of Ayatollah Rouhani hospital of Babol were collected. These samples were analyzed and microbial agents were identified by direct examination and culture. The results were compared with an initial diagnosis from a physician. Out of the 84 patients clinically diagnosed for fungal infection, 67 cases (79.8%) were laboratory-approved, while the rate of bacterial- infection was 33 cases (66%). Our findings showed that the initial diagnosis of from a physician along with laboratory tests is necessary for accurate diagnosis and treatment of ear infections.

Urinary tract infection is the most prevalent infection in the human societies. This study aims to identify G. vaginalis in the urinary samples of the patients suffered from UTI by culture and PCR techniques. 200 patients with urinary tract infection signs have entered in this study. urine sample was cultured in the blood agar medium, a small quantity of the sample used for the purpose of molecular technique. In order to analyze the data, SPSS software and chi square (x2) test has been used. Of 200 studied patients, G. vaginalis was isolated and identified in 6 (3%) and 29 (14.5%) patients with the culture and PCR techniques, respectively. The sequencing of the samples confirmed phenotyping identification. On the basis of the results obtained from both two techniques, it can be expressed that PCR technique is more appropriate to identify G. vaginalis. The achieved results show that G. vaginalis is regarded as one the agents causing the occurrence of urine infection. Yet, in many medical diagnosis laboratories, the bacterium is undetectable due to hardly growth. Also, considering the conducted techniques, PCR method is of a higher accuracy and speed.

L-Glutaminase is a therapeutic enzyme found in various microbial source have been considered in the cancer therapy. Sampling was carried out from the shores of the Persian Gulf. After identifying and performing specific biochemical tests, marine Streptomyces was isolated and DNA extraction was performed. Through the PCR test, the strains of streptomycin with the L-glutamine enzyme gene were identified. The L-gluta gene was positively transmitted to the host bacterium Escherichia coli via a vector and cloned through the TA technique, and the Real Time PCR technique was used to measure the expression of genes in E. coli origami. The software clustalX and Mega5 were used to draw the phylogenetic tree. Out of 12 Streptomyces isolates, 58.3% of isolates were carried L-gluta gene. After cloning the L-glutaminase gene by colony selection (blue / white), the cloned strains were isolated. The real-time PCR test showed a successful expression of the L-gluta gene on the cloned strains. Phylogenetic results with the neighbor joining (NJ) method show that, Streptomyces species with bootstrap values 99% located in a clade which indicated their close relatedness. The results of this study showed that the Persian Gulf is one of the high potential sources with the production of secondary metabolites and useful antimicrobial products that can be used as a useful source of various biological products such as L-glutaminase.

Fusarium oxysporum f.sp. lycopersici is an important disease agent of tomato which causes wilt and seedling. The present study was performed to evaluate the antifungal effect of Achillea millefolium, Salvia verticillata and Ziziphora clinopodioides extracts and their abilities to inhibit the fungus. For this, methanol extracts of reference plants was extracted and tested in concentrations ranging from 1 , 1.5 and 2 mg/ml on mycelial growth of Fusarium oxysporum. The same extracts were then tested for antifungal activity in vivo in the greenhouse on inoculated tomato plants. Z. clinopodioides demonstrated highest antifungal activity against mycelial growth of F. oxysporum strain that recorded 77.1%, 62.03% and 61.99% at 2, 1.5 and 1 (mg/ml), respectively. the MIC value for of Z. clinopodioides against F. oxysporum was 3.125 mg/ml followed A. millefolium and S. verticillata extract having 6.25 mg/ml. The MFC of extracts was found to be 6.25 mg/ml in Z. clinopodioides and 12.5 mg/ml for A. millefolium and S. verticillate. In greenhouse experiment employing methanol extracts of three plant species showed an increase in the mean plant height and also fresh and dry weight of root and shoot with the consequent reduction in the disease symptoms of the tomato seedlings. Overall, the results showed significant growth inhibition activity of Z. clinopodioides methanol extract against F. oxysporum in both in vitro and greenhouse condition. Although the extracts of A. millefolium and S. verticillata which had no effect in vitro assays, in greenhouse conditions, these plants showed considerable antifungal activity.

In the last decade Enterococcus spp. has become one of the most important nosocomial pathogens. The prevalence of multi-resistant strains of Enterococcus faecium and Enterococcus faecalis responsible for hospital-acquired infections is associated with their ability to acquire and share antimicrobial resistance genes contained in Mobile Genetic Elements (MGE). This study investigated the distribution of antibiotic resistance in Enterococcus spp. isolated from clinical patients in the University Hospital "Luigi Vanvitelli" of Naples, Italy. The aim of the present study was to monitor the antimicrobial drug resistance and spread of nosocomial infection, to allow the optimal choice of antibiotic therapy. From January 2017 to December 2018, 351 Enterococcus spp. isolates were collected from different clinical samples, at the University Hospital “Luigi Vanvitelli”. Bacteria identification was made using MALDI-TOF technology (Bruker Daltonics, Bremen, Germany). Susceptibility to 9 antibiotics was tested using BD Phoenix (Becton, Dickinson and Company). Data were analyzed using the statistical software SPSS v.22.0 (IBM SPSS Inc., New York, USA). Among the 351 collected samples, 88 (25.1%) were identified as Enterococcus faecium and 263 (74.9%) were as Enterococcus faecalis. Enterococcus faecalis showed the highest resistance rate to Tetracycline (73,5%) and Erythromycin (88,6%) than the Enterococcus faecium. The Enterococcus faecium has showed increase in resistance rates against Ciprofloxacin and Imipenem. Persistent surveillance of antimicrobial patterns was essential to adopt the empirical treatment guideline to treat infection caused by Enterococcus spp.

Acinetobacter baumannii has emerged as an important hospital pathogen worldwide especially in the burn ward. The aim of this study is to determine the pattern of antibiotic susceptibility and genotyping of A. baumannii isolated from clinical and environmental samples of Shahid Motahari hospital in Tehran using MLVA method. In this study, 173 clinical and 28 environmental isolates of A. baumannii were collected from Shahid Motahari hospital within a 9-month period (2018-2019). The isolates were confirmed by biochemical and molecular tests with OXA-51 primer. Antibiotic sensitivity was performed by the disc diffusion method according CLSI M100-S21 guidelines. MLVA-PCR was performed with six STR markers including Abaum-3530, Abaum-3002, Abaum-2240, Abaum-1988, Abaum-826, and Abaum-2396. Out of 201 tested strains for antibiotic sensitivity, 127 (63.2%) and 35 (17.5%) of isolates of the strains were multidrug-resistant (MDR) and extensively drug-resistant (XRD). Microsatellite typing of 201 A. baumannii isolates showed 197 genotypes in four clusters. The Hunter-Gaston diversity index (HGDI) of six markers (STRs) for all isolates was 0.9169. The progressive increase in A. baumannii infections and antibiotic resistance in hospitals demands some measures for rapid description of the typing of isolates and identification of sources of infection. Our results indicated that MLVA a method based on PCR was more effective for typing of clinical and environmental strains of A. baumannii. These findings highlight the importance of international resistance against different antibiotics as well as molecular epidemiological control of A. baumannii isolates with XDR and MDR characteristics.

Activated Sludge (AS) processes are the most widely used biological processes in Wastewater Treatment Plants (WWTPs). Controlling wastewater is important for reducing overall water pollution.Therefore the main objective of this study is to evaluate the performance of the Activated sludge system in removal microbial indicators from the wastewater sludge .This research was conducted on for 4 months in two seasons .The sludge Samples were taken from the return Activated sludge line. Microbial indicators including Fecal Coliform (FC), Total Coliform (TC), parasitic egg and some parameters such as pH, temperature examined for the comparison between aerobic digester and lime stabilization sludges. The lime stabilization is capable of decreasing large quantities of pathogenic bacteria. The results indicated that Fecal Coliform and parasitic egg decreased to 976MPN/gr.ds and 0MPN/4gr.ds respectively. When limed sludge reached the pH of 12.5 .The remaining parasitic egg from lime stabilization of the sludge were 203 MPN/4gr.ds after two months. The average removal efficiency of TSS, TC, FC and parasitic egg were 80.8%, 99% and 100% and 74% respectively. The optimum ratio of lime was identified 245(gr/kg.Ts).The lime stabilization could archived to class B of USEPA category. So at a pH higher than 12 treated sludge with lime meet the guideline for pathogen reduction of class B regarding parasitic egg and has a higher hygienic effect on sludge pathogens and more cost effective than aerobic stabilization when sludge used to inferiority soil improvement , modify acid soils, disposal in the forest but was not used as agricultural fertilizer.

Genital Mycoplasma infections can have a negative effect on men's reproductive health and lead to infertility. The aim of this study was to investigate the presence of Mycoplasma huminis and Ureaplasma urealyticum in semen samples of infertile men using culture and molecular techniques and comparison the sensitivity and specifity of these methods. This descriptive-analytic and cross-sectional study carried out on infertile men referred to the Royan Institute of Infertility Treatment Center. The Rapid Test was done using the Mycofast Rapid kit (ELItech, France). DNA extraction from semen was achieved, and then PCR test was performed by the IFG kit (Iranian technology gene). PCR products were transferred to agarose gel 1.5% and electrophoresed. The bands created were transferred to the photographic device and the result was observed. In this study, 147 samples of leukocytospermia were detected in M.hominis and U.urealyticum .Thirty seven samples (25.2%) were positive for U.urealyticum positive in the culture method, and positive samples were positive for M. huminisis Of 147 samples, 28 samples (19 Percent) was positive for U.urealyticum and 13 samples (8.8%) were positive for M.hominis. Considering that Iranian kits were used for PCR in the present study and there was no significant difference in M.hominis, it is believed that the Iranian PCR kit has a problem or to prove this French method and replace this method with rapid molecular culture with the method Hard and costly PCRs need more examples to prove this technique.