Polydopamine contained hydrogel nanocomposites with combined antimicrobial and antioxidant properties for accelerated wound healing.

Overproduction of reactive oxygen species (ROS) in infected wounds induces a tremendous inflammatory reaction to delay wound healing. To address this problem, we designed a multifunctional polyacrylamide/PVA-based hydrogel containing synthesized poly(1-glycidyl-3-butylimidazolium salicylate) (polyGBImSal) and fabricated polydopamine-coated polyphenolic nanosheet (PDA@PNS) for wound dressing. The PDA@PNS particles were designed to induce I) antioxidant and anti-inflammatory features through ROS-scavenging and II) cell adhesive properties by the existing polydopamine into the hydrogels. The poly(ionic liquid)-based polyGBImSal was designed to allocate effective hydrogel antimicrobial activity. The fabricated hydrogel nanocomposites showed excellent properties in the swelling ratio, cell adhesiveness, protein adsorption, and anti-inflammatory, proving their general performance for application in wound healing. Furthermore, these hydrogels showed high antimicrobial activity (over 95 %) against three common wound-infecting pathogenic microbes: Escherichia coli, Staphylococcus aureus, and Candida albicans. The healing process of full-thickness dermal wounds in rats was accelerated by applying hydrogel nanocomposites with 0.5 wt% of PDA@PNS and 28 wt% of polyGBImSal. The wound closure contraction attained full closure, reaching 100 %, after 14 days, contrasted with the control group employing commercial wound dressing (Tegaderm), which achieved a closure rate of 68 % within the equivalent timeframe. These results make these hydrogel nanocomposites promising candidates for multifunctional wound dressing applications.

Tracing the Antibacterial Performance of Bis-Imidazolium-based Ionic Liquid Derivatives.

Ionic liquid (IL) cationic species have recently captivated the attention of pharmacists, biochemists, and biomedical scientists as promising antibacterial agents to deal with the multidrug resistance bacteria crisis. The structure and functional groups of ILs influence their physiochemical properties and biological activities. However, a comprehensive study is required to fully understand the details of the antibacterial activity of ILs carrying various functional groups. Herein, dicationic ILs (DCILs) are reported based on imidazolium rings as efficient antibacterial agents. The DCILs carried various functionalities such as 2-hydroxybutyl (DCIL-1), 2-hydroxy-3-isopropoxypropyl (DCIL-2), 2-hydroxy-3-(methacryloyloxy)propyl (DCIL-3), 2-hydroxy-2-phenylethyl (DCIL-4), and 2-hydroxy-3-phenoxypropyl (DCIL-5). The structure-antibacterial activity relationships of the DCILs against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) were comprehensively studied through antibacterial tests, morphology analysis, and adhesion tests. The experimental assays revealed an antibacterial efficacy order of DCIL-5 > DCIL-1 > DCIL-4 > DCIL-2 > DCIL-3. The all-atom molecular dynamics (MD) simulation showed a deep permeation of the hydrophobic -OPh functional group of DCIL-5 through the E. coli membrane model in agreement with the experimental observations. Current findings assist scientists in designing new task-specific DCILs for effective interactions with biological membranes for different applications.

High frequency of SCCmec type IV and multidrug-resistant SCCmec type I among hospital acquired methicillin-resistant Staphylococcus aureus isolates in Birjand Imam Reza Hospital, Iran.

Background and Objectives: The ever-increasing of antibiotic resistance in methicillin-resistant Staphylococcus aureus (MRSA) has become a major threat to public health worldwide. Molecular typing is used to determine the source of MRSA infections as well as to control and prevent the spread of these pathogens. The present study aimed to investigate the characteristics of staphylococcal cassette chromosome mec (SCCmec) types and antibiotic resistance of community- acquired (CA-) and hospital acquired (HA-) MRSA isolates. Materials and Methods: In this cross-sectional study, the antibiotic susceptibility patterns of 109 clinical S. aureus isolates were determined by the Kirby-Bauer disk-diffusion and microdilution broth methods. MRSA isolates were confirmed using the polymerase chain reaction (PCR) method for the detection of the mecA gene. SCCmec typing was performed by a multiplex PCR assay among MRSA isolates. Results: The prevalence of MRSA isolates was 39.4%. Linezolid, vancomycin, and ceftaroline were the most effective agents against MRSA isolates. The incidence of multidrug-resistant (MDR) and resistance to most antibiotics were significantly higher in MRSA than methicillin-susceptible S. aureus (MSSA) isolates (P<0.05). SCCmec types I, III, and IV were identified in 27.9%, 23.3%, and 37.2% of MRSA isolates, respectively. SCCmec type I and IV were the most prevalent SCCmec types in HA-MRSA isolates (each was 32.4%). While SCCmec type IV (66.7%) was the most frequently SCCmec type associated with CA-MRSA isolates. Conclusion: Our findings demonstrated a high rate of MDR among MRSA isolates. The high existence of SCCmec type IV was reported among the HA-MRSA isolates, which can indicate the spread of MRSA community isolates to hospital settings. Therefore, appropriate antibiotic stewardship plans and microbiological surveillance initiatives must be implemented in healthcare facilities to monitor and limit the spread of these resistant bugs.

Effect of Atkins versus a low-fat diet on gut microbiota, and cardiometabolic markers in obese women following an energy-restricted diet: Randomized, crossover trial.

BACKGROUND AND AIMS: There is controversy about effects of the Atkins diet on cardiometabolic markers in previous studies. No study compared effects of Atkins versus a low-fat diet on gut microbiota in obese women during a weight-loss program up to date. METHODS AND RESULTS: A 6-week, randomized, crossover trial was conducted. Twenty-four healthy women with obesity (BMI≥30 kg/m2) were randomly assigned to receive the Atkins (55%, 25%, and 20% of total daily calories from fat, protein, and carbohydrates), or low-fat (20%, 15%, and 65% of total daily calories from fat, protein, and carbohydrates) diets while following a weight-loss program. Vegetable oils were used as the main source of dietary fat. Dietary groups were switched after two weeks of washout period with a weight maintenance low-fat diet. The effects of the two diets did not differ for the most endpoints. However, Gut Actinobacteria residency and serum total antioxidant capacity significantly increased in the Akins diet group compared with the low-fat one (p = 0.02 and p = 0.04). Adjusting for all parameters, gut Actinobacteria residency 1.48- and 2.5-folds decreased the serum LDL.C/HDL.C ratio and non-HDL.C levels (95%CI: 3.1, -0.22; p = 0.03 and -0.07, -0.002; p = 0.04), respectively. Decrease in gut Proteobacteria residency showed a significant reduction in serum total oxidant status (95%CI: 7.4, -0.07; p = 0.04). CONCLUSIONS: The Atkins diet, based on vegetable oils, alters gut microbiota composition, atherogenic and antioxidant parameters. REGISTRATION NUMBER FOR CLINICAL TRIAL: IRCT20200929048876N3.

Antimicrobial Susceptibility, Serotyping, and Molecular Characterization of Antibiotic Resistance Genes in Listeria monocytogenes Isolated from Pregnant Women with a History of Abortion.

BACKGROUND: Listeria monocytogenes show high mortality among pregnant women and newborns. This study aimed to detect L. monocytogenes in pregnant women with a history of abortion and assess the serotypes, antibiotic susceptibility patterns, and its resistance genes. METHODS: Overall, 400 vaginal swabs were taken from pregnant women with a history of abortion in the past few years in a tertiary care hospital in Tehran, Iran, during 2015-2018. Antibiotics susceptibility to a panel of 10 antibiotics was determined using the standard disk diffusion method and the isolates serotyped by the agglutination method. The antimicrobial-resistant isolates were also screened for the presence of tetM, ermB and dfrD genes by PCR. RESULTS: Overall, 22 L. monocytogenes isolates were identified. High rates of resistance were observed for trimethoprim (50%; n=11), sulphamethoxazole (50%; n=11), tetracycline (45.45%; n=10) and gentamicin (36.36%; n=8). From 22 L. monocytogenes isolates, 13 (59.10 %), 5 (22.73%), 3 (13.63%) and 1 (4.54%) belonged to serotypes 4b, 1/2a, 1/2b, and 3c, respectively. The genetic determinant tetM was detected in 70% of the tetracycline-resistant isolates. Out of 11 trimethoprim-resistant isolates, 27.27% isolates contained dfrD. Moreover, the ermB gene was found in 83.33% of the erythromycin-resistant isolates. CONCLUSION: Ampicillin and partly penicillin consider to be suitable antimicrobial agents to treat human listeriosis. Moreover, due to resistance against many antibiotics, it is necessary to continue monitoring and managing antimicrobial resistance.

The Sensitivity and Specificity of Loop-Mediated Isothermal Amplification and PCR Methods in Detection of Foodborne Microorganisms: A Systematic Review and Meta-Analysis.

BACKGROUND: The loop-mediated isothermal amplification (LAMP) method is frequently used for identifying many microorganisms. The present review aimed to evaluate the sensitivity and specificity of LAMP method for detection of food-borne bacteria and to compare these features with those of polymerase chain reaction (PCR), as an alternative molecular diagnostic procedure, and with cultivation method, as the gold standard method. METHODS: The literature was searched in electronic databases (PubMed, Scopus, Web of Science, and EMBASE) for recruiting publications within Jan 2000 to Jul 2021. We used the combinations of keywords including foodborne disease, LAMP, PCR, Loop-mediated isothermal amplification, and polymerase chain reaction. Meta-analysis was used to adjust the correlation and heterogeneity between the studies. The efficiency of the methods was presented by negative likelihood ratio, positive likelihood ratio, sensitivity, specificity, and odds ratio using forest plots. A P-value less than 0.05 was considered as statistical significance cut off. The confidence intervals were presented at the 95% interval. RESULTS: Overall, 23 relevant studies were analyzed. The sensitivities of LAMP and PCR methods were estimated to be 96.6% (95% CI: 95.0-97.7) and 95.6% (95%CI: 91.5-97.8), respectively. The specificities of LAMP and PCR were also estimated to be 97.6% (95%CI: 92.6-99.3) and 98.7% (95%CI: 96.5-99.5), respectively. CONCLUSION: The specificities of LAMP and PCR assays were determined by comparing their results with cultivation method as the gold standard. Overall, the specificity of both PCR and LAMP methods was low for detection of fastidious bacteria. Nevertheless, LAMP and PCR methods have acceptable specificities and sensitivities, and their application in clinical practice necessitates more studies.

Reduced Efflux Pumps Expression of Pseudomonas Aeruginosa with Satureja Khuzistanica Essential Oil.

BACKGROUND: Efflux pumps such as MexEF-OprN and mexXY-OprM play an important role in the resistance of Pseudomonas Aeruginosa (P. aeruginosa) to antibiotics. The present study aimed to assess the reduced expression of efflux pump genes of P. aeruginosa with Satureja Khuzistanica essential oil (SKEO). METHODS: The present cross-sectional study was conducted in 2016 at the Microbiology Laboratory of Baqiyatallah University of Medical Sciences, Tehran, Iran. The disk diffusion method was used for susceptibility testing of gentamicin and norfloxacin. Minimum inhibitory concentration (MIC) was determined for gentamicin and norfloxacin. The antibacterial efficacy of SKEO was defined by determining the MIC values using the microdilution method. In vitro, the synergistic interaction of SKEO combined with gentamicin or norfloxacin was examined via checkerboard assay and defined as a fractional inhibitory concentration index. The reverse transcription-polymerase chain reaction technique was used to measure changes in the expression of the efflux pump genes. The data were analyzed using SPSS software version 16.0, and P<0.05 was considered statistically significant. RESULTS: The MIC values of SKEO were in the range of 6 to 12 µg/mL. In the presence of sub-inhibitory concentrations (1.16 to 2 MIC) of SKEO, synergistic effects were revealed using the checkerboard method. The effect of norfloxacin and gentamicin increased up to 8-fold. The expression of mexY and mexE was reduced after treatment with SKEO. CONCLUSION: SKEO reduced the expression of efflux pumps and the MIC values of norfloxacin and gentamicin in vitro.

High Frequency of AdeA, AdeB and AdeC Genes among Acinetobacter baumannii Isolates.

BACKGROUND: Efflux pumps are involved in resistance of Acinetobacter baumannii isolates to antimicrobial agents. AdeABC efflux pump is one of the RND superfamily efflux pump and consists of adeA (membrane fusion), adeB (multidrug transporter) and adeC (outer membrane) genes. In this study, the frequency of adeA, adeB and adeC genes among A. baumannii isolates with resistance to erythromycin, trimethoprim, meropenem and imipenem was investigated. METHODS: Overall, 79 strains of A. baumannii were isolated from patients admitted to two major hospitals in Tehran during 2016. Antibiotic susceptibility testing was determined by disc diffusion and microdilution methods according to Clinical and Laboratory Standards Institute (CLSI) guideline. The presence of adeA, adeB and adeC genes was also determined using Multiplex PCR assay. RESULTS: The highest and the lowest resistance among A. baumannii isolates were to trimethoprim (93%) and erythromycin (53%), respectively. The frequency of adeA, adeB and adeC genes was 96.2%, 96.2% and 91.1 % respectively. There was a significant relationship between imipenem resistance and presence of efflux pump genes (P<0.05). CONCLUSION: According to the high prevalence of the AdeABC efflux system genes, it may involve in resistance of clinical isolates of A. baumannii to imipenem, especially.

Ferritin Degradation by Pneumococcal HtrA, RadA and ClpP Serine Proteases : A Probable Way For Releasing and Acquisition Of Iron.

PURPOSE: Iron is a necessary element for the growth of bacteria; however, there are limited iron sources known for these microorganisms yet. Intracellular iron is stored as ferritin from, which releases iron in a gradual and controlled manner. The present study aimed to characterize ferritin-binding proteins (FBPs) of Streptococcus pneumoniae. MATERIAL AND METHODS: S. pneumoniae species were cultured in BHI broth containing ferritin (1094 ng/mL) for 4h at 37°C. Ferritin level was measured using ELISA assay. Bacterial proteome was electrophoresed on SDS-PAGE and then transferred on PVDF nitrocellulose membrane. Afterward, the PVDF membranes were incubated with a ferritin solution. Identification of ferritin binding proteins was performed using anti-ferritin monoclonal antibody conjugated with HRP enzyme. Molecular docking was used to assess the interaction between pneumococcal proteases and FBPs applying phenylmethylsulfonyl fluoride (PMSF) as a protease inhibitor. RESULTS: No FBPs were identified in S. pneumoniae proteome. Moreover, ferritin levels have significantly (p<0.05) decreased following the growth of S. pneumoniae in ferritin-rich BHI medium. Also, molecular docking showed that RadA protease, ClpP hydrolase, and HtrA protease can potentially interact with PMSF protease inhibitors. On the other hand, the addition of the PMSF to the culture of S. pneumoniae prevented the reduction of ferritin, which indicates a potential role of RadA, ClpP, and HtrA proteases in ferritin degradation. CONCLUSION: Our results suggest that S. pneumoniae produces no FBPs and also cannot directly use ferritin as an iron source. However, ferritin may be degraded through a protease-mediated mode.

Psp140: an immunodominant antigen in the supernatant of Streptococcus pneumoniae culture.

BACKGROUND AND OBJECTIVES: Streptococcus pneumoniae causes many lethal infections. Due to its reduced sensitivity to commonly used antibiotics, development of new strategies against pneumococcal infections seems to be necessary. We aimed to investigate immunodominant antigens in S. pneumoniae culture supernatant in order to develop novel targets for pneumococcal vaccines. MATERIALS AND METHODS: In this study S. pneumoniae ATCC49619 was sub-cultured into BHI broth from overnight culture at 37°C for 4 h. The supernatant proteins were precipitated using acetone precipitation method. A rabbit was intramuscularly immunized with alum adjuvant and 100 µg pneumococcal supernatant proteins, 6 times at 14 days' intervals to produce hyperimmune serum. ELISA assay was performed to determine the antibody level response to pneumococcal secretory proteins. Then dot blot applied for rapid evaluation of hyperimmune serum reactivity to pneumococcus supernatant proteins. The western blot was also used to determine the interaction of supernatant proteins with immunogenic rabbit's hyperimmune-serum. RESULTS: According to the western blot analysis, the immunodominant protein had 140KDa molecular weight and designated as pneumococcal secretory protein140 (Psp140). CONCLUSION: The Psp140 protein in the supernatant of S. pneumoniae culture is an immunodominant protein and it is likely related to pneumococcal secretory protein or surface exposed protein which released into culture supernatant during bacterial growth.

Association of human papilloma virus (HPV) infection with oncological outcomes in urothelial bladder cancer.

BACKGROUND: Bladder cancer is one of the leading causes of cancer death in adults worldwide. There are various risk factors described for the bladder cancer development including genetic background as well as environmental exposure. Currently, infectious agents such as human papilloma virus (HPV) has also been linked to bladder cancer risk. The current study aimed to evaluate the potential correlation between HPV infection and the oncological outcome in urothelial bladder cancer. METHODS: Totally 106 tissue samples of histopathologically confirmed transitional cell carcinoma (TCC) of the urinary bladder were included in this study. The presence of high risk (types 16 and 18) and low risk (types 11 and 6) types of HPV was evaluated using polymerase chain reaction (PCR) followed by in situ hybridization. RESULTS: Out of 106 bladder cancer patients, a total of 24 cases (22.6%) were positive HPV infection. The most common type of HPV detected was type 16 followed by types 11 and 18, and 6. According to independent T-test results, there was a significant association between mean age and HPV infection (P = 0.015). Moreover, our findings showed a significant relation between infection with HPV and tumor stage, tumor grade, muscle invasion of the tumor, as well as tumor recurrence. The results of Chi-square Test indicated that there is significant statistical association between types of HPV and tumor grade (P-Value = 0.044). CONCLUSION: Our findings indicated that a family history of cancer and HPV infection can be potential independent predictive factors for tumor recurrence in bladder cancer. Overall, the results of this study strongly indicate a significant relationship between HPV infection and an aggravated outcome of the disease and a higher risk of recurrence in patients with bladder cancer.

Autolysin (lytA) recombinant protein: a potential target for developing vaccines against pneumococcal infections.

PURPOSE: N-acetylmuramoyl-l-alanine amidase known as lytA, is an immunogenic protein that plays an important role in the pathogenesis of Streptococcus pneumoniae. It is highly conserved among S. pneumoniae strains and is absent among other Streptococcus species. In the present study, the level of antibodies against the lytA recombinant protein was evaluated in healthy individuals' sera. MATERIALS AND METHODS: DNA was extracted from S. pneumoniae ATCC 49619 to amplify lytA gene by polymerase chain reaction assay. The lytA amplicon and pET28a vector were separately double digested using Nde-1 and Xho1 restriction enzymes and then ligated together with ligase enzyme. The recombinant plasmid was expressed in Escherichia coli BL21 strain and the lytA recombinant protein purified using nickel-nitrilotriacetic acid affinity chromatography. Western blot was carried to detect lytA recombinant protein. Sixty healthy individual's sera (at three age groups: group 1, <2; group 2, 2-40; and group 3, 60-90 years old) were collected and the titers of anti-lytA antibodies were determined. RESULTS: The lytA gene was highly expressed in E. coli BL21 host. The recombinant lytA protein was purified and confirmed by western blotting. Tukey test analysis showed that there were no significant differences among the age groups considering the anti-lytA titer of 10. However, at the anti-lytA titer of 60, significant differences were observed between group 1 vs. group 2 (p<0.001); group 1 vs. group 3 (p=0.003), and group 2 vs. group 3 (p=0.024). CONCLUSION: The lytA protein seems to be a highly immunogenic antigen and a potential target for developing vaccines against pneumococcal infections.

Molecular Epidemiology of Carbapenemase-Producing Pseudomonas aeruginosa Isolated from an Iranian University Hospital: Evidence for Spread of High-Risk Clones.

PURPOSE: Given the importance of treatment failure due to multidrug-resistant (MDR) strains, studies on population structure of these organisms are necessary to improve control strategies. Accordingly, the current study aimed to determine the prevalence of carbapenem-resistant P. aeruginosa (CRPA) at a teaching referral hospital in Iran and to analyz their molecular clonality by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) for epidemiological purposes. METHODS: In this study, modified Hodge test (MHT) and double-disk synergy test (DDST) were used for carbapenemase production and metallo-ß-lactamases (MBLs) screening, respectively. All P. aeruginosa isolates were tested for antimicrobial resistance. Moreover, MBL genes (blaIMP, blaVIM, blaSPM, blaNDM) were detected by multiplex PCR assay. RESULTS: Among 68 P. aeruginosa clinical isolates, 38 (55.88%) isolates were CRPA. Antibiotic susceptibility testing revealed that most of these isolates were MDR. PFGE analyses showed 5 common types and 27 single types among CRPA isolates. MLST analysis revealed three major clusters (MLST-sequence types (STs): 235, 357, and 861) among them. The 30 non-CRPA isolates corresponded mainly to MLST-STs 253, 360, and 446. CONCLUSION: Our results showed that internationally distributed MLST-STs with widely genomic diversity have spread in our hospital, and clonal expansion of MDR strains of P. aeruginosa was described as well.

Molecular docking of the pneumococcal main autolysin (LytA) and deoxycholate ligand.

In the Streptococcus pneumoniae, the N-acetylmuramoyl-l-alanine amidase known as LytA protein is a main autolysin and in the presence of sodium deoxycholate, it activates and breaks S. pneumoniae cell wall. In the present study, the interaction between the LytA protein and deoxycholate as ligand was investigated. The Lyt A protein was retrieved from PDB databank and energetically minimized by Molegro Virtual Docker. The binding sites of LytA protein were detected and molecular docking carried out using MolDock algorithm. Finally, the number of hydrogen and electrostatic bonds were obtained for each predicted pose. A total of 5 binding sites predicted on LytA protein. The number of 5 predicted poses for each binding site also detected and molecular docking showed that all the poses have interactions (by H bonds) with deoxycholate. The interaction of the LytA protein with the deoxycholate ligand reveal five binding sites, which are involved in deoxycholate substrate recognition.

Comparison of Reverse Transcription Loop-Mediated Isothermal Amplification Method with SYBR Green Real-Time RT-PCR and Direct Fluorescent Antibody Test for Diagnosis of Rabies.

Rabies as an endemic disease in most Asian and African countries, especially in remote areas, and requires a reliable diagnostic method. This study aimed to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of rabies virus RNA in the brain samples, compared to SYBR Green real time RT-PCR test as a molecular technique and direct fluorescent antibody test as a serological method. In this study, RT-LAMP was developed to diagnose rabies. Six primers were designed based on the nucleoprotein (N) of rabies virus. The sensitivity and specificity of SYBR Green real-time RT-PCR and RT-LAMP methods were also determined.RT-LAMP was optimized at 58 ℃ for 60 min. The sensitivity and specificity of RT-LAMP and SYBR Green real-time RT-PCR were 91.2% and 84.2%, and 94.12% and 88.9%, respectively. The slight difference between the sensitivity and specificity of RT-LAMP and that of SYBR Green Real-Time RT-PCR demonstrated that RT-LAMP could be used as a reliable and cost-effective method for the diagnosis of rabies.

Serological evidence of leptospirosis in Iran; A systematic review and meta-analysis.

Leptospirosis is a neglected infectious zoonotic disease that affects domestic animals and wildlife as well as humans. Although leptospirosis is known as an endemic disease in Iran, there is no accurate information on the overall prevalence of this disease in humans and animals. The aim of this systematic review and meta-analysis was to estimate the seroprevalence of leptospirosis among human and domestic and wild animals in Iran. A systematic review of English and Persian articles (since 1998 to December 2017) was conducted using Google Scholar, Medline/PubMed, Science Direct, Scopus, Web of science and Iranian databases Iranmedex, Scientific Information Database (SID), Magiran, and IRANDOC. Search terms include leptospirosis, Leptospira, serology, seroprevalence, seroepidemiology, serological, Iran, cow, goat, sheep, camel, dog, cat, equine, donkey, horse, mule and rodent. In Eventually 66 articles were selected to analyze based on inclusion criteria. Seroprevalence of leptospirosis in human was 27.84% (95% CI: 13.22-22.47) and 19.71% (95% CI: 6.78-32.65%) based on ELISA and MAT, respectively. The pooled prevalence of leptospirosis in cow, sheep, goat and camel was 26.62% (95% CI: 18.76-34.48), 17.38% (95% CI: 13.32-21.43), 12.18% (95% CI: 9.96-14.41) and 22.68% (95% CI: 18.97-26.40), respectively. The prevalence of leptospirosis in horse, donkey, and mule was 19.99% (95% CI: 13.32-26.68), 40.59% (95% CI: 33.20-47.97) and 9.10% (95% CI: 2.90-15.30), respectively. The prevalence in dog and cat were estimated 14.63% (95% CI: 3.49-25.77) and 14.44% (95% CI: 3.25-25.65), respectively. The prevalence of seropositivity in rodents was estimated 20.96% (95% CI: 10.62-31.30). This study is a very comprehensive report on the status of leptospirosis in Iran. Based on our results, leptospirosis has considerable seroprevalence among human and animals in Iran. This high seroprevalence of leptospirosis showed should be given more attention for this disease in Iran and thus health measures must be taken to diagnosis, control and prevent it.

Evaluation of (GTG) 5-PCR for Genotyping of Klebsiella pneumonia Strains Isolated from Patients with Urinary Tract Infections.

BACKGROUND: Klebsiella pneumonia is one of the common causes of pneumonia and bacteremia in intensive care patients. The present study was aimed to determine the capability of (GTG) 5-PCR assay for molecular typing of K. pneumonia strains isolated from patients with urinary tract infections. METHODS: In this descriptive-sectional study, K. pneumoniae strains were collected from hospitalized patients with urinary tract infection in Baqiyatallah Hospital, Tehran, Iran during 2017-2018. Isolates were identified by conventional microbiological tests. Bacterial DNA was extracted using boiling method and (GTG) 5-PCR assay was used for subtyping of the isolates. For clustering of isolates, dendrogram was generated according to the un-weighted pair group method with arithmetic (UPGMA). RESULTS: Overall, 88 K. pneumoniae isolates were isolated and subjected to the molecular typing study. The (GTG) 5-PCR assay was able to differentiate the K. pneumoniae strains into 9 clusters including G1-G9. Genotype clusters G4 and G9 consist of highest (26) and lowest (1) number isolate, respectively. CONCLUSION: The K. pneumonia strains isolated under the study belonged to various clones and the (GTG) 5-PCR assay as simple and rapid method can be a powerful tool for molecular typing of K. pneumoniae strains.

Rapid Detection of Mycoplasma pneumoniae by Loop-Mediated Isothermal Amplification (LAMP) in Clinical Respiratory Specimens.

BACKGROUND: Mycoplasma pneumoniae is a common cause of community-acquired pneumonia (CAP) worldwide, especially among children and debilitated populations. The present study aimed to investigate a loop-mediated isothermal amplification (LAMP) technique for rapid detection of M. pneumoniae in clinical specimens collected from patients with pneumonia. METHODS: Throat swabs were collected from 110 outpatients who suffered from pneumonia. Throat swab samples were obtained from patients referred to the hospital outpatient clinics of Tehran University hospitals, Iran in 2017. The presence of M. pneumoniae in the clinical specimens was evaluated by LAMP, PCR and culture methods. Sensitivity and specificity of the LAMP and PCR assays were also determined. RESULTS: Out of 110 specimens, LAMP assay detected M. pneumoniae in 35 specimens. Detection limit of the LAMP assay was determined to be 33fg /µL or ∼ 40 genome copies/reaction. Moreover, no cross-reaction with genomic DNA from other bacteria was observed. Only 25 specimens were positive by the culture method. The congruence between LAMP assay and culture method was 'substantial' (Ï°=0.77). Specificity and sensitivity of LAMP assay were 88.2%, 100% in compare with culture. However, the congruence between LAMP assay and PCR assay was 'almost perfect' (Ï°=0.86). Specificity and sensitivity of LAMP assay were 92.5%, 100% in compare with PCR. CONCLUSION: Overall, the LAMP assay is a rapid and cost-efficient laboratory test in comparison to other methods including PCR and culture. Therefore, the LAMP method can be applied in identification of M. pneumoniae isolates in respiratory specimens.

Molecular Cloning and Immunogenicity Evaluation of PpiC, GelE, and VS87_01105 Proteins of Enterococcus faecalis as Vaccine Candidates

Background: Among the enterococci strains, Enterococcus faecalis is considered as one of the important nosocomial pathogens affecting immunocompromised patients. In this study, the immunogenicity of PpiC, GelE, and VS87_01105 proteins against enterococcal infection was investigated in a mice model. Methods: The genes encoding these proteins were cloned into pET21a expression vector, and the recombinant proteins were produced. Mice and rabbits were immunized with the purified recombinant proteins, and subsequently, mice were challenged with E. faecalis for the evaluation of their survival and bacterial clearances. The antibody responses to recombinant proteins were determined by ELISA assay, and opsonophagocytic activities of the antibodies were also measured. Passive immunization was performed using purified antibodies. Mice were challenged, and their survival and bacterial clearance were determined. Results: Immunized mice with PpiC, GelE, and VS87_01105 recombinant proteins showed 80%, 70%, and 40% survival rate, respectively. The survival rates among passively immunized mice that received 500 µg of IgG fraction in 100 µl PBS buffer of each of anti-PpiC, anti-GelE, and anti-VS87_01105 were 60%, 50%, and 20%, respectively. The rates of opsonization with anti-PpiC, anti-GelE, and anti-VS87_01105 antibodies at 1/10 dilution were 77%, 64%, and 23%, respectively. Conclusion: Based on our findings, PpiC, and GelE proteins can protect the mice against E. faecalis ATCC 29212 and effectively induce a protective antibody response. Thus, these proteins could be used as an additional therapeutic tool against enterococcal infections. Further studies to determine the role of PpiC in ligand binding and demonstration of epitope mapping may establish a credible target for vaccination.