Antimicrobial and antibiofilm effects of cyclic dipeptide-rich fraction from Lactobacillus plantarum loaded on graphene oxide nanosheets.

Introduction: One effective method to combat bacterial infections is by using bacteria itself as a weapon. Lactobacillus is a type of fermenting bacterium that has probiotic properties and has demonstrated antimicrobial benefits against other bacteria. Cyclodipeptides (CDPs), present in the supernatant of Lactobacillus, possess several antimicrobial properties. Methods: In this study, the CDP fraction was isolated from the supernatant of Lactobacillus plantarum (L. plantarum). This fraction was then loaded onto graphene oxide nanosheets (GO NSs). The study assessed the substance's ability to inhibit bacterial growth by using the minimum inhibitory concentration (MIC) method on A. baumannii and S. aureus strains that were obtained from clinical samples. To determine the substance's impact on biofilm formation, the microtiter plate method was used. Moreover, the checkerboard technique was employed to explore the potential synergistic effects of these two substances. Results and discussion: According to the study, the minimum inhibitory concentration (MIC) of the desired compound was found to be 1.25 mg/mL against S. aureus and 2.5 mg/mL against A. baumannii. Furthermore, at a concentration of 10 mg/mL, the compound prevented 81.6% (p < 0.01) of biofilm production in A. baumannii, while at a concentration of 1.25 mg/mL, it prevented 47.5% (p < 0.05) of biofilm production in S. aureus. The study also explored the synergistic properties of two compounds using the checkerboard method. Conclusion: In general, we found that GO NSs possess antimicrobial properties and enhance cyclodipeptides' activity against S. aureus and A. baumannii.

Investigating the effect of Fusobacterium nucleatum on the aggressive behavior of cancer-associated fibroblasts in colorectal cancer.

Fusobacterium nucleatum, (F. nucleatum) as a known factor in inducing oncogenic, invasive, and inflammatory responses, can lead to an increase in the incidence and progression of colorectal cancer (CRC). Cancer-associated fibroblasts (CAF) are also one of the key components of the tumor microenvironment (TME), which lead to resistance to treatment, metastasis, and disease recurrence with their markers, secretions, and functions. This study aimed to investigate the effect of F. nucleatum on the invasive phenotype and function of fibroblast cells isolated from normal and cancerous colorectal tissue. F. nucleatum bacteria were isolated from deep periodontal pockets and confirmed by various tests. CAF cells from tumor tissue and normal fibroblasts (NF) from a distance of 10 cm of tumor tissue were isolated from 5 patients by the explant method and were exposed to secretions and ghosts of F. nucleatum. The expression level of two markers, fibroblast activation protein (FAP), and α-smooth muscle actin (α-SMA), and the amount of production of two cytokines TGF-ß and IL-6 from fibroblast cells were measured by flow cytometry and ELISA test, respectively before and after exposure to different bacterial components. The expression of the FAP marker was significantly higher in CAF cells compared to NF cells (P < 0.05). Also, the expression of IL-6 in CAF cells was higher than that of NF cells. In investigating the effect of bacterial components on the function of fibroblastic cells, after comparing the amount of IL-6 produced between the normal tissue of each patient and his tumoral tissue under 4 treated conditions, it was found that the amount of IL-6 production from the CAF cells of patients in the control group, treated with heat-killed ghosts and treated with paraformaldehyde-fixed ghosts had a significant increase compared to NF cells (P < 0.05). Due to the significant increase in FAP marker expression in fibroblast cells of tumor tissue compared to normal tissue, it seems that FAP can be used as a very good therapeutic marker, especially in patients with high levels of CAF cells. Various components of F. nucleatum could affect fibroblast cells differentially and at least part of the effect of this bacterium in the TME is mediated by CAF cells.

Antimicrobial efficacy of antiplaque agents of common toothpastes against Porphyromonas gingivalis and Streptococcus oralis: An in vitro study.

Background: This study was designed to evaluate the antimicrobial activity of common gum protection and antiplaque toothpastes against Porphyromonas gingivalis (P. gingivalis) and Streptococcus oralis (S. oralis) as important periodontal pathogens. Materials and Methods: This experimental study investigated the antimicrobial activity of 15 commonly used toothpastes from different companies on the two common types of periopathogens, S. oralis and P. gingivalis . The antimicrobial activity of toothpaste was evaluated at three concentrations of 100%, 50%, and 25% and analyzed by agar well diffusion plate method and zone of inhibition. The obtained data were compared and statistically analyzed by SPSS software using one-way ANOVA and the least significant difference post hoc tests (α = 0.05). Results: One-way ANOVA showed that the mean diameter of the two-bacterial zone of inhibition was significantly different at 100%, 50%, and 25% concentrations of toothpastes (P < 0.001). In general, the mean diameter of the zone of inhibition was greater at 100% concentration than the other two concentrations in all toothpastes. The highest zone of inhibition of the S. oralis was in the toothpastes containing tin. Further, the highest zone of inhibition of P. gingivalis was found in the triclosan-containing toothpastes. Conclusion: Toothpastes containing triclosan had the most antimicrobial activity against P. gingivalis . Moreover, toothpastes containing tin compounds had the most antimicrobial effect against S. oralis .

Enhanced antibacterial properties of polyvinyl alcohol/starch/chitosan films with NiO-CuO nanoparticles for food packaging.

Packaging is very important to maintain the quality of food and prevent the growth of microbes. Therefore, the use of food packaging with antimicrobial properties protects the food from the growth of microorganisms. In this study, antibacterial nanocomposite films of polyvinyl alcohol/starch/chitosan (PVA/ST/CS) together with nickel oxide-copper oxide nanoparticles (NiO-CuONPs) are prepared for food packaging. NiO-CuONPs were synthesized by the co-precipitation method, and structural characterization of nanoparticles (NPs) was carried out by XRD, FTIR, and SEM techniques. Composites of PVA/ST/CS, containing different percentages of NPs, were prepared by casting and characterized by FTIR and FESEM. The mechanical properties, diffusion barrier, and thermal stability were determined. The nanoparticles have a round structure with an average size of 6.7 ± 1.2 nm. The cross-section of PVA/ST/CS film is dense, uniform, and without cracks. In the mechanical tests, the addition of NPs up to 1% improved the mechanical properties (TS = 31.94 MPa), while 2% of NPs lowered TS to 14.76 MPa. The fibroblast cells toxicity and the films antibacterial activity were also examined. The films displayed stronger antibacterial effects against Gram-positive bacteria (Staphylococcus aureus) compared to Gram-negative bacteria (Escherichia coli). Furthermore, these films have no toxicity to fibroblast cells and the survival rate of these cells in contact with the films is more than 84%. Therefore, this film is recommended for food packaging due to its excellent mechanical and barrier properties, good antibacterial activity, and non-toxicity.

Preparation and characterization of gelatin/chitosan nanocomposite reinforced by NiO nanoparticles as an active food packaging.

Food packaging with antibacterial properties has attracted much attention recently. In this study, nickel oxide nanoparticles (NiONPs) were synthesized by co-precipitation and then gelatin/chitosan polymer films (GEL/CS) with different percentages of NiONPs, bio-nanocomposites, were prepared by casting. Morphology, crystal microstructure, molecular interactions and thermal stabilities of the NPs and the composite films were characterized by FESEM, XRD, FTIR and TGA, respectively. The bio-nanocomposite films exhibited excellent barrier, thermal and mechanical properties by addition of an optimized content of NPs. For example, the tensile strength (TS) of the GEL/CS film without NPs was 23.83 MPa and increased to 30.13 MPa by incorporation of 1% NPs. The antibacterial properties and toxicity of the films were investigated. These films show good antibacterial behavior against Gram-positive (Staphylococcus aureus) bacteria compared to Gram-negative (Escherichia coli) bacteria. Furthermore, the films were found to be non-toxic to fibroblast cells that came into contact with the films, with a survival rate of more than 88%. Therefore, these films can be applied for food packaging due to their excellent mechanical, barrier, and antibacterial properties.

Antimicrobial photodynamic therapy with dendrosomal curcumin and blue laser against Porphyromonas gingivalis.

BACKGROUND: Periodontitis is a chronic inflammatory disease that leads to the loss of tooth-supporting structures. Porphyromonas gingivalis is one of the main pathogens responsible for periodontitis. Because of the limitations of antibiotic use, various alternative approaches have been developed. Antimicrobial photodynamic therapy uses photosensitizers and light to eliminate pathogens. Curcumin is a promising photosensitizer, but has low bioavailability and water solubility. However, dendrosomes can efficiently encapsulate curcumin, overcoming these obstacles. This study aimed to evaluate the efficacy of photodynamic therapy with blue laser and dendrosomal curcumin against Porphyromonas gingivalis. METHODS: In this in vitro experiment, the minimum inhibitory concentration (MIC) of dendrosomal curcumin was determined using a serial dilution approach. Porphyromonas gingivalis suspensions were subjected to blue laser irradiation (447 nm, output power 100 mW) for 30 to 180 s. Finally, several subMIC dendrosomal curcumin concentrations and blue laser irradiation periods were applied to the bacterial suspensions. The negative control group received no therapy, whereas the positive control group was treated with 0.2% chlorhexidine. Consequently, the colony count of each group was calculated. RESULTS: Treatment of Porphyromonas gingivalis with dendrosomal Curcumin at concentrations of 8-250 µg/mL significantly reduced bacterial growth compared to untreated group. 90 second exposure to a blue laser (31.8 J/cm2) completely inhibited the growth of Porphyromonas gingivalis. Blue laser irradiation for 60 s (21.2 J/cm2) markedly reduced bacterial growth but did not completely prevent its survival. Photodynamic therapy using dendrosomal curcumin at concentrations of 2-4 µg/mL and irradiation for 30-90 s resulted in complete eradication of Porphyromonas gingivalis compared to controls (P < 0.05). CONCLUSION: The reduction in survival of Porphyromonas gingivalis following photodynamic therapy with dendrosomal curcumin and blue laser indicates that this technique could be a useful approach to eradicate Porphyromonas gingivalis infections.

Determination of Virulence Factors and Resistance Profile of Methicillin-Resistant Staphylococcus aureus Strains among Different Types of spa, agr, and SCCmec.

In order to restrict the spread of methicillin-resistant S. aureus (MRSA) in hospitals, it is necessary to characterize isolates rapidly and precisely. The objective of this study was to determine virulence factors and resistance profiles of MRSA strains among spa, agr, and SCCmec types. In total, 55 MRSA isolates were collected from clinical specimens. The MRSA isolates were characterized by antimicrobial susceptibility testing, virulence genes, agr typing, spa typing, and SCCmec typing. According to our findings, all MRSA strains were resistant to cefoxitin; 88% and 86.7% of which were resistant to erythromycin and clindamycin, respectively. Type II agr was predominant with 54.54% frequency. Among 27 different spa types, type t030 was most frequently (25.45%). Most MRSA isolates (63.3%) were SCCmec type III. The pvl and tst genes were found in 25.3% and 32.7% of MRSA isolates, respectively. Among the MRSA strains, ermA, ermB, and ermC were present in 50%, 33.3%, and 57.3% of cases, respectively. In addition, 43 of the 55 MRSA strains (78%) harbored aminoglycoside resistance genes. The results of our study revealed that the MRSA rate in our region is dramatically high. Better infection control guidelines in hospitals, as well as ongoing epidemiological surveillance studies, could be strongly suggested for effective prevention of the spread of MRSA to inpatients.

Antibiotic resistance pattern of Helicobacter pylori strains isolated from patients in Isfahan, Iran.

Background: The objective of this study was to evaluate the antibiotic resistance pattern of Helicobacter pylori strains isolated from patients in Isfahan province. Materials and Methods: Gastric antrum biopsy specimens of patients undergoing endoscopy were cultured. The samples with the growth of H. pylori underwent antibiotic susceptibility test by disk diffusion method. Reaults: Of 96 samples, 50 samples (53%) were positive for H. pylori. The rates of antibiotic resistance were as follows: amoxicillin, 6%; azithromycin, 20%; furazolidone, 22%; levofloxacin, 16%; metronidazole, 20%; rifampin, 12%; and tetracycline, 22%. Conclusion: H. pylori strains in our area have high rates of resistance to azithromycin, levofloxacin, metronidazole, tetracycline, and furazolidone.

Identification of two major direct repeat unit clusters, 8i and 11ce, among methicillin resistant Staphylococcus aureus strains: the emergence of novel dru types and repeats.

BACKGROUND: The changing epidemiology and decreasing susceptibility to first-line antibiotics, such as vancomycin and linezolid, leave clinicians with few therapeutic options for MRSA infections. This study aimed to conduct an epidemiology study and characterize MRSA isolates. METHODS: A total of 150 MRSA isolates were collected from clinical specimens. Antimicrobial susceptibility was determined using the disk diffusion method. Resistance and major virulence genes were screened using the polymerase chain reaction. The SCCmec and dru typing were used to conduct molecular epidemiology. The BioNumerics tandem-repeat sequence typing plug-in tool was utilized for dru type cluster analysis. We constructed a minimum spanning tree using the similarity matrix of the DSI model. RESULTS: We discovered 24 dru types among the 55 dru sequenced MRSA isolates. Additionally, eight new dru types were discovered and added to the dru typing database. Two dru clusters (8i, 11ce) and nine single dru types were identified in 55 dru sequenced MRSA isolates. The two dru clusters, 8i and 11ce, accounted for 46 MRSA isolates (83.63%). The most common one of the nine singles dru types in this study was dt9bd, which belonged to the SCCmec types of IX. CONCLUSIONS: Given that two clusters account for the majority of strains in our study, we can conclude that the genetic origin of these strains is the same. Therefore, the spread of these strains can be prevented with effective MRSA monitoring in hospitals and communities.

Ocular Surface Microbial Flora and Photorefractive Keratectomy.

Purpose: To assess the influence of photorefractive keratectomy (PRK) on ocular surface microbial flora. Methods: A prospective study was conducted on patients who underwent PRK. The samples were taken from the inferior conjunctival fornix using a sterile swab, immediately before surgery, and then within three months following the PRK. The samples were tested using three culture mediums including blood agar, chocolate agar, and eosin methylene blue agar. Results: Thirty-five eyes of 35 patients including 19 females (54.3%) with a mean age of 24 ± 3.2 years were enrolled. The culture-positive rate was 15/35 eyes (42.9%) preoperative and 17/35 (48.6%) postoperative samples (P=0.47). The most common microorganisms isolated from preoperative samples were coagulase-negative Staphylococcus (CoNS) spp. in 14 (40%) samples, followed by Streptococcus spp. in 2 (5.7%), and Staphylococcus aureus in one (2.9%). Postoperative microorganisms isolated from conjunctival samples were CoNS spp. in 15 (42.9%), Streptococcus spp. in 3 (8.6%), and Staphylococcus aureus in one (2.9%), and Corynebacterium spp. in one (2.9%). Conclusion: This study indicated that there is not any remarkable difference in microorganisms isolated from conjunctival samples three months after PRK.

Genetic Diversity of Helicobacter pylori Isolates from Patients with Gastric Diseases in Isfahan.

Background: Helicobacter pylori (H. pylori), a spiral-shaped bacterium colonizing the human stomach, is generally acquired in childhood. This pathogen is highly diverse and can be used as genetic markers for predict the history of human migrations. This study aimed to determine the genetic diversity of H. pylori isolates from patients with dyspepsia by the multi-locus sequence typing (MLST) and update data on the prevalence of H. pylori among Iranian dyspeptic patients. Materials and Methods: In this descriptive cross-sectional study, 165 gastric biopsy specimens were obtained from patients with dyspepsia referred to Dr. Shariati Hospital of Isfahan, Iran, from April to July 2018. The status of H. pylori infection was determined by FISH in paraffin-embedded biopsy specimens. MLST of seven housekeeping genes was performed for 20 H. pylori isolates. The phylogenetic tree was plotted using CLC v8 and iTol software. Results: The overall prevalence of H. pylori infection was 53.3%. In the results of the analysis of MLST, a total of 14 new STs were recorded. The results of the global analysis showed that all the isolates, with a wide diversity, have a genetic affinity with members of the European population, such as Italy and Russia, and are in the hpEurope haplotype. Conclusion: Given the high prevalence of H. pylori infection in this region, early and accurate identification of patients seems necessary. Sequence analysis and determination of the origin of the phylogeny of strains can be effective in clinical management and monitoring of risk factors for chronic and recurrence of infection.

Comparison of antimicrobial effect of several decontaminating methods on contaminated Titanium discs.

Background: Decontaminating the implant surface, exposed to bacterial biofilm, is a concern in the treatment of peri-implant inflammatory disease. The aim of this study was to compare the effect of several methods on reduction of the bacterial load, colonized on the surfaces of titanium discs. Materials and Methods: In this in vivo study, seven titanium discs with Sandblasted, Large-grit, acid-etched (SLA) surface were placed in the mouth of each of ten patients with chronic periodontitis by an intra-oral maxillary splint for 24 h. In each patient, the contaminated discs, except for the negative control ones, were randomly treated by one of the six antiseptic methods including sterile normal saline, plastic curette, air polisher, hydrogen peroxide, 980 nm diode laser, and Er-YAG laser. A spectrophotometer was used to measure Optical Density (OD) in case of aerobic microorganisms. Colony-Forming Units (CFUs) were used for anaerobic bacteria. Data were analyzed through Kruskal-Wallis and Mann-Whitney Tests at a significance level of α =0.05 by SPSS software. Results: Statistical analysis showed a significant decrease in OD of aerobic bacteria among the seven groups during a 0-24 h time interval (P < 0.001). Furthermore, these tests showed a significant difference in the CFU (P < 0.001) for anaerobic bacteria after 48 h. Conclusion: The results of this study showed that all of the adopted methods significantly reduced microbial colonies on the surfaces of titanium discs with SLA surface. Er: YAG laser and normal saline had the highest and the lowest effects, respectively.

Comparison of the Prevalence of Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) among Staphylococcus aureus Isolates in a Burn Unit with Non-Burning Units.

BACKGROUND: Staphylococcus aureus (S. aureus) is one of the most important pathogens in burn infections colonized in the nose and increase the risk of infections. METHODS: Overall, 85 S. aureus isolates were isolated from clinical and nasal hospitalized patients and health care workers (HCWs) in a burn unit and non-burn units in Isfahan from June 2016 and September 2016. Genes encoding penicillin-binding protein 2a (mecA) and adhesive surface proteins, including fibronectin-binding proteins (fnbA,fnbB), fibrinogen binding protein (fib), laminin-binding protein(eno), collagen binding protein (cna), elastin binding protein (ebps), intracellular adhesion operon (icaA and icaD) were detected using PCR method. RESULTS: The rate of methicillin-resistant S. aureus (MRSA) among burn and non-burn isolates were 62% (18/29) and 25% (14/56), respectively. The most prevalent MSCRAMMs genes in burn units were eno (86%) and fib (66%). The most common gene pattern in burn center was icaA+fib+eno. The frequency of icaD, fib and ebpS was higher in clinical samples than nasal samples. No relation was found between the MSCRAMMs genes in the burn unit and non-burn units. CONCLUSION: The high prevalence of MRSA in burn center can be a new challenge for clinicians. The higher frequency of icaD, fib and ebpS in clinical isolates than nasal isolates may reflect the important role of these genes in colonization and pathogenesis of S. aureus.

Comparison of the Effects of Povidone-Iodine 5%, Polyhexamethylene Biguanide, and Chlorhexidine as a Preoperative Antiseptic in Endophthalmitis Prophylaxis in Patients Undergoing Phacoemulsification Cataract Surgery.

BACKGROUND: This study aims to compare the efficacy and toxicity of povidone-iodine (PI) 5%, polyhexamethylene biguanide (PHMB) 0.02%, and chlorhexidine 0.02% in patients undergoing phacoemulsification cataract surgery. MATERIALS AND METHODS: This single-center, randomized study was done on 330 patients who referred to Feiz hospital in Isfahan and scheduled for cataract surgery. They were assigned randomly to 1 of 3 groups of 110 eyes who received 1 drop of PI 5% in group 1, 1 drop of PHMB 0.02% in group 2 and 1 drop of chlorhexidine 0.02% in group 3. Pre-operative Cultures samples were obtained without any topical application and it was repeated 5 min after use of antiseptic solutions. Cultures were obtained from the inferior conjunctival fornix, using sterile culture swabs while avoiding contact to the eyelids and lashes. RESULTS: The numbers of colony-forming units (CFUs) did not differ significantly among the three groups (P = 0.149 and P = 0.260, respectively). After the intervention, CFUs numbers in the three groups were decreased with a significant difference in both blood and chocolate agars (P = 0.304 and P = 0.136, respectively). Of the 317 eyes, 108 (34.1%) showed no bacterial growth in the pre-preparation period, which was similar in the three groups. Staphylococcus epidermidis was the most common isolated bacteria. Conjunctival injection was significantly different among studied groups (P = 0.0001), five patients in iodine group had severe conjunctival injection and no one in the other group. SPE was significantly fewer in chlorhexidine group than PHMB and iodine groups (P = 0.0001). CONCLUSION: Pretreatment with 5% Povidone-Iodine (PVI) for at least 15 min or repeated applications over 10 min is effective in the reduction of conjunctival organisms, and results in less postoperative endophthalmitis.

Molecular Assessment of Resistance to Clarithromycin in Helicobacter pylori Strains Isolated from Patients with Dyspepsia by Fluorescent In Situ Hybridization in the Center of Iran.

BACKGROUND AND AIMS: Helicobacter pylori is a common infectious bacterium mostly found in gastroduodenal diseases. The increased prevalence of clarithromycin-resistant H. pylori strains is a major challenge in the successful treatment of infections caused by this organism. The present study is aimed at detecting the clarithromycin resistance pattern of H. pylori strains isolated from gastric biopsies and evaluating point mutations of the 23S rRNA gene. Patients and methods. In the present descriptive cross-sectional study, 165 patients with gastrointestinal disorders, who were referred to the Endoscopy Center of Dr. Shariati Hospital of Isfahan, Iran, were enrolled from April to July 2018. H. pylori infection was diagnosed by culture, and susceptibility of the isolates to clarithromycin was assessed by the E-test. Minimum inhibitory concentration (MIC) values were obtained based on EUCAST recommendations. Also, fluorescence in situ hybridization (FISH) was used to determine point mutations associated with clarithromycin resistance. RESULTS: By using culturing, H. pylori was isolated from 50.3% (83/165) gastric biopsy specimens. The overall frequency of resistance to clarithromycin was 25.3% (21/83) by the E-test. In the resistance genotypic analysis, 19 isolates had mutations. The prevalence of A2143G and A2144G mutations was 68.4% (13/19) and 31.5% (6/19), respectively. A2143C mutation was not tracked in any isolate. Two isolates with MIC > 0.5 µg/mL had no mutations that could be related to other mechanisms of resistance. CONCLUSION: As presented in the study, the high prevalence of clarithromycin-resistant H. pylori due to point mutations of the 23S rRNA gene indicates the necessity of revising the standard treatment regimen based on antibiotic susceptibility pattern of each region.

Identification and molecular characterization of mycobacteria isolated from animal sources in a developing country.

The essential role of animals in the transmission of infectious diseases has long been recognized. Apart from zoonosis due to Mycobacterium bovis in domestic cattle, acquired mycobacterial zoonosis from animals are vastly under-reported worldwide. This is partly the result of not recognizing that animals can be the source of zoonotic nontuberculous mycobacteria (NTM) infection. The present study intended to be a contribution to the knowledge of somewhat neglected role of animals in harboring, maintenance and dissemination of NTM in the environment. A total of 326 samples from 250 animals were collected and analyzed for the presence of mycobacteria using standard protocols. The preliminary identification and Runyon's classification of isolates were performed by conventional tests. The PCR amplification of a 228 bp fragment of 65-kDa heat shock protein (hsp) gene was applied for the genus identification and the partial sequence analysis of 16S rRNA was applied for the species identification. In total 32 isolates including 26 rapidly growing and 6 slowly growing mycobacteria were recovered from 250 animal samples (12.8%). The isolates recovered from 21 (65.60%) fish, 8 (25%) insects and 3 (9.4%) house cats, dogs and mice. M. fortuitum was the most frequent Mycobacterium spp (13 isolates; 40.6% of all isolates), followed by M. abscessus-chelonae-M. saopaulense group, (5 isolates; 15.6% of all isolates), M. iranicum (3 isolates; 9.4% of all isolates),and M. marinum, M. terrae complex and M. chlorophenolicum (2 isolates each; 18.8% of all isolates), and the single isolates of M. mucogenicum, M. neoaurum, M. conceptionense, M. virginiense, and M. gordonae (5 isolates; 15.6% of all isolates). The current study indicates that a variety of animals can be a permanent or transient source of mycobacterial agents. This ensures the life cycle of the bacteria and the chance of their survival in the environment, which may pose a potential threat to human health.

Investigation of the antibacterial effect of laser irradiation and chemical agent on human oral biofilms contaminated titanium discs.

INTRODUCTION: A main challenge in treatment of peri-implant disease is the effective decontamination of the implant surface. This challenge has always been a problem, associated with the treatment of these diseases with regard to the difficulty in removing and eliminating bacterial biofilm from the surface of dental implants, especially rough surfaces. The aim of this in-vivo study was to evaluate the effect of five different antimicrobial methods in reducing bacteria adhering to titanium surfaces. MATERIALS AND METHODS: In the present in-vivo study, the contaminated discs, except for the negative control group, randomly underwent one of five treatments: Erbium: Yattrium Aluminum Garnet (Er-YAG) laser, plastic curette, 0.12% chlorhexidine, aPDT, and 810 nm diode laser. A pectrophotometer was used to measure Optical Density (OD) in case of aerobic microorganisms. Colony-Forming Units (CFUs) were used for anaerobic bacteria. Then, all the analyses were carried out at a significance level of α = 0.05 through SPSS software. FINDINGS: One-way analysis of variance (ANOVA) of aerobic bacteria showed a significant difference among 6 groups in terms of OD variations during a 0-24 h time interval (P < 0.001). The results of Kruskal-Wallis test were used to investigate the effect of study methods on anaerobic bacteria after 48 h, and the results showed a significant difference among 6 groups in terms of CFUs (P < 0.001). CONCLUSION: The results of the present study showed that all five mechanicals (plastic curette), chemical (CHX), laser (810 nm diode and Er: YAG), and aPDT methods could reduce oral biofilms from roughed surfaces of titanium discs. Er: YAG laser and plastic curette had the highest and the lowest effects respectively.

Clonal Diversity in Multi Drug Resistant (MDR) Enterococci Isolated from Fecal Normal Flora.

Enterococci are Gram positive and catalase- negative cocci that are found in the gastrointestinal tract of mammals and birds, and are readily isolated from soil, surface and waters. The aim of this study was to discriminate between Enterococcus isolates based on repetitive element sequence based -PCR (Rep-PCR) with the BOXA2R primer and their antibiotics profile. Enterococci isolates were obtained from 180 fecal samples. The isolates were identified by biochemical reaction and specific identification was confirmed by PCR with species specific primers. All isolates were subjected to Rep typing and antimicrobial susceptibility tests. Rep-PCR analysis of 180 isolates revealed 93 REP types with forty-five single types (ST1 to ST45) and forty-eight common types (CT1 to 48). Antibiotic susceptibility tests exhibited that 53 (29.4%), 43 (23.8%), 11 (6.1%) and 9 (5%) were resistant to erythromycin, tetracycline, gentamicin and ciprofloxacin respectively but among the isolates, sixteen were multi drug resistant (MDR). These MDR isolates showed 11 Rep types with seven single types and four common types. In addition, 81.2% of MDR isolates were from male subjects and the average age of these persons was more than fifty years. This study showed that 56.2% of MDR isolates were homogeneous with 95 % similarity, and high rate of resistance to tetracycline and erythromycin (81.2%) were observed in these isolates. The concern about these normal flora isolates are the pathogenic potential of these bacteria through the horizontal transfer of antibiotic resistance and virulence genes.

PCR-based identification of methicillin-resistant Staphylococcus aureus strains and their antibiotic resistance profiles.

OBJECTIVE: To evaluated the PCR for mecA gene compared with the conventional oxacillin disk diffusion method for methicillin-resistant Staphylococcus aureus (S. aureus) identification. METHODS: A total of 292 S. aureus strains were isolated from various clinical specimens obtained from hospitalized patients. Susceptibility test to several antimicrobial agents was performed by disk diffusion agar according to Clinical and Laboratory Standards Institute guidelines. The PCR amplification of the mecA gene was carried out in all the clinical isolates. RESULTS: Among antibiotics used in our study, penicillin showed the least anti-staphylococcal activity and vancomycin was the most effective. The rate of methicillin-resistant S. aureus prevalence determined by oxacillin disk diffusion method was 47.6%; whereas, 45.1% of S. aureus isolates were mecA- positive in the PCR assay. CONCLUSIONS: This study is suggestive that the PCR for detection of mecA gene is a fast, accurate and valuable diagnostic tool, particularly in hospitals in areas where methicillin-resistant S. aureus is endemic.

The comparison of Staphylococcus aureus types 5 and 8 with respect to methicillin resistance in patients admitted to Al-Zahra Hospital by PCR.

BACKGROUND: Staphylococcus aureus is a common human pathogen in community- and hospital-acquired infection, and its capsule is involved in pathogenesis. The predominance of 2 capsular polysaccharides types 5 and 8, on the surface of clinical isolates, led to the development of conjugate vaccine (Staph VAX) based on capsular polysacchrides types 5 and 8 conjugated to a carrier protein. The aim of this study was to determine the prevalence of capsular polysaccharides types 5 and 8 Staphylococcus aureus strains among isolates and their comparison with respect to methicillin resistance. MATERIALS AND METHODS: We studied the capsular genotypes of 193 isolates that encompassed both hospital- and community-acquired infection in Al-Zahra Hospital of Isfahan city from 2008 to 2009. Cap5 and 8 genes were detected by PCR method. Methicillin resistance was determined by PCR (mecA) and disk diffusion methods as well. RESULT: In this population (193 cases), most of the clinical isolates (73%) expressed capsular polysaccharide type 5 (24%) and 8 (49%), whereas 27% were non-typeable. The prevalence of MRSA in type 8 was 67.9%, whereas MRSA isolates in the capsular genotype 5 were 22.2%. CONCLUSION: This study Staphylococcus aureus confirms that the prevalence of capsular polysaccharide types (5 and 8) are predominant, and Staphylococcus aureus type 8 is more resistant to methicillin compared to type 5.