Iron-tannic acid nano-coating: A promising treatment approach for enhancing Lactococcus lactis antibiotic resistance.

The objective of this study was to explore a novel methodology for the synthesis of nanocoated probiotics following their collection and cultivation under optimized conditions, in light of their significant contribution to human health. Probiotics are instrumental in sustaining immune health by modulating the gastrointestinal microbiota and facilitating digestion. However, the equilibrium they maintain can be adversely affected by antibiotic treatments. It is critical to investigate the vulnerability of probiotics to antibiotics, considering the potential implications. This research aimed to assess whether nanoparticle coating could augment the probiotics' resistance to antibiotic influence. A strain of Lactococcus lactis (L. lactis) was isolated, cultured, and comprehensively characterized utilizing state-of-the-art methodologies, including the VITEK® 2 compact system, VITEK® MS, and 16S rRNA gene sequencing. The nanoparticle coating was performed using iron (III) chloride hexahydrate and tannic acid, followed by an evaluation of the probiotics' resistance to a range of antibiotics. The analysis through scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrated a partial nanoparticle coating of the probiotics, which was further supported by UV/Vis spectroscopy findings, suggesting enhanced resistance to standard antibiotics. The results revealed that this strain possesses a unique protein profile and is genetically similar to strains identified in various other countries. Moreover, nano-encapsulation notably increased the strain's resistance to a spectrum of standard antibiotics, including Benzylpenicillin, Teicoplanin, Oxacillin, Vancomycin, Tetracycline, Rifampicin, Erythromycin, and Clindamycin. These findings imply that nanoparticle-coated probiotics may effectively counteract the detrimental effects of extended antibiotic therapy, thus preserving their viability and beneficial influence on gastrointestinal health.

Evaluation of stability of (1R,2 S)-(-)-2-methylamino-1-phenyl-1-propanol hydrochloride in plasma and urine samples-inoculated with Escherichia coli using high-performance liquid chromatography (HPLC).

The preservation of drug stability in biological evidence during the processes of collection and storage poses a substantial obstacle to the progress of forensic investigations. In conjunction with other constituents, the microorganisms present in the samples play a vital role in this investigation. The present investigation employed the high-performance liquid chromatography (HPLC) technique to assess the stability of (1R,2 S)-(-)-2-methylamino-1-phenyl-1-propanol hydrochloride in plasma and urine samples that were inoculated with Escherichia coli. These samples were subjected to storage conditions of 37 °C for 48 h and - 20 °C for a duration of 6 months. Minimal inhibitory concentration (MIC) and Minimal bactericidal concentration (MBC) of MPPH against E. coli were determined using microdilution method. The stability of MPPH in plasma and urine samples inoculated with E. coli was investigated using HPLC method. The results showed the MIC and MBC of MPPH were 87.5 ± 25 ppm and 175 ± 50 ppm, respectively. While MPPH remained stable in plasma for 48 h at 37 °C, it showed a notable decrease of about 11% in stability when stored in urine for the same period and temperature. From the beginning of the first month, a decrease in the stability of the compound appeared in all samples that were stored at - 20 °C, and the decrease reached 7% for plasma samples and about 11% for urine samples. The decrease in the stability reached its peak in the sixth month, reaching more than 30% and 70% of plasma and urine samples preserved at - 20 °C. This work concluded that E. coli can negatively affect the stability of MPPH in plasma and urine samples. This may lead to incorrect conclusions regarding the analysis of biological samples in criminal cases.

Synthesis of Schiff Bases Based on Chitosan and Heterocyclic Moiety: Evaluation of Antimicrobial Activity.

Schiff bases of chitosan (CS) were prepared by reaction of four different heterocyclic compounds, namely, 1,3-dimethyl-2,4,6-trioxohexahydropyrimidine-5-carbaldehyde (M1), 3-acetyl-2H-chromen-2-one (M2), 5-chloro-3-methyl-1-phenyl-1H-pyrazole-4-carbaldehyde (M3), and 4-oxo-4H-chromene-3-carbaldehyde (M4), with CS using thermal and ultrasound approaches. CS Schiff base formation was confirmed by using FT-IR, XRD, and TGA. Characteristic data show that amino groups in chitosan reacted with the functional group in the heterocyclic compound to form the Schiff base. CS Schiff bases show thermal stability more than pure CS. The antimicrobial activity of Schiff bases was tested against +ve Gram bacteria and -ve Gram bacteria. The result shows that Schiff bases prepared by temperature and ultrasound methods possess high antimicrobial activity against +ve Gram bacteria and -ve Gram bacteria; in comparison, Schiff bases produced by the ultrasound method have higher antimicrobial activity. The Schiff base (CSM4U), prepared by the ultrasound method by reaction of CS with 4-oxo-4H-chromene-3-carbaldehyde, exhibited higher antimicrobial activity than Gentamicin as an antibacterial agent. The inhibition range caused by CSM4U was between 19 and 27 mm. Moreover, CSM4U also acted as an antifungal agent, causing an inhibition zone of 21 mm for both Candida albicans and Candida tropicalis, which was higher than that of Terbinafine.

The transmutation of Escherichia coli ATCC 25922 to small colony variants (SCVs) E. coli strain as a result of exposure to gentamicin.

BACKGROUND: Small colony variants (SCVs) are biotypes of bacteria that have a size of approximately one-tenth or less of the wild types and has distinct characteristics comparing to the wild type strains. Clinical SCVs are usually associated with persistent infection and require a long-term treatment program with antibiotics. In Saudi Arabia, there are few studies about SCVs Escherichia coli for this reason, this study is aimed to investigate the ability of gentamicin to mutate E. coli ATCC 25922 to produce small SCVs and investigate the genotypes and phenotypes changes and stress tolerance comparing to clinical SCVs E. coli and normal clinical E. coli Isolated from blood samples. METHODS: In this investigation, four clinical blood samples were collected ted from patients and the cultivation and isolation were carried out in KFMC between December 2019 and February 2021. The identification of positive blood culture samples was done using phoenix MD. Non-SCV E. coli ATCC25922 were mutated to SCV using exposure to increasing gradual concentrations of gentamicin at 100-generation intervals. Biochemical features and susceptibility to standard antibiotics using automated Phoenix MD 50 and. The survival assays were done using several stresses including heat shock, low pH, high osmotic pressure, and oxidative pressure. Virulence genes screening included the detection of genes that encoded to α-haemolysin, CS12 fimbriae, F17-like fimbrial adhesion, P-related fimbriae, yersiniabactin siderophore system, P-fimbriae, aerobactin, iron-regulated genes using PCR and gel electrophoresis. RESULTS: The data from the mutating E. coli ATCC 25922 small colony test with gentamicin revealed that the first emergence of the multidrug resistance (MDR) SCV E. coli strain occurred at generation number 250, corresponding to a gentamicin concentration of 57 g/ml. Pathogenicity islands detection revealed that all tested E. coli strains have PAI IV 536 genes on their chromosomes furthermore, mutated SCV E. coli ATCC 25922 acquired PAII CFT073 and PAI IV 536. Survival tests showed no significant differences changes in tolerance of mutated SCVs comparing to parental strain. CONCLUSION: The present work concluded that gentamicin sub-MIC concentration gradual exposure can induce mutation responsible for SCV formation and evolving of MDR E. coli strains. The mutated SCVs evolved high-level aminoglycoside resistance for gentamicin and resistance to amikacin, it also developed resistance to 2 cephalosporin antibiotics cefuroxime, and cephalothin and a resistance to aztreonam.

Prevalence of Multidrug-Resistant Bacteria in Healthcare-Associated Bloodstream Infections at Hospitals in Riyadh, Saudi Arabia.

Bloodstream infection (BSI) prevalence in hospitalized patients has increased owing to the spread of antibiotic-resistant pathogens; moreover, antimicrobial resistance in bacteria is a global problem. Here, BSIs are investigated in several patients at a hospital in Saudi Arabia, and the resistance of bacterial isolates to widely used drugs is determined. Throughout 2020, bacteria isolated from patients were identified and subjected to antibiotic susceptibility testing. In total, 1125 bacterial isolates were obtained from 1039 patients; among them, gram-positive bacteria were significantly more abundant than gram-negative bacteria. The most prevalent bacteria were Staphylococcus epidermidis and Klebsiella pneumoniae. Notably, gram-negative bacteria were mainly isolated from adult patients, and 20.63% of the gram-positive isolates were from pediatric patients, which was significantly higher than the corresponding percentages in elders and adults. The gram-positive isolates were mainly resistant to cephalothin, oxacillin, amoxicillin-clavulanate, and erythromycin and susceptible to penicillin, gentamicin, ciprofloxacin, and vancomycin. Additionally, the gram-negative isolates were mainly resistant to ampicillin, cephalothin, and amoxicillin-clavulanate and susceptible to amikacin, ertapenem, aztreonam, colistin, and trimethoprim-sulfamethoxazole. Consequently, the high prevalence of infective multidrug-resistant bacteria may account as a significant health issue; it is considered a hazard in Riyadh hospitals and must be prevented at all costs.

Stability of (1R,2S)-(-)-Ephedrine hydrochloride in Candida albicans-inoculated urine and blood samples.

The stability of drugs in biological evidence during collection and storage is of particular concern in forensic investigations. Microbes in the samples and other elements are an essential component of these investigations. In the current study, the HPLC method was used to examine the stability of (1R, 2S)-(-)-Ephedrine hydrochloride in plasma and urine samples inoculated with C. albicans after storage at 37 °C for 48 h and -20 °C for six months. In the stability experiment, MIC50% of (1R, 2S)-(-)-Ephedrine hydrochloride was applied according to MIC and MFC that were determined in this work. This drug had MIC and MFC of 50 and 100 ppm, respectively. In HPLC analysis, the standard (1R, 2S)-(-)-Ephedrine hydrochloride had a retention time of 1.63 and was used to identify this drug in samples that had or had not been exposed to C. albicans. The findings demonstrated that within 48 h at 37 °C, C. albicans had an impact on the drug concentration (10% and more than 15%, respectively, were lost in plasma and urine samples inoculated with C. albicans). In plasma samples, the drug content remained stable at -20 °C for three months, although it degraded in urine samples after one month. In plasma and urine samples, the concentration reduction had surpassed 70% and 50% by the sixth month, respectively. The results of this investigation show that C. albicans can change the stability of (1R, 2S)-(-)-Ephedrine hydrochloride in plasma and urine samples that contain MIC50% of Ephedrine hydrochloride.

Biosynthesis of silver nanoparticles (Ag-NPs) using Senna alexandrina grown in Saudi Arabia and their bioactivity against multidrug-resistant pathogens and cancer cells.

There is no doubt that the risk of drug-resistant pathogens and cancer diseases is on the rise. So, the goal of this study was to find out how effective silver nanoparticles (Ag-NPs) made by Senna alexandrina are at fighting these threats. In this work, S. alexandrina collected from Medina, Saudi Arabia was used and the biosynthesis method was applied to produce the Ag-NPs. The characterization of Ag-NPs was done using different analytical techniques, including UV spectroscopy, FT-IR, TEM, and XRD analysis. The MIC, MBC, and MTT protocols were applied to confirm the bioactivity of the Ag-NPs as antibacterial and anticancer bioagents. The findings reported indicating that the aqueous extract of S. alexandrina leaves, grown naturally in Saudi Arabia, is ideal for the production of bioactive Ag-NPs. The hydroxyl, aliphatic, alkene, N-H bend of primary amines, C-H bonds, and C-O bonds of alcohol were detected in this product. The small, sphere-shaped particles (4-7 nm) were the most prevalent among the bioactive Ag-NPs produced in this work. These nanoparticles inhibited some important multidrug-resistant pathogens (MDRPs) (Escherichia coli, Acinetobacter baumanii/haemolyticus, Staphylococcus epidermidis, and Methicillin-resistant Staphylococcus aureus (MRSA)), as well as their ability to inhibit breast cancer cells (MCF-7 cells). The MIC of Ag-NPs ranged from 0.03 to 0.6 mg/mL, while their MBC ranged from 0.06 to 2.5 mg/mL. Anticancer activity test showed that IC50 of the Ag-NPs against tested breast cancer cells was 61.9 ± 3.8 µg/mL. According to the current results, biosynthesis using S. alexandrina leaves grown naturally in Saudi Arabia was an ideal technique for producing bioactive Ag-NPs that could be used to combat a variety of MDRPs and cancer diseases.

High molecular weight hyaluronic acid reduces the growth and biofilm formation of the oral pathogen Porphyromonas gingivalis.

Background: Porphyromonas gingivalis (P. gingivalis) is viewed as a keystone microorganism in the pathogenesis of periodontal and peri-implant diseases. Hyaluronic acid (HA) is believed to exert antimicrobial activity. The aim of this study is to assess the in-vitro growth and biofilm formation of P. gingivalis under HA and compare the effect of HA to that of azithromycin (AZM) and chlorhexidine (CHX). Materials and methods: In each material, the minimum inhibitory concentration (MIC), 50% MIC, 25% MIC, and 12.5% MIC were tested. The growth of P. gingivalis was evaluated by absorbance spectrophotometry after 48 h. A biofilm inhibition assay was performed on a 72-hour culture by washing planktonic bacterial cells, fixing and staining adherent cells, and measuring the variation in stain concentrations relative to the untreated control using absorbance spectrophotometry. Results: The results show that the overall growth of P. gingivalis after 48 h was 0.048 ± 0.030, 0.008 ± 0.013, and 0.073 ± 0.071 under HA, AZM, and CHX, respectively, while the untreated control reached 0.236 ± 0.039. HA was also able to significantly reduce the biofilm formation of P. gingivalis by 64.30 % ± 22.39, while AZM and CHX reduced biofilm formation by 91.16 %±12.58 and 88.35 %±17.11, respectively. Conclusions: High molecular-weight HA significantly inhibited the growth of P. gingivalis. The overall effect of HA on the growth of P. gingivalis was similar to that of CHX but less than that of AZM. HA was also able to significantly reduce the biofilm formation of P. gingivalis. However, the ability of HA to prevent the biofilm formation of P. gingivalis was generally less than that of both AZM and CHX.

Biochemical Profile by GC-MS of Fungal Biomass Produced from the Ascospores of Tirmania nivea as a Natural Renewable Resource.

The edible fruiting bodies of desert truffles are seasonally collected and consumed in many regions of the world. Although they are very expensive, they are bought and sold as a result of considerable scientific reports confirming their health and nutritional benefits. This study aimed to conduct laboratory production of the fungal biomass of Tirmania nivea as a natural renewable resource of many active biological compounds using an artificial growth medium. The T. nivea collected from Hafar Al-Batin, which is north of Saudi Arabia, and their ascospores were harvested and used to produce fungal biomass in potato dextrose broth. The cultivation was conducted using a shaking incubator at 25 °C for two weeks at 200 rpm. The crud extracts of the fungal biomass and mycelium-free broth were prepared using ethyl acetate, methanol and hexane. Preliminary gas chromatography-mass spectrometry (GC-MS) analysis and their biological activity as antimicrobial agents were investigated. The results showed that the crude extracts have biological activity against mold, yeast and bacteria. The preliminary GC-MS analysis reported that the fungal biomass and extracellular metabolites in the growth medium are industrial renewable resources of several biological compounds that could be used as antifungal, antibacterial, antiviral, anticancer, antioxidant, anti-trypanosomal and anti-inflammatory agents.

Partially purified actinomycetes compounds enhance the intracellular damages in multi-drug resistant P. aeruginosa and K. pneumoniae.

Based on the excellent nutrient level, the current study was focused on isolation and anti-bacterial activity of the actinomycetes from marine mangrove soil samples. As result, 10 different strains of actinomycetes strains were identified on actinomycetes isolation agar plates. The identified strains were shown with white, clear, uncontaminated well matured spore producing ability. Based on the initial observation, the isolated colonies were actinomycetes. The partially extracted crude compound shown excellent anti-bacterial activity against P. aeruginosa and K. pneumoniae with 15 mm and 13 mm zone of inhibitions were observed at 500 µL concentrations. The minimum inhibition concentration result was also confirmed the 500 µL concentration against both the tested concentration with high inhibition rate. Then, the intracellular damages, decreased cell growth of the crude actinomycetes extract treated bacterial strains were clearly observed by confocal laser scanning electron microscope. The extracellular damages of bacterial cell wall and shape of the both the pathogens were clearly shown by scanning electron microscope. Therefore, all the results were clearly supported to the partially extracted crude compound and it has excellent anti-bacterial activity against tested multi drug resistant bacteria.

Isolation and molecular identification of biofilm producing P. aeruginosa and K. pneumoniae from urinary tract infections patient urine sample.

BACKGROUND: Recent years, multi drug resistant pathogens and their pathogenicity were increased worldwide due to unauthorized consumption of antibiotics. In addition, correlation between multi drug resistant bacteria and biofilm formation is heightened due to the production of more virulence behavior. There is no better identification methods are available for detection of biofilm producing gram negative bacteria. MATERIALS AND METHODS: In this research work, multi drug resistant strains of Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) were identified based on the specific antibiotics and third generation cephalosporin discs by disc diffusion assay. Subsequently, biofilm forming ability of selected pathogens were identified tissue culture plate and tube test. Based on the multi-drug resistant ability and biofilm production, the molecular identification of P. aeruginosa and K. pneumoniae were confirmed by PCR using universal primers. RESULTS AND CONCLUSIONS: No zone of inhibition present around the discs of muller hinton agar plates were confirm, selected P. aeruginosa and K. pneumoniae strains were multi drug resistant pathogens. Performed third generation cephalosporin antibiotics were also highly sensitive to selected pathogens of P. aeruginosa and K. pneumoniae. Further, biofilm forming ability of selected P. aeruginosa and K. pneumoniae was confirmed by tissue culture plate and tube methods. Finally, molecular identification of P. aeruginosa and K. pneumoniae was named as P. aeruginosa and K. pneumoniae. Our result was conclude, selected P. aeruginosa and K. pneumoniae as biofilm producing pathogens and also highly resistant to current antibiotics.

Piperacillin/tazobactum and cefotaxime decrease the effect of beta lactamase production in multi-drug resistant K. pneumoniae.

BACKGROUND: Worldwide, multi-drug resistant Klebsiella pneumoniae (K. pneumonia) and their virulence's were contributed more in the multi-drug resistant effect. According to the World Health organization report, it is an emerging thread in developing countries and also comes under first ever critical list. In this context, the current study was concentrated on detection of extended spectrum beta lactamase (ESBL) producing strain and their antimicrobial susceptibility study of K. pneumoniae. MATERIALS AND METHODS: Firstly, the multi-drug resistant effect of the K. pneumoniae was identified from specific CLSI guidelines recommended antibiotics by disc diffusion method. Consecutively, the primary ESBL identification test was performed using ceftazidime and cefotaxime, followed by double disc combination and phenotypic confirmation tests using ceftazidime/clavulanic acid and cefotaxime/clavulanic acid. Finally, the minimum inhibition concentration of some important sensitive antibiotics were performed against selected K. pneumoniae was confirmed by micro broth dilution method. RESULTS AND CONCLUSIONS: The current result was most favorable to selected K. pneumoniae with more multi drug resistant characteristic nature. All the performed antibiotics were almost more sensitive to selected K. pneumoniae. The effective antibiotics of piperacillin was also exhibited more resistant effect against tested bacteria and it cleaved the bacterial enzyme clearly. The present result of primary ESBL identification test result was exhibited with ≤22 mm and ≤27 mm against ceftazidime and cefotaxime were observed respectively. Followed result of double disc combination and phenotypic confirmation experiments results were clearly stated that the selected K. pneumoniae was ESBL producer. The ceftazidime, cefotaxime and ceftazidime/clavulanic acid and cefotaxime/clavulanic acid were exhibited with merged zones and ≥5 mm zones around the combination disc when compared with disc alone were observed. All the ESBL detection test results were clearly indicated that the selected K. pneumoniae strain was ESBL producer.

A synergic action of colistin, imipenem, and silver nanoparticles against pandrug-resistant Acinetobacter baumannii isolated from patients.

BACKGROUND: The upgrowth and rapid prevalence of pandrug-resistant Acinetobacter baumannii strains that have a pathogenic activity to cause several infections are of considerable influence on the health of communities worldwide. No infections by these bacterial strains were recorded before 1998, and currently, the numbers are on the rise. METHODS: The A. baumannii strains were isolated from male and female patients in Medical Microbiology Department, King Fahd Medical City (KFMC) in Riyadh, Saudi Arabia between 1/1/2020 to 29/12/2020. The statistical analysis was performed base on sex, age, source of samples, and response to commercially available antibiotics. The A. baumannii strains that resisted all the antibiotics including colistin and imipenem were selected for the synergic test. RESULTS: The data showed that 62.28%, 77.07% of 342 A. baumannii strains were isolated from males and patients over 35 years of age. A. baumannii strains (pandrug-A. baumannii) that can resist all tested antibiotics were 8.19%. The major source of the A. baumannii isolates was the respiratory system (>50%). Among all isolates (N = 342), azidothymidine-resistant A. baumannii strains were more than 85%. There is a statistically significant difference (P < 0.05) in the number of colistin-resistant A. baumannii strains isolated from males comparing with the female. The combinations of colistin and silver nanoparticles or imipenem and silver nanoparticles resulted in synergistic action led to reduction of MICs of colistin, imipenem, and silver nanoparticles (more than four-fold reduction). Also, the combinations of colistin and imipenem had high synergistic action. CONCLUSION: The pandrug-resistant A. baumannii strains may represent a current and future threat that must be fought, and the synergy action of antibiotics and nanoparticles may be one of the available, rapid, and easy strategies to confront this global problem.

The extreme drug resistance (XDR) Staphylococcus aureus strains among patients: A retrospective study.

The objective of the present work was to observe and profile various antibiotic resistant strains of Staphylococcus aureus and highlight the need for continuous surveillance. Data regarding antibiotic-resistant S. aureus strains isolated and identified at the Medical Microbiology Department, King Khalid Hospital, Riyadh was obtained. Bacterial isolates were collected from several sites of infections in patients and an evaluation of susceptibility were carried out using a fully automated Vitek2 system. Relative frequency (%), odds ratios and Ward's minimum variance were calculated. The results showed that wounds were a source of more than 40% of the S. aureus (MRSA) strains that have ability to resist methicillin, and more than 45% of the methicillin-susceptible S. aureus (non-MRSA) strains. 40% of the isolates were MRSA (N = 251), and all MRSA strains were sensitive to vancomycin, daptomycin, teicoplanin, tigecycline, nitrofurantoin, and itraconazole while all non-MRSA (N = 338) strains were sensitive to vancomycin, cefoxitin, daptomycin, gentamicin, oxacillin, teicoplanin, tigecycline, and mupirocin. Strength of association between antibiotic-resistant S. aureus strains and source of samples (site of infection) was established. The study concluded that S. aureus strains had developed resistance towards 20 (for non-MRSA) and 22 (for MRSA) of the antibiotics tested. All MRSA strains were non-sensitive to amoxicillin/clavulanate, ampicillin cefoxitin, cefazolin, imipenem, oxacillin, and penicillin.

Prevalence of Escherichia coli strains resistance to antibiotics in wound infections and raw milk.

Antibiotic-resistant Escherichia coli strains including extended-spectrum ß-lactamase (ESBL) isolates are globally widespread in medical, food, and environmental sources. Some of these strains are considered the most pathogenic bacteria in humans. The present work examined the predominance of antibiotic resistance in E. coli strains in wound infections comparing with E. coli strains isolated from a raw milk as a potential source of those strains. The wound infections included abdomen, anus, arm, back, buttock, chest, foot, hand, head, leg, lung, mouth, neck, penis, thigh, toe, and vagina infections. In total, 161 and 153 isolates identified as E. coli were obtained from wound infections and raw milk, respectively. A Vitek 2 system innovated by bioMérieux, France was applied to perform the identification and susceptibility tests. The E. coli isolates that have ability to produce ESBL were detected by an ESBL panel and NO45 card (bioMérieux). Over half of the E. coli were from abdomen, back, and buttock wound infections. More than 50%of the E. coli isolates obtained from wound infections were resistant to cefazolin, ampicillin, cefuroxime, ciprofloxacin, mezlocillin, moxifloxacin, piperacillin, and tetracycline; 70% of the isolates from wound infections and 0% of the isolates from raw milk were E. coli isolates produced ESBL. The data showed that the strains resistance to multi-antibiotic and produced ESBL are more widespread among wound infections than in raw milk.

Antibiotic-resistant Staphylococcus epidermidis isolated from patients and healthy students comparing with antibiotic-resistant bacteria isolated from pasteurized milk.

Antibiotic-resistant Staphylococci are a global issue affecting humans, animals, and numerous natural environments. Antibiotic-resistant Staphylococcus epidermidis is an opportunistic pathogen frequently isolated from patients and healthy individuals. This study aimed to examine the antibiotic resistance of S. epidermidis isolated from patients, healthy students and compare the results with antibiotic-resistant bacteria isolated from pasteurized milk. Clinical strain isolation was performed in several hospitals in the Riyadh. Skin swabs from 100 healthy undergraduate candidate students were obtained at King Saud University. The pasteurized milk samples were obtained from local market (company, X). After isolation, identification and susceptibility tests were performed using an automated system. A multiplex tuf gene-based PCR assay was used to confirm identification. Biofilm production and biofilm-related gene expression were studied. S. epidermidis represented 17% of clinical bacterial isolates, and 1.7% of isolates obtained from healthy students were multiantibiotic-resistant. All patient strains were teicoplanin- and vancomycin-susceptible, while all student strains were gentamicin-, levofloxacin-, moxifloxacin-, and trimethoprim/sulfamethoxazole-susceptible. All the bacteria isolated from pasteurized milk were benzylpenicillin and oxacillin-resistant strains. Of the S. epidermidis strains, 91% could produce biofilms, and mecA, icaADBR, ica-ADB, ica-AD, ica-A only, and ica-C only were expressed in 83, 17.1, 25.7, 37.1, 20, and 0% of the strains, respectively. This work demonstrates that S. epidermidis can be accurately identified using a multiplex tuf-based assay, and that multiantibiotic-resistant S. epidermidis strains are widespread amongst patients and healthy students.

Anti-cancer, anti-biofilm, and anti-inflammatory properties of hen's albumen: A photodynamic approach.

The albumen plays a major role in the protection of eggs against microorganisms. It contains an arsenal of natural antimicrobial molecules and antibacterial proteins, including the well-known ovotransferrin and lysozyme, which exert their activities against a range of bacteria. In the present study, the hen's albumen extract treated with the dried insect body of blister beetle M. pustulata was assessed for antibacterial, antibiofilm, anti-inflammatory and anti-proliferative activity. The zone of inhibition against Gram positive E. faecalis and S. aureus was 10.8 mm and 12.1 mm respectively at 100 µg mL-1. However, it was 13.6 mm and 15.3 mm for Gram negative P. aeruginosa and P. vulgaris respectively. The biofilm of tested bacteria was significantly inhibited at 100 µg mL-1. The hydrophobicity of bacterial biofilms was considerably condensed after treatment with the hen's albumen extracts at 100 µg mL-1. The anti-inflammatory activity of hen's albumen extracts was confirmed by the inhibition of cyclooxygenase (COX) enzyme to 84.91% at 100 µg mL-1 with the relative IC50 of 8.26 µg mL-1. The albumen extract effectively inhibited the viability (23.61%) of HepG2 hepatic cancer cells at 100 µg mL-1. The anti-proliferative activity of the albumen extracts was further revealed by the induction of HepG2 apoptotic cell morphology. This study concludes that the hen's albumen extract treated with M. pustulata is a natural therapeutic agent to treat biofilm associated clinical bacteria, inflammations and human hepatic cancer cells.

Correction to: Brevibacillus laterosporus isolated from the digestive tract of honeybees has high antimicrobial activity and promotes growth and productivity of honeybee's colonies.

The original publication of this paper contains a mistake. The correct affiliation no. 3 is shown in this paper.

Brevibacillus laterosporus isolated from the digestive tract of honeybees has high antimicrobial activity and promotes growth and productivity of honeybee's colonies.

The development of novel antimicrobial drugs, as well as the discovery of novel compounds able to promote honeybee's growth, represents major challenges for modern entomology. The main aim of this study was to investigate whether Brevibacillus laterosporus isolated from the digestive tract of Saudi honeybees, Apis mellifera, was able to stimulate colony strength parameters of honeybees and to evaluate its ability to produce antimicrobial agents. Honeybees were collected in Dirab, Riyadh Region, Saudi Arabia, and microorganisms were isolated and identified by 16S ribosomal RNA analysis. Microscopic identification of the microorganism in its native state was facilitated by atomic force microscopy at high-resolution imaging. Active biological compounds were produced by submerged fermentation with B. laterosporus. The fermented broth was subjected to extraction and purification, and then semi-pure compounds were analyzed by gas chromatography-mass spectrometry. The effectiveness of the crude extract and semi-pure compounds as antimicrobial agents was evaluated by susceptibility assays. More than 22% of the microorganisms isolated from the digestive tract of healthy honeybees have been identified as B. laterosporus, this kind of species has a unique shape and morphological structure. The cyclic dipeptide cyclo(Leu-Pro) produced by B. laterosporus showed biological activity against several pathogenic microorganisms. Furthermore, the total counts of workers, closed brood, and open brood, as well as the production of bee pollen and honey, were better in honeybees treated with a B. laterosporus suspension. The data indicated that the B. laterosporus strain isolated from a healthy honeybee might be a novel probiotic and a producer of important biological compounds.

Effects of Piper cubeba L. essential oil on methicillin-resistant Staphylococcus aureus: an AFM and TEM study.

The increasing prevalence of antibiotic-resistant bacteria is creating a real challenge for health care systems worldwide, making the development of novel antibiotics a necessity. In addition to the development of new antibiotics, there is an urgent need for in-depth characterization of the mechanisms of bacterial resistance toward new drugs. Here, we used essential oils extracted in our laboratory from Piper cubeba against methicillin-resistant Staphylococcus aureus ATCC 43300, one of the most prominent antibiotic-resistant bacteria. Effects of the essential oils extracted from P cubeba on bacteria were mainly evaluated using 2 powerful microscopy techniques: atomic force microscopy and transmission electron microscopy. High-resolution atomic force microscopy images of the cells were obtained close to their native environment by immobilizing the cells on porous Polyether sulfone membranes, which were prepared in our laboratory with a wide range and distribution of pore sizes and depth. Inhibition zones (mm) and minimum inhibitory concentrations were determined. Two different concentrations of the oil were used to treat the cells: 50 µg/mL minimum inhibitory concentration and 25 µg/mL. The 50 µg/mL oil solution caused severe damage to the bacterial cells at microscopic levels while the 25 µg/mL solution showed no effects compared to the control. However, at nanoscopic levels, the 25 µg/mL oil solution caused significant changes in the cell wall, which could potentially impair bacterial activities. These results were also confirmed by transmission electron microscopy micrographs. Our results indicate that the extract has a good biological activity against methicillin- and oxacillin-resistant S aureus and that it acts on the cell wall and plasma (cytoplasmic) membrane.