Department of Biomedical Dental Science, Microbiology and Immunology Division, Collage of Dentistry, Imam Abdulrahman bin Faisal University, Dammam, Saudi Arabia
Dr. Doaa AlEraky
Department of Biomedical Dental Science, Microbiology and Immunology Division, Collage of Dentistry, Imam Abdulrahman bin Faisal University, Dammam, Saudi Arabia
Candida auris is a globally-emerging pathogen that is correlated to nosocomial infections and high mortality rates, causing major outbreaks in hospitals and serious public health concerns worldwide. This study investigated the antifungal activity of silver nanoparticles (AgNPs) on clinical isolates of C. auris. A total of eight clinical isolates were collected from blood, urine, ear swab, and groin. C. auris was confirmed by MALDI-TOF MS, and gene sequencing. All isolates confirmed as C. auris were subjected to antimicrobial agents, including amphotericin B, fluconazole, caspofungin, voriconazole, micafungin, and flucytosine. A serial dilution of a silver nanoparticles solution was prepared to test antifungal susceptibility testing under planktonic conditions. Moreover, an antibiofilm activity assay was determined using a colony-forming assay and a cell viability assay by a live–dead yeast kit. Significant antifungal and antibiofilm activity of AgNPs was detected against all isolates; MIC was <6.25 μg/mL, the range of MFC was from 6.25 to 12.5 μg/mL for all isolates, and the highest value of IC50 was 3.2 μg/mL. Silver nanomaterials could represent a possible antimicrobial agent to prevent outbreaks caused by C. auris infections.
Candida auris is an emerging multi-drug resistant pathogen with high mortality rate; nosocomial infections have been reported worldwide, causing a major challenge for clinicians and microbiological laboratories. The study aims to describe new cases of C. auris and detect drug resistance-associated mutations of C. auris by the sequencing of ERG11 and FKS1 genes. A total of six specimens were collected from blood, urine, ear swab, and groin screening samples. Isolates were incubated for 48 h on Sabouraud Dextrose agar (SDA) at 42 °C, then confirmed by MALDI-TOF MS. Furthermore, antifungal susceptibility testing was performed using the Vitek 2 system to detect Minimum Inhibitory Concentrations (MICs) of six antifungals. Sequences of 18S rRNA gene and ITS regions from isolates and phylogenetic analysis were performed. Gene sequencing was analysed to detect drug resistance-associated mutations by FKS1 and ERG11 genes sequencing. All C. auris isolates were confirmed by MALDI-TOF MS, and evolutionary analyses using sequences of 18S rRNA gene and ITS region. Antifungal susceptibility testing showed that all isolates were resistant to fluconazole. Sequencing of ERG11 and FKS1 genes from the isolates revealed the presence of two (F132Y and K143R) drug resistance-associated mutations in ERG11, however, FKS1 gene was devoid of mutations. The study sheds light on a public health threat of an emerging pathogen, and the hospital implemented strict contact screening and infection control precautions to prevent C. auris infection. Finally, there is a critical need to monitor the antifungal resistance in different geographical areas and implementation of efficient guidelines for treatment.
Poly(methyl methacrylate) (PMMA) is a commonly used material, as it is biocompatible and relatively cheap. However, its mechanical properties and weak antibiofilm activity are major concerns. With the development of new technology, 3D-printed resins are emerging as replacements for PMMA. Few studies have investigated the antibiofilm activity of 3D-printed resins. Therefore, this study aimed to investigate the antibiofilm activity and surface roughness of a 3D-printed denture base resin modified with different concentrations of zirconium dioxide nanoparticles (ZrO2 NPs). A total of 60 resin disc specimens (15 × 2 mm) were fabricated and divided into six groups (n = 10). The groups comprised a heat-polymerized resin (PMMA) group, an unmodified 3D-printed resin (NextDent) group, and four 3D-printed resin groups that were modified with ZrO2 NPs at various concentrations (0.5 wt%, 1 wt%, 3 wt%, and 5 wt%). All specimens were polished using a conventional method and then placed in a thermocycler machine for 5000 cycles. Surface roughness (Ra, µm) was measured using a non-contact profilometer. The adhesion of Candida albicans (C. albicans) was measured using a fungal adhesion assay that consisted of a colony forming unit assay and a cell proliferation assay. The data were analyzed using Shapiro–Wilk and Kruskal–Wallis tests. A Mann–Whitney U test was used for pairwise comparison, and p-values of less than 0.05 were considered statistically significant. The lowest Ra value (0.88 ± 0.087 µm) was recorded for the PMMA group. In comparison to the PMMA group, the 3% ZrO2 NPs 3D-printed group showed a significant increase in Ra (p < 0.025). For the 3D-printed resins, significant differences were found between the groups with 0% vs. 3% ZrO2 NPs and 3% vs. 5% ZrO2 NPs (p < 0.025). The highest Ra value (0.96 ± 0.06 µm) was recorded for the 3% ZrO2 NPs group, and the lowest Ra values (0.91 ± 0.03 µm) were recorded for the 0.5% and 5% ZrO2 NPs groups. In terms of antifungal activity, the cell proliferation assay showed a significant decrease in the C. albicans count for the 0.5% ZrO2 NPs group when compared with PMMA and all other groups of 3D-printed resins. The group with the lowest concentration of ZrO2 NPs (0.5%) showed the lowest level of C. albicans adhesion of all the tested groups and showed the lowest Candida count (0.29 ± 0.03). The addition of ZrO2 NPs in low concentrations did not affect the surface roughness of the 3D-printed resins. These 3D-printed resins with low concentrations of nanocomposites could be used as possible materials for the prevention and treatment of denture stomatitis, due to their antibiofilm activities.
Clostridium perfringens is a spore-forming, Gram-positive anaerobic pathogen that causes several disorders in humans and animals. A multidrug-resistant Clostridium strain was isolated from the fecal sample of a patient who was clinically suspected of gastrointestinal infection and had a recent history of antibiotic exposure and diarrhea. The strain was identified by 16s rRNA sequencing as Clostridium perfringens. The strain’s pathogenesis was analyzed through its complete genome, specifically antimicrobial resistance-related genes. The Clostridium perfringens IRMC2505A genome contains 19 (Alr, Ddl, dxr, EF-G, EF-Tu, folA, Dfr, folP, gyrA, gyrB, Iso-tRNA, kasA, MurA, rho, rpoB, rpoC, S10p, and S12p) antibiotic-susceptible genetic species according to the k-mer-based detection of antimicrobial resistance genes. Genome mapping using CARD and VFDB databases revealed significant (p-value = 1 × 10−26) genes with aligned reads against antibiotic-resistant genes or virulence factors, including phospholipase C, perfringolysin O, collagenase, hyaluronidase, alpha-clostripain, exo-alpha-sialidase, and sialidase activity. In conclusion, this is the first report on C. perfringens from Saudi Arabia that conducted whole genome sequencing of IRMC2505A and confirmed the strain as an MDR bacterium with several virulence factors. Developing control strategies requires a detailed understanding of the epidemiology of C. perfringens, its virulence factors, and regional antimicrobial resistance patterns.
23 August 2021 in Saudi Journal of Biological Sciences
This study aims to assess differences in biofilm bacterial composition between patients with low and high caries. Patients without a medical problem and with no history of antibiotic use, mouth wash or fluoride application in the previous 3 months were recruited. Caries was recorded at cavitation level; score was calculated by a national mean (dmft of 4.8 and DMFT of 2.7). Pooled biofilm samples were collected from mesial, distal, buccal, lingual, and occlusal surfaces. Based on caries experience, individuals were classified into low and high caries and both groups were compared regarding bacteria identified using 16S rRNA gene sequencing, and molecular phylogenetic analysis of the isolates was performed. A total of twenty seven randomly selected samples with low (n = 13) and high (n = 14) caries. Identification of oral bacteria was performed using 16S rRNA sequence, Rothia mucilaginosa and R. aeria were identified in low caries individuals, while R. dentocariosa was detected in high caries individuals. Two Streptococcus spp. were identified only in low caries S. salivarius and S. gordonii whereas S. sanguinis, S. mitis, S. sinensis, S. rubneri, S. vestibularis, S. cristatus and S. massiliensis were identified only in individuals with high caries. This study revealed the absence of R. mucilaginosa in the high caries subjects and its coexistence with the low caries subjects. Streptococcus mutans was insignificant contributor of caries among samples, while, Streptococcus sanguinis was the main constituent of high caries Saudi patients.
Objective. To evaluate the antimicrobial effects of different concentrations of zirconium dioxide nanoparticles (nano-ZrO2) reinforcement of poly(methyl) methacrylate (PMMA) on surface roughness and C. albicans biofilm. Methods. 20 heat-polymerized acrylic resin discs were conventionally made and divided into 4 groups (n = 5) according to nano-ZrO2 concentration: control (0% filler) and 3 experimental groups (2.5% (Z2.5), 5.0% (Z5.0), and 7.5% (Z7.5)). An optical profilometer was used for surface roughness evaluation, followed by Candida adherence assay. Specimens were sterilized, then immersed in cultured yeast (C. albicans), and incubated at 37°C for 48 hours. After that, discs were rinsed before extracting the clustered pellets of Candida. The attached C. albicans was counted using the direct method after spreading on agar media and incubating for 48 hours. Statistical analysis was performed using one-way ANOVA and Tukey’s post hoc test at α = 0.05. Results. Surface roughness was significantly increased with all modified groups compared with control ( ), which showed the lowest roughness value (0.027 ± 0.004 μm). There was no significant difference in the roughness value among reinforced groups (2.5, 5.0, and 7.5%) ( ), with Z7.5 showing the highest roughness value (0.042 ± 0.004 μm). Candida count was reduced as the nano-ZrO2 increased but not significantly ( ). Conclusions. The addition of different concentrations of nano-ZrO2 particles to PMMA increased the surface roughness compared with control; in contrast, insignificant reduction of C. albicans biofilm was detected.
About Me
Doaa Mostafa AlEraky, MSc, MBA, Ph.D, FHEA, is a Microbiologist, Course Director and E-Learning Co-ordinator at the Biomedical Dental Science, College of Dentistry, University of Imam Abdulrahman Al Faisal, Saudi Arabia Current research interests: Oral Microbiology Reverse vaccinology Hospital-Acquired Infections Innovative approaches in Teaching and Assessment of Higher Education.
Doaa Mostafa AlEraky, MSc, MBA, Ph.D, FHEA, is a Microbiologist, Course Director and E-Learning Co-ordinator at the Biomedical Dental Science, College of Dentistry, University of Imam Abdulrahman Al Faisal, Saudi Arabia Current research interests: Oral Microbiology Reverse vaccinology Hospital-Acquired Infections Innovative approaches in Teaching and Assessment of Higher Education.