Methods: Whole-genome sequencing (WGS) was carried out on 13 CrAb isolates from clinical samples in Malaysia from 2011 to 2016.
Results: Endotracheal aspirate was the dominant clinical sample source (n = 6), and only one isolate was obtained from wound swab. A total of 6 sequence types (STs) of the Oxford scheme were identified, including 4 reported STs and 2 novel STs. Eleven isolates were classified into clonal complex 92 (CC92/ICII), among which ST195 and ST208 were the most prevalent STs. All 13 CrAb isolates harbored multiple β-lactamase genes. blaOXA-23 (n = 13) and blaOXA-66 (n = 11) were the dominant carbapenemase gene families found in these isolates. All isolates harbor blaADC, blaOXA-51-like, and blaOXA-23-like genes. blaTEM (n = 7), blaNDM-1 (n = 3), blaCARB-8 (n = 1), and blaPER-3 (n = 1) are amongst other β-lactamase genes found in this study. ISAba1 was found upstream to blaOXA-23 (n = 13), blaOXA-66 (n = 1), and blaADC (n = 11). All blaNDM-1 isolates had ISAba125 (mobile genetic element) upstream to the genes. All isolates were positive for Tn2006/2008 and Tn2009 but were negative for Tn2007.
Conclusion: Most of the isolates were grouped under the CC92 clonal complex which belongs to international clonal lineage 2. These findings predict that carriage of carbapenem-resistant genes possibly constitutes the underlying basis of high level of international clone II prevalence. Therefore, molecular surveillance and antimicrobial stewardship are essential in implementing policies to prevent and control the spread of CrAb in hospital settings.
PATIENTS AND METHODS: Ninety-seven clinical strains of T. marneffei were received from various Malaysian hospitals from the year 2020 until 2022. Their identities were determined using microscopic, macroscopic and molecular methods. Next, the susceptibility of yeast and mold forms of each isolate against amphotericin B, itraconazole, voriconazole, posaconazole, ketoconazole, isavuconazole, terbinafine, caspofungin and micafungin were tested according to the broth microdilution according to the Clinical and Laboratory Standards Institute (CLSI) M38 and M27 guidelines. The geometric means of minimal inhibitory concentration (GM MIC), MIC50, and MIC90 were determined for each antifungal. Additionally, Wilcoxon signed-rank test was used to compare the significant difference of GM MICs for each antifungal, GM MIC, MIC50 and MIC90 for the combined nine antifungals against different growth forms of T. marneffei. The significance was set at p<0.05.
RESULTS: Micafungin had the highest GM MIC, MIC50 and MIC90 for mold form of T. marneffei. For yeast form, amphotericin B achieved the highest GM MIC and MIC50 while micafungin achieved the highest MIC90. However, the GM MIC, MIC50 and MIC90 of terbinafine and azole antifungals on T. marneffei were similar to each other, namely between 0.03 and 0.60µg/mL. The difference of GM MIC of all tested antifungals except caspofungin and micafungin was insignificant. Overall, GM MIC, MIC50 and MIC90 of the combined nine antifungals against two growth forms were insignificant.
CONCLUSION: The findings suggested either yeast or mold form can be used in the susceptibility testing of T. marneffei against amphotericin B, itraconazole, voriconazole, posaconazole, ketoconazole, isavuconazole and terbinafine.
METHODS: Thirty-six mandibular premolar teeth with an average surface area of 64.49 mm2 were prepared to receive CAM/CAM fabricated endocrowns. Samples were divided randomly and equally into groups of lithium disilicate with 2 mm intracoronal depth (LD2), lithium disilicate with 4 mm intracoronal depth (LD4), polymer infiltrated ceramic network with 2 mm intracoronal depth (PICN2) and polymer infiltrated ceramic network with 4 mm intracoronal depth (PICN4). All endocrowns were cemented using ParaCore resin cement with 14N pressure and cured for 20 seconds. Fifty measurements of absolute marginal discrepancy (AMD) were done using a stereomicroscope after cementation. After 24 hours, all samples were subjected to thermocycling before the retention test. This involved using a universal testing machine with a crosshead speed of 0.5 mm/min and applying a load of 500N. The maximum force to detach the crown was recorded in newtons and the mode of failure was identified.
RESULTS: Two-way ANOVA revealed that the AMD for PICN was statistically significantly better than lithium disilicate (p=0.01). No statistically significant difference was detected in the AMD between the two intracoronal depths (p=0.72). PICN and endocrowns with 4 mm intracoronal depth had statistically significant better retention (p<0.05). 72.22% of the sample suffered from cohesive failures and 10 LD endocrowns suffered adhesive failures.
CONCLUSIONS: Within the limitations of this study, we found that different materials and intracoronal depths can indeed influence the retention of CAD/CAM fabricated endocrowns. Based on the controlled setting findings, PICN was found to have better retention and better marginal adaptation than similar lithium disilicate premolar endocrowns.