A novel small-molecule compound S-342-3 effectively inhibits the biofilm formation of Staphylococcus aureus.

IMPORTANCE: Biofilms are an important virulence factor in Staphylococcus aureus and are characterized by a structured microbial community consisting of bacterial cells and a secreted extracellular polymeric matrix. Inhibition of biofilm formation is an effective measure to control S. aureus infection. Here, we have synthesized a small molecule compound S-342-3, which exhibits potent inhibition of biofilm formation in both MRSA and MSSA. Further investigations revealed that S-342-3 exerts inhibitory effects on biofilm formation by reducing the production of polysaccharide intercellular adhesin and preventing bacterial adhesion. Our study has confirmed that the inhibitory effect of S-342-3 on biofilm is achieved by downregulating the expression of genes responsible for biofilm formation. In addition, S-342-3 is non-toxic to Galleria mellonella larvae and A549 cells. Consequently, this study demonstrates the efficacy of a biologically safe compound S-342-3 in inhibiting biofilm formation in S. aureus, thereby providing a promising antibiofilm agent for further research.

Polymyxin B and fusidic acid, a novel potent synergistic combination against Klebsiella pneumoniae and Escherichia coli isolates with polymyxin B resistance.

The increasing prevalence of multidrug-resistant (MDR) Gram-negative bacteria and comparatively limited options of antibiotics pose a major threat to public health worldwide. Polymyxin B is the last resort against extensively resistant Gram-negative bacterial infections. However, a large number of Gram-negative bacteria exhibited high-level resistance to Polymyxin B, bringing challenges for antimicrobial chemotherapy. Combination therapies using polymyxins and other antibiotics are recommended to treat multidrug-resistant pathogens. In this study, we selected Gram-negative bacterial strains, including Klebsiella pneumoniae and Escherichia coli, to explore whether fusidic acid and polymyxin B have a synergistic killing effect. Through broth microdilution, we observed that minimum inhibitory concentrations (MICs) against polymyxin B in the isolates tested were significantly reduced by the addition of fusidic acid. Notably, chequerboard analysis indicated a synergistic effect between polymyxin B and fusidic acid. In addition, subsequent time-kill experiments showed that the combination of polymyxin B and fusidic acid was more effective than a single drug in killing bacteria. Finally, our investigation utilizing the murine model revealed a higher survival rate in the combination therapy group compared to the monotherapy group. Our research findings provide evidence of the synergistic effect between polymyxin B and fusidic acid. Fusidic acid was shown to increase the sensitivity of multi-drug resistant E. coli and K. pneumoniae to polymyxin B, thereby enhancing its bactericidal activity. This study provides new insights into a potential strategy for overcoming polymyxin B resistance, however, further investigations are required to evaluate their feasibility in real clinical settings.

Molecular Characteristics and Gonococcal Genetic Island Carrying Status of Thirty-Seven Neisseria gonorrhoeae Isolates in Eastern China.

Purpose: To investigate the carrying situation of N. gonorrhoeae genetic island (GGI), and to understand the existence of GGI of different multilocus sequence types (MLST), so as to provide evidence for epidemiology. Methods: From January 2018 to December 2020, a total of 37 clinical isolates of N. gonorrhoeae were collected. Resistance to tetracycline, ß-lactam, and azithromycin were measured. Genes in GGI (atlA, traG, and traH) were amplified via polymerase chain reaction (PCR). All clinical isolates were subjected to N. gonorrhoeae MLST. Results: The GGI of N. gonorrhoeae were widespread, and the positive detection rates of atlA, traG and traH were all 81.08% (30/37). In this study, atlA, traG and traH were always detected positive together. No significant difference in the positive rate of the GGI between the azithromycin-sensitive and the resistance groups or between the ß-lactam positive and negative groups (P > 0.05) was found; however, there was a significant difference between the high-level tetracycline-resistant group and the non-high-level resistant group (P < 0.05), with the carrier rates being 60.00% and 94.45%, respectively. Among the 37 isolates studied, 12 distinct MLST were determined, while MLST ST8123 occurred most frequently, accounting for 18.91% (7/37), followed by ST1928, ST7367 and ST7822, all 13.51% (5/37). Conclusion: N. gonorrhoeae typed as ST1928, ST1901, ST1588 and ST7822, the GGI were all positive. These four types are more likely to become highly virulent strains.

The Dissemination of Fusidic Acid Resistance Among Staphylococcus epidermidis Clinical Isolates in Wenzhou, China.

Purpose: Fusidic acid (FA), a potent steroidal antibiotic, is used topically to treat skin and soft tissue infections (SSTIs) caused by Staphylococci. The aim of this study is to report the prevalence of fusidic acid resistance among Staphylococcus epidermidis clinical isolates from a tertiary hospital in Wenzhou, east China. Methods: The antibiotic susceptibility of S. epidermidis isolates was determined by disc diffusion method and agar dilution method. Then, FA-resistant S. epidermidis isolates were characterized by multi-locus sequence typing, SCCmec typing and pulsed-field gel electrophoresis. Results: In the present study, the 55 (7.7%) FA-resistant S. epidermidis among 711 S. epidermidis clinical isolates were isolated from different parts of 53 patients. Fifty-five FA-resistant S. epidermidis isolates with FA MIC values ranged from 4 to 32 µg/mL. Among them, 50 (90.9%) were identified as methicillin-resistant Staphylococcus epidermidis (MRSE), in which mecA were positive. Meanwhile, the positive rates of fusB and fusC genes among FA-resistant S. epidermidis isolates were 85.5% (47/55) and 7.3% (4/55), respectively. All 55 isolates mentioned above were susceptible to vancomycin. More than 50% of FA-resistant isolates were resistant to non-ß-lactam antimicrobials including erythromycin (80.0%, 44/55), clindamycin (65.5%, 36/55), ciprofloxacin (63.6%, 35/55) and sulfamethoxazole (63.6%, 35/55). A total of 14 sequence types (STs) were identified among the 55 FA-resistant S. epidermidis isolates, of which, ST2 (24/55, 43.6%) was the most predominant type. And the eBURST analysis showed that CC2, CC5 and CC247 accounted for 43.6% (24/55), 27.3% (15/55) and 14.5% (5/55), respectively. Meanwhile, a total of four SCCmec types (I, III, IV, V) were identified among the 55 FA-resistant S. epidermidis. Furthermore, the pulsed field gel electrophoresis divided the 55 isolates into 20 types, namely A-T. Q-type strains were most prevalent, accounting for 30.9% (17/55). Conclusion: Taken together, the dissemination of S. epidermidis ST2 clone with FA resistance can cause trouble in controlling S. epidermidis infections.

Antimicrobial Resistance And Molecular Characteristics Among Neisseria gonorrhoeae Clinical Isolates In A Chinese Tertiary Hospital.

PURPOSE: The resistance of N. gonorrhoeae to antimicrobial agents has been increasing year by year due to the overuse of antibiotics. The primary aims of the present study were to investigate the molecular characteristics of the clinical isolates of Neisseria gonorrhoeae and the resistance to azithromycin in a Chinese tertiary hospital. METHODS: From January 2014 to May 2017, a total of 55 clinical isolates of N. gonorrhoeae were collected. Genes associated with azithromycin resistance (AZM-R), including mutations in 23S rRNA alleles, the mtrR promoter and coding regions, and rplD and rplV were evaluated by PCR and DNA sequencing. All clinical isolates were subjected to N. gonorrhoeae multiantigen sequence typing (NG-MAST), while the AZM-R isolates were further characterized by multilocus sequence typing (MLST). RESULTS: The AZM-R rate in this study was 23.64% (13/55), and a single (A)-nucleotide deletion mutation in the mtrR promoter region, a G45D mutation in the mtrR coding region, a point mutation in rplD, and an A2047G mutation in 23S rRNA alleles were detected in 13, 4, 3 and 4 isolates, respectively; no mutations were found in rplV. There was no significant difference in the mtrR coding region mutation rate between the azithromycin-sensitive and AZM-R groups (P > 0.05); however, there was a significant difference in the mutation rate of the mtrR promoter region (P < 0.05). Among the 55 isolates studied, 43 distinct NG-MAST were determined, while the AZM-R isolates were allocated into 10 distinct MLST/NG-MAST combinations. All three isolates with high-level AZM-R belonged to the sequence types (STs) NG-MAST ST1866 and MLST ST10899. CONCLUSION: N. gonorrhoeae clinical isolates from Wenzhou, eastern China, showed considerable genetic diversity. Measures should be implemented to monitor the spread of the NG-MAST ST1866 and MLST ST10899 N. gonorrhoeae clones, which exhibit high-level AZM-R in eastern China.

Low prevalence of mupirocin resistance among Staphylococcus aureus clinical isolates from a Chinese tertiary hospital.

Between September 2013 and March 2016, 26 (1.95 %) of 1333 Staphylococcus aureus clinical isolates from a Chinese hospital were found to be resistant to mupirocin, including 18 (1.35 %) with high-level mupirocin resistance and 8 (0.6 %) with low-level mupirocin resistance. Among the 18 isolates with high-level mupirocin resistance, 17 were associated with plasmid-mediated mupA. Meanwhile, the 8 isolates with low-level mupirocin resistance were shown to have a V588F mutation in ileS. A total of 14 sequence types (STs) and 18 spa types were identified. All four isolates with t062 belonged to ST965. Three ST5-MRSA-SCCmec II were linked to t311, which was not previously reported. Furthermore, ST764-MRSA-SCCmec II-t002, exclusively found in Japan before, was identified in this study. In conclusion, we observed relatively low prevalence of mupirocin resistance among S. aureus with considerable heterogeneity in East China. Newly emerging MRSA clones with high-level mupirocin resistance should be of concern.