Objective: To investigate the in vitro interference of cefotaxime at subinhibitory con-centrations [sub-minimal inhibitory concentrations (MIC)] on biofilm formation by nontypeable Haemophilus influenzae (NTHi). Methods: The interference of subinhibitory concentrations of cefotaxime on biofilm formation of the clinical strong-biofilm forming isolates of NTHi was evaluated by a microtiter plate biofilm formation assay. The effect of sub-MIC cefotaxime on bacterial cell-surface hydrophobicity was determined using a standard microbial adhesion to n-hexadecane test. Additionally, the effects on bacterial adherence to human fibronectin and expression of bacterial adhesins were also investigated. Results: Subinhibitory concentrations of cefotaxime, both at 0.1× and 0.5× MIC levels, efficiently reduced the NTHi biofilm formation, and this effect was independent of decreasing bacterial viability. Sub-MIC cefotaxime also decreased bacterial cell-surface hydrophobicity and reduced adherence to human fibronectin. Inhibition in the P2 and P6 gene expressions upon exposure to sub-MIC cefotaxime was also noted. Conclusions: Taken together, our results indicate that sub-MIC cefotaxime interferes with the formation of NTHi biofilm, and this effect is feasibly related to the interference with cell-surface hydrophobicity, fibronectin-binding activity as well as alteration of the P2 and P6 gene expression. The findings of the present study therefore provide a rationale for the use of subinhibitory concentrations of cefotaxime for treatment of NTHi-related diseases.

Objective: To investigate the molecular epidemiology and antimicrobial resistance patterns of methicillin-resistant Staphylococcus aureus (MRSA) among healthcare workers and patients. Methods: MRSA isolates were recovered from nasal swabs collected at a tertiary care hospital of Nepal and confirmed on the basis of Gram staining, conventional biochemical tests, and PCR amplification of mecA gene. PCRs were also used for detection of the different resistance genes and staphylococcal cassette chromosome (SCC) mec types. Antibiotic susceptibility patterns of isolates were assessed by disc diffusion method and minimum inhibitory concentrations were determined by E-test. Results: A total of 29 MRSA were isolated from 536 nasal swabs (5.4%) of health care workers and patients at a tertiary care hospital in Nepal. All isolates were susceptible to amikacin, gentamicin, vancomycin (minimal inhibitory concentrations<2μg/mL), tigecy-cline, tetracycline, nitrofurantoin, rifampicin, quinupristin-dalfopristin, and linezolid. Among the 29 MRSA isolates, resistance to erythromycin (72%), ciprofloxacin (75%), co-trimoxazole (62%), clindamycin (10%), and chloramphenicol (10%) was found, and fifteen isolates (51%) exhibited high-level mupirocin resistance (minimal inhibitory concentrations>1024μg/mL). Fourteen isolates were found harboring the mupA gene and one isolate was found carrying the novel mupB gene. High prevalence (68%) of SCCmec I type was found, followed by SCCmec V (13%) and SCCmec III (3%) among all the MRSA isolates. Conclusions: We found the emergence of SCCmec type Ⅰ with high-level mupirocin resistance among MRSA in Nepal. Data also suggest that MRSA SCCmec type V strain has spread from the community to the hospital.

Objective To investigate the in vitro interference of cefotaxime at subinhibitory concentrations [sub-minimal inhibitory concentrations (MIC)] on biofilm formation by nontypeable Haemophilus influenzae (NTHi). Methods The interference of subinhibitory concentrations of cefotaxime on biofilm formation of the clinical strong-biofilm forming isolates of NTHi was evaluated by a microtiter plate biofilm formation assay. The effect of sub-MIC cefotaxime on bacterial cell-surface hydrophobicity was determined using a standard microbial adhesion to n-hexadecane test. Additionally, the effects on bacterial adherence to human fibronectin and expression of bacterial adhesins were also investigated. Results Subinhibitory concentrations of cefotaxime, both at 0.1× and 0.5× MIC levels, efficiently reduced the NTHi biofilm formation, and this effect was independent of decreasing bacterial viability. Sub-MIC cefotaxime also decreased bacterial cell-surface hydrophobicity and reduced adherence to human fibronectin. Inhibition in the P2 and P6 gene expressions upon exposure to sub-MIC cefotaxime was also noted. Conclusions Taken together, our results indicate that sub-MIC cefotaxime interferes with the formation of NTHi biofilm, and this effect is feasibly related to the interference with cell-surface hydrophobicity, fibronectin-binding activity as well as alteration of the P2 and P6 gene expression. The findings of the present study therefore provide a rationale for the use of subinhibitory concentrations of cefotaxime for treatment of NTHi-related diseases.

Objective To investigate the molecular epidemiology and antimicrobial resistance patterns of methicillin-resistant Staphylococcus aureus (MRSA) among healthcare workers and patients. Methods MRSA isolates were recovered from nasal swabs collected at a tertiary care hospital of Nepal and confirmed on the basis of Gram staining, conventional biochemical tests, and PCR amplification of mecA gene. PCRs were also used for detection of the different resistance genes and staphylococcal cassette chromosome (SCC) mec types. Antibiotic susceptibility patterns of isolates were assessed by disc diffusion method and minimum inhibitory concentrations were determined by E-test. Results A total of 29 MRSA were isolated from 536 nasal swabs (5.4%) of health care workers and patients at a tertiary care hospital in Nepal. All isolates were susceptible to amikacin, gentamicin, vancomycin (minimal inhibitory concentrations < 2 μg/mL), tigecycline, tetracycline, nitrofurantoin, rifampicin, quinupristin-dalfopristin, and linezolid. Among the 29 MRSA isolates, resistance to erythromycin (72%), ciprofloxacin (75%), co-trimoxazole (62%), clindamycin (10%), and chloramphenicol (10%) was found, and fifteen isolates (51%) exhibited high-level mupirocin resistance (minimal inhibitory concentrations > 1 024 μg/mL). Fourteen isolates were found harboring the mupA gene and one isolate was found carrying the novel mupB gene. High prevalence (68%) of SCCmec I type was found, followed by SCCmec V (13%) and SCCmec III (3%) among all the MRSA isolates. Conclusions We found the emergence of SCCmec type I with high-level mupirocin resistance among MRSA in Nepal. Data also suggest that MRSA SCCmec type V strain has spread from the community to the hospital.