In vitro and in vivo antimicrobial potential of lithium complex against multi-drug resistant Acinetobacter baumannii.

IMPORTANCE: Multi-drug resistance (MDR) by virtue of evolving resistance and virulence mechanisms among A. baumannii is a global concern which is responsible for lethal hospital-acquired infections. Therefore, it is crucial to develop new therapeutics against it. Metal complexes are compact structures with diverse mechanisms that the pathogens cannot evade easily which make them a strong drug candidate. In this study, we assessed the in vitro and in vivo efficacy of lithium complex {[Li(phen)2 sal]} against biofilm-forming MDR A. baumannii. The lithium complex displayed strong antimicrobial activity and reduced the pre-formed mature biofilm which is key barrier for antimicrobial action. Moreover, it employs oxidative stress as one of its mode of actions and causes cellular rupturing. Lithium complex was non-toxic and was significantly effective to overcome pneumonia in mice model. These results highlight the untapped potential of metal complexes that can be explored and utilized for combating notorious A. baumannii infections.

Antimicrobial and biofilm inhibiting potential of an amide derivative [N-(2', 4'-dinitrophenyl)-3ß-hydroxyurs-12-en-28-carbonamide] of ursolic acid by modulating membrane potential and quorum sensing against colistin resistant Acinetobacter baumannii.

Acinetobacter baumannii is Gram-negative, an opportunistic pathogen responsible for life-threatening ventilator-associated pneumonia. World Health Organization (WHO) enlisted it as a priority pathogen for which therapeutic options need speculations. Biofilm further benefits this pathogen and aids 100-1000 folds more resistant against antimicrobials and the host immune system. In this study, ursolic acid (1) and its amide derivatives (2-4) explored for their antimicrobial and antibiofilm potential against colistin-resistant A. baumannii (CRAB) reference and clinical strains. Viability, crystal violet, microscopic, and gene expression assays further detailed the active compounds' antimicrobial and biofilm inhibition potential. Compound 4 [N-(2',4'-dinitrophenyl)-3ß-hydroxyurs-12-en-28-carbonamide)], a synthetic amide derivate of ursolic acid significantly inhibits bacterial growth with MIC in the range of 78-156 µg/mL against CRAB isolates. This compound failed to completely kill the CRAB isolates even at 500 µg/mL concentration, suggesting the compound's anti-virulence and bacteriostatic nature. Short and prolonged exposure of 4 inhibited or delayed the bacterial growth at sub MIC, MIC, and 2× MIC, as evident in time-kill and post-antibacterial assay. It significantly inhibited and eradicated >70% of biofilm formation at MIC and sub MIC levels compared to colistin required in high concentrations. Microscopic analysis showed disintegrated biofilm after treatment with the 4 further strengthened its antibiofilm potential. Atomic force microscopy (AFM) hinted the membrane disrupting effect of 4 at MIC's. Further it was confirmed by DiBAC4 using fluorescence-activating cells sorting (FACS), suggesting a depolarized membrane at MIC. Gene expression analysis also supported our data as it showed reduced expression of biofilm-forming (bap) and quorum sensing (abaR) genes after treatment with sub MIC of 4. The results suggest that 4 significantly inhibit bacterial growth and biofilm mode of colistin-resistant A. baumannii. Thus, further studies are required to decipher the complete mechanism of action to develop 4 as a new pharmacophore against A. baumannii.

Flow Injection-High Resolution-Electrospray Ionization-Mass Spectrometry (FI-HR-ESI-MS) Method for the Screening of Antimicrobial Pharmaceutical Drugs and Compounds against Klebsiella pneumoniae.

The development of drug resistant microorganism is a global threat. Therefore, screening of more compounds for antimicrobial potentials is needed. Hence, a rapid method was developed for the screening of antimicrobial drugs and compounds against Klebsiella pneumoniae using Flow Injection Analysis-High Resolution-Mass Spectrometry. The method was optimized for bacterial culture time and concentration of drugs. IC50 values for the drugs were calculated from the percent intensity of 704.5207 m/z of K. pneumoniae at 5 hrs incubation. This mass was proposed as diacylglycerophosphoethanolamine and observed as a potential biomarker of K. pneumoniae for the evaluation of inhibition potential of antimicrobial drugs and compounds. The calculated values for half maximal inhibitory concentration of cefixime, gentamicin and enrofloxacin were 0.052, 0.028 and 0.042 µg/mL, respectively. Ten compounds were also screened against the developed method, among them one compound (RSE-6) was found to be active with IC50 value of 45.08 µg/mL. The obtained results were further compared with MIC values, obtained from micro dilution and Alamar blue assay after 24 hrs incubation. In comparison to these methods, developed method is sensitive, reproducible, rapid and robust for the determination of IC50 value or inhibition potential of the drugs and compounds even at early incubation period of 5 hours.