Phage-based therapy against biofilm producers in gram-negative ESKAPE pathogens.

Persistent antibiotic use results in the rise of antimicrobial resistance with limited or no choice for multidrug-resistant (MDR) and extensively drug resistant (XDR) bacteria. This necessitates a need for alternative therapy to effectively combat clinical pathogens that are resistant to last resort antibiotics. The study investigates hospital sewage as a potential source of bacteriophages to control resistant bacterial pathogens. Eighty-one samples were screened for phages against selected clinical pathogens. Totally, 10 phages were isolated against A. baumannii, 5 phages against K. pneumoniae, and 16 phages were obtained against P. aeruginosa. The novel phages were observed to be strain-specific with complete bacterial growth inhibition of up to 6 h as monotherapy without antibiotics. Phage plus colistin combinations reduced the minimum-biofilm eradication concentration of colistin up to 16 folds. Notably, a cocktail of phages exhibited maximum efficacy with complete killing at 0.5-1 µg/ml colistin concentrations. Thus, phages specific to clinical strains have a higher edge in treating nosocomial pathogens with their proven anti-biofilm efficacy. In addition, analysis of phage genomes revealed close phylogenetic relations with phages reported from Europe, China, and other neighbouring countries. This study serves as a reference and can be extended to other antibiotics and phage types to assess optimum synergistic combinations to combat various drug resistant pathogens in the ongoing AMR crisis.

Genotyping of Acinetobacter baumannii in Nosocomial Outbreak and Surveillance.

Acinetobacter baumannii is considered to be an important nosocomial pathogen responsible for various outbreaks that have resulted in a need for effective epidemiological typing methods. Different typing methods are available for A. baumannii epidemiological studies. Currently, the phenotypic typing methods are not being used and replaced by various molecular methods. In this chapter, two important epidemiological typing methods, pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST), are discussed.

Antimicrobial Susceptibility Testing Methods for Acinetobacter spp.

Serial twofold dilution methods are most commonly used to identify the antimicrobial activities of antibiotics. This can be achieved by different methods like broth dilution or agar dilution. Though these methods are simple, they can be influenced by various experimental factors and result in discrepancy. The following protocol has been validated for Acinetobacter species, including A. baumannii. It is important to include appropriate control strains to determine the minimum inhibitory concentration values and to compare the experiment results.