Sub-inhibitory concentrations of colistin and imipenem impact the expression of biofilm-associated genes in Acinetobacter baumannii.

Acinetobacter baumannii is an opportunistic pathogen that is responsible for nosocomial infections. Imipenem and colistin are drugs that are commonly used to treat severe infections caused by A. baumannii, such as sepsis, ventilator-associated pneumonia, and bacteremia. However, some strains of A. baumannii have become resistant to these drugs, which is a concern for public health. Biofilms produced by A. baumannii increase their resistance to antibiotics and the cells within the inner layers of biofilm are exposed to sub-inhibitory concentrations (sub-MICs) of antibiotics. There is limited information available regarding how the genes of A. baumannii are linked to biofilm formation when the bacteria are exposed to sub-MICs of imipenem and colistin. Thus, this study's objective was to explore this relationship by examining the genes involved in biofilm formation in A. baumannii when exposed to low levels of imipenem and colistin. The study found that exposing an isolate of A. baumannii to low levels of these drugs caused changes in their drug susceptibility pattern. The relative gene expression profiles of the biofilm-associated genes exhibited a change in their expression profile during short-term and long-term exposure. This study highlights the potential consequences of overuse and misuse of antibiotics, which can help bacteria become resistant to these drugs.

Comparison of Antibiofilm Activity of Pseudomonas aeruginosa Phages on Isolates from Wounds of Diabetic and Non-Diabetic Patients.

The persistence of organisms as biofilms and the increase in antimicrobial resistance has raised the need for alternative strategies. The study objective was to compare the ability of isolated bacteriophages to remove in vitro biofilms formed by Pseudomonas aeruginosa isolated from the environment with those isolated from diabetic and non-diabetic wounds. P. aeruginosa were isolated from clinical and environmental sites, and antimicrobial susceptibility was tested. Bacteriophages were isolated and characterized based on plaque morphology and host range. A reduction in the viable count assayed the lytic ability of candidate phages. The crystal violet method was used to determine the residual biofilm after 24 h of phage treatment on 72-h-old biofilms. The statistical significance of phage treatment was tested by one-way ANOVA. Of 35 clinical isolates, 17 showed resistance to 1 antibiotic at least, and 7 were multidrug resistant. Nineteen environmental isolates and 11 clinical isolates were drug-sensitive. Nine phages showed 91.2% host coverage, including multidrug-resistant isolates. Phages eradicated 85% of biofilms formed by environmental isolates compared to 58% of biofilms of diabetic isolates and 56% of biofilms of non-diabetic isolates. Clinical isolates are susceptible to phage infection in planktonic form. Biofilms of P. aeruginosa isolated from diabetic wounds and non-diabetic wounds resist removal by phages compared to biofilms formed by environmental isolates. All phages were efficient in dispersing PAO1 biofilms. However, there was a significant difference in their ability to disperse PAO1 biofilms across the different surfaces tested. Partial eradication of biofilm by phages can aid in complementing antibiotics that are unable to penetrate biofilms in a clinical set-up.

Glucose-Induced Enhanced Virulence in Strains of Multidrug-Resistant Pseudomonas aeruginosa Isolated from Diabetic Patients.

Pseudomonas aeruginosa is known for its metabolic versatility and uses a variety of substrates; interestingly, glucose is not the favored carbon source. Although glucose is not readily utilized by them, there is a possibility that the increased susceptibility of diabetics to infections with P. aeruginosa is related to the effect of glucose on the expression of virulence genes. The curiosity in understanding the effect of glucose on virulence gene expression in P. aeruginosa and the lacuna of studies in this field prompted us to undertake the current investigation. It included the quantification of various virulence factors and their gene expression upon supplementation with glucose in clinical MDR P. aeruginosa isolates recovered from diabetics. Interestingly, the study observed a remarkable difference in the virulence attributes in the isolates with and without glucose supplementation. External glucose was found to be modulating the QS gene expression, thus altering the elaboration of other virulence factors. Variations in the gene expressions induced by glucose partly explain the increased susceptibility of diabetic patients to P. aeruginosa infections.

Characterisation of broad-spectrum phiKZ like jumbo phage and its utilisation in controlling multidrug-resistant Pseudomonas aeruginosa isolates.

The emergence of highly virulent multidrug-resistant P. aeruginosa has become increasingly evident among hospital-acquired infections and has raised the need for alternative therapies. Phage therapy can be one such alternative to antibiotic therapy to combat multidrug-resistant pathogenic bacteria, but this requires the availability of phages with a broad host range. In this study, isolation and molecular characterisation of P. aeruginosa specific phages were carried out. A total of 17 phages isolated showed different spectra of activity and efficiency of lysis against 82 isolates of P. aeruginosa obtained from clinical samples (n = 13), hospital effluent (n = 46) and fish processing plant effluent (n = 23). Antibiotic susceptibility test results revealed multi-drug resistance in 61 of the total 82 isolates. Three new jumbo lytic P. aeruginosa specific broad host range phages were isolated and characterised in this present study belonged to the family Myoviridae (order Caudovirales). The genetic analysis of ɸU5 revealed that phage has a genome size of 282.6 kbp with 373 putative open reading frames (ORFs), and its genetic architecture is similar to phiKZ like jumbo phages infecting P. aeruginosa. The bacteriophages isolated in this study had lytic ability against biofilm-forming and multidrug-resistant P. aeruginosa and could be candidates for further studies towards phage therapy.

A rare occurrence of multidrug-resistant environmental Acinetobacter baumannii strains from the soil of Mangaluru, India.

Over the last decade, Acinetobacter baumannii has emerged as one of the main causes of infections acquired in the hospital setting. Outbreaks associated with this pathogen are caused mainly due to contamination and transmission in hospital territories. However, the natural habitats of A. baumannii of clinical significance still remain unclear. In this study, we highlight the isolation and identification of multidrug-resistant environmental strains of A. baumannii from the soil of Mangaluru city. All the recovered isolates were biofilm formers, and two isolates were multidrug-resistant and showed resistance to fluoroquinolone, aminoglycosides, sulfonamide, tetracycline, and carbapenems. In addition, they exhibited protease activity, and produced phospholipase C and siderophore. To the best of our knowledge, this is the first study to isolate and identify drug-resistant strains of A. baumannii from the soil.

Non-thermal Plasma Treatment of ESKAPE Pathogens: A Review.

The acronym ESKAPE refers to a group of bacteria consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. They are important in human medicine as pathogens that show increasing resistance to commonly used antibiotics; thus, the search for new effective bactericidal agents is still topical. One of the possible alternatives is the use of non-thermal plasma (NTP), a partially ionized gas with the energy stored particularly in the free electrons, which has antimicrobial and anti-biofilm effects. Its mechanism of action includes the formation of pores in the bacterial membranes; therefore, resistance toward it is not developed. This paper focuses on the current overview of literature describing the use of NTP as a new promising tool against ESKAPE bacteria, both in planktonic and biofilm forms. Thus, it points to the fact that NTP treatment can be used for the decontamination of different types of liquids, medical materials, and devices or even surfaces used in various industries. In summary, the use of diverse experimental setups leads to very different efficiencies in inactivation. However, Gram-positive bacteria appear less susceptible compared to Gram-negative ones, in general.

Modulation of quorum sensing-associated virulence in bacteria: carbohydrate as a key factor.

Quorum sensing (QS) is a method of inter-cellular communication that permits bacteria to dispense information about cell density and to synchronize the gene expression accordingly. Gram-positive and Gram-negative bacteria utilize distinct quorum sensing mechanisms for effective pathogenesis. Virulence factor production by pathogenic bacteria is one of the important traits that is under the control of QS. A growing body of evidence has indicated the role of the nutritional environment notably by carbohydrates in dictating the QS-associated virulence gene regulation. The modulation of QS by carbohydrates mitigates the survival and establishment of the pathogen within its host which in turn leads to an increase in morbidity and mortality. This mini-review throws light on the predilection of pathogenic bacteria to rapidly regulate its QS-linked virulence gene expression based on the changing nutrient levels that assist them in prospering within diverse niches.

Isolation and identification of quorum sensing antagonist from Cinnamomum verum leaves against Pseudomonas aeruginosa.

PURPOSE: The study aimed at isolating and identifying potential anti-quorum sensing (QS) compounds from Cinnamomum verum leaves against Pseudomonas aeruginosa. METHODOLOGY: Isolation of anti-QS compounds from C. verum leaf ethanol extract was carried out by column chromatography. The bioactive fraction was analysed by UV, IR, and GCMS spectroscopy. Various virulence assays were performed to assess the QS quenching ability of the purified compounds. In vivo toxicity of the purified compounds was examined in zebrafish model. The expression of the virulence genes was evaluated by qPCR analysis and in silico assessment was accomplished to check the binding ability of the compounds with the autoinducer molecule. KEY FINDINGS: The QS inhibitors isolated and identified showed a remarkable ability in reducing the production of elastase, pyocyanin, swarming motility and biofilm formation in P. aeruginosa. In the presence of the characterized compounds, the expression of virulence genes of P. aeruginosa was significantly reduced. Toxicity studies in zebrafish model indicated no effects on development and organogenesis at a concentration below 100 mg/l. Further, in silico analysis demonstrated the binding efficiency of the anti-QS compounds to AHL molecules, thus proving the QS quenching ability of the isolated compounds. SIGNIFICANCE: To the best of our knowledge this is the first report of isolation of anti-QS compounds from C. verum leaves against P. aeruginosa. The identified compounds qualify as potential QS antagonists. Further studies on these compounds can pave way for an effective and attractive anti-pathogenic therapy, to overcome the emergence of antibiotic resistance in bacteria.