A study on twitching motility dynamics in Ralstonia solanacearum microcolonies by live imaging.

Ralstonia solanacearum is a rod-shaped phytopathogenic bacterium that causes lethal wilt disease in many plants. On solid agar growth medium, in the early hour of the growth of the bacterial colony, the type IV pili-mediated twitching motility, which is important for its virulence and biofilm formation, is prominently observed under the microscope. In this study, we have done a detailed observation of twitching motility in R. solanacearum colony. In the beginning, twitching motility in the microcolonies was observed as a density-dependent phenomenon that influences the shape of the microcolonies. No such phenomenon was observed in Escherichia coli, where twitching motility is absent. In the early phase of colony growth, twitching motility exhibited by the cells at the peripheral region of the colony was more prominent than the cells toward the center of the colony. Using time-lapse photography and merging the obtained photomicrographs into a video, twitching motility was observed as an intermittent phenomenon that progresses in layers in all directions as finger-like projections at the peripheral region of a bacterial colony. Each layer of bacteria twitches on top of the other and produces a multilayered film-like appearance. We found that the duration between the emergence of each layer diminishes progressively as the colony becomes older. This study on twitching motility demonstrates distinctly heterogeneity among the cells within a colony regarding their dynamics and the influence of microcolonies on each other regarding their morphology.

Nanoremediation strategies to address environmental problems.

A rapid rise in population, extensive anthropogenic activities including agricultural practices, up-scaled industrialization, massive deforestation, etc. are the leading causes of environmental degradation. Such uncontrolled and unabated practices have affected the quality of environment (water, soil, and air) synergistically by accumulating huge quantities of organic and inorganic pollutants in it. Environmental contamination is posing a threat to the existing life on the Earth, therefore, demands the development of sustainable environmental remediation approaches. The conventional physiochemical remediation approaches are laborious, expensive, and time-consuming. In this regard, nanoremediation has emerged as an innovative, rapid, economical, sustainable, and reliable approach to remediate various environmental pollutants and minimize or attenuate the risks associated with them. Owing to their unique properties such as high surface area to volume ratio, enhanced reactivity, tunable physical parameters, versatility, etc. nanoscale objects have gained attention in environmental clean-up practices. The current review highlights the role of nanoscale objects in the remediation of environmental contaminants to minimize their impact on human, plant, and animal health; and air, water, and soil quality. The aim of the review is to provide information about the applications of nanoscale objects in dye degradation, wastewater management, heavy metal and crude oil remediation, and mitigation of gaseous pollutants including greenhouse gases.

Microliter spotting and micro-colony observation: A rapid and simple approach for counting bacterial colony forming units.

For enumerating viable bacteria, traditional dilution plating to count colony forming units (CFUs) has always been the preferred method in microbiology owing to its simplicity, albeit being laborious and time-consuming. Similar CFU counts can be obtained by quantifying growing micro-colonies in conjunction with the benefits of a microscope. Here, we employed a simple method of five to ten microliter spotting of a diluted bacterial culture multiple times on a single Petri dish followed by determining CFU by counting micro-colonies using a phase-contrast microscope. In this method, the CFU of an Escherichia coli culture can be estimated within a four-hour period after spotting. Further, within a ten-hour period after spotting, CFU in a culture of Ralstonia solanacearum, a bacterium with a generation time of around 2 h, can be estimated. The CFU number determined by micro-colonies observed for 106-fold dilutions or lower is similar to that obtained by the dilution plating method for 107-fold dilutions or lower. Micro-colony numbers observed in the early hours of growth (2 h in case of E. coli and 8 h in case of R. solanacearum) were found to remain consistent at later hours (4 h in case of E. coli and 10 h in case of R. solanacearum), where the visibility of the colonies was better due to a noticeable increase in the size of the colonies. This suggested that micro-colonies observed in the early hours indeed represent the bacterial number in the culture. Practical applications to this counting method were employed in studying the rifampicin-resistant mutation rate as well as performing a fluctuation test in E. coli. The spotting method described here to enumerate bacterial CFU results in reduction of labour, time and resources.

Genomics, Proteomics, and Metabolomics Approaches to Improve Abiotic Stress Tolerance in Tomato Plant.

To explore changes in proteins and metabolites under stress circumstances, genomics, proteomics, and metabolomics methods are used. In-depth research over the previous ten years has gradually revealed the fundamental processes of plants' responses to environmental stress. Abiotic stresses, which include temperature extremes, water scarcity, and metal toxicity brought on by human activity and urbanization, are a major cause for concern, since they can result in unsustainable warming trends and drastically lower crop yields. Furthermore, there is an emerging reliance on agrochemicals. Stress is responsible for physiological transformations such as the formation of reactive oxygen, stomatal opening and closure, cytosolic calcium ion concentrations, metabolite profiles and their dynamic changes, expression of stress-responsive genes, activation of potassium channels, etc. Research regarding abiotic stresses is lacking because defense feedbacks to abiotic factors necessitate regulating the changes that activate multiple genes and pathways that are not properly explored. It is clear from the involvement of these genes that plant stress response and adaptation are complicated processes. Targeting the multigenicity of plant abiotic stress responses caused by genomic sequences, transcripts, protein organization and interactions, stress-specific and cellular transcriptome collections, and mutant screens can be the first step in an integrative approach. Therefore, in this review, we focused on the genomes, proteomics, and metabolomics of tomatoes under abiotic stress.

Quorum Quenching: A Drug Discovery Approach Against Pseudomonas aeruginosa.

Pseudomonas aeruginosa, a ubiquitous opportunistic and nosocomial biofilm-forming pathogen with complex, interconnected and hierarchical nature of QS systems (Las, Rhl, PQS, and IQS), is posing the biggest challenge to the healthcare sector and have made current chemotherapies incapable. Conventional antibiotics designed to intercept the biochemical or physiological processes precisely of planktonic microorganisms exert extreme selective pressure and develop resistance against them thereby emphasizing the development of alternative therapeutic approaches. Additionally, quorum sensing induced pathogenic microbial biofilms and production of virulence factors have intensified the pathogenicity, drug resistance, recurrence of infections, hospital visits, morbidity, and mortality many-folds. In this regard, QS could be a potential druggable target and the discovery of QS inhibiting agents as an anti-virulent measure could serve as an alternative therapeutic approach to conventional antibiotics. Quorum quenching (QQ) is a preferred strategy to combat microbial infections since it attenuates the pathogenicity of microbes and enhances the microbial biofilm susceptibility to antibiotics, thus qualifying as a suitable target for drug discovery. This review discusses the QS-induced pathogenicity of P. aeruginosa, the hierarchical QS systems, and QS inhibition as a drug discovery approach to complement classical antibiotic strategy.

Microbial biofilm: A matter of grave concern for human health and food industry.

Pathogenic microorganisms have adapted different strategies during the course of time to invade host defense mechanisms and overcome the effect of potent antibiotics. The formation of biofilm on both biotic and abiotic surfaces by microorganisms is one such strategy to resist and survive even in presence of antibiotics and other adverse environmental conditions. Biofilm is a safe home of microorganisms embedded within self-produced extracellular polymeric substances comprising of polysaccharides, extracellular proteins, nucleic acid, and water. It is because of this adaptation strategy that pathogenic microorganisms are taking a heavy toll on the health and life of organisms. In this review, we discuss the colonization of pathogenic microorganisms on tissues and medically implanted devices in human beings. We also focus on food spoilage, disease outbreaks, biofilm-associated deaths, burden on economy, and other major concerns of biofilm-forming pathogenic microorganisms in food industries like dairy, poultry, ready-to-eat food, meat, and aquaculture.