Genome comparison and physiological characterization of eight Streptococcus thermophilus strains isolated from Italian dairy products

Eight Streptococcus thermophilus strains of dairy origin isolated in Italy were chosen to investigate autochthonous bacterial diversity in this important technological species. In the present study a comparative analysis of all the 17 S. thermophilus genomes publicly available was performed to identify the core and the variable genes, which vary among strains from 196 to 265. Additionally, correlation between the isolation site and the genetic distance was investigated at genomic level. Results highlight that the phylogenetic reconstruction differs from the geographical strain distribution. Moreover, strain M17PTZA496 has a genome of 2.15 Mbp, notably larger than that of the others, determined by lateral gene transfer (including phage-mediated incorporation) and duplication events. Important technological characters, such as growth kinetics, bacteriocin production, acidification kinetics and surface adhesion capability were studied in all the Italian strains. Results indicate a wide range of variability in adhesion properties that significantly clustered strains into four groups. Genomic differences among strains in relation to these characters were identified but a clear correlation between genotype and phenotype was not always found since most of the genomic modifications arise from single nucleotide polymorphisms. This research represents a step forward in the identification of strains-specific functions in Streptococcus thermophilus and it has also the potential to provide valuable information to predict strain specific behaviors in industrial processes

The business of invention: considering project management in the arts and industry

Project management has well developed theoretical constructs and is becom- ing increasingly well established in core strategy beyond the industrial and corporate sec- tors from which it first emerged. With a concurrent increase in the significance of innova- tion, project managing for creativity is an area of research and enquiry of considerable sig- nificance. Notionally occupying polar opposite cultural positions in terms of perspectives and processes of creativity, project management in the arts is widely considered to vary significantly from corporate strategy and process. If business were to be more generally characterised by ‘organisation’ and discipline, the arts are more commonly celebrated for disorganisation, indiscipline, and the fundamental challenge to organisation itself. Consid- ering both the confluences and variations between established project management theory in business and practice in the arts, this text introduces theoretical constructs pertaining to creative processes and highlights areas for consideration in the understanding and further development of project management theory

Combination Therapy Strategy of Quorum Quenching Enzyme and Quorum Sensing Inhibitor in Suppressing Multiple Quorum Sensing Pathways of P. <i>aeruginosa</i>

Abstract The threat of antibiotic resistant bacteria has called for alternative antimicrobial strategies that would mitigate the increase of classical resistance mechanism. Many bacteria employ quorum sensing (QS) to govern the production of virulence factors and formation of drug-resistant biofilms. Targeting the mechanism of QS has proven to be a functional alternative to conventional antibiotic control of infections. However, the presence of multiple QS systems in individual bacterial species poses a challenge to this approach. Quorum sensing inhibitors (QSI) and quorum quenching enzymes (QQE) have been both investigated for their QS interfering capabilities. Here, we first simulated the combination effect of QQE and QSI in blocking bacterial QS. The effect was next validated by experiments using AiiA as QQE and G1 as QSI on Pseudomonas aeruginosa LasR/I and RhlR/I QS circuits. Combination of QQE and QSI almost completely blocked the P. aeruginosa las and rhl QS systems. Our findings provide a potential chemical biology application strategy for bacterial QS disruption

Effects of Osmotic Stress on Rhamnolipid Synthesis and Time-Course Production of Cell-To-Cell Signal Molecules by Pseudomonas aeruginosa

Biosynthesis of biosurfactant rhamnolipids by Pseudomonas aeruginosa depends on two hierarchical quorum sensing systems, LasRI and RhlRI, which synthesize and sense the signal molecules N-(3-oxododecanoyl)-L-homoserine lactone (3OC12-HSL) and N-butyryl-L-homoserine lactone (C4-HSL), respectively. The Pseudomonas Quinolone Signal (PQS) is a third cell-to-cell signal molecule connecting these two systems, and its precursor, 2-heptyl-4-quinolone (HHQ), also constitutes a signal. The chronology of the production of signal molecules and rhamnolipids was determined during growth in PPGAS medium. Hyperosmotic condition (0.5 M NaCl) moderately affected growth, and led to intra-cellular accumulation of compatible solutes. Production of signal molecules was delayed and their highest concentrations were 2.5 to 5 fold lower than in NaCl-free PPGAS, except for HHQ, the highest concentration of which was increased. The presence of NaCl prevented rhamnolipid synthesis. When the osmoprotectant glycine betaine was added to PPGAS/NaCl medium, it was imported by the cells without being metabolized. This did not improve growth, but reestablished the time-courses of HSL and HHQ accumulation and fully or partially restored the HSL and PQS levels. It also partially restored rhamnolipid production. Quantification of mRNAs encoding enzymes involved in HSL, PQS, and rhamnolipid biosyntheses confirmed the effect of hyperosmotic stress and glycine betaine at the gene expression level

Multi-Focused Laboratory Experiments Based on Quorum Sensing and Quorum Quenching for Acquiring Microbial Physiology Concepts

After a time away from the classrooms and laboratories due to the global pandemic, the return to the teaching activities during the semester represented a challenge to both teachers and students. Our particular situation in a Microbial Physiology course was the necessity of imparting in a shorter time, laboratory practices that usually take longer. This article describes a two-week long laboratory exercise that covers several concepts in an interrelated way: conjugation as a gene transfer mechanism, regulation of microbial physiology, production of secondary metabolites, degradation of macromolecules and biofilm formation. Utilizing a Quorum Quenching (QQ) strategy, the Quorum Sensing (QS) system of Pseudomonas aeruginosa is first attenuated. Then, phenotypes regulated by QS are evidenced. QS is a regulatory mechanism of the microbial physiology that relays on signal molecules. QS is related in P. aeruginosa to several virulence factors, some of which are exploited in the laboratory practices presented in this work. QQ is phenomenon by which QS is interrupted or attenuated. We utilized a QQ approach based on the enzymatic degradation of the P. aeruginosa QS signals in order to put in evidence QS-regulated traits that are relevant for our Microbial Physiology course.Fil: Torres, Mariela Analía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Valdez, Alejandra Leonor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Olea, Carolina de Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Nieto Peñalver, Carlos Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentin

Role of the quorum sensing mechanism in the response of pseudomonas aeruginosa to lethal and sublethal UVA irradiation

The role of quorum sensing (QS) in the response of Pseudomonas aeruginosa to UVA radiation was investigated in the PAO1 strain and derivatives defective in the synthesis of the QS signals 3OC12-HSL (lasI strain), C4-HSL (rhlI strain) or both (lasI rhlI strain). Cell viability measurements demonstrated that the double mutant was significantly more sensitive to UVA than single mutants, which in turn showed reduced cell survival with regard to the PAO1 strain. Irradiation under nitrogen atmosphere and chemiluminescence measurements indicated the oxidative nature of the UVA-induced damage. The activity of the antioxidant enzymes catalase and superoxide dismutase was assayed in these strains before and after irradiation, and a strong correlation between catalase levels and UVA sensitivity was observed. When a sublethal UVA dose was applied to PAO1, a growth delay was observed and this mechanism depended on the rhl system. Moreover, low doses of UVA irradiation triplicated the level of C4-HSL in log PAO1 cells. It is concluded that QS is fundamental in the defense against the toxic effects of UVA in P. aeruginosa. The induction of the QS system by UVA independently of cell density could function as an adaptative strategy to withstand this hostile environmental condition.Fil: Costa, Cristina Susana. Comisión Nacional de Energía Atómica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia de Radiobiología (Centro Atómico Constituyentes); ArgentinaFil: Pezzoni, Magdalena. Comisión Nacional de Energía Atómica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia de Radiobiología (Centro Atómico Constituyentes); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fernández, Rubén O.. Comisión Nacional de Energía Atómica; ArgentinaFil: Pizarro, Ramón Augusto. Comisión Nacional de Energía Atómica. Gerencia de Area de Aplicaciones de la Tecnología Nuclear. Gerencia de Radiobiología (Centro Atómico Constituyentes); Argentin

Bacteria that inhibit quorum sensing decrease biofilm formation and virulence in Pseudomonas aeruginosa PAO1

In this study, we investigated the biotherapeutic potential of previously isolated quorum quenching (QQ) bacteria. Some of them produce and secrete small compounds that inhibit quorum sensing (QS), others quench QS by enzymatic degradation of N-acylhomoserine lactones (AHLs). The supernatant of cultures of these isolates was tested for inhibitory properties against P.aeruginosa PAO1 biofilms. Most isolates had a moderate effect on biofilm formation, as shown by viability staining and/or staining of the biofilm biomass. A substantial part of the isolates reduced P.aeruginosa elastase production in a concentration-dependent manner. Using Caenorhabditis elegans as an in vivo model system for virulence testing, we found that some of the isolates were able to increase survival of P.aeruginosa PAO1 and Burkholderia cenocepacia LGM16656-infected nematodes when co-administered with the pathogen. Altogether, these data indicate that some QQ bacteria, or the active compounds they produce, could be useful to attenuate virulence of P.aeruginosa PAO1 and possibly also other Gram-negative pathogens that use AHLs to regulate the production of virulence factors

A PoW-less Bitcoin with Certified Byzantine Consensus

Distributed Ledger Technologies (DLTs), when managed by a few trusted validators, require most but not all of the machinery available in public DLTs. In this work, we explore one possible way to profit from this state of affairs. We devise a combination of a modified Practical Byzantine Fault Tolerant (PBFT) protocol and a revised Flexible Round-Optimized Schnorr Threshold Signatures (FROST) scheme, and then we inject the resulting proof-of-authority consensus algorithm into Bitcoin (chosen for the reliability, openness, and liveliness it brings in), replacing its PoW machinery. The combined protocol may operate as a modern, safe foundation for digital payment systems and Central Bank Digital Currencies (CBDC)