Endodontic irrigants from a comprehensive perspective.

This review article explores the fundamental principles of modern endodontics with a focus on root canal cleaning and shaping. It reviews commonly used endodontic irrigant, namely sodium hypochlorite (NaOCl), herbal extracts, chlorhexidine (CHX), and chelating agents, highlighting their properties, applications, and potential drawbacks. NaOCl, a key antimicrobial agent, demonstrates effectiveness against various microorganisms but poses challenges such as high cytotoxicity. Herbal extracts, gaining recognition in endodontics, present an alternative with potential advantages in preserving dentin integrity. CHX, known for its broad-spectrum antimicrobial activity, is discussed in both liquid and gel formulations, emphasizing its role in reducing smear layer formation and preserving hybrid layer durability. Chelating agents, specifically ethylenediaminetetraacetic acid and citric acid, play a vital role in removing the smear layer, enhancing dentin permeability, and facilitating the penetration of antimicrobial agents. The review article underscores the importance of careful application and consideration of each irrigant's properties to ensure safe and effective endodontic procedures. It serves as a valuable guide for clinicians in selecting appropriate irrigants based on specific treatment requirements.

Competitive fitness and stability of ammonium-excreting Azotobacter vinelandii strains in the soil.

Non-symbiotic N2-fixation would greatly increase the versatility of N-biofertilizers for sustainable agriculture. Genetic modification of diazotrophic bacteria has successfully enhanced NH4+ release. In this study, we compared the competitive fitness of A. vinelandii mutant strains, which allowed us to analyze the burden of NH4+ release under a broad dynamic range. Long-term competition assays under regular culture conditions confirmed a large burden for NH4+ release, exclusion by the wt strain, phenotypic instability, and loss of the ability to release NH4+. In contrast, co-inoculation in mild autoclaved soil showed a much longer co-existence with the wt strain and a stable NH4+ release phenotype. All genetically modified strains increased the N content and changed its chemical speciation in the soil. This study contributes one step forward towards bridging a knowledge gap between molecular biology laboratory research and the incorporation of N from the air into the soil in a molecular species suitable for plant nutrition, a crucial requirement for developing improved bacterial inoculants for economic and environmentally sustainable agriculture. KEY POINTS: • Genetic engineering for NH4+ excretion imposes a fitness burden on the culture medium • Large phenotypic instability for NH4+-excreting bacteria in culture medium • Lower fitness burden and phenotypic instability for NH4+-excreting bacteria in soil.

A novel genetic engineering platform for the effective management of biological contaminants for the production of microalgae.

Microalgal cultivation that takes advantage of solar energy is one of the most cost-effective systems for the biotechnological production of biofuels, and a range of high value products, including pharmaceuticals, fertilizers and feed. However, one of the main constraints for the cultivation of microalgae is the potential contamination with biological pollutants, such as bacteria, fungi, zooplankton or other undesirable microalgae. In closed bioreactors, the control of contamination requires the sterilization of the media, containers and all materials, which increases the cost of production, whereas open pond systems severely limits the number of species that can be cultivated under extreme environmental conditions to prevent contaminations. Here, we report the metabolic engineering of Chlamydomonas reinhardtii to use phosphite as its sole phosphorus source by expressing the ptxD gene from Pseudomonas stutzeri WM88, which encodes a phosphite oxidoreductase able to oxidize phosphite into phosphate using NAD as a cofactor. Engineered C. reinhardtii lines are capable of becoming the dominant species in a mixed culture when fertilized with phosphite as a sole phosphorus source. Our results represent a new platform for the production of microalgae, potentially useful for both closed photobioreactors and open pond systems without the need for using sterile conditions nor antibiotics or herbicides to prevent contamination with biological pollutants.

Metabolic engineering of ammonium release for nitrogen-fixing multispecies microbial cell-factories.

The biological nitrogen fixation carried out by some Bacteria and Archaea is one of the most attractive alternatives to synthetic nitrogen fertilizers. In this study we compared the effect of controlling the maximum activation state of the Azotobacter vinelandii glutamine synthase by a point mutation at the active site (D49S mutation) and impairing the ammonium-dependent homeostatic control of nitrogen-fixation genes expression by the ΔnifL mutation on ammonium release by the cells. Strains bearing the single D49S mutation were more efficient ammonium producers under carbon/energy limiting conditions and sustained microalgae growth at the expense of atmospheric N2 in synthetic microalgae-bacteria consortia. Ammonium delivery by the different strains had implications for the microalga׳s cell-size distribution. It was uncovered an extensive cross regulation between nitrogen fixation and assimilation that extends current knowledge on this key metabolic pathway and might represent valuable hints for further improvements of versatile N2-fixing microbial-cell factories.

High lipid productivity of an Ankistrodesmus-Rhizobium artificial consortium.

Microalgae have great potential as alternative productive platforms for sustainable production of bioenergy, food, feed and other commodities. Process optimization to realize the claimed potential often comprises strains selection and improvement and also developing of more efficient cultivation, harvesting and downstream processing technology. In this work we show that inoculation with the bacterium Rhizobium strain 10II resulted in increments of up to 30% in chlorophyll, biomass and lipids accumulation of the oleaginous microalgae Ankistrodesmus sp. strain SP2-15. Inoculated cultures have reached a high lipid productivity of up to 112 mg L(-1) d(-1) after optimization. The resulting biomass presented significant levels of Ω3 fatty acids including stearidonic acid, suggesting potential as an alternative land-based source of essential fatty acids.

Genetic engineering of multispecies microbial cell factories as an alternative for bioenergy production.

There is currently much interest in developing technology to use microlgae or cyanobacteria for the production of bioenergy and biomaterials. Here, we summarize some remarkable achievements in strains improvement by traditional genetic engineering and discuss common drawbacks for further progress. We present general knowledge on natural microalgal-bacterial mutualistic interactions and discuss the potential of recent developments in genetic engineering of multispecies microbial cell factories. This synthetic biology approach would rely on the assembly of complex metabolic networks from optimized metabolic modules such as photosynthetic or nitrogen-fixing parts.

Bioprospecting for fast growing and biomass characterization of oleaginous microalgae from South-Eastern Buenos Aires, Argentina.

As part of pioneering efforts to assess the potential of native microalgae as biofuel feedstock in South-Eastern Buenos Aires, 34 monoalgal cultures (corresponding to the Phylum Chlorophyta) were established and 21 were selected for further growth and biomass composition characterization. Novel RNA sequences in the ITS1-5.8S-ITS2 region were identified. Some strains showed desirable traits as biodiesel feedstock such as (i) apparent maximal doubling times of 6h, (ii) lipids accumulation of up to 43% of their dry biomass, (iii) high ration of mono-unsaturated to poly-unsaturated fatty acids, (iv) high response to CO(2) supplementation, and (v) complete sedimentation in 4h. Data of the outdoors performance of some strains suggested they might represent valuable resources for future research towards the regional development of the technology for microalgae-based biofuels.

Association with an ammonium-excreting bacterium allows diazotrophic culture of oil-rich eukaryotic microalgae.

Concerns regarding the depletion of the world's reserves of oil and global climate change have promoted an intensification of research and development toward the production of biofuels and other alternative sources of energy during the last years. There is currently much interest in developing the technology for third-generation biofuels from microalgal biomass mainly because of its potential for high yields and reduced land use changes in comparison with biofuels derived from plant feedstocks. Regardless of the nature of the feedstock, the use of fertilizers, especially nitrogen, entails a potential economic and environmental drawback for the sustainability of biofuel production. In this work, we have studied the possibility of nitrogen biofertilization by diazotrophic bacteria applied to cultured microalgae as a promising feedstock for next-generation biofuels. We have obtained an Azotobacter vinelandii mutant strain that accumulates several times more ammonium in culture medium than wild-type cells. The ammonium excreted by the mutant cells is bioavailable to promote the growth of nondiazotrophic microalgae. Moreover, this synthetic symbiosis was able to produce an oil-rich microalgal biomass using both carbon and nitrogen from the air. This work provides a proof of concept that artificial symbiosis may be considered an alternative strategy for the low-N-intensive cultivation of microalgae for the sustainable production of next-generation biofuels and other bioproducts.