A method to assess bioavailability of antibiotics in anthropogenic polluted ecosystems by using a bacterial fitness test

Antibiotics released in the environment exert a selective pressure on the resident microbiota. It is well accepted that the mere measurement of antibiotics does not reflect the actual bioavailability. In fact, antibiotics can be adsorbed or complexed to particles and/or chemicals in water and soil. Bioavailable concentrations of antibiotics in soil and water are subjected to great uncertainty, therefore biological assays are increasingly recognized as that allow an indirect determination of the residual antibiotic activity. Here we propose how a fitness test for bacteria can be used to qualitatively assess the bioavailability of a specific antibiotic in the environment. The findings show that by using a pair of resistant and sensitive bacterial strains, the resulting fitness can indirectly reflect antibiotic bioavailability. Hence, this test can be used as a complementary assay to other biological and chemical tests to assess bioavailability of antibiotics

Second generation bioethanol production: on the use of pulp and paper industry wastes as feedstock

Due to the health and environment impacts of fossil fuels utilization, biofuels have been investigated as a potential alternative renewable source of energy. Bioethanol is currently the most produced biofuel, mainly of first generation, resulting in food-fuel competition. Second generation bioethanol is produced from lignocellulosic biomass, but a costly and difficult pretreatment is required. The pulp and paper industry has the biggest income of biomass for non-food-chain production, and, simultaneously generates a high amount of residues. According to the circular economy model, these residues, rich in monosaccharides, or even in polysaccharides besides lignin, can be utilized as a proper feedstock for second generation bioethanol production. Biorefineries can be integrated in the existing pulp and paper industrial plants by exploiting the high level of technology and also the infrastructures and logistics that are required to fractionate and handle woody biomass. This would contribute to the diversification of products and the increase of profitability of pulp and paper industry with additional environmental benefits. This work reviews the literature supporting the feasibility of producing ethanol from Kraft pulp, spent sulfite liquor, and pulp and paper sludge, presenting and discussing the practical attempt of biorefineries implementation in pulp and paper mills for bioethanol production.publishe

Two-stage aeration fermentation strategy to improve bioethanol production by scheffersomyces stipitis

Hardwood spent sulfite liquor (HSSL) is a by-product from pulp industry with a high concentration of pentose sugars, besides some hexoses suitable for bioethanol production by Scheffersomyces stipitis. The establishment of optimal aeration process conditions that results in specific microaerophilic conditions required by S. stipitis is the main challenge for ethanol production. The present study aimed to improve the ethanol production from HSSL by S. stipitis through a two-stage aeration fermentation. Experiments with controlled dissolved oxygen tension (DOT) in the first stage and oxygen restriction in the second stage were carried out. The best results were obtained with DOT control at 50% in the first stage, where the increase of oxygen availability provided faster growth and higher biomass yield, and no oxygen supply with an agitation rate of 250 rpm, in the second stage allowed a successful induction of ethanol production. Fermentation using 60% of HSSL (v/v) as substrate for S. stipitis provided a maximum specific growth rate of 0.07 h−1 , an ethanol productivity of 0.04 g L h−1 and an ethanol yield of 0.39 g g−1 , respectively. This work showed a successful two-stage aeration strategy as a promising aeration alternative for bioethanol production from HSSL by S. stipitis.publishe

Effect of anthropogenic pollution on the fitness of tetracycline sensitive Shigella flexneri in Thames river water

Urban rivers may be source of antibiotics contamination that could support spread of antibiotic resistant bacteria (ARB) to the population. It is important to understand to what extent the presence of pollutants in urban rivers influences fitness of ARB. In an exercise to estimate this contribution, microcosms were generated from Thames river (London, UK) from different locations: upstream and downstream the city center. The concentration of the polycyclic aromatic hydrocarbons (PAHs) benzo(a)pyrene, pyrene and phenantrene was found to be 128, 171 and 128 times higher in downstream sector when compared to upstream sector, respectively. Filtered microcosms for each sector were enriched with tetracycline at lethal (10 μg/mL) and sub-lethal (10 ng/mL) concentrations and the fitness of an isogenic pair of Shigella flexneri 2a YSH6000 (tetR) and S. flexneri 2a 1363 (tetS) was then measured. In the presence of selective pressure in upstream microcosms, the resistant strain outcompeted the sensitive one, as expected. In contrast, sensitive S. flexneri tetS was found to significantly compete with resistant S. flexneri tetR at lethal concentrations of tetracycline in downstream microcosms, where levels of PAHs were the highest. Further experiments showed that PAHs rendered the resistant S. flexneri tetR ∼20% more sensitive to tetracycline. Sensitive S. flexneri tetS strain was able to persist at lethal concentration of tetracycline in downstream microcosms, at higher concentrations of PAHs. Our findings suggest that in a polluted river sensitive S. flexneri cells may still thrive in presence of selective pressure. Fitness tests provide an additional tool to measure bioavailability

Ethanol production from hydrolyzed kraft pulp by mono- and co-cultures of yeasts: the challenge of C6 and C5 sugars consumption

Second-generation bioethanol production’s main bottleneck is the need for a costly and technically di cult pretreatment due to the recalcitrance of lignocellulosic biomass (LCB). Chemical pulping can be considered as a LCB pretreatment since it removes lignin and targets hemicelluloses to some extent. Chemical pulps could be used to produce ethanol. The present study aimed to investigate the batch ethanol production from unbleached Kraft pulp of Eucalyptus globulus by separate hydrolysis and fermentation (SHF). Enzymatic hydrolysis of the pulp resulted in a glucose yield of 96.1 3.6% and a xylose yield of 94.0 7.1%. In an Erlenmeyer flask, fermentation of the hydrolysate using Saccharomyces cerevisiae showed better results than Sche ersomyces stipitis. At both the Erlenmeyer flask and bioreactor scale, co-cultures of S. cerevisiae and S. stipitis did not show significant improvements in the fermentation performance. The best result was provided by S. cerevisiae alone in a bioreactor, which fermented the Kraft pulp hydrolysate with an ethanol yield of 0.433 g g1 and a volumetric ethanol productivity of 0.733 g L1 h1, and a maximum ethanol concentration of 19.24 g L1 was attained. Bioethanol production using the SHF of unbleached Kraft pulp of E. globulus provides a high yield and productivity.publishe

Time-coded neurotransmitter release at excitatory and inhibitory synapses.

Communication between neurons at chemical synapses is regulated by hundreds of different proteins that control the release of neurotransmitter that is packaged in vesicles, transported to an active zone, and released when an input spike occurs. Neurotransmitter can also be released asynchronously, that is, after a delay following the spike, or spontaneously in the absence of a stimulus. The mechanisms underlying asynchronous and spontaneous neurotransmitter release remain elusive. Here, we describe a model of the exocytotic cycle of vesicles at excitatory and inhibitory synapses that accounts for all modes of vesicle release as well as short-term synaptic plasticity (STSP). For asynchronous release, the model predicts a delayed inertial protein unbinding associated with the SNARE complex assembly immediately after vesicle priming. Experiments are proposed to test the model's molecular predictions for differential exocytosis. The simplicity of the model will also facilitate large-scale simulations of neural circuits

Getting Value from Pulp and Paper Industry Wastes: On the Way to Sustainability and Circular Economy

The pulp and paper industry is recognized as a well-established sector, which throughout its process, generates a vast amount of waste streams with the capacity to be valorized. Typically, these residues are burned for energy purposes, but their use as substrates for biological processes could be a more efficient and sustainable alternative. With this aim, it is essential to identify and characterize each type of waste to determine its biotechnological potential. In this context, this research highlights possible alternatives with lower environmental impact and higher revenues. The bio-based pathway should be a promising alternative for the valorization of pulp and paper industry wastes, in particular for bioproduct production such as bioethanol, polyhydroxyalkanoates (PHA), and biogas. This article focuses on state of the art regarding the identification and characterization of these wastes, their main applied deconstruction technologies and the valorization pathways reported for the production of the abovementioned bioproductspublishe

Enzymatic Potential of Filamentous Fungi as a Biological Pretreatment for Acidogenic Fermentation of Coffee Waste

This work was developed within the scope of the project CICECO-Aveiro Institute of Materials (UIDB/50011/2020, UIDP/50011/2020, and LA/P/0006/2020), and the Associate Laboratory for Green Chemistry-LAQV (UIDB/50006/2020 and UIDP/50006/2020). It was financed by national funds through the FCT/MCTES (PIDDAC) and, when appropriate, co-financed by FEDER under the PT2020 Partnership Agreement. Paulo C. Lemos acknowledges the support of FCT/MCTES for contract IF/01054/2014/CP1224/CT0005 and Joana Pereira thanks FCT/MCTES for her Ph.D. grant SFRH/BD/130003/2017.Spent coffee grounds (SCGs) are a promising substrate that can be valorized by biotechnological processes, such as for short-chain organic acid (SCOA) production, but their complex structure implies the application of a pretreatment step to increase their biodegradability. Physicochemical pretreatments are widely studied but have multiple drawbacks. An alternative is the application of biological pretreatments that include using fungi Trametes versicolor and Paecilomyces variotii that naturally can degrade complex substrates such as SCGs. This study intended to compare acidic and basic hydrolysis and supercritical CO 2 extraction with the application of these fungi. The highest concentration of SCOAs, 2.52 gCOD/L, was achieved after the acidification of SCGs pretreated with acid hydrolysis, but a very similar result, 2.44 gCOD/L, was obtained after submerged fermentation of SCGs by T. versicolor. This pretreatment also resulted in the best acidification degree, 48%, a very promising result compared to the 13% obtained with the control, untreated SCGs, highlighting the potential of biological pretreatments.publishersversionpublishe