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In this study, bacterial isolates C1-4-7, D2-4-6, and M1-4-11 from Antarctic soil were phenotypically and genotypically characterized, and their antibacterial spectrum and that of cell-free culture supernatant were investigated. Finally, the effect of temperature and culture medium on the production of antimicrobial compounds was investigated. The three bacteria were identified as different strains of the genus Pseudomonas. The three bacteria were multi-drug resistant to antibiotics. They exhibited different patterns of growth inhibition of pathogenic bacteria. M1-4-11 was remarkable for inhibiting the entire set of pathogenic bacteria tested. All three bacteria demonstrated optimal production of antimicrobial compounds at 15 °C and 18 °C. Among the culture media studied, Nutrient broth would be the most suitable to promote the production of antimicrobial compounds. The thermostability exhibited by the antimicrobial molecules secreted, their size of less than 10 kDa, and their protein nature would indicate that these molecules are bacteriocin-like compounds.
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Aquaculture is a growing industry worldwide, but it faces challenges related to animal health. These challenges include infections by parasites, bacteria, and viral pathogens. These harmful pathogens have devastating effects on the industry, despite efforts to control them through vaccination and antimicrobial treatments. Unfortunately, these measures have proven insufficient to address the sanitary problems, resulting in greater environmental impact due to the excessive use of antimicrobials. In recent years, probiotics have emerged as a promising solution to enhance the performance of the immune system against parasitic, bacterial, and viral pathogens in various species, including mammals, birds, and fish. Some probiotics have been genetically engineered to express and deliver immunomodulatory molecules. These promote selective therapeutic effects and specific immunization against specific pathogens. This review aims to summarize recent research on the use of probiotics in fish aquaculture, with a particular emphasis on genetically modified probiotics. In particular, we focus on the advantages of using these microorganisms and highlight the main barriers hindering their widespread application in the aquaculture industry.
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A completely biobased composite material was developed using a matrix of natural resin extracted from the Elaegia pastoensis Mora plant, commonly known as Mopa-Mopa or "Barniz de Pasto", reinforced with fibers extracted from plantain rachis agricultural residues. A solvent process, involving grinding, distillation, filtration, and drying stages, was implemented to extract the resin from the plant bud. To obtain the resin from the plant bud, the vegetable material was ground and then dissolved in a water-alcohol blend, followed by distillation, filtration, and grinding until the powdered resin was ready for use in the preparation of the biocomposite. Likewise, using mechanical techniques, the plantain rachis fibers were extracted and worked in their native condition and with a previous alkalinization surface treatment. Finally, the biocomposite material was developed with and without incorporating stearic acid, which was included to reduce the material's moisture absorption. Ultimately stearic acid was used as an additive to reduce biocomposite moisture absorption. The tensile mechanical results showed that the Mopa-Mopa resin reached a maximum strength of 20 MPa, which decreased with the incorporation of the additive to 12 MPa, indicating its plasticization effect. Likewise, slight decreases in moisture absorption were also evidenced with the incorporation of stearic acid. With the inclusion of rachis plantain fibers in their native state, a reduction in the tensile mechanical properties was found, proportional to the amount added. On the other hand, with the alkalinization treatment of the fibers, the behavior was the opposite, evidencing increases in tensile strength, indicating that the fiber modification improved the interfacial adhesion with the Mopa-Mopa matrix. On the other hand, the evaluation of the moisture absorption of the biocomposite material evidenced, as expected, that the absorption level was favored by the relative humidity used in the conditioning (47, 77, and 97%), which also had an impact on the decrease of the mechanical tensile properties, being this was slightly counteracted by the inclusion of stearic acid in the formulation of the material.
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The conformational changes of poly(maleic anhydride-alt-styrene) (PSMA) modified with different amino acids (PSMA-Aa) were studied in an aqueous medium as a function of ionic strength and pH. The specific viscosity of PSMA-Aa decreased with increasing salt concentration due to a more compact conformation. There was a decrease in surface tension with increasing concentrations of the modified polyelectrolyte having a greater effect for the PSMA modified with l-phenylalanine at pH 7.0, demonstrating a greater surface-active character. The conformational changes were also confirmed by molecular dynamics studies, indicating that PSMA-Aa exhibits a compact structure at pH 4.0 and a more extended structure at pH 7.0. On the other hand, the conformational changes of PSMA-Aa were related to its biological response, where the higher surface-active character of the PSMA modified with l-phenylalanine correlates very well with the higher hemolytic activity observed in red blood cells, in which the surface-active capacity supports lytic potency in erythrocytes. The cytocompatibility assays indicated that there were no significant cytotoxic effects of the PSMA-Aa. Additionally, in solvent-accessible surface area studies, it was shown that the carboxylate groups of the PSMA modified with l-phenylalanine are more exposed to the solvent at pH 7.0 and high salt concentrations, which correlates with lower fluorescence intensity, reflecting a loss of mitochondrial membrane potential. It is concluded that the study of the conformational changes in PE modified with amino acids is essential for their use as biomaterials and relevant to understanding the possible effects of PE modified with amino acids in biological systems.
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Aminoácidos , Anidridos Maleicos , Humanos , Anidridos Maleicos/química , Poliestirenos/química , Água , Fenilalanina , Hemólise , SolventesRESUMO
This work studied the effect of cellulose nanocrystal (NCC) content on the biodegradation kinetics of PLA-based multiscale cellulosic biocomposites (PLAMCBs). To facilitate biodegradation, the materials were subjected to thermo-oxidation before composting. Biodegradation was carried out for 180 days under controlled thermophilic composting conditions according to the ASTM D 5338 standard. A first-order model based on Monod's kinetics under limiting substrate conditions was used to study the effect of cellulose nanocrystal (NCC) content on the biodegradation kinetics of multiscale composite materials. It was found that thermo-oxidation at 70 °C for 160 h increased the biodegradability of PLA. Also, it was found that the incorporation of cellulosic fibrous reinforcements increased the biodegradability of PLA by promoting hydrolysis during the first stage of composting. Likewise, it was found that partial substitution of micro cellulose (MFC) by cellulose nanocrystals (NCCs) increased the biodegradability of the biocomposite. This increase was more evident as the NCC content increased, which was attributed to the fact that the incorporation of cellulose nanocrystals facilitated the entry of water into the material and therefore promoted the hydrolytic degradation of the most recalcitrant fraction of PLA from the bulk and not only by surface erosion.
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Butyrate is a microbiota-produced metabolite, sensed by host short-chain fatty acid receptors FFAR2 (Gpr43), FFAR3 (Gpr41), HCAR2 (Gpr109A), and Histone deacetylase (HDAC) that promotes microbiota-host crosstalk. Butyrate influences energy uptake, developmental and immune response in mammals. This microbial metabolite is produced by around 79 anaerobic genera present in the mammalian gut, yet little is known about the role of butyrate in the host-microbiota interaction in salmonid fish. To further our knowledge of this interaction, we analyzed the intestinal microbiota and genome of Atlantic salmon (Salmo salar), searching for butyrate-producing genera and host butyrate receptors. We identified Firmicutes, Proteobacteria, and Actinobacteria as the main butyrate-producing bacteria in the salmon gut microbiota. In the Atlantic salmon genome, we identified an expansion of genes orthologous to FFAR2 and HCAR2 receptors, and class I and IIa HDACs that are sensitive to butyrate. In addition, we determined the expression levels of orthologous of HCAR2 in the gut, spleen, and head-kidney, and FFAR2 in RTgutGC cells. The effect of butyrate on the Atlantic salmon immune response was evaluated by analyzing the pro and anti-inflammatory cytokines response in vitro in SHK-1 cells by RT-qPCR. Butyrate decreased the expression of the pro-inflammatory cytokine IL-1ß and increased anti-inflammatory IL-10 and TGF-ß cytokines. Butyrate also reduced the expression of interferon-alpha, Mx, and PKR, and decreased the viral load at a higher concentration (4 mM) in cells treated with this molecule before the infection with Infectious Pancreatic Necrosis Virus (IPNV) by mechanisms independent of FFAR2, FFAR3 and HCAR2 expression that probably inhibit HDAC. Moreover, butyrate modified phosphorylation of cytoplasmic proteins in RTgutGC cells. Our data allow us to infer that Atlantic salmon have the ability to sense butyrate produced by their gut microbiota via different specific targets, through which butyrate modulates the immune response of pro and anti-inflammatory cytokines and the antiviral response.
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Introduction: Children with cancer have a higher risk of adverse outcomes during critical illness than general pediatric populations. In Low- and middle-income countries, lack of resources can further negatively impact outcomes in critically ill children with cancer. Methods: In this study, we describe the outcomes of a large cohort of children with cancer including mortality and resource utilization. We performed a retrospective review of all patients admitted to our PICU between December 12th, 2013 and December 31st, 2019. Outcomes were defined as recovery or death and resource utilization was described via use of critical care interventions, Length of stay as well as PICU- and Mechanical Ventilation- free days. Results: Overall mortality was 6.9% while mortality in the unplanned admissions was 9.1%. This remained lower than expected mortality based on PIM2 scoring. Type of PICU admission, Neurological Deterioration as a cause of PICU admission, and PIM2 were significant as risk factors in univariate analysis, but only PIM2 remained significant in the multivariate analysis. Discussion: Our Study shows that high survival rates are achievable for children with cancer with critical illness in resource-limited settings with provision of high-quality critical care. Organizational and clinical practice facilitating quality improvement and early identification and management of critical illness may attenuate the impact of known risk factors for mortality in this population.
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Natural fiber-reinforced composite (NFRC) filaments for 3D printing were fabricated using polylactic acid (PLA) reinforced with 1-5 wt% henequen flour comprising particles with sizes between 90-250 µm. The flour was obtained from natural henequen fibers. NFRCs and pristine PLA specimens were printed with a 0° raster angle for tension tests. The results showed that the NFRCs' measured density, porosity, and degree of crystallinity increased with flour content. The tensile tests showed that the NFRC Young's modulus was lower than that of the printed pristine PLA. For 1 wt% flour content, the NFRCs' maximum stress and strain to failure were higher than those of the printed PLA, which was attributed to the henequen fibers acting as reinforcement and delaying crack growth. However, for 2 wt% and higher flour contents, the NFRCs' maximum stress was lower than that of the printed PLA. Microscopic characterization after testing showed an increase in voids and defects, with the increase in flour content attributed to particle agglomeration. For 1 wt% flour content, the NFRCs were also printed with raster angles of ±45° and 90° for comparison; the highest tensile properties were obtained with a 0° raster angle. Finally, adding 3 wt% content of maleic anhydride to the NFRC with 1 wt% flour content slightly increased the maximum stress. The results presented herein warrant further research to fully understand the mechanical properties of printed NFRCs made of PLA reinforced with natural henequen fibers.
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Carotenoids are highly important in pigmentation, and its content in farmed crustaceans and fish correlates to their market value. These pigments also have a nutritional role in aquaculture where they are routinely added as a marine animal food supplement to ensure fish development and health. However, there is little information about carotenoids obtained from Antarctic bacteria and its use for pigmentation improvement and flesh quality in aquaculture. This study identified carotenoids produced by Antarctic soil bacteria. The pigmented strain (CN7) was isolated on modified Luria-Bertani (LB) media and incubated at 4 °C. This Gram-negative bacillus was identified by 16S rRNA analysis as Flavobacterium segetis. Pigment extract characterization was performed through high-performance liquid chromatography (HPLC) and identification with liquid chromatography-mass spectrometry (LC-MS). HPLC analyses revealed that this bacterium produces several pigments in the carotenoid absorption range (six peaks). LC-MS confirms the presence of one main peak corresponding to lutein or zeaxanthin (an isomer of lutein) and several other carotenoid pigments and intermediaries in a lower quantity. Therefore, we propose CN7 strain as an alternative model to produce beneficial carotenoid pigments with potential nutritional applications in aquaculture.
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In salmon farming, viruses are responsible for outbreaks that produce significant economic losses for which there is a lack of control tools other than vaccines. Type I interferon has been successfully used for treating some chronic viral infections in humans. However, its application in salmonids depends on the proper design of a vehicle that allows its massive administration, ideally orally. In mammals, administration of recombinant probiotics capable of expressing cytokines has shown local and systemic therapeutic effects. In this work, we evaluate the use of Lactococcus lactis as a type I Interferon expression system in Atlantic salmon, and we analyze its ability to stimulate the antiviral immune response against IPNV, in vivo and in vitro. The interferon expressed in L. lactis, even though it was located mainly in the bacterial cytoplasm, was functional, stimulating Mx and PKR expression in CHSE-214 cells, and reducing the IPNV viral load in SHK-1 cells. In vivo, the oral administration of this L. lactis producer of Interferon I increases Mx and PKR expression, mainly in the spleen, and to a lesser extent, in the head kidney. The oral administration of this strain also reduces the IPNV viral load in Atlantic salmon specimens challenged with this pathogen. Our results show that oral administration of L. lactis producing Interferon I induces systemic effects in Atlantic salmon, allowing to stimulate the antiviral immune response. This probiotic could have effects against a wide variety of viruses that infect Atlantic salmon and also be effective in other salmonids due to the high identity among their type I interferons.