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This study investigated crotamine (CTA), a peptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, known for its exceptional cell penetration potential. The objective was to explore the antibacterial and antifungal activity of CTA, its ability to inhibit efflux pumps and evaluate the effectiveness of its pharmacological combination with antibiotics and antifungals. In microbiological assays, CTA in combination with antibiotics was tested against strains of S. aureus and the inhibition of NorA, Tet(K) and MepA efflux pumps was also evaluated. CTA alone did not present clinically relevant direct antibacterial action, presenting MIC > 209.7 µM against strains S. aureus 1199B, IS-58, K2068. The standard efflux pump inhibitor CCCP showed significant effects in all negative relationships to assay reproducibility. Against the S. aureus 1199B strain, CTA (20.5 µM) associated with norfloxacin diluted 10 × (320.67 µM) showed a potentiating effect, in relation to the control. Against the S. aureus IS-58 strain, the CTA associated with tetracycline did not show a significant combinatorial effect, either with 2304 or 230.4 µM tetracycline. CTA at a concentration of 2.05 µM associated with ciprofloxacin at a concentration of 309.4 µM showed a significant potentiating effect. In association with EtBr, CTA at concentrations of 2.05 and 20.5 µM potentiated the effect in all strains tested, reducing the prevention of NorA, Tet(K) and MepA efflux pumps. In the C. albicans strain, a potentiating effect of fluconazole (334.3 µM) was observed when combined with CTA (2.05 µM). Against the C. tropicalis strain, a significant effect was also observed in the association of fluconazole 334.3 µM, where CTA 2.05 µM considerably reduced fungal growth and decreased the potentiation of fluconazole. Against the C. krusei strain, no significant potentiating effect of fluconazole was obtained by CTA. Our results indicate that CTA in pharmacological combination potentiates the effects of antibiotics and antifungal. This represents a new and promising antimicrobial strategy for treating a wide variety of infections.

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Thermolysin (TLN) is a microbial highly-priced thermostable metallo-endoprotease with complementary substrate specificity to those of proteases widely used in science and industry for protein digestion and milk-clotting. This study is the first to immobilize TLN on aminated superparamagnetic nanoparticles (Fe3O4@silica-NH2) aiming for higher stability, recoverability, reusability, and applicability in proteolysis and as a microbial rennet-like milk-clotting enzyme. The nanobiocatalyst developed (Fe3O4@silica-TLN) displays hydrolytic activity on a synthetic TLN substrate and, apparently, was fully recovered from reaction media by magnetic decantation. More importantly, Fe3O4@silica-TLN retains TLN catalytic properties in the presence of calcium ions even after exposure to 60 °C for 48 h, storage at 4 °C for 80 days and room temperature for 42 days, use in proteolyses, and in milk-clotting for up to 11 cycles. Its proteolytic activity on bovine milk casein in 24 h furnished 84 peptides, of which 29 are potentially bioactive. Also, Fe3O4@silica-TLN catalyzed the digestion of bovine serum albumin. In conclusion, Fe3O4@silica-TLN showed to be a new, less autolytic, thermostable, non-toxic, magnetically-separable, and reusable nanobiocatalyst with highly attractive properties for both science (peptide/protein chemistry and structure, proteomic studies, and the search for new bioactive peptides) and food industry (cheese manufacture).

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Microbial resistance to drugs continues to be a global public health issue that demands substantial investment in research and development of new antimicrobial agents. Essential oils (EO) have demonstrated satisfactory and safe antimicrobial action, being used in pharmaceutical, cosmetic, and food formulations. In order to improve solubility, availability, and biological action, EO have been converted into nanoemulsions (NE). This review identified scientific evidence corroborating the antimicrobial action of nanoemulsions of essential oils (NEEO) against antibiotic-resistant pathogens. Using integrative review methodology, eleven scientific articles evaluating the antibacterial or antifungal assessment of NEEO were selected. The synthesis of evidence indicates that NEEO are effective in combating multidrug-resistant microorganisms and in the formation of their biofilms. Factors such as NE droplet size, chemical composition of essential oils, and the association of NE with antibiotics are discussed. Furthermore, NEEO showed satisfactory results in vitro and in vivo evaluations against resistant clinical isolates, making them promising for the development of new antimicrobial and antivirulence drugs.

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Soil microbial traits and functions play a central role in soil organic carbon (SOC) dynamics. However, at the macroscale (regional to global) it is still unresolved whether (i) specific environmental attributes (e.g., climate, geology, soil types) or (ii) microbial community composition drive key microbial traits and functions directly. To address this knowledge gap, we used 33 grassland topsoils (0-10 cm) from a geoclimatic gradient in Chile. First, we incubated the soils for 1 week in favorable standardized conditions and quantified a wide range of soil microbial traits and functions such as microbial biomass carbon (MBC), enzyme kinetics, microbial respiration, growth rates as well as carbon use efficiency (CUE). Second, we characterized climatic and physicochemical properties as well as bacterial and fungal community composition of the soils. We then applied regression analysis to investigate how strongly the measured microbial traits and functions were linked with the environmental setting versus microbial community composition. We show that environmental attributes (predominantly the amount of soil organic matter) determined patterns of MBC along the gradient, which in turn explained microbial respiration and growth rates. However, respiration and growth normalized for MBC (i.e., specific respiration and growth) were more linked to microbial community composition than environmental attributes. Notably, both specific respiration and growth followed distinct trends and were related to different parts of the microbial community, which in turn resulted in strong effects on microbial CUE. We conclude that even at the macroscale, CUE is the result of physiologically decoupled aspects of microbial metabolism, which in turn is partially determined by microbial community composition. The environmental setting and microbial community composition affect different microbial traits and functions, and therefore both factors need to be considered in the context of macroscale SOC dynamics.

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Introduction: The aim of the research was to demonstrate the efficiency of microorganisms and the effectiveness of biodegradation techniques on Low-density polyethylene (LDPE) plastics. The research question was: What is the efficiency of LDPE-degrading microorganisms and the effectiveness of biodegradation techniques? Methods: The systematic review was based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Articles were obtained from Scopus, Web of Science (WOS), Embase, and Google Scholar. The DeCS/Mesh search terms were: Low-density polyethylene, efficiency, biodegradation, microbial consortia, fungi, bacteria. Inclusion criteria were: scientific articles that included bacteria, fungi, and microbial consortia reported as LDPE degraders that report the percentage of weight loss; articles published from January 2010 to October 2022, and publications in Spanish and English with open access. Exclusion criteria were: studies that do not report gravimetry, the biodegradation time of LDPE, and the genus or species of the polyethylene-degrading microorganism. Results: Out of 483 studies found, 50 were included in this Systematic Review (SR). The most frequent study techniques were scanning electron microscopy (SEM), gravimetry, and fourier transform infrared spectroscopy (FTIR), and in the case of microorganisms, the most studied belonged to the genus Pseudomonas, Bacillus, and Aspergillus. Regarding the isolation place, the most frequent mentioned in the reviewed articles were landfill soil and sanitary landfill soil. The efficiency of LDPE-degrading microorganisms was higher in bacteria such as Enterobacter spp., Pantoea spp., Pseudomonas spp., Escherichia coli, and Bacillus spp., which obtained a range of DE of 9.00-70.00%, 24.00-64%, 1.15 - 61.00%, 45.00%, and 1.5-40% with DT of 4-150, 120, 4-150, 30, and 30-120 days, respectively; in the case of fungi, the main microorganisms are Neopestalotiopsis phangngaensis, Colletotrichum fructicola, and Thyrostroma jaczewskii with efficiencies of 54.34, 48.78, and 46.34%, in 90 days, respectively; and the most efficient microbial consortia were from Enterobacter spp. and Pantoea sp. with 38.00 - 81.00%, in 120 days; and, Pseudomonas protegens, Stenotrophomonas sp., B. vallismortis and Paenibacillus sp. with 55. 00 - 75.00% in 120 days. Conclusions: The most efficient microorganisms in LDPE degradation are Enterobacter spp., Pantoea spp., Pseudomonas spp., Escherichia coli, and Bacillus spp.; in fungi Neopestalotiopsis phangngaensis, Colletotrichum fructicola, and Thyrostroma jaczewskii; and in microbial consortia, those formed by Enterobacter spp. and Pantoea sp., and that of P. protegens, Stenotrophomonas sp., B. vallismortis and Paenibacillus sp.; and the most effective techniques used in LDPE biodegradation are SEM, gravimetry, and FTIR.

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The activities of microbiomes in river sediments play an important role in sustaining ecosystem functions by driving many biogeochemical cycles. However, river ecosystems are frequently affected by anthropogenic activities, which may lead to microbial biodiversity loss and/or changes in ecosystem functions and related services. While parts of the Atlantic Forest biome stretching along much of the eastern coast of South America are protected by governmental conservation efforts, an estimated 89% of these areas in Brazil are under threat. This adds urgency to the characterization of prokaryotic communities in this vast and highly diverse biome. Here, we present prokaryotic sediment communities in the tropical Juliana River system at three sites, an upstream site near the river source in the mountains (Source) to a site in the middle reaches (Valley) and an estuarine site near the urban center of Ituberá (Mangrove). The diversity and composition of the communities were compared at these sites, along with environmental conditions, the former by using qualitative and quantitative analyses of 16S rRNA gene amplicons. While the communities included distinct populations at each site, a suite of core taxa accounted for the majority of the populations at all sites. Prokaryote diversity was highest in the sediments of the Mangrove site and lowest at the Valley site. The highest number of genera exclusive to a given site was found at the Source site, followed by the Mangrove site, which contained some archaeal genera not present at the freshwater sites. Copper (Cu) concentrations were related to differences in communities among sites, but none of the other environmental factors we determined was found to have a significant influence. This may be partly due to an urban imprint on the Mangrove site by providing organic carbon and nutrients via domestic effluents.

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The use of natural and sustainable additives, that are less aggressive to the environment, is a trend in the food industry. Rhamnolipids (RL) biosurfactants have shown potential for controlling food pathogens however, due to the presence of free carboxyl groups, the pH and ionic strength may influence the properties of such surfactants. In this study, we describe the antimicrobial activity of RL under different pH values and NaCl concentrations, towards both planktonic and biofilms of Listeria monocytogenes. RL were effective at pH 5.0 and the addition of 5 % NaCl improved the bactericidal efficacy for planktonic and sessile cells. The effect of NaCl was more pronounced at pH above 6 showing a significant increase in RL antimicrobial activity. At pH 7.0 planktonic population was eradicated by RL only when salt was present whereas biofilm viability was decreased by 5 log with MBIC varying from > 2500.0 mg/L (RL) to 39.0 mg/L (RL + 5 % NaCl). Larger vesicular and lamellar RL self-assembly structures were predominant when NaCl was present, suggesting their association with the antimicrobial activity observed. The pH and ionic strength of the medium are important parameters to be considered for the development of RL-based strategies to control L. monocytogenes.

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Protists can endure challenging environments sustaining key ecosystem processes of the microbial food webs even under aridic or hypersaline conditions. We studied the diversity of protists at different latitudes of the Atacama Desert by massive sequencing of the hypervariable region V9 of the 18S rRNA gene from soils and microbial mats collected in the Andes. The main protist groups in soils detected in active stage through cDNA were cercozoans, ciliates, and kinetoplastids, while the diversity of protists was higher including diatoms and amoebae in the microbial mat detected solely through DNA. Co-occurrence networks from soils indicated similar assemblages dominated by amplicon sequence variants (ASVs) identified as Rhogostoma, Euplotes, and Neobodo. Microbial mat networks, on the other hand, were structured by ASVs classified as raphid-pennate diatoms and amoebae from the genera Hartmannella and Vannella, mostly negatively correlated to flagellates and microalgae. Additionally, our phylogenetic inferences of ASVs classified as Euplotes, Neobodo, and Rhogostoma were supported by sequence data of strains isolated during this study. Our results represent the first snapshot of the diversity patterns of culturable and unculturable protists and putative keystone taxa detected at remote habitats from the Atacama Desert.

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The study aims to assess the impact of oven-drying and decontamination on crude protein concentration and in vitro crude protein digestibility of yellow mealworms. Two kilograms of 12-wk-old mealworm larvae were subjected to freezing prior to the drying process. Approximately 1.5 kg of mealworm larvae were divided into 3 groups and exposed to oven-drying at temperatures of 50 °C for 36 h, 60 °C, and 70 °C for 24 h each. At intervals of 2 h, sets of 3 replicates were withdrawn to record water loss. Consistent weight stabilization was observed at 36 h for 50 °C (T50), 18 h for 60 °C (T60), and 14 h for 70 °C (T70). The remaining 0.5 kg of mealworm larvae was divided and dried under treatments T50, T60, and T70. Each treatment was then split into 2 portions, with one portion subjected to 90 °C for 15 min (denoted as T50-90, T60-90, T70-90) to eliminate microbial contamination. The 6 treatments were then used to determine concentrations of dry matter, crude ash, crude protein, pre-caecal protein digestibility, and dry matter residues after neutral detergent fiber, acid detergent fiber, and acid detergent lignin treatments. No interaction was observed between drying and decontamination treatments (P > 0.17). Pre-caecal crude protein digestibility increased with decreasing temperature (T50: 58% crude protein; T60: 51% crude protein; T70: 50% crude protein). Therefore, lower temperatures for longer times preserve crude protein digestibility. These findings are crucial for understanding how drying temperature and time impact protein bioavailability.

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This study presents comprehensive insights into the microbiological profile across all concentrated chicken broth processing stages, utilizing a combination of amplicon sequencing based on metataxonomic and culturing techniques. Samples were systematically collected throughout the production chain, with each batch yielding 10 samples per day across eight different dates. These samples underwent thorough analysis, including 16S rRNA and ITS sequencing (n = 30), culture-dependent microbiological tests (n = 40), and physical-chemical characterization (n = 10). Culturing analysis revealed the absence of Listeria monocytogenes and Salmonella spp. at any stage of processing, counts of various microorganisms such as molds, yeasts, Enterobacteria, and others remained below detection limits. Notably, spore counts of selected bacterial groups were observed post-processing, indicating the persistence of certain species, including Bacillus cereus and Clostridium perfringens, albeit in low counts. Furthermore, the study identified a diverse array of bacterial and fungal species throughout the processing chain, with notable occurrence of spore-forming bacteria. The presence of spore-forming bacteria in the final product, despite thermal processing, suggests the need for enhanced strategies to mitigate their introduction and persistence in the processing premises. Thus, this study offers valuable insights into microbial dynamics and diversity through processing concentrated chicken broth.

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