Optimizing Lactic Acid Bacteria Proportions in Sourdough to Enhance Antifungal Activity and Quality of Partially and Fully Baked Bread.

The organic acids produced by lactic acid bacteria (LAB) during the fermentation of sourdoughs have the ability to reduce the growth of different molds. However, this ability depends on the LAB used. For this reason, in this study, the proportions of different LAB were optimized to obtain aqueous extracts (AEs) from sourdough to reduce fungal growth in vitro, control the acetic acid concentration, and obtain a specific lactic to acetic acid ratio. In addition, the optimized mixtures were used to formulate partially baked bread (PBB) and evaluate the mold growth and bread quality during refrigerated storage. Using a simplex-lattice mixture design, various combinations of Lactiplantibacillus plantarum, Lacticaseibacillus casei, and Lactobacillus acidophilus were evaluated for their ability to produce organic acids and inhibit mold growth. The mixture containing only Lpb. plantarum significantly reduced the growth rates and extended the lag time of Penicillium chrysogenum and P. corylophilum compared with the control. The AEs' pH values ranged from 3.50 to 3.04. Organic acid analysis revealed that using Lpb. plantarum yielded higher acetic acid concentrations than when using mixed LAB. This suggests that LAB-specific interactions significantly influence organic acid production during fermentation. The reduced radial growth rates and extended lag times for both molds compared to the control confirmed the antifungal properties of the AEs from the sourdoughs. Statistical analyses of the mixture design using polynomial models demonstrated a good fit for the analyzed responses. Two optimized LAB mixtures were identified that maximized mold lag time, targeted the desired acetic acid concentration, and balanced the lactic to acetic acid ratio. The addition of sourdough with optimized LAB mixtures to PBB resulted in a longer shelf life (21 days) and adequately maintained product quality characteristics during storage. PBB was subjected to complete baking and sensory evaluation. The overall acceptability was slightly higher in the control without sourdough (7.50), followed by bread formulated with the optimized sourdoughs (ranging from 6.78 to 7.10), but the difference was not statistically significant (p > 0.05). The sensory analysis results indicated that the optimization was used to successfully formulate a sourdough bread with a sensory profile closely resembling that of a nonsupplemented one. The designed LAB mixtures can effectively enhance sourdough bread's antifungal properties and quality, providing a promising approach for extending bread shelf life while maintaining desirable sensory attributes.

Employment of pqqE gene as molecular marker for the traceability of Gram negative phosphate solubilizing bacteria associated to plants.

Insoluble phosphorous compounds solubilization by soil bacteria is of great relevance since it puts available the phosphorus to be used by plants. The production of organic acids is the main microbiological mechanism by which insoluble inorganic phosphorus compounds are solubilized. In Gram negative bacteria, gluconic acid is synthesized by the activity of the holoenzyme glucose dehydrogenase-pyrroloquinoline quinine named GDH-PQQ. The use of marker genes is a very useful tool to evaluate the persistence of the introduced bacteria and allow to follow-up the effect of biotic and abiotic factors on these beneficial microorganisms in the soil. In previous studies we detected the presence of the pqqE gene in a great percentage of both non-culturable and culturable native soil bacteria. The objective of this study was to analyze the phylogeny of the sequence of pqqE gene and its potential for the study of phosphate solubilizing bacteria from pure and mixed bacterial cultures and rhizospheric soil samples. For this, the presence of the pqqE gene in the genome of phosphate solubilizing bacteria that belong to several bacteria was determined by PCR. Also, this gene was analyzed from mixed bacterial cultures and rhizospheric soil associated to peanut plants inoculated or not with phosphate solubilizing bacteria. For this, degenerate primers designed from several bacterial genera and specific primers for the genus Pseudomonas spp., designed in this study, were used. DNA template used from simple or mixed bacterial cultures and from rhizospheric soil samples was obtained using two different DNA extraction techniques. Results indicated that pqqE gene amplification product was found in the genome of all Gram negative phosphate solubilizing bacteria analyzed. It was possible to detect this gene in the DNA obtained from mixed cultures where these bacteria grew in interaction with other microorganisms and in that obtained from rhizospheric soil samples inoculated or not with these bacteria. The phylogenetic analysis indicated that pqqE gene is a conserved gene within related genera. In conclusion, pqqE gene could be a potential marker for the study of phosphate solubilizing bacterial populations.

Peptide PaDBS1R6 has potent antibacterial activity on clinical bacterial isolates and integrates an immunomodulatory peptide fragment within its sequence.

BACKGROUND: Resistant infectious diseases caused by gram-negative bacteria are among the most serious worldwide health problems. Antimicrobial peptides (AMPs) have been explored as promising antibacterial, antibiofilm, and anti-infective candidates to address these health challenges. MAJOR CONCLUSIONS: Here we report the potent antibacterial effect of the peptide PaDBS1R6 on clinical bacterial isolates and identify an immunomodulatory peptide fragment incorporated within it. PaDBS1R6 was evaluated against Acinetobacter baumannii and Escherichia coli clinical isolates and had minimal inhibitory concentration (MIC) values from 8 to 32 µmol L-1. It had a rapid bactericidal effect, with eradication showing within 3 min of incubation, depending on the bacterial strain tested. In addition, PaDBS1R6 inhibited biofilm formation for A. baumannii and E. coli and was non-toxic toward healthy mammalian cells. These findings are explained by the preference of PaDBS1R6 for anionic membranes over neutral membranes, as assessed by surface plasmon resonance assays and molecular dynamics simulations. Considering its potent antibacterial activity, PaDBS1R6 was used as a template for sliding-window fr agmentation studies (window size = 10 residues). Among the sliding-window fragments, PaDBS1R6F8, PaDBS1R6F9, and PaDBS1R6F10 were ineffective against any of the bacterial strains tested. Additional biological assays were conducted, including nitric oxide (NO) modulation and wound scratch assays, and the R6F8 peptide fragment was found to be active in modulating NO levels, as well as having strong wound healing properties. GENERAL SIGNIFICANCE: This study proposes a new concept whereby peptides with different biological properties can be derived by the screening of fragments from within potent AMPs.

The fungus Aphanoascella galapagosensis affects bacterial diversity of Galapagos giant tortoise carapaces.

AIMS: This study aimed to describe the bacterial microbiome associated with the carapace of three species of Galapagos giant tortoises (Chelonoidis porteri, Chelonoidis donfaustoi, and Chelonoidis vandenburghi) and determine the potential effect of the whitish lesions caused by the fungus Aphanoascella galapagosensis. METHODS AND RESULTS: We used Oxford Nanopore's MinION to evaluate the external bacterial microbiome associated with the carapaces from the aforementioned species. Taxonomic assignment was carried out by Bugseq and the bacterial communities were compared between carapaces with and without lesions using a NMDS with Bray-Curtis as the dissimilarity index. We found four genera of bacteria that were ubiquitous throughout all individuals, suggesting the presence of shared taxa. The results also displayed a significant difference in the microbiome between carapaces with and without lesions, and for species-carapace interaction, but not among species. CONCLUSIONS: This study establishes a baseline of the bacterial diversity of the carapace within three Galapagos giant tortoise species, showcasing the presence of a distinctive microbial community. Furthermore, our findings suggest a significant influence of the fungus Aphanoascella galapagosensis on the bacterial populations inhabiting the carapace of these reptiles.

Challenges and trends in Gram-negative bacterial infections in critically neonates: A seven-and-a-half-year observational study.

BACKGROUND: Analyze the incidence, risk factors, and fatality rates of bloodstream infections by Gram-negative bacteria (GNB-BSIs) in a Neonatal Intensive Care Unit. METHODS: This study employs a retrospective cohort design utilizing records of neonates admitted to the Neonatal Intensive Care Unit between January 2015 and June 2022. RESULTS: Among 1,495 neonates, 5.2% developed GNB-BSIs. The average incidence of infection per 1,000 patient-days was 2.9. Primary risk factors for infection that included preceeding carbapenem use were significant risk factors (odds ratio=514.4; P < .01) and fourth-generation cephalosporins (odds ratio=66; P < .01). Among the 85 GNB, 75.3% were fermenters, and 24.7% were non-fermenters. Of the isolates, 14.1% produced extended-spectrum beta-lactamase, and 2.3% carbapenem-resistant. Infection correlated with prolonged hospital stays (10-39days) and increased mortality (10%-29.9%). CONCLUSIONS: The high incidence of GNB-BSIs was exacerbated by the preceeding use of broad-spectrum antimicrobials, increasing the presence of multidrug-resistant isolates and fatality rates. These findings emphasize the importance of active surveillance.

Comprehensive dynamics of bacterial and fungal diversity throughout concentrated chicken broth processing: Integrating culture-dependent and independent techniques.

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.

Discovery of antimicrobial peptides clostrisin and cellulosin from Clostridium: insights into their structures, co-localized biosynthetic gene clusters, and antibiotic activity.

Antimicrobial resistance presents a substantial threat to global public health, demanding urgent attention and action. This study focuses on lanthipeptides, ribosomally encoded peptides that display significant structural diversity and hold promising potential as antibiotics. Genome mining was employed to locate biosynthetic gene clusters (BGCs) containing class II lanthipeptide synthetases encoded by lanM genes. A phylogenetic study analyzing homologous sequences of functional LanM sequences revealed a unique evolutionary clade of 17 LanM proteins associated with 12 Clostridium bacterial genomes. In silico exploration identified nine complete BGCs, including one super-cluster containing two co-localized operons from Clostridium cellulovorans 743B, that encode for two new peptides named clostrisin and cellulosin. Each operon was heterologously expressed in Escherichia coli. Molecular weights associated with the expected post-translational modifications of the purified lanthipeptide were confirmed by MS-MS/MS analysis for cellulosin, while clostrisin was not post-translationally modified. Both peptides demonstrated antimicrobial activity against multidrug-resistant bacteria, such as a clinical strain of Staphylococcus epidermidis MIQ43 and Pseudomonas aeruginosa PA14. This is the first report of lanthipeptides from the Clostridium genus produced with its native biosynthetic machinery, as well as chemically and biologically characterized. This study showcases the immense potential of genome mining in identifying new RiPP synthetases and associated bioactive peptides.

Quinoa snack elaborated with Lactiplantibacillus plantarum CRL 1964 sourdough increases the mineral bioavailability in mice.

BACKGROUND: Consumption of pseudocereal-based foods decreased in phytate concentration can provide better nutrition concerning mineral bioavailability. This study aimed to evaluate the mineral bioavailability of quinoa sourdough-based snacks in a murine model. The mice were divided into five groups. One group was fed with basal snacks; three control groups received quinoa-based snacks made from non-fermented dough, dough without inoculum, and chemically acidified dough; and the test group (GF) received quinoa snacks elaborated from sourdough fermented by a phytase-positive strain, Lactiplantibacillus plantarum CRL 1964. Food intake, body weight, and mineral concentration in blood and organs (liver, kidney, and femur) were determined. RESULTS: Food consumption increased during the feeding period and had the highest (16.2-24.5%) consumption in the GF group. Body weight also increased during the 6-weeks of trial. The GF group showed higher (6.0-10.2%) body weight compared with the other groups from the fifth week. The concentrations of iron, zinc, calcium, magnesium, and phosphorus in blood, iron and phosphorus in the liver, manganese and magnesium in the kidney, and calcium and phosphorus in the femur increased significantly (1.1-2.7-fold) in the GF group compared to the control groups. CONCLUSION: The diet that includes quinoa snacks elaborated with sourdough fermented by phytase-positive strain L. plantarum CRL 1964 increased the concentrations of minerals in the blood, liver, kidney, and femur of mice, counteracting the antinutritional effects of phytate. This study demonstrates that the diminution in phytate content and the consequent biofortification in minerals are a suitable tool for producing novel foods. © 2024 Society of Chemical Industry.

Response of sweet potato cultivars to Bacillus velezensis T149-19 and Bacillus safensis T052-76 used as biofertilizers.

The global market of sweet potato (Ipomoea batatas (L.) Lam.) is continuously growing and, consequently, demands greater productivity from the agricultural sector. The use of biofertilizers facilitates plant growth by making essential nutrients available to crops or providing resistance against different abiotic and biotic factors. The strains Bacillus safensis T052-76 and Bacillus velezensis T149-19 have previously been inoculated in the sweet potato cultivar Ourinho, showing positive effects on plant shoot growth and inhibiting the phytopathogen Plenodomus destruens. To elucidate the effects of these strains on sweet potato growth, four different cultivars of sweet potato were selected: Capivara, IAPAR 69, Rosinha de Verdan and Roxa. The plants were grown in pots in a greenhouse and inoculated with the combined strains according to a randomized block design. A control (without the inoculation of both strains) was also used. A slight positive effect of the inoculation of the two Bacillus strains was observed on the aerial parts of some of the cultivars. An increase in the fresh weight of the sweet potatoes of the inoculated plants was obtained, varying from 2.7 to 11.4 %. The number of sweet potatoes obtained from the inoculated cultivars IAPAR 69 and Roxa increased 15.2 % and 16.7 %, respectively. The rhizosphere soil of each cultivar was further sampled for DNA extraction, and the 16S rRNA gene metabarcoding technique was used to determine how the introduction of these Bacillus strains influenced the rhizosphere bacterial community. The bacterial communities of the four different cultivars were dominated by Actinobacteria, Proteobacteria and Firmicutes. Nonmetric multidimensional scaling (NMDS) revealed that the rhizosphere bacterial communities of plants inoculated with Bacillus strains were more similar to each other than to the bacterial communities of uninoculated plants. This study highlights the contribution of these Bacillus strains to the promotion of sweet potato growth.

Neuroprotective Effects Exerted by a Combination of Selected Lactic Acid Bacteria in a Mouse Parkinsonism Model under Levodopa-Benserazide Treatment.

Alterations of the microbiota-gut-brain axis has been associated with intestinal and neuronal inflammation in Parkinson's disease (PD). The aim of this work was to study some mechanisms associated with the neuroprotective effect of a combination (MIX) of lactic acid bacteria (LAB) composed by Lactiplantibacillus plantarum CRL2130 (riboflavin overproducing strain), Streptococcus thermophilus CRL808 (folate producer strain), and CRL807 (immunomodulatory strain) in cell cultures and in a chronic model of parkinsonism induced with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in aged mice, and under levodopa-benserazide treatment. In vitro, N2a differentiated neurons were exposed to the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or with conditioned media from BV-2 cells exposed to the bacterial extracts. In vivo, motor skills, tyrosine hydrolase (TH) in brain and cytokine concentrations in serum and in brain were evaluated. The study of the faecal microbiota and the histology of the small intestine was also performed. The results showed that the neuroprotective effect associated with LAB MIX administration did not interfere with levodopa-benserazide treatment. This effect could be associated with the antioxidant and immunomodulatory potential of the LAB selected in the MIX, and was associated with the significant improvement in the motor tests and a higher number of TH + cells in the brain. In addition, LAB MIX administration was associated with modulation of the immune response. LAB administration decreased intestinal damage with an increase in the villus length /crypt depth ratio. Finally, the administration of the LAB MIX in combination with levodopa-benserazide treatment was able to partially revert the intestinal dysbiosis observed in the model, showing greater similarity to the profiles of healthy controls, and highlighting the increase in the Lactobacillaceae family. Different mechanisms of action would be related to the protective effect of the selected LAB combination which has the potential to be evaluated as an adjuvant for conventional PD therapies.

Efficiency of Microorganisms and Effectiveness of Biodegradation Techniques on LDPE Plastics: A Systematic Review.

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.

Diversity of endophytic bacteria with antimicrobial potential isolated from marine macroalgae from Yacila and Cangrejos beaches, Piura-Peru.

Endophytic bacteria found in marine macroalgae have been studied for their potential antimicrobial activity, consequently, they could serve as a valuable source of bioactive compounds to control pathogenic bacteria, yeasts, and fungi. Algae endophytic bacteria were isolated from Caulerpa sp., Ulva sp., Ahnfeltiopsis sp., and Chondracantus chamissoi from Yacila and Cangrejo Beaches (Piura, Peru). Antimicrobial assays against pathogenic bacteria were evaluated using cross-culture, over-plate, and volatile organic compound tests. Afterward, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of selected crude extracts were determined, also ITS molecular analysis, antifungal activity, and PCR of iturin, fengycin, and surfactin genes were performed for bacteria strains exhibiting better activity. Forty-six algae endophytic bacteria were isolated from algae. Ten strains inhibited gram-positive pathogenic bacteria (Enterococcus faecalis, Staphylococcus epidermidis, S. aureus, and Listeria monocytogenes), and 12 inhibited gram-negative bacteria (Escherichia coli and Salmonella enteric sv typhimurium). Bacteria with better activity belong to Bacillus sp., Kluyvera ascorbata, Pantoea agglomerans, Leclercia adecarboxylata, and Enterobacter sp., which only four showed antifungal activities against Candida albicans, C. tropicalis, Colletotrichium sp., Fusarium sp., Fusarium oxysporum, and Alternaria sp. Furthermore, K. ascorbata YAFE21 and Bacillus sp. YCFE4 exhibited iturin and fengycin genes. The results indicate that the algae endophytic bacteria found in this study, particularly K. ascorbata YAFE21, Bacillus sp. YCFR6, L. adecarboxylata CUFE2, Bacillus sp. YUFE8, Enterobacter sp. YAFL1, and P. agglomerans YAFL6, could be investigated as potential producers of antimicrobial compounds due to their broad activity against various microorganisms.

Calidad higiénico-sanitaria y prácticas de manufactura de alimentos en un comedor estudiantil en México / Hygienic ­ sanitary quality and food manufacturing practices in a student cafeteria in Mexico / Qualidade higiênico-sanitária e práticas de fabricação de alimentos em uma refeitório estudantil no México

Introducción: El apego a las normas oficiales sanitarias sirve para prevenir riesgos a la salud humana. Objetivo: Evaluar la calidad higiénico-sanitaria y las buenas prácticas de manufactura de alimentos (BPMA) de un comedor estudiantil en México. Materiales y métodos: Estudio cuasiexperimental y analítico. Durante el año 2020, se realizaron pruebas bacteriológicas a muestras de alimentos, agua, superficies y manos de manipuladores de alimentos, además de también evaluar las BPMA. Conforme a las normas oficiales sanitarias vigentes en México, se recolectaron 57 muestras, se aislaron y se lograron identificar patógenos. Las BPMA se valoraron en 20 manipuladores, antes y después de una intervención educativa de 10 semanas de duración y se utilizó la prueba t con α=0,05. Resultados: Más del 50 % de las muestras resultaron con microorganismos de riesgo para la salud, como Escherichia coli, Staphylococcus aureus, Pseudomonas, Acinetobacter baumanni complex y Coliformes totales. Las evaluaciones, antes y después de la intervención educativa de BPMA, evidenciaron diferencias estadísticamente significativas en el número de aciertos (p≤0,05). Conclusiones: La calidad higiénico-sanitaria del comedor analizado representó riesgo para la salud de los estudiantes, lo cual tuvo relación con la primera evaluación de las BPMA entre los manipuladores, las cuales mejoraron después de la intervención.

Introduction: Adherence to official health standards is essential to prevent human health risks. Objective: To assess the hygienic-sanitary quality and good food manufacturing practices (GMP) in a student cafeteria in Mexico. Materials and methods: Quasi-experimental and analytical study. During 2020, bacteriological tests were carried out on samples taken from food, water, surfaces, and hands of food handlers. In addition, GMP were evaluated. Based on the current Mexican official health regulations, 57 samples were collected to isolate and identify pathogens. GMP were assessed in 20 food handlers before and after a 10-week training intervention and a test was used with α=0.05. Results: More than 50% of samples were found to have microorganisms associated with health risks, including Escherichia coli, Staphylococcus aureus, Pseudomonas, Acinetobacter baumanni complex and total Coliforms. The analyses before and after the GMP training intervention showed statistically significant differences in terms of the presence of these pathogens (p≤0.05). Conclusions: The hygienic-sanitary quality of the analyzed cafeteria turned out to be a risk for the health of students, which was related to the first assessment of GMP in food handlers. Consequently, the results improved after the intervention.

Introdução: A adesão às normas sanitárias oficiais serve para prevenir riscos à saúde humana. Objetivo: Avaliar a qualidade higiênico-sanitária e as boas práticas de fabricação de alimentos (BPMA) de um refeitorio estudantil no México. Materiais e métodos: Estudo quase-experimental e analítico. Durante 2020, foram realizados testes bacteriológicos em amostras de alimentos, água, superfícies e mãos de manipuladores de alimentos, além de avaliação de BPMA. De acordo com as normas sanitárias oficiais em vigor no México, foram coletadas e isoladas 57 amostras e identificados patógenos. Os BPMA foram avaliados em 20 manipuladores, antes e após uma intervenção educativa de 10 semanas e foi utilizado o teste t com α=0,05. Resultados: Verificou-se que mais de 50% das amostras continham microrganismos de risco à saúde, como Escherichia coli, Staphylococcus aureus, Pseudomonas, complexo Acinetobacter baumanni e Coliformes totais. As avaliações, antes e após a intervenção educativa BPMA, apresentaram diferenças estatisticamente significativas no número de acertos (p≤0,05). Conclusões: A qualidade higiênico-sanitária do refeitório analisado representou um risco para a saúde dos alunos, o que esteve relacionado à primeira avaliação do BPMA entre os manipuladores, que melhorou após a intervenção.

Identificación de microorganismos entéricos en áreas de preparación y consumo de alimentos / Identification of enteric microorganisms in food preparation and consumption areas / Identificação de microrganismos entéricos em áreas de preparação e consumo de alimentos

La contaminación de las áreas de preparación al entrar en contacto con los alimentos crudos o cocinados, es por esto que una de las principales causas de la contaminación de las superficies inertes es la inadecuada manipulación de los alimentos a la hora de ser preparados. Con el objetivo de controlar la aplicación de normas de higiene en las áreas de preparación y consumo de alimentos mediante análisis microbiológicos para disminuir los riesgos de contaminación alimentaria. Esta investigación es de carácter descriptivo, en la cual se realizó una inspección visual del establecimiento con el propósito de evaluar las condiciones higiénicas sanitarias, mediante la aplicación de la Guía Técnica para el Análisis Microbiológico de Superficies en contacto con Alimentos y Bebidas. Para el análisis microbiológico de las muestras se emplearon las técnicas de inoculación, método de estriado, aislamiento bacteriano, tinción diferencial y utilización de las pruebas bioquímicas como: TSI, SIM, Citrato de Simmons, Urea, Lisina, Catalasa y Oxidasa, además de la utilización de medios de cultivo selectivo y diferencial como agar EMB y agar MacConkey para la identificación de bacterias entéricas como: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. Los resultados arrojaron que la frecuencia bacteriana de las superficies inertes de los restaurantes en el área de preparación de alimentos (mesón y tabla de picar) tienen presencia de bacterias: Salmonella con mayor frecuencia; E. coli, Klebsiella pneumoniae y Pseudomonas aeruginosa de mediana frecuencia y de baja para Shigella, y en el área de consumo de alimentos (mesas) la bacteria de mayor frecuencia es la E. coli y Shigella, la Klebsiella pneumoniae de mediana y Pseudomona aeruginosa se encuentra en baja frecuencia. Se llegó a la conclusión que las superficies inertes tanto en el área de preparación como en el área de consumo de alimentos se encuentran contaminados por lo que hay un riesgo de infección alimentaria para los comensales de la Universidad Técnica de Machala.

Contamination of preparation areas when coming into contact with raw or cooked foods, which is why one of the main causes of contamination of inert surfaces is inadequate handling of food when it is being prepared. With the aim of controlling the application of hygiene standards in the areas of food preparation and consumption through microbiological analysis to reduce the risks of food contamination. This research is descriptive in nature, in which a visual inspection of the establishment was carried out with the purpose of evaluating the sanitary and hygienic conditions, through the application of the Technical Guide for the Microbiological Analysis of Surfaces in Contact with Food and Beverages. For the microbiological analysis of the samples, inoculation techniques, streaking method, bacterial isolation, differential staining and use of biochemical tests such as: TSI, SIM, Simmons Citrate, Urea, Lysine, Catalase and Oxidase, in addition to use of selective and differential culture media such as EMB agar and MacConkey agar for the identification of enteric bacteria such as: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. The results showed that the bacterial frequency of the inert surfaces of the restaurants in the food preparation area (counter and cutting board) have the presence of bacteria: Salmonella more frequently; E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa of medium frequency and low frequency for Shigella, and in the food consumption area (tables) the most frequent bacteria are E. coli and Shigella, Klebsiella pneumoniae of medium and Pseudomona aeruginosa It is at low frequency. It was concluded that the inert surfaces in both the preparation area and the food consumption area are contaminated, so there is a risk of food infection for diners at the Technical University of Machala

Contaminação das áreas de preparo ao entrar em contato com alimentos crus ou cozidos, por isso uma das principais causas de contaminação de superfícies inertes é o manuseio inadequado dos alimentos no momento do preparo. Com o objetivo de controlar a aplicação de padrões de higiene nas áreas de preparação e consumo de alimentos através de análises microbiológicas para reduzir os riscos de contaminação alimentar. Esta pesquisa é de natureza descritiva, na qual foi realizada uma inspeção visual do estabelecimento com a finalidade de avaliar as condições sanitárias e higiênicas, por meio da aplicação do Guia Técnico para Análise Microbiológica de Superfícies em Contato com Alimentos e Bebidas. Para a análise microbiológica das amostras foram utilizadas técnicas de inoculação, método de estrias, isolamento bacteriano, coloração diferencial e utilização de testes bioquímicos como: TSI, SIM, Citrato de Simmons, Ureia, Lisina, Catalase e Oxidase, além de utilização de testes seletivos e diferenciais. meios de cultura como ágar EMB e ágar MacConkey para identificação de bactérias entéricas como: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. Os resultados mostraram que a frequência bacteriana das superfícies inertes dos restaurantes na área de preparo de alimentos (balcão e tábua de corte) apresentam com maior frequência a presença de bactérias: Salmonella; E. coli, Klebsiella pneumoniae e Pseudomonas aeruginosa de média frequência e baixa frequência para Shigella, e na área de consumo alimentar (tabelas) as bactérias mais frequentes são E. coli e Shigella, Klebsiella pneumoniae de média e Pseudomona aeruginosa Está em baixa frequência. Concluiu-se que as superfícies inertes tanto na área de preparação como na área de consumo de alimentos estão contaminadas, pelo que existe risco de infecção alimentar para os comensais da Universidade Técnica de Machala

Identificación de microorganismos entéricos en áreas de preparación y consumo de alimentos

La contaminación de las áreas de preparación al entrar en contacto con los alimentos crudos o cocinados, es por esto que una de las principales causas de la contaminación de las superficies inertes es la inadecuada manipulación de los alimentos a la hora de ser preparados. Con el objetivo de controlar la aplicación de normas de higiene en las áreas de preparación y consumo de alimentos mediante análisis microbiológicos para disminuir los riesgos de contaminación alimentaria. Esta investigación es de carácter descriptivo, en la cual se realizó una inspección visual del establecimiento con el propósito de evaluar las condiciones higiénicas sanitarias, mediante la aplicación de la Guía Técnica para el Análisis Microbiológico de Superficies en contacto con Alimentos y Bebidas. Para el análisis microbiológico de las muestras se emplearon las técnicas de inoculación, método de estriado, aislamiento bacteriano, tinción diferencial y utilización de las pruebas bioquímicas como: TSI, SIM, Citrato de Simmons, Urea, Lisina, Catalasa y Oxidasa, además de la utilización de medios de cultivo selectivo y diferencial como agar EMB y agar MacConkey para la identificación de bacterias entéricas como: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. Los resultados arrojaron que la frecuencia bacteriana de las superficies inertes de los restaurantes en el área de preparación de alimentos (mesón y tabla de picar) tienen presencia de bacterias: Salmonella con mayor frecuencia; E. coli, Klebsiella pneumoniae y Pseudomonas aeruginosa de mediana frecuencia y de baja para Shigella, y en el área de consumo de alimentos (mesas) la bacteria de mayor frecuencia es la E. coli y Shigella, la Klebsiella pneumoniae de mediana y Pseudomona aeruginosa se encuentra en baja frecuencia. Se llegó a la conclusión que las superficies inertes tanto en el área de preparación como en el área de consumo de alimentos se encuentran contaminados por lo que hay un riesgo de infección alimentaria para los comensales de la Universidad Técnica de Machala.

Contamination of preparation areas when coming into contact with raw or cooked foods, which is why one of the main causes of contamination of inert surfaces is inadequate handling of food when it is being prepared. With the aim of controlling the application of hygiene standards in the areas of food preparation and consumption through microbiological analysis to reduce the risks of food contamination. This research is descriptive in nature, in which a visual inspection of the establishment was carried out with the purpose of evaluating the sanitary and hygienic conditions, through the application of the Technical Guide for the Microbiological Analysis of Surfaces in Contact with Food and Beverages. For the microbiological analysis of the samples, inoculation techniques, streaking method, bacterial isolation, differential staining and use of biochemical tests such as: TSI, SIM, Simmons Citrate, Urea, Lysine, Catalase and Oxidase, in addition to use of selective and differential culture media such as EMB agar and MacConkey agar for the identification of enteric bacteria such as: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. The results showed that the bacterial frequency of the inert surfaces of the restaurants in the food preparation area (counter and cutting board) have the presence of bacteria: Salmonella more frequently; E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa of medium frequency and low frequency for Shigella, and in the food consumption area (tables) the most frequent bacteria are E. coli and Shigella, Klebsiella pneumoniae of medium and Pseudomona aeruginosa It is at low frequency. It was concluded that the inert surfaces in both the preparation area and the food consumption area are contaminated, so there is a risk of food infection for diners at the Technical University of Machala.

Contaminação das áreas de preparo ao entrar em contato com alimentos crus ou cozidos, por isso uma das principais causas de contaminação de superfícies inertes é o manuseio inadequado dos alimentos no momento do preparo. Com o objetivo de controlar a aplicação de padrões de higiene nas áreas de preparação e consumo de alimentos através de análises microbiológicas para reduzir os riscos de contaminação alimentar. Esta pesquisa é de natureza descritiva, na qual foi realizada uma inspeção visual do estabelecimento com a finalidade de avaliar as condições sanitárias e higiênicas, por meio da aplicação do Guia Técnico para Análise Microbiológica de Superfícies em Contato com Alimentos e Bebidas. Para a análise microbiológica das amostras foram utilizadas técnicas de inoculação, método de estrias, isolamento bacteriano, coloração diferencial e utilização de testes bioquímicos como: TSI, SIM, Citrato de Simmons, Ureia, Lisina, Catalase e Oxidase, além de utilização de testes seletivos e diferenciais. meios de cultura como ágar EMB e ágar MacConkey para identificação de bactérias entéricas como: E. coli, Salmonella, Klebsiella pneumoniae, Shigella, Pseudomona aeruginosa. Os resultados mostraram que a frequência bacteriana das superfícies inertes dos restaurantes na área de preparo de alimentos (balcão e tábua de corte) apresentam com maior frequência a presença de bactérias: Salmonella; E. coli, Klebsiella pneumoniae e Pseudomonas aeruginosa de média frequência e baixa frequência para Shigella, e na área de consumo alimentar (tabelas) as bactérias mais frequentes são E. coli e Shigella, Klebsiella pneumoniae de média e Pseudomona aeruginosa Está em baixa frequência . Concluiu-se que as superfícies inertes tanto na área de preparação como na área de consumo de alimentos estão contaminadas, pelo que existe risco de infecção alimentar para os comensais da Universidade Técnica de Machala.

Environmental stressor assessment of hydrocarbonoclastic bacteria biofilms from a marine oil spill.

The environmental and economic impact of an oil spill can be significant. Biotechnologies applied during a marine oil spill involve bioaugmentation with immobilised or encapsulated indigenous hydrocarbonoclastic species selected under laboratory conditions to improve degradation rates. The environmental factors that act as stressors and impact the effectiveness of hydrocarbon removal are one of the challenges associated with these applications. Understanding how native microbes react to environmental stresses is necessary for effective bioaugmentation. Herein, Micrococcus luteus and M. yunnanensis isolated from a marine oil spill mooring system showed hydrocarbonoclastic activity on Maya crude oil in a short time by means of total petroleum hydrocarbons (TPH) at 144 h: M. luteus up to 98.79 % and M. yunnanensis 97.77 % removal. The assessment of Micrococcus biofilms at different temperature (30 °C and 50 °C), pH (5, 6, 7, 8, 9), salinity (30, 50, 60, 70, 80 g/L), and crude oil concentration (1, 5, 15, 25, 35 %) showed different response to the stressors depending on the strain. According to response surface analysis, the main effect was temperature > salinity > hydrocarbon concentration. The hydrocarbonoclastic biofilm architecture was characterised using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Subtle but significant differences were observed: pili in M. luteus by SEM and the topographical differences measured by AFM Power Spectral Density (PSD) analysis, roughness was higher in M. luteus than in M. yunnanensis. In all three domains of life, the Universal Stress Protein (Usp) is crucial for stress adaptation. Herein, the uspA gene expression was analysed in Micrococcus biofilm under environmental stressors. The uspA expression increased up to 2.5-fold in M. luteus biofilms at 30 °C, and 1.3-fold at 50 °C. The highest uspA expression was recorded in M. yunnanensis biofilms at 50 °C with 2.5 and 3-fold with salinities of 50, 60, and 80 g/L at hydrocarbon concentrations of 15, 25, and 35 %. M. yunnanensis biofilms showed greater resilience than M. luteus biofilms when exposed to harsh environmental stressors. M. yunnanensis biofilms were thicker than M. luteus biofilms. Both biofilm responses to environmental stressors through uspA gene expression were consistent with the behaviours observed in the response surface analyses. The uspA gene is a suitable biomarker for assessing environmental stressors of potential microorganisms for bioremediation of marine oil spills and for biosensing the ecophysiological status of native microbiota in a marine petroleum environment.

Burden and Management of Multi-Drug Resistant Organism Infections in Solid Organ Transplant Recipients Across the World: A Narrative Review.

Solid organ transplant (SOT) recipients are particularly susceptible to infections caused by multidrug-resistant organisms (MDRO) and are often the first to be affected by an emerging resistant pathogen. Unfortunately, their prevalence and impact on morbidity and mortality according to the type of graft is not systematically reported from high-as well as from low and middle-income countries (HIC and LMIC). Thus, epidemiology on MDRO in SOT recipients could be subjected to reporting bias. In addition, screening practices and diagnostic resources may vary between countries, as well as the availability of new drugs. In this review, we aimed to depict the burden of main Gram-negative MDRO in SOT patients across HIC and LMIC and to provide an overview of current diagnostic and therapeutic resources.

Metabarcoding expands knowledge on diversity and ecology of rare actinobacteria in the Brazilian Cerrado.

Rare and unknown actinobacteria from unexplored environments have the potential to produce new bioactive molecules. This study aimed to use 16 s rRNA metabarcoding to determine the composition of the actinobacterial community, particularly focusing on rare and undescribed species, in a nature reserve within the Brazilian Cerrado called Sete Cidades National Park. Since this is an inaccessible area without due legal authorization, it is understudied, and, therefore, its diversity and biotechnological potential are not yet fully understood, and it may harbor species with groundbreaking genetic potential. In total, 543 operational taxonomic units (OTUs) across 14 phyla were detected, with Actinobacteria (41.2%), Proteobacteria (26.5%), and Acidobacteria (14.3%) being the most abundant. Within Actinobacteria, 107 OTUs were found, primarily from the families Mycobacteriaceae, Pseudonocardiaceae, and Streptomycetaceae. Mycobacterium and Streptomyces were the predominant genera across all samples. Seventeen rare OTUs with relative abundance < 0.1% were identified, with 82.3% found in only one sample yet 25.5% detected in all units. Notable rare and transient genera included Salinibacterium, Nocardia, Actinomycetospora_01, Saccharopolyspora, Sporichthya, and Nonomuraea. The high diversity and distribution of Actinobacteria OTUs indicate the area's potential for discovering new rare species. Intensified prospection on underexplored environments and characterization of their actinobacterial diversity could lead to the discovery of new species capable of generating innovative natural products.

Inoculation with Bacillus megaterium CNPMS B119 and Bacillus subtilis CNPMS B2084 improve P-acquisition and maize yield in Brazil.

Phosphorus (P) is a critical nutrient for plant growth, yet its uptake is often hindered by soil factors like clay minerals and metal oxides such as aluminum (Al), iron (Fe), and calcium (Ca), which bind P and limit its availability. Phosphate-solubilizing bacteria (PSB) have the unique ability to convert insoluble P into a soluble form, thereby fostering plant growth. This study aimed to assess the efficacy of inoculation of Bacillus megaterium B119 (rhizospheric) and B. subtilis B2084 (endophytic) via seed treatment in enhancing maize yield, grain P content, and enzyme activities across two distinct soil types in field conditions. Additionally, we investigated various mechanisms contributing to plant growth promotion, compatibility with commercial inoculants, and the maize root adhesion profile of these strains. During five crop seasons in two experimental areas in Brazil, Sete Lagoas-MG and Santo Antônio de Goiás-GO, single inoculations with either B119 or B2084 were implemented in three seasons, while a co-inoculation with both strains was applied in two seasons. All treatments received P fertilizer according to plot recommendations, except for control. Both the Bacillus strains exhibited plant growth-promoting properties relevant to P dynamics, including phosphate solubilization and mineralization, production of indole-3-acetic acid (IAA)-like molecules, siderophores, exopolysaccharides (EPS), biofilms, and phosphatases, with no antagonism observed with Azospirillum and Bradyrizhobium. Strain B2084 displayed superior maize root adhesion compared to B119. In field trials, single inoculations with either B119 or B2084 resulted in increased maize grain yield, with relative average productivities of 22 and 16% in Sete Lagoas and 6 and 3% in Santo Antônio de Goiás, respectively. Co-inoculation proved more effective, with an average yield increase of 24% in Sete Lagoas and 11% in Santo Antônio de Goiás compared to the non-inoculated control. Across all seasons, accumulated grain P content correlated with yield, and soil P availability in the rhizosphere increased after co-inoculation in Santo Antônio de Goiás. These findings complement previous research efforts and have led to the validation and registration of the first Brazilian inoculant formulated with Bacillus strains for maize, effectively enhancing and P grain content.

Comprehensive characterization of γ-aminobutyric acid (GABA) production by Levilactobacillus brevis CRL 2013: insights from physiology, genomics, and proteomics.

Introduction: Levilactobacillus brevis CRL 2013, a plant-derived lactic acid bacterium (LAB) with immunomodulatory properties, has emerged as an efficient producer of γ-aminobutyric acid (GABA). Notably, not all LAB possess the ability to produce GABA, highlighting the importance of specific genetic and environmental conditions for GABA synthesis. This study aimed to elucidate the intriguing GABA-producing machinery of L. brevis CRL 2013 and support its potential for safe application through comprehensive genome analysis. Methods: A comprehensive genome analysis of L. brevis CRL 2013 was performed to identify the presence of antibiotic resistance genes, virulence markers, and genes associated with the glutamate decarboxylase system, which is essential for GABA biosynthesis. Then, an optimized chemically defined culture medium (CDM) was supplemented with monosodium glutamate (MSG) and yeast extract (YE) to analyze their influence on GABA production. Proteomic and transcriptional analyses were conducted to assess changes in protein and gene expression related to GABA production. Results: The absence of antibiotic resistance genes and virulence markers in the genome of L. brevis CRL 2013 supports its safety for potential probiotic applications. Genes encoding the glutamate decarboxylase system, including two gad genes (gadA and gadB) and the glutamate antiporter gene (gadC), were identified. The gadB gene is located adjacent to gadC, while gadA resides separately on the chromosome. The transcriptional regulator gadR was found upstream of gadC, with transcriptional analyses demonstrating cotranscription of gadR with gadC. Although MSG supplementation alone did not activate GABA synthesis, the addition of YE significantly enhanced GABA production in the optimized CDM containing glutamate. Proteomic analysis revealed minimal differences between MSG-supplemented and non-supplemented CDM cultures, whereas YE supplementation resulted in significant proteomic changes, including upregulation of GadB. Transcriptional analysis confirmed increased expression of gadB and gadR upon YE supplementation, supporting its role in activating GABA production. Conclusion: These findings provide valuable insights into the influence of nutrient composition on GABA production. Furthermore, they unveil the potential of L. brevis CRL 2013 as a safe, nonpathogenic strain with valuable biotechnological traits which can be further leveraged for its probiotic potential in the food industry.