ABSTRACT Objective: To evaluate the antimicrobial activity of soyabean and pumpkin seeds methanolic extract against oral microbiome. Material and Methods: An in vitro study compared the antimicrobial activity of methanolic extract of soyabean and pumpkin seed along with 0.2% chlorhexidine (positive control) by Agar well diffusion method against Streptococcus mutans, Streptococcus pyogenes, Staphylococcus aureus, Lactobacillus and Candida albicans. Suitable dilutions of each seed extract determined Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Qualitative phytochemical analysis and Thin Layer Chromatography (TLC) was done to determine the phytochemical components. Results: Both the methanolic extracts showed the highest zone of inhibition, 26±0.31 mm and 24±0.82 mm, followed by 22±0.87 mm and 21±0.55 mm against Streptococcus mutans and Streptococcus pyogenes, respectively. The zone of inhibition was higher at concentrations in both extracts, which was not statistically significant (p>0.05). MIC values fluctuated from 1.56 to 4.5 mg/ml and 3.5 to 5.0 mg/ml for soyabean and pumpkin seed extract, respectively. Phytochemical analysis showed the presence of alkaloids, glycosides, terpenoids, tannins, and flavonoids. Conclusion: Both extracts were effective against S. mutans and S. pyogenes and contain compounds with therapeutic potential. Objective microbiome Methods 02 0 2 0.2 positive control aureus albicans (MIC MBC. MBC . (MBC) TLC (TLC components Results 26031 26 31 26±0.3 24082 24 82 24±0.8 22087 22 87 22±0.8 21055 21 55 21±0.5 respectively p>0.05. p005 p p>0.05 05 (p>0.05) 156 1 56 1.5 45 4 5 4. mgml mg ml 35 3 3. 50 5. alkaloids glycosides terpenoids tannins flavonoids Conclusion S potential 0. (MBC 2603 26±0. 2408 8 24±0. 2208 22±0. 2105 21±0. p00 p>0.0 (p>0.05 15 1. 260 26±0 240 24±0 220 22±0 210 21±0 p0 p>0. (p>0.0 26± 24± 22± 21± p>0 (p>0. p> (p>0 (p> (p

Resumen El objetivo de este trabajo fue seleccionar microorganismos promisorios como agentes de control biológico (ACB). Se visitaron predios florihortícolas ubicados en Buenos Aires, Argentina, de los cuales se obtuvo un total de 41 muestras de suelo. Se utilizaron las técnicas de insecto trampa y diluciones seriadas de suelo para la obtención de aislados de hongos entomopatógenos y hongos del género Trichoderma respectivamente. Se obtu vieron un total de 20 aislados, cinco pertenecientes al género Metarhizium y 15 aislados correspondientes al género Trichoderma. Los aislados fueron liofilizados y depositados como cultivos de referencia en la Colección Micológica del Centro de Estudios Parasi tológicos y de Vectores (CEPAVE). Se realizaron estudios de cultivos duales de los aislados recolectados frente a los patógenos Botrytis cinerea Pers. (1797) y Sclerotinia sclerotiorum (Lib.) de Bary (1884). Se seleccionaron 11 aislados para la realización de estudios de promoción de crecimiento en plantas de tomate (Solanum lycopersicum L.). Los aislados de Metarhizium taii Liang y Liu (1991) CEP-722, CEP-723 y de Trichoderma afroharzianum Chaverri, Rocha, Degenkolb y Druzhinina (2015) CEP-753 y CEP-754, identificados molecularmente por medio de la amplificación de las zonas ITS y TEF1α, presentaron los mejores resultados en las pruebas de cultivo dual y promoción de crecimiento. Se espera avanzar en estudios posteriores que evalúen la virulencia de cepas de hongos en insectos.

Abstract This work aimed to select promising microorganisms as biological control agents (BCA). Forty-one soil samples were obtained from florihorticultural farms located in Buenos Aires, Argentina. Insect trap techniques and soil serial dilutions were used to obtain isolates of entomopathogenic fungi and fungi of genera Trichoderma, respectively. A total of 20 isolates included five Metarhizium and 15 Trichoderma. The isolates were lyophilized and deposited as reference cultures in the Mycological Collection of the Centro de Estudios Parasitológicos y de Vectores (CEPAVE). We performed dual culture studies of the isolates collected against the pathogens Botrytis cinerea Pers. (1797) and Sclerotinia sclerotiorum (Lib.) de Bary (1884). Eleven isolates were selected for growth promotion studies in tomato plants (Solanum lycopersicum L.). The isolates of Metarhizium taii Liang & Liu (1991) CEP-722, CEP-723 Trichoderma afroharzianum Chaverri, Rocha, Degenkolb & Druzhinina (2015) CEP-753 and CEP-754, molecularly identified by amplification of the ITS and TEF1α zones, presented the best results in the dual culture and growth promotion tests. Subsequent studies will evaluate virulence of fungal strains in insects.

Resumen Tritrichomonas foetus es un protozoo parásito de transmisión sexual que provoca abortos en el ganado. Los nitroimidazoles, como el metronidazol, pueden ayudar en el tratamiento de la tricomonosis bovina, pero no se aconseja su uso. Los extractos de plantas pueden tener un efecto antiparasitario. El estudio actual tuvo como objetivo evaluar la actividad del aceite de Cymbopogon nardus contra T. fetus como alternativa al metronidazol. Se incubaron trofo zoítos de T. fetus en medio de cultivo con diluciones seriadas de aceite de C. nardus. La cito toxicidad se evaluó a las 24 horas. El aceite de C. nardus mató las células de T. fetus. La concen tración efectiva media máxima (CE50) fue de 0,4 μg/ml. Estos hallazgos sugieren que el aceite de C. nardus podría servir para el descubrimiento y aislamiento de fitofármacos para el tratamiento de la tricomoniasis bovina y de otras enfermedades protozoarias.

Abstract Tritrichomonas foetus, a sexually transmitted parasitic protozoan, causes abortion in cattle. Nitroimidazoles, such as metronidazole, treat bovine trichomonosis, but their use is precluded. Plant extracts might have antiparasitic effects. This study aimed to assess Cymbopogon nardus oil activity against T. foetus as an alternative to metronidazole. T. foetus trophozoites were incubated in culture medium containing serial dilutions of C. nardus oil. Cytotoxicity was assessed 24 hours later. C. nardus oil killed T. foetus cells. Half maximal effective concentration (EC50) was 0.4 μg/mL. These findings suggest that C. nardus oil could be exploited for discovery and compound isolation of plant-derived phytopharmaceuticals for bovine trichomoniasis and other protozoan diseases.

RESUMEN Introducción: La producción de etanol a partir de caña de azúcar genera subproductos importantes, especialmente la vinaza, que presenta desafíos ambientales. Objetivo: Evaluar el uso de la vinaza como diluyente en la fermentación de melaza para la reducción del consumo de agua y mejorar la sostenibilidad del proceso de producción de etanol. Materiales y Métodos: Se utilizó melaza de la Empresa Agroindustrial Azucarera “Heriberto Duquesne” en Villa Clara, Cuba, y la vinaza se extrajo de la columna de destilación. Se probaron cinco tratamientos, con diluciones de vinaza entre el 10% y el 50%. Las fermentaciones se realizaron por duplicado y monitoreando el brix y el pH durante 40 horas. Resultados y Discusión: Las mejores eficiencias de fermentación se lograron con un 10% y 20% de vinaza, obteniendo eficiencias del 77,67% y 77,28%, respectivamente. Las concentraciones más altas de vinaza resultaron en una disminución del pH y en una menor producción de etanol. Se determinó que la tasa óptima de recirculación de vinaza es del 18%, alcanzando una eficiencia del 78,1%. Esta recirculación redujo el consumo de agua en aproximadamente un 20%, lo que equivale a un ahorro diario de 13,7 m³ de agua fresca. Conclusiones: El estudio concluye que el uso de vinaza como diluyente en la fermentación de melaza puede reducir el consumo de agua sin afectar negativamente la eficiencia de la fermentación. Es factible sustituir hasta un 20% del agua por vinaza para una producción de etanol más sostenible.

ABSTRACT Introduction: Ethanol production from sugarcane generates significant by-products, particularly vinasse, which poses environmental challenges. Objective: To evaluate the use of vinasse as a diluent in molasses fermentation to reduce water consumption and enhance the sustainability of the ethanol production process. Materials and Methods: Molasses from the “Heriberto Duquesne” Sugar Agroindustrial Company in Villa Clara, Cuba, was used, and the vinasse was extracted from the distillation column. Five treatments were tested, with vinasse dilutions between 10% to 50%. Fermentations were conducted in duplicate and monitoring the brix and pH for 40 hours. Results and Discussion: The best fermentation efficiencies were achieved with 10% and 20% vinasse, resulting in 77.67% and 77.28% efficiencies, respectively. The highest vinasse concentrations resulted in a decrease in pH and lower ethanol production. The optimal vinasse recirculation rate was determined to be 18%, reaching an efficiency of 78.1%. This recirculation reduced water consumption by approximately 20%, which is equivalent to a daily saving of 13.7 m³ of fresh water. Conclusions: The study concludes that using vinasse as a diluent in molasses fermentation can reduce water usage without negatively affecting fermentation efficiency. It is feasible to replace up to 20% of water with vinasse for more sustainable ethanol production.

The selection of an underground mining method stems from a multivariate analysis that considers geomechanical, geological, economic, and operational parameters. Even after identifying the most compatible method, there is no assurance that the mining company will achieve the best performance. The geological complexity of some deposits requires adaptations of methods described in the literature to obtain more selectivity and reduce mining waste. There are some studies on geometry of underground structures, but a methodology that describes an adaptation of a room-and-pillar mining method for ore bodies with down-dip varying from 20° to 25° is novel. The present work aimed to reduce dilution by adapting the traditional room-and-pillar mining method (TRP) to inclined ore bodies. This new method is entitled short-hole room-and-pillar (SHRP). The equations that measure the dilution are defined according to the geometry of stopes and openings. The results comprise comparative analyses of the operational and planned dilutions to measure the performance of the SHRP method. The average operating dilution of the SHRP method was more than five times lower than the planned dilution according to the TRP method. Low operational dilution indicates high selectivity of the method and its potential to reduce underground mining tailings.

RESUMO A correlação canônica é usada para correlacionar grupos de variáveis distintas. O objetivo deste estudo foi utilizar análise de correlação canônica para avaliar a interação entre atributos de qualidade da água causadores de entupimento e o desempenho hidráulico de gotejadores não autocompensantes operando com diluições de efluente de aquicultura em água de poço. O experimento foi conduzido em delineamento inteiramente casualizado, em esquema de parcelas subsubdivididas, com três repetições, com cinco diluições nas parcelas (Diluição 1: 100% de efluente de aquicultura; Diluição 2: 75% de efluente de aquicultura + 25% de água de poço; Diluição 3 : 50% efluente de aquicultura + 50% água de poço; Diluição 4: 25% efluente de aquicultura + 75% água de poço; e Diluição 5: 100% água de poço), três emissores nas subparcelas (ST (1,6 L h-1) , SL (1,6 L h-1) e NJ (1,7 L h-1)), e três tempos de operação nas subsubparcelas (início do experimento, 80 e 160 horas, em que 0 horas de operação significa o início das avaliações). As análises estatísticas realizadas foram matriz de correlação e análise canônica. A análise de correlação canônica indicou que quando as concentrações dos atributos aumentam, o desempenho hidráulico dos gotejadores diminui. A interação desse grupo de atributos indica que o entupimento dos gotejadores que operam com as cinco diluições de efluente de aquicultura em água de poço ocorre pela ação combinada de agentes físicos, químicos e biológicos. O gotejador ST foi o menos sensível ao grupo de atributos de qualidade das diluições, enquanto o mais sensível foi o gotejador SL. distintas casualizado subsubdivididas repetições 1 100 2 75 25 50 4 5 poço, poço) 1,6 16 6 (1, h1 h h-1 1,7 17 7 h1, h-1)) 8 avaliações. avaliações . avaliações) aumentam diminui físicos biológicos 10 1, (1 h- (

ABSTRACT The canonical correlation is used to correlate groups of distinct variables. The objective of this study was to use canonical correlation analysis to evaluate the interaction between water quality attributes clogging-causing and the hydraulic performance of non-pressure compensating drippers operating with dilutions of aquaculture effluent in well water. The experiment was in a completely randomized design in a split-split-plot scheme with three replicates, with five dilutions in the plots (Dilution 1: 100% aquaculture effluent; Dilution 2: 75% aquaculture effluent + 25% well water; Dilution 3: 50% aquaculture effluent + 50% well water; Dilution 4: 25% aquaculture effluent + 75% well water; and Dilution 5: 100% well water), three emitters in the sub-plots (ST (1.6 L h-1), SL (1.6 L h-1) and NJ (1.7 L h-1)), and three operating times in the sub-subplots (0, 80 and 160 hours, in which 0 hours operating time means the start of evaluations). The statistical analyses performed were correlation matrix and canonical analysis. The canonical correlation analysis indicated that when the concentrations of attributes increase, the hydraulic performance of the drippers decreases. The interaction of this group of attributes indicates that the clogging of drippers operating with the five dilutions of aquaculture effluent in well water occurs by the combined action of physical, chemical, and biological agents. The ST dripper was the least sensitive to the group of quality attributes of the dilutions, whereas the most sensitive was the SL dripper. variables cloggingcausing causing nonpressure non pressure splitsplitplot split plot replicates 1 100 2 75 25 3 50 4 5 water, , water) subplots sub 1.6 16 6 (1. h1, h1 h h-1 1.7 17 7 h-1)) subsubplots 0, (0 8 evaluations. evaluations . evaluations) increase decreases physical chemical agents 10 1. (1 h- (

Abstract: Soil nitrogen is crucial for agriculture, but it is often limited, affecting crop yields. Deficiency requires synthetic fertilizers, but their improper use results in environmental damage and high costs. Bacteria of the genus Rhizobium, symbionts of legumes, offer a sustainable solution by fixing nitrogen, thus reducing dependence on fertilizers. This research determined the most probable number (MPN) of cells of Rhizobium spp. from two commercial biofertilizers of Ecuadorian and Mexican origin under greenhouse conditions. For this, direct inoculation with serial dilutions (10-1 to 10-10) was performed in pots with steam-sterilized pumice where Blue Lake variety snap bean (Phaseolus vulgaris L.) plants were germinated. The following morphological indicators were evaluated at 45 days after sowing (DAS): leaf area, plant wet weight, plant height, and number of flowers, determining statistical differences between the type of biofertilizer and the concentration of each dilution. The experiment followed a randomized complete block design with a split-plot arrangement, with three replicates per dilution, considering temperature fluctuations in the study area. The MPN at 95% confidence was 4.45x107 rhizobia g-1 of pumice at a 10-5 dilution for the Mexican biofertilizer, and 1.48x105 rhizobia g-1 of pumice at a 10-4 dilution for the Ecuadorian biofertilizer. The estimated optimal dilution for both products was 10-8.

Resumen: El nitrógeno en el suelo es crucial para la agricultura, pero a menudo es limitante, afectando el rendimiento de los cultivos. La deficiencia requiere fertilizantes sintéticos, pero su uso inadecuado causa daños ambientales y costos elevados. Las bacterias del género Rhizobium, simbiontes de leguminosas, ofrecen una solución al fijar nitrógeno de manera sostenible, reduciendo la dependencia de fertilizantes. Esta investigación determinó el número más probable (NMP) de células de Rhizobium spp. procedente de dos biofertilizantes comerciales de origen ecuatoriano y mexicano en condiciones de invernadero. Para esto, se realizó la inoculación directa con disoluciones seriadas (10-1 a 10-10) en macetas con pomina esterilizada al vapor, donde se germinaron plantas de frijol ejotero (Phaseolus vulgaris L.) de la variedad Blue Lake. A los 45 días después de siembra (DDS) se evaluaron los siguientes indicadores morfológicos: área foliar, peso de la planta en húmedo, altura de planta y número de flores, determinándose diferencias estadísticas entre el tipo de biofertilizante y la concentración de cada dilución. El experimento se adecuó a un diseño de bloques completos al azar con arreglo de parcela dividida con tres repeticiones por dilución, en consideración de las fluctuaciones de temperatura presentes en la zona de estudio. El NMP, al 95% de confianza, fue de 4,45x107 rizobios g-1 de pomina en una dilución 10-5 para el biofertilizante mexicano, y 1,48x105 rizobios g-1 de pomina en una dilución 10-4 para el biofertilizante ecuatoriano. La dilución óptima estimada para ambos productos fue de 10-8.

Resumen El objetivo de este trabajo de investigación fue evaluar el efecto antimicrobiano de 2 fuentes de aceite frente a Staphylococcus aureus y Coliformes Fecales. Para la extracción el aceite se aplicó el método arrastre de vapor en donde se utilizó 5000g de muestra (muña, Hojas de eucalipto), en lo cual el rendimiento presento: muña 50ml = 0.9268%, eucalipto 52ml = 0.9717%, se le realizo una caracterización fisicoquímica cuyo resultado fue: en aceite de muña: índice de peróxido: 0.75 (Meq/Kg), Índice de Iodo: 6.87 (IV), índice de Acidez:1.78 (%AGL), Índice de refracción: 1.486 (n), densidad: 0.898 (Kg/L). En hojas de eucalipto: índice de peróxido: 0.68 (Meq/Kg), Índice de Iodo: 8.08 (IV), índice de Acidez: 1.82 (%AGL), Índice de refracción: 1.495 (n), densidad: 0.845 (Kg/L). Se determinó los halos de inhibición en milímetros (mm), con aceite de muña en diluciones a 25, 50 y 75% frente a Coliformes Fecales, en donde fue al 25%: 9.7950mm, 50%: 10.7467mm, 75%: 13.2700mm, frente a Staphylococcus aureus, al 25%: 9.9600mm, 50%: 10.7408mm, 75%: 13.1533mm; con dilución de aceite esencial de Eucalipto frente a Coliformes Fecales al 25%: 12.0983mm, 50%: 13.2900mm, 75%: 14.5875mm; después frente a Staphylococcus aureus en 25%: 11.7200mm, 50%: 13.5608mm, 75%: 14.3767. Se consiguió establecer el efecto inhibitorio en 2 fuentes de aceites frente a Coliformes Fecales y Staphylococcus aureus, por lo cual se concluye que a más dilución del aceite más inhibición microbiana, estas especien ya que cuenta con sus respectivos principios activos como monoterpenos, alcoholes, cetonas y óxidos terpenicos que impiden el crecimiento microbiano.

Abstract The objective of this research work was to evaluate the antimicrobial effect of 2 oil sources against Staphylococcus aureus and Fecal Coliforms. For the oil extraction, the steam drag method was applied where 5000g of sample was used (muña, Eucalyptus leaves), in which the yield presented: muña 50ml = 0.9268%, eucalyptus 52ml = 0.9717%, a Physicochemical characterization whose result was: in muña oil: peroxide index: 0.75 (Meq / Kg), Iodine index: 6.87 (IV), Acidity index: 1.78 (% FFA), Refractive index: 1.486 (n), density: 0.898 (Kg / L). In eucalyptus leaves: peroxide index: 0.68 (Meq / Kg), Iodine index: 8.08 (IV), Acidity index: 1.82 (% FFA), Refractive index: 1.495 (n), density: 0.845 (Kg / L). Inhibition halos were determined in millimeters (mm), with muña oil in dilutions at 25, 50 and 75% against Fecal Coliforms, where it was at 25%: 9.7950mm, 50%: 10.7467mm, 75%: 13.2700 mm, against Staphylococcus aureus, 25%: 9.9600mm, 50%: 10.7408mm, 75%: 13.1533mm; with dilution of Eucalyptus essential oil against Fecal Coliforms at 25%: 12.0983mm, 50%: 13.2900mm, 75%: 14.5875mm; then against Staphylococcus aureus in 25%: 11.7200mm, 50%: 13.5608mm, 75%: 14.3767. It was possible to establish the inhibitory effect in 2 sources of oils against Fecal Coliforms and Staphylococcus aureus, for which it is concluded that the more dilution of the oil, more microbial inhibition, these species have their respective active principles such as monoterpenes, alcohols, ketones and terpenic oxides that prevent microbial growth.

Resumo Os biopreparados representam uma importante alternativa ao uso de agroquímicos sintéticos. Entre eles, o esterco de urtiga (Urtica sp.) é utilizado como bioestimulante e para o controle de pragas e doenças, embora não existe informação dos seus efeitos sobre os organismos não-branco do agroecossistema. Apresenta-se como objetivo do trabalho estudar os efeitos letais de diferentes diluções de esterco de ortiga sobre os nematóides modelo de vida livre Caenorhabditis elegans e Panagrellus redivivus (organismos chave na estrutura edáfica). Realizaram-se cinco bioensaios de toxicidade em meio líquido sobre C. elegans e um sobre P. redivivus, utilizando três estercos com distintas condições de elaboração e tempo de almacenamento. O esterco demostrou ter efeitos letais sobre os indivíduos de C. elegans dependendo da dilução de exposição. A informação obtida sobre a ecotoxicidade dos biopreparados pode contribuir à optimização das práticas hortícolas.

Abstract Biopreparations represent an important alternative to the use of synthetic agrochemicals. Among them, nettle slurry (Urtica sp.) is used as a biostimulant and in the control of pests and diseases. However, there is no information on its effects on non-target organisms of the agroecosystem. The objective of this work was to study the lethal effects of different dilutions of nettle slurry on the free-living model nematodes Caenorhabditis elegans and Panagrellus redivivus (key organisms in the soil structure). Five toxicity bioassays were carried out in a liquid medium on C. elegans and one on P. redivivus. Three nettle slurries with different processing conditions and storage times were used. The slurry proved to have lethal effects on C. elegans individuals depending on the dilution of exposure. The information obtained on the ecotoxicity of biopreparations can contribute to the optimization of horticultural practices.

Resumen Los biopreparados representan una importante alternativa al uso de agroquímicos sintéticos. Entre ellos, el purín de ortiga (Urtica sp.) es utilizado como bioestimulante y para el control de plagas y enfermedades, aunque no existe información acerca de sus efectos sobre los organismos no-blanco del agroecosistema. Se plantea como objetivo del trabajo estudiar los efectos letales de diferentes diluciones de purín de ortiga sobre los nematodos modelo de vida libre Caenorhabditis elegans y Panagrellus redivivus (organismos clave en la estructura edáfica). Se realizaron cinco bioensayos de toxicidad en medio líquido sobre C. elegans y uno sobre P. redivivus, utilizándose tres purines con distintas condiciones de elaboración y tiempo de almacenamiento. El purín demostró tener efectos letales sobre los individuos de C. elegans dependiendo de la dilución de exposición. La información obtenida sobre la ecotoxicidad de los biopreparados puede contribuir a la optimización de las prácticas hortícolas.

Abstract Indian major carps are the widely consumed fish species of Pakistan, being a cheap source of proteins and unsaturated fatty acids, they are good for cardiovascular health. Water pollution due to discharge of untreated industrial waste water into water bodies contaminates this precious source of nutrients. The present study therefore, was aimed to assess deterioration of fatty acid profile of three Indian major carp species due to different concentrations of industrial wastes. The water samples were collected from the river Chenab at the site where it receives industrial wastewater via Chakbandi drain. After exposure to 1.5%, 3.0%, and 4.5% dilutions of collected water in different aquaria it was observed that proportion of unsaturated fatty acids in selected fish species were decreased significantly as the intensity of the dose increased (P < 0.05). Conversely the level of saturated fatty acids increased with the increasing dose of treatment (P < 0.05). These findings suggest that untreated wastewater not only deteriorate the fatty acid profile of aquatic animals but also these toxic substances can reach human body through fish meat and pose further health hazards. Therefore, it is highly recommended that industrial effluents should be treated before they are dumped into water bodies.

Resumo As carpas indianas são as espécies de peixes mais consumidas no Paquistão, sendo uma fonte barata de proteínas e de ácidos graxos insaturados e boa para a saúde cardiovascular. A poluição da água por causa do descarte de resíduos industriais não tratados em corpos d’água contamina essa preciosa fonte de nutrientes. Portanto, o presente estudo teve como objetivo avaliar a deterioração do perfil de ácidos graxos de três principais espécies de carpas indianas em diferentes concentrações de resíduos industriais. As amostras de água foram coletadas do rio Chenab no local onde recebe esgoto industrial via dreno de Chakbandi. Após a exposição a diluições de 1,5%, 3% e 4,5% da água coletada em diferentes aquários, foi observado que a proporção de ácidos graxos insaturados em espécies de peixes selecionadas diminuiu significativamente com o aumento da intensidade da dose (P < 0,05). Por outro lado, o nível de ácidos graxos saturados aumentou com a elevação da dose de tratamento (P < 0,05). Essas descobertas sugerem que águas residuais não tratadas não apenas deterioram o perfil de ácidos graxos dos animais aquáticos, mas também essas substâncias tóxicas podem atingir o corpo humano por meio da carne de peixe e representar mais riscos à saúde. Portanto, é recomendável que os efluentes industriais sejam tratados antes de serem despejados em corpos d’água.

ABSTRACT Introduction: Dichrostachys cinerea L. (marabou) is a plant that grows in Cuba, medicinal properties of this plant are being studied. The resistance of yeasts of the Candida genus to current available synthetic antifungals is increasingly greater. This is why, new compounds of plant origin are being searched for the effective treatment of infections caused by this germ. Objective: To evaluate the in vitro antifungal activity of Dichrostachys cinerea L against a strain of Candida albicans. Methods: An in vitro a cross-sectional analytic observational study was carried out to evaluate the antifungal activity of fluid extracts of D. cinerea L leaves and stems using the broth macro-dilution method and alcohol as the reference substance. Results: Through the process was shown the antifungal activity of the fluid extract of Dichrostachys cinerea L. stems at 50% up to the 1/32 dilution; it was determined as the minimum inhibitory concentration. The leaves fluid extract at 30% failed to inhibit the growth of the Candida albicans ATCC 10231 strain. Conclusions: The antifungal activity of the fluid extract at 50% of the leaves was effective, but not the pharmaceutical preparation at 30%. The minimum inhibitory concentration of the fluid extract of leaves at 50% was determined and it was shown that it exceeded that of alcohol at 50% in three dilutions against Candida albicans.

RESUMEN Introducción: La Dichrostachys cinerea L. (marabú) es una planta que crece en Cuba, de la que se estudian propiedades medicinales. La resistencia de levaduras del género Candida a los antifúngicos sintéticos disponibles en la actualidad es cada vez mayor, por lo que se buscan nuevos compuestos de origen vegetal que puedan ser eficaces en el tratamiento de infecciones causadas por este germen. Objetivo: Evaluar la actividad antifúngica in vitro de Dichrostachys cinerea L. contra una cepa de Candida albicans. Métodos: Se realizó un estudio observacional analítico transversal in vitro para evaluar la actividad antifúngica de extractos fluidos de hojas y de tallos de D. cinerea L mediante el método de macrodilución en caldo y como sustancia de referencia el alcohol. Resultados: A través del proceso se mostró la actividad antifúngica del extracto fluido de tallos de D. cinerea L. al 50 % hasta la dilución 1/32, determinada como la concentración mínima inhibitoria; el extracto fluido de hojas al 30 % no logró inhibir el crecimiento de la cepa de Candida albicans ATCC 10231. Conclusiones: La actividad antifúngica del extracto fluido al 50 % de las hojas de Dichrostachys cinerea L. fue efectiva, no así el preparado farmacéutico al 30 %. Se determinó la concentración mínima inhibitoria del extracto fluido de hojas al 50 % y se demostró que ésta superó a la del alcohol al 50 % en tres diluciones contra la Candida albicans.

ABSTRACT To evaluate the damage of oil exploration to ecosystems, ecotoxicological tests are a fundamental tool in environmental risk programs. The products used constitute xenobiotics that can be potentially toxic to soil biota and cause severe metabolic alterations in organisms, which affect structural and functional changes in terrestrial ecosystems. The objective of this research was to carry out a toxicological diagnosis of a water-based drilling fluid for use in oil exploration in Cuba. Toxicological procedures recommended by the Organization for Economic Cooperation and Development (OECD) were used, with two terrestrial contamination bioindicators, earthworms of the species Eisenia andrei and lettuce seeds (Lactuca sativa). The fluid was prepared by making a 1:9 dilutions (sludge: water) (v/v) and the dilutions to be tested were made from the fraction of suspended solids (FSS). As a toxicity criterion, the mean lethal concentration (LC50) was calculated from the mortalities of the bioindicators, with the Statgraphics Centurion XV statistical program, according to the Probit method. The values were compared with the current federal codes (CFR) that classify a toxic fluid when the LC50 is less than 3%. The LC50 for the bioassay with worms was 36.95% and 30.1% for lettuce seeds. According to established criteria, this fluid is non-toxic for the survival of terrestrial biota. These results support the use of this drilling fluid responsibly and with respect to the environment.

RESUMEN Para evaluar los daños de la exploración petrolera a los ecosistemas, los ensayos de ecotoxicológicos son una herramienta fundamental en los programas de riesgo ambiental. Los productos utilizados constituyen xenobióticos que pueden ser potencialmente tóxicos para la biota del suelo y ocasionar severas alteraciones metabólicas en los organismos, que repercuten en cambios estructurales y funcionales en los ecosistemas terrestres. El objetivo de esta investigación fue realizar un diagnóstico toxicológico a un fluido de perforación base agua para su uso en la exploración petrolera en Cuba. Se utilizaron procedimientos toxicológicos recomendadados por la Organización para la Cooperación Económica y el Desarrollo (OECD), con dos bioindicadores de contaminación terrestres, lombrices de tierra de la especie Eisenia andrei y semillas de lechuga (Lactuca sativa). El fluido se preparó realizando una dilución 1:9 (lodo: agua) (v/v) y a partir de la fracción de sólidos suspendidos (FSS) se realizaron las diluciones a ensayar. Como criterio de toxicidad se calculó la concentración letal media (CL50) a partir de las mortalidades de los bioindicadores, con el programa estadístico Statgraphics Centurion XV, según el método Probit. Los valores se compararon con los códigos federales (CFR) vigentes que cataloga a un fluido tóxico cuando la CL50 es menor que 3 %. La CL50 para el bioensayo con lombrices fue de 36.95 % y de 30.1 % para las semillas de lechuga; según los criterios establecidos este fluido es no tóxico para la supervivencia de la biota terrestre. Estos resultados apoyan el uso de este fluido de perforación con la responsabilidad y respecto al medio ambiente.

Abstract Immunodiagnostic tests for detecting dengue virus infections encounter challenges related to cross-reactivity with other related flaviviruses. Our research focuses on the development of a synthetic multiepitope antigen tailored for dengue immunodiagnostics. Selected dengue epitopes involved structural linearity and dissimilarity from the proteomes of Zika and Yellow fever viruses which served for computationally modeling the three-dimensional protein structure, resulting in the design of two proteins: rDME-C and rDME-BR. Both proteins consist of seven epitopes, separated by the GPGPG linker, and a carboxy-terminal 6 × -histidine tag. The molecular weights of the final proteins rDME-C and rDME-BR are 16.83 kDa and 16.80 kDa, respectively, both with an isoelectric point of 6.35. The distinguishing factor between the two proteins lies in the origin of their epitope sequences, where rDME-C is based on the reference dengue proteome, while rDME-BR utilizes sequences from prevalent Dengue genotypes in Brazil from 2008 to 2019. PyMol analysis revealed exposure of epitopes in the secondary structure. Successful expression of the antigens was achieved in soluble form and fluorescence experiments indicated a disordered structure. In subsequent testing, rDME-BR and rDME-C antigens were assessed using an indirect Elisa protocol against Dengue infected serum, previously examined with a commercial diagnostic test. Optimal concentrations for antigens were determined at 10 µg/mL for rDME-BR and 30 µg/mL for rDME-C, with serum dilutions ranging from 1:50 to 1:100. Both antigens effectively detected IgM and IgG antibodies in Dengue fever patients, with rDME-BR exhibiting higher sensitivity. Our in-house test showed a sensitivity of 77.3 % and 82.6 % and a specificity of 89.4 % and 71.4 % for rDME-C and rDEM-BR antigens. No cross-reactivity was observed with serum from Zika-infected mice but with COVID-19 serum samples. Our findings underscore the utility of synthetic biology in crafting Dengue-specific multiepitope proteins and hold promise for precise clinical diagnosis and monitoring responses to emerging Dengue vaccines. crossreactivity cross reactivity flaviviruses immunodiagnostics threedimensional three dimensional structure rDMEC rDME C rDMEBR. rDMEBR BR. BR linker carboxyterminal carboxy terminal histidine tag 1683 16 83 16.8 1680 80 respectively 635 35 6.35 proteome 200 2019 testing 1 µgmL µg mL 3 rDMEC, C, 150 50 1:5 1100. 1100 100. 100 1:100 patients inhouse house 773 77 77. 826 82 82. 894 89 4 89. 714 71 71. rDEMBR rDEM Zikainfected COVID19 COVID 19 COVID-1 samples Denguespecific specific vaccines 168 8 16. 63 6.3 20 201 15 5 1: 110 1:10 7 COVID1 COVID- 6. 2 11 1:1

Resumo Entre os produtos da colmeia de abelhas sem ferrão, o mel se destaca por possuir propriedades físico-químicas diferentes do mel produzido pela abelha Apis mellifera. Seu sabor é mais ácido e menos doce, além de naturalmente conter maior porcentagem de água. O mel é um dos produtos mais frequentemente adulterados comercialmente. Dessa forma, este trabalho teve como objetivo verificar se os testes qualitativos realizados para o mel de A. mellifera também são válidos para o mel de abelhas sem ferrão das espécies Tetragonisca angustula, Melipona quadrifasciata e Tetragona elongata, e se é possível identificar suas adulterações mais comuns: adulterações do mel de abelhas sem ferrão com xarope de milho, açúcar invertido e mel de A. mellifera foram simuladas e testadas com as reações de Lugol, Fiehe e Lund. Para essas três análises, também foi testada a redução do volume da amostra. O teste de Lund não foi efetivo com as amostras de mel das abelhas sem ferrão, pois elas apresentam um maior teor de água, e resultados confiáveis não puderam ser obtidos. Para os outros dois testes, a redução do volume da amostra utilizada foi eficiente. O teste de Fiehe detectou adulteração apenas com xarope de milho. O teste de Lugol detectou adulterações com xarope de milho e açúcar invertido em todas as diluições utilizadas e para todas as três espécies de abelhas sem ferrão. As adulterações por adição de água ou mel de A. mellifera não foram detectadas em nenhum dos testes. Portanto, é possível utilizar o teste qualitativo da reação de Lugol para uma avaliação inicial da qualidade do mel. Como é um teste rápido e de baixo custo, ele permite o desenvolvimento de cadeias produtivas para o mel de abelhas sem ferrão. Assim, a detecção de mel inautêntico pode ser feita por uma abordagem combinada de testes qualitativos como triagem inicial, seguida de testes quantitativos, se necessário. físicoquímicas físico químicas doce comercialmente forma A angustula elongata comuns análises obtidos eficiente Portanto custo Assim quantitativos necessário

Abstract Among the hive products of stingless bees (SB), honey is distinguished because it has different physicochemical properties than the honey produced by Apis mellifera. Its taste is more acidic and less sweet, and it naturally contains a higher percentage of water. Honey is one of the most frequently adulterated products marketed. Therefore, this work aimed to verify if the qualitative tests performed for A. mellifera honey are also valid for SB honey from Tetragonisca angustula, Melipona quadrifasciata, and Tetragona elongata and if they can detect the most common adulterations. Adulterations of SB honey with corn syrup, inverted sugar, and A. mellifera honey were simulated and tested with Lugol, Fiehe, and Lund reactions. For these three analyses, sample volume reduction was also tested. The Lund test did not work well with honey samples from SB because they have a higher water content, and reliable results could not be obtained. For the other two tests, the sample volume reduction used was efficient. The Fiehe test detected adulteration with corn syrup only. The Lugol test detected corn syrup and inverted sugar adulterations in all dilutions for all three SB species. No adulteration by added water or honey from A. mellifera was detected in any test. Therefore, using the qualitative Lugol’s reaction test to evaluate SB honey quality is reasonably affordable. Since it is a rapid and inexpensive test, it allows the development of production chains for SB honey. Thus, detecting inauthentic honey can be done by combining qualitative tests as the first screening, followed by quantitative tests if necessary. SB, , (SB) sweet marketed Therefore A angustula quadrifasciata reactions analyses content obtained efficient only species Lugols s affordable Thus screening necessary (SB

RESUMO: Técnicas colorimétricas, como sais de tetrazólio e resazurina, são indicadores de viabilidade (atividade respiratória) de fungos, bactérias e sementes, amplamente utilizados em diferentes áreas de pesquisa. O objetivo deste estudo foi desenvolver um protocolo com dois métodos, qualitativo e quantitativo, de triagem para detecção da viabilidade de Fusarium semitectum em sementes de soja, utilizando 2,3,5-trifenil cloreto de tetrazólio (TZ). O delineamento experimental utilizado foi inteiramente casualizado (DIC) em arranjo fatorial triplo (tempos de incubação, concentrações de TZ e diluições seriadas de inóculo) com níveis de 3, 5 e 5 de cada fator, respectivamente, com 3 repetições em ambos os métodos. Para o método qualitativo, a sensibilidade foi mais satisfatória no tempo de 72h, na concentração mínima de TZ de 0,1 mg mL-1 e com a concentração de inóculo 10⁻¹. No método quantitativo (espectrofotometria) não foi possível quantificar as Unidades Formadoras de Colônias (UFC) em placas, sendo a Densidade Óptica (DO) interferida pela estrutura do micélio no microtubo, embora seja uma metodologia reprodutível está condicionada pela morfologia do fungo. O presente estudo permitiu fácil visualização das estruturas de F. semitectum em sementes de soja, a partir do TZ, porém não foi possível quantificar a densidade de inóculo devido a baixa produção de esporos precisando de maiores ajustes na metodologia. RESUMO colorimétricas resazurina atividade respiratória fungos pesquisa métodos soja 2,3,5trifenil 235trifenil trifenil 2,3,5 2 TZ. . (TZ) DIC (DIC tempos incubação fator respectivamente 72h h 01 0 1 0, mL1 mL mL- 10¹ 10 ¹ 10⁻¹ espectrofotometria (espectrofotometria UFC (UFC placas DO (DO microtubo fungo F 5trifenil 235 2,3, (TZ 10⁻ 23 2,3 2,

ABSTRACT: Colorimetric techniques, such as tetrazolium salts and resazurin, are indicators of viability (respiratory activity) of fungi, bacteria, and seeds widely used in different areas of research. The objective of this study was to develop a protocol with two screening methods, qualitative and quantitative, to detect the viability of Fusarium semitectum in soybean seeds using 2,3,5-triphenyl tetrazolium chloride (TZ). The experimental design was completely randomized in a triple factorial arrangement (incubation times, TZ concentrations, and inoculum serial dilutions) with levels of 3, 5, and 5 of each factor, respectively, and 3 replications in both methods. For the qualitative method, sensitivity was more satisfactory at 72 hours, at the minimum TZ concentration of 0.1 mg mL-1, and with the inoculum concentration of 10-¹. In the quantitative method (spectrophotometry), it was not possible to quantify the colony forming units (CFU) on plates, with the Optical Density (OD) being interfered with by structure of the mecelium in the microtube. Although it is a reproducible methodology, it is conditioned by fungal morphology. The present study allowed easy visualization of the structures of F. semitectum in soybean seeds, based on the TZ. However, it was not possible to quantify the inoculum density, due to low spore production, requiring greater adjustments in the methodology. ABSTRACT techniques resazurin respiratory activity fungi bacteria research methods 2,3,5triphenyl 235triphenyl triphenyl 2,3,5 2 . (TZ) incubation times concentrations dilutions factor respectively 7 hours 01 0 1 0. mL1, mL1 mL 1, mL-1 10¹. 10¹ 10 ¹. ¹ 10-¹ spectrophotometry, spectrophotometry , (spectrophotometry) CFU (CFU plates OD (OD microtube methodology morphology F However density production 5triphenyl 235 2,3, (TZ mL- 10- (spectrophotometry 23 2,3 2,