Characterization of a Clostridioides difficile ST-293 isolate from a recurrent infection in Argentina

Resumen Clostridioides difficile es un patógeno esporulado oportunista responsable de diarrea asociada a antibióticos en humanos. C. difficile produce 2 toxinas principales: TcdAy TcdB, además de la toxina binaria (CDT), también asociada a la virulencia. Este estudio buscó caracterizar el aislamiento ALCD3, involucrado en un episodio de recurrencia de una infección nosocomial. La caracterización molecular mostró que dicho aislamiento pertenece al toxinotipo 0/v y el análisis por MLST demostró un perfil alélico adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 y tpi:1, lo cual corresponde al ST293 (MLST clado 1). Durante el crecimiento, el aislamiento ALCD3 mostró un incremento temprano de la tasa de esporulación y valores máximos de formas termorresistentes luego de 2 días de incubación. Tanto la cinética de esporulación como la producción de formas termorresistentes fueron más rápidas en el aislamiento ALCD3 que en la cepa de referencia VPI 10463. La germinación en presencia del germinante natural taurocolato fue más rápida en el aislamiento ALCD3 que en la cepa VPI 10463, lo que indica que aquel comienza la hidrólisis del córtex antes. También, el co-germinante glicina indujo una rápida liberación de ácido dipicolínico en ALCD3. Estos hallazgos indican que el aislamiento ALCD3 es particularmente eficiente en la esporulación y en la germinación. El presente trabajo representa el primer informe de la circulación de C. difficile ST293 en Argentina. La habilidad del aislamiento ALCD3 para producir toxinas y su alta capacidad de esporulación/germinación son características claves compatibles con un alto potencial de diseminación e inducción de infecciones recurrentes.

Abstract Clostridioides difficile is an opportunistic spore-forming pathogen responsible for antibiotic-associated diarrhea in humans. C. difficile produces two main toxins: TcdA and TcdB as well as a third toxin named binary toxin (CDT) that is also involved in virulence. The present study aimed at characterizing the C. difficile isolate ALCD3 involved in a relapse episode of nosocomial infection. Molecular characterization showed that isolate ALCD3 belongs to tox-inotype 0/v and the MLST analysis demonstrated allelic profile adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 and tpi:1 which corresponds to ST293 (MLST clade: 1). During growth, isolate ALCD3 showed an early increase in the sporulation ratio as well as maximal values of heat resis-tant forms after 2 days of incubation. Both sporulation kinetics and production of heat resistant forms were faster for isolate ALCD3 than for the reference strain VPI 10463. Germination in the presence of the natural germinant taurocholate was faster for isolate ALCD3 than for strain VPI 10463, which indicates that isolate ALCD3 starts cortex hydrolysis earlier than strain VPI 10463. Furthermore, the co-germinant glycine, induces rapid release of dipicolinic acid (DPA) in isolate ALCD3. These findings indicate that isolate ALCD3 is particularly efficient in both sporulation and germination. The present work represents the first report of the circulation of C. difficile ST293 in Argentina. The ability of isolate ALCD3 to produce toxins and its high sporulation/germination capacity are key features compatible with a microorganism with high dissemination potential and the possibility of inducing recurrent infections.

Kinetics Approach on the Evolution of the Nutritive Properties, Antinutritonal Factors and Free Radical Scavenging Capacity of DPPH during Germination of Two Local Legume Varieties (Phaseolus vulgaris and Vigna unguiculata)

Aims: To improve the processes of pre-treatment of legumes for their nutritional valorization. Study Design: Original research. Place and Duration of Study: This study took place at the Laboratory of Biotechnology, Agriculture and Valorization of Biological Resources, Félix Houphouët-Boigny University between February and July 2022. Methodology: Red beans and cowpeas purchased on the local market of Adjamé were subjected, after unitary operation of sorting and washing, to a two-factor design of experiment: seed/water ratio and the soaking time, in order to identify the ideal ratio and soaking time to well reduce phytates. Seeds resulting from this pre-treatment were germinated for 72 h and some key nutrient and functional parameters were evaluated. Results: The ratio 8/9 and soaking time of 18 hours allowed a maximum reduction of 62 and 66.6% of phytate and 72.83 and 67.48% of tanins in cowpea and red beans, respectively. Protein content of these 72 hours germinated pre-treated seeds decreased very slightly and finally remained at high level of 22.02 and 23.13 g/100 g for cowpea and red bean, while reducing sugar levels increased significantly throughout germination to a maximum of 8.19 and 8.13 mg/100 g. Regarding functional and antioxidant properties, a maximum increase in total polyphenols (49.08 and 68.314 mg/100 g) and total flavonoids (13.75 and 39.67 mg/100 g) was observed after 48 h of germination for cowpea and red beans, respectively. Furthermore, this improvement in phenolic content led to a significant improvement in the free radical scavenging capacity of DPPH of 24.50 and 46.38 %. It should also be noted that the germinated red bean showed better nutritional value than the germinated cowpea. Conclusion: This approach of pre-processing germinated legumes at seed-to-water ratio of 8/9 soaked for 18 hours, providing functional foods with guaranteed nutritional value, appears to be a way to improve local diets

Filtrates from cultures of endophytic fungi isolated from leaves of Copaifera oblongifolia (Fabaceae) affect germination and seedling development differently / Fungos endofíticos isolados de folhas de Copaifera oblongifolia (Fabaceae) afetam diferentemente a germinação e o desenvolvimento das plântulas

Abstract Interactions between endophytic fungi (EFs) and their host plants range from positive to neutral to negative. The results of such interactions can vary depending on the organ of the infected host plant. EFs isolated from the leaves of some species of plants have potential for use as agents to inhibit seed germination and control invasive plants. The objectives of this study were to identify EFs present in the leaves of Copaifera oblongifolia and to evaluate the role of these fungi in seed germination and seedling development. A total of 11 species of EFs were isolated, which were identified using the internal transcribed spacers (ITS) sequence of the nuclear ribosomal DNA. The isolated species of EFs are generalists and probably are transmitted horizontally. Laboratory tests revealed that filtrates of these fungal isolates differently affect seed germination and seedling development of C. oblongifolia. The species Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum and Phomopsis sp. negatively affected seed germination, with N. parvum standing out for its negative effects, inhibiting seedling germination and survival in 89 and 222%, respectively. In addition, Cochliobolus intermedius negatively affected seedling development. Thus, the combined use of N. parvum and C. intermedius, or products from the metabolism of these microorganisms, in the control of invasive plants deserves attention from future studies.

Resumo As interações entre fungos endofíticos (FEs) e suas plantas hospedeiras variam de positivas, neutras a negativas. Os resultados destas interações podem variar dependendo do órgão da planta hospedeira infectada. FEs isolados de folhas de algumas espécies de plantas têm potencial para serem usados como agentes inibidores da germinação de sementes e no controle de plantas invasoras. Os objetivos deste estudo foram identificar os FEs presentes nas folhas de Copaifera oblongifolia e avaliar o papel destes fungos na germinação das sementes e no desenvolvimento das plântulas. Um total de 11 espécies de FEs foi isolado das folhas de C. oblongifolia e identificado através da sequência dos espaçadores internos transcritos do DNA ribossomal nuclear. As espécies de FEs isoladas são generalistas e provavelmente devem ser transmitidas horizontalmente. Os resultados dos testes de germinação mostraram que filtrados destes isolados fúngicos podem afetar diferentemente a germinação das sementes e o desenvolvimento das plântulas de C. oblongifolia. As espécies Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum e Phomopsis sp. afetaram negativamente a germinação das sementes de C. oblongifolia. Dentre estas espécies devemos destacar que N. parvum reduziu a germinação e a sobrevivência das plântulas em 89 e 222%, respectivamente. Além disso, Cochiliobolus intermedius afetou negativamente o desenvolvimento das plântulas. Assim, o uso combinado de N. parvum e C. intermedius, ou de produtos do metabolismo destas espécies de fungos, têm potencial para serem usados no manejo de plantas invasoras.

Efeito da germinação e da desidratação no valor nutritivo, compostos bioativos e atividade antioxidante de farinhas de leguminosas / Germination and dehydration effects on nutritional value, bioactive compounds and antioxidant activity of legume flours / Efectos de la germinación y la deshidratación sobre el valor nutritivo, los compuestos bioactivos y la actividad antioxidante de harinas de leguminosas

Os objetivos deste trabalho foram avaliar o efeito da germinação no teor de macronutrientes e da desidratação em compostos bioativos, nutrientes e atividade antioxidante de farinhas das leguminosas Phaseolus lunatus L. e Cajanus cajan (L. Millsp.) germinadas. As condições de germinação foram controladas em laboratório e as amostras foram submetidas à desidratação térmica (55°C) e liofilização. Foram realizadas as análises de composição centesimal, fenólicos totais flavonoides, atividade antioxidante (DPPH, FRAP), perfil de ácidos graxos (CG-FID) e de vitâmeros de tocoferol (HPLC- DAD). A germinação das leguminosas promoveu aumento de proteínas, lipídios e carboidratos. O processamento térmico para desidratação contribuiu para perdas de proteínas, lipídios, cinzas, e maior atividade antioxidantes em ambas as espécies. O perfil de ácidos graxos e vitâmeros de tocoferol mostraram-se com variações que dependem do tipo de leguminosa e condições de processamento, podendo ser positivas ou negativas. Farinhas de leguminosas germinadas pode ser uma alternativa para a inserção na dieta ou em formulações alimentícias e as condições de processamento devem ser monitoradas com intuito de otimiza propriedades nutritivas e funcionais.

The objectives of this work were to evaluate the effect of germination on macronutrient content and dehydration on bioactive compounds, nutrients and antioxidant activity of flours from germinated Phaseolus lunatus L. and Cajanus cajan (L. Millsp.) legumes. The germination conditions were controlled in the laboratory and the samples were submitted to thermal dehydration (55°C) and lyophilization. Analyses were carried out for centesimal composition, total phenolic flavonoids, antioxidant activity (DPPH, FRAP), fatty acid profile (GC-FID) and tocopherol vitamers (HPLC-DAD). Germination of the legumes promoted an increase in proteins, lipids and carbohydrates. Thermal processing for dehydration contributed to losses of proteins, lipids, ash, and higher antioxidant activity in both species. The profile of fatty acids and tocopherol vitamers showed variations that depend on the type of legume and processing conditions, and can be positive or negative. Sprouted legume flours can be an alternative for insertion in the diet or in food formulations and the processing conditions must be monitored in order to optimize nutritional and functional properties.

Los objetivos de este trabajo fueron evaluar el efecto de la germinación sobre el contenido en macronutrientes y de la deshidratación sobre los compuestos bioactivos, los nutrientes y la actividad antioxidante de harinas de leguminosas germinadas de Phaseolus lunatus L. y Cajanus cajan (L. Millsp.). Las condiciones de germinación se controlaron en el laboratorio y las muestras se sometieron a deshidratación térmica (55°C) y liofilización. Se realizaron análisis de composición centesimal, flavonoides fenólicos totales, actividad antioxidante (DPPH, FRAP), perfil de ácidos grasos (GC-FID) y vitaminas tocoferol (HPLC-DAD). La germinación de las leguminosas promovió un aumento de proteínas, lípidos y carbohidratos. El procesado térmico para la deshidratación contribuyó a pérdidas de proteínas, lípidos, cenizas y a una mayor actividad antioxidante en ambas especies. El perfil de ácidos grasos y vitaminas tocoferol mostró variaciones que dependen del tipo de legumbre y de las condiciones de procesado, pudiendo ser positivas o negativas. Las harinas de leguminosas germinadas pueden ser una alternativa para su inserción en la dieta o en formulaciones alimentarias y las condiciones de procesado deben ser monitorizadas para optimizar las propiedades nutricionales y funcionales.

Germination of fruits eaten by the maned wolf Chrysocyon brachyurus (Illiger, 1815) (Carnivora, Canidae) / Germinação de frutos consumidos pelo lobo-guará Chrysocyon brachyurus (Illiger, 1815) (Carnivora, Canidae)

Abstract Zoochory is a fundamental process that can be the main mechanism for seed and plant dispersal for many species. Mammals of the Carnivora order are among the most important dispersing agents; however, little is known regarding the role of canids as seed dispersers. Although the maned wolf (Chrysocyon brachyurus) has a potentially important role in seed dispersal, given its relatively high consumption of fruits, few studies have investigated the germination rate of ingested seeds. Here, we used seeds removed from the feces of two captive specimens (maned wolf treatment) and those directly collected from unconsumed fruits (control) in germination essays to evaluate the germination rate and emergence velocity index (IVE). We used mature fruits from five species of trees in the Cerrado and Atlantic Forest in a 5 (species) × 2 (method of seed collection) factorial arrangement. The passage of seeds through the digestive tract of the maned wolf favored the germination of Genipa americana and Psidium guajava, delayed germination of Psidium cattleianum, and maintained the germination of Plinia cauliflora and Ficus obtusifolia. Our results revealed that germination occurred for all tested plant species consumed by the maned wolf; therefore, this canid species has high dispersal potential and can be an important ally in the restoration of the Cerrado and Atlantic Forest-Cerrado contact zone ecosystems.

Resumo A zoocoria é um processo fundamental para muitas espécies vegetais, podendo ser o principal mecanismo de dispersão de sementes e plantas. Os mamíferos da ordem Carnivora estão entre os agentes dispersores mais importantes, contudo, pouca atenção tem sido dada ao papel dos canídeos como dispersores de sementes. Embora o lobo-guará (Chrysocyon brachyurus) tenha um papel potencialmente importante na dispersão de sementes dado o seu consumo relativamente elevado de frutos, poucos estudos investigaram a taxa de germinação de sementes ingeridas. Aqui utilizamos sementes retiradas de fezes de dois exemplares em cativeiro (tratamento do lobo-guará) e diretamente retiradas de frutos não consumidos (controle) em ensaios de germinação para avaliar a taxa de germinação e o índice de velocidade de emergência (IVE). Utilizamos frutos maduros de cinco espécies de árvores que ocorrem no Cerrado e Mata Atlântica formando um arranjo fatorial de 5 (espécie) × 2 (forma de coleta de sementes). A passagem das sementes pelo trato digestivo do lobo-guará favoreceu a germinação de duas espécies, Genipa americana e Psidium guajava, atrasou a germinação de uma espécie, Psidium cattleianum e não afetou a germinação de duas espécies, Plinia cauliflora e Ficus obtusifolia. Com base nos nossos resultados, particularmente a descoberta de que todas as espécies vegetais testadas consumidas pelo lobo-guará germinaram, concluímos que esta espécie de canídeo tem um elevado potencial de dispersão, sendo um importante aliado na restauração de ecossistemas do Cerrado e da zona de contato entre Cerrado e Mata Atlântica.

Filtrates from cultures of endophytic fungi isolated from leaves of Copaifera oblongifolia (Fabaceae) affect germination and seedling development differently / Fungos endofíticos isolados de folhas de Copaifera oblongifolia (Fabaceae) afetam diferentemente a germinação e o desenvolvimento das plântulas

Interactions between endophytic fungi (EFs) and their host plants range from positive to neutral to negative. The results of such interactions can vary depending on the organ of the infected host plant. EFs isolated from the leaves of some species of plants have potential for use as agents to inhibit seed germination and control invasive plants. The objectives of this study were to identify EFs present in the leaves of Copaifera oblongifolia and to evaluate the role of these fungi in seed germination and seedling development. A total of 11 species of EFs were isolated, which were identified using the internal transcribed spacers (ITS) sequence of the nuclear ribosomal DNA. The isolated species of EFs are generalists and probably are transmitted horizontally. Laboratory tests revealed that filtrates of these fungal isolates differently affect seed germination and seedling development of C. oblongifolia. The species Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum and Phomopsis sp. negatively affected seed germination, with N. parvum standing out for its negative effects, inhibiting seedling germination and survival in 89 and 222%, respectively. In addition, Cochliobolus intermedius negatively affected seedling development. Thus, the combined use of N. parvum and C. intermedius, or products from the metabolism of these microorganisms, in the control of invasive plants deserves attention from future studies.

As interações entre fungos endofíticos (FEs) e suas plantas hospedeiras variam de positivas, neutras a negativas. Os resultados destas interações podem variar dependendo do órgão da planta hospedeira infectada. FEs isolados de folhas de algumas espécies de plantas têm potencial para serem usados como agentes inibidores da germinação de sementes e no controle de plantas invasoras. Os objetivos deste estudo foram identificar os FEs presentes nas folhas de Copaifera oblongifolia e avaliar o papel destes fungos na germinação das sementes e no desenvolvimento das plântulas. Um total de 11 espécies de FEs foi isolado das folhas de C. oblongifolia e identificado através da sequência dos espaçadores internos transcritos do DNA ribossomal nuclear. As espécies de FEs isoladas são generalistas e provavelmente devem ser transmitidas horizontalmente. Os resultados dos testes de germinação mostraram que filtrados destes isolados fúngicos podem afetar diferentemente a germinação das sementes e o desenvolvimento das plântulas de C. oblongifolia. As espécies Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum e Phomopsis sp. afetaram negativamente a germinação das sementes de C. oblongifolia. Dentre estas espécies devemos destacar que N. parvum reduziu a germinação e a sobrevivência das plântulas em 89 e 222%, respectivamente. Além disso, Cochiliobolus intermedius afetou negativamente o desenvolvimento das plântulas. Assim, o uso combinado de N. parvum e C. intermedius, ou de produtos do metabolismo destas espécies de fungos, têm potencial para serem usados no manejo de plantas invasoras.

Filtrates from cultures of endophytic fungi isolated from leaves of Copaifera oblongifolia (Fabaceae) affect germination and seedling development differently

Abstract Interactions between endophytic fungi (EFs) and their host plants range from positive to neutral to negative. The results of such interactions can vary depending on the organ of the infected host plant. EFs isolated from the leaves of some species of plants have potential for use as agents to inhibit seed germination and control invasive plants. The objectives of this study were to identify EFs present in the leaves of Copaifera oblongifolia and to evaluate the role of these fungi in seed germination and seedling development. A total of 11 species of EFs were isolated, which were identified using the internal transcribed spacers (ITS) sequence of the nuclear ribosomal DNA. The isolated species of EFs are generalists and probably are transmitted horizontally. Laboratory tests revealed that filtrates of these fungal isolates differently affect seed germination and seedling development of C. oblongifolia. The species Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum and Phomopsis sp. negatively affected seed germination, with N. parvum standing out for its negative effects, inhibiting seedling germination and survival in 89 and 222%, respectively. In addition, Cochliobolus intermedius negatively affected seedling development. Thus, the combined use of N. parvum and C. intermedius, or products from the metabolism of these microorganisms, in the control of invasive plants deserves attention from future studies.

Resumo As interações entre fungos endofíticos (FEs) e suas plantas hospedeiras variam de positivas, neutras a negativas. Os resultados destas interações podem variar dependendo do órgão da planta hospedeira infectada. FEs isolados de folhas de algumas espécies de plantas têm potencial para serem usados como agentes inibidores da germinação de sementes e no controle de plantas invasoras. Os objetivos deste estudo foram identificar os FEs presentes nas folhas de Copaifera oblongifolia e avaliar o papel destes fungos na germinação das sementes e no desenvolvimento das plântulas. Um total de 11 espécies de FEs foi isolado das folhas de C. oblongifolia e identificado através da sequência dos espaçadores internos transcritos do DNA ribossomal nuclear. As espécies de FEs isoladas são generalistas e provavelmente devem ser transmitidas horizontalmente. Os resultados dos testes de germinação mostraram que filtrados destes isolados fúngicos podem afetar diferentemente a germinação das sementes e o desenvolvimento das plântulas de C. oblongifolia. As espécies Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum e Phomopsis sp. afetaram negativamente a germinação das sementes de C. oblongifolia. Dentre estas espécies devemos destacar que N. parvum reduziu a germinação e a sobrevivência das plântulas em 89 e 222%, respectivamente. Além disso, Cochiliobolus intermedius afetou negativamente o desenvolvimento das plântulas. Assim, o uso combinado de N. parvum e C. intermedius, ou de produtos do metabolismo destas espécies de fungos, têm potencial para serem usados no manejo de plantas invasoras.

Decolourization and Detoxification of Azo Dye, Malachite Green by Pseudomonas monteilii Strain RZT1, a Bacterium Isolated from Textile Wastewater

Discharge of textile industrial effluent without proper treatment has become a severe hazard for the animal health and environment worldwide. Therefore, this study was designed to isolate azo dye-degrading bacteria from textile wastewater and evaluate their ability to biodegrade reactive dyes into non-toxic products. The potent bacterial strain which was isolated from textile wastewater was identified as Pseudomonas monteilii strain RZT1 on the basis of 16S rDNA sequence. The isolated bacterial strain exhibited good decolorization ability with yeast extract supplementation as cosubstrate in static conditions for Malachite Green dye. The optimal condition for the decolorization of Malachite Green dye by P. monteilii strain RZT1 were at pH 7.0 and 28癈. Decolorization rates of Malachite Green dye by P. monteilii strain RZT1 were varied with initial dye concentration as follow: 84.8%, 75.4%, 63.4% and 45.5% decolorization for 100ppm, 200ppm, 300ppm and 400ppm initial dye concentration respectively. We investigated the effects of dyes used in the textile industry on the seed germination of Five crops - Rice (Oryza sativa), Wheat (Triticum aestivum L.), Khesari (Lathyrus sativus), Mustard (Brassica nigra) and Bitter Melon (Momordica charantia). It was found that textile dye Malachite Green had negative effect on seed germination and seedling growth in test cultures. The harmful effects of dye on seed germination and early seedling growth parameters were augmented with increase of dye concentration. Interestingly, treatment of the Malachite Green dye with isolated bacteria reduced the adverse effects of that dye on seed germination and seedling growth. Thus, it indicated the potentiality of P. monteilii strain RZT1 for bioremediation of textile effluents into a non-toxic form for plants.

Impacto de la germinación en los compuestos bioactivos de dos ecotipos de kiwicha de grano color negro y rosado (Amaranthus caudatus L.)

Las investigaciones en kiwicha de grano color negro y rosado (Amaranthus caudatus L.) son muy limitadas ocasionando la subutilización de estos ecotipos debido al desconocimiento de su potencial alimentario. Estos granos nativos deberían ser rescatados y valorados como una opción de alimento nutritivo empleando tecnologías sencillas como la germinación para mejorar sus cualidades nutricionales. El objetivo de la investigación fue determinar el impacto del periodo de germinación en los compuestos bioactivos de dos ecotipos de kiwicha de "grano color negro" y "grano color rosado". En un diseño experimental y comparativo se estudiaron los ecotipos de kiwicha (negro y rosado) y periodos de germinación (0, 24, 48 y 72 h). Se determinó: compuestos fenólicos totales, betalaínas, capacidad antioxidante, proteínas, grasas, cenizas y humedad. Al aumentar el periodo de germinación se incrementaron progresivamente los compuestos fenólicos totales, betalaínas, capacidad antioxidante, proteínas y cenizas en ambos granos de kiwicha; y disminuyeron gradualmente el contenido de grasas y humedad, comparado con el periodo cero (granos sin germinar). La exposición a periodos de germinación logró mayores contenidos de compuestos fenólicos totales, capacidad antioxidante, proteínas y cenizas en granos de kiwicha de color negro en comparación a granos de color rosado; sin embargo, los granos de color rosado obtuvieron contenidos de betalaínas y humedades superiores a los granos de color negro.

Research on black and pink-colored kiwicha grains (Amaranthus caudatus L.) is very limited, resulting in the underutilization of these ecotypes due to the lack of knowledge of their food potential. These native grains should be rescued and valued as a nutritious food option using simple technologies such as germination to improve their nutritional qualities. The objective of the research was to determine the impact of the germination period on the bioactive compounds of two ecotypes of kiwicha "black-colored grain" and "pink-colored grain". In an experimental and comparative design, kiwicha ecotypes (black and pink) and germination periods (0, 24, 48 and 72 h) were studied. The following were determined: total phenolic compounds, betalains, antioxidant capacity, protein, fat, ash and moisture. As the germination period increased, total phenolic compounds, betalains, antioxidant capacity, proteins and ashes progressively increased in both kiwicha grains; and fat and moisture content gradually decreased, compared to the zero period (ungerminated grains). Exposure to germination periods resulted in higher contents of total phenolic compounds, antioxidant capacity, proteins and ashes in black-colored kiwicha grains compared to pink-colored grains; however, pink-colored grains obtained higher betalain and moisture content than black-colored grains.

Conservación ex situ y propagación in vitro de germoplasma de Neltuma alba Griseb / Germoplasm ex situ conservation and in vitro propagation of Neltuma alba Griseb

RESUMEN Neltuma alba Griseb es una especie emblemática de la Ecorregión Chaqueña, por el valor de su madera y frutos. Los resultados de la presente investigación, dan cuenta de que las semillas de algarrobo blanco pueden ser críoconservadas sin afectar el poder y la energía para germinar. Las semillas fueron conservadas por tres meses a Temperatura ambiente (24° y 30°C, Congelamiento (-18°C) y Nitrógeno líquido (-196°C). Posteriormente, fueron escarificadas mecánicamente y sembradas en tierra negra y aserrín (3:1). Los resultados fueron sometidos a pruebas no paramétricas de Kruskal-Wallis, analizándose poder, energía y vigor germinativo, y no se detectaron diferencias significativas entre tratamientos. Para la regeneración in vitro en medio de cultivo MS suplementados con 3 mg L-1 IBA y 0,05 mg L-1 CIN, se utilizaron segmentos nodales de 2 cm de longitud obtenidos de plántulas provenientes de semillas. Los porcentajes de enraizamiento obtenidos fueron elevados (87-90%). No existiendo diferencias significativas entre los tratamientos ensayados. El almacenamiento de semillas a 24°-30°C resulta ser el más apropiado para alcanzar altos porcentajes de germinación

ABSTRACT Neltuma alba Gris is an emblematic specie of Chaco Ecoregion for its wood and fruits value. This research show that algarrobo blanco's seeds can cryopreserve without affecting germination power and energy. The seeds were kept for three months at 24°-30°C, (-18°C) and (-196°C). Later, they were mechanically scarified and sown in black soil and sawdust (3: 1). The results were subjected to non-parametric Kruskal-Wallis tests, analyzing germination power, energy and vigor. The test did not detect significant differences between treatments. The nodal segments (2 cm long) obtained from plants of seeds were used for in vitro regeneration in MS culture medium supplemented with 3 mg L-1 IBA and 0.05 mg L-1 CIN, The rooting percentages obtained were high (87-90%). There are no significant differences between treatments tested. The storage of seeds at 24°-30°C is the most approppiate to rich high percentages of germination.

Determination of resource mobilization during seedling growth of palmyra palm, Borassus flabellifer L

Asian palmyra palm (Borassus flabellifer L.) is a multipurpose tree with year round products as food for the dependent society. Besides edible items the tree offers around 801 useful goods which are economically important. Such a plant’s seedling biology has not been fully understood. Hence, the present study was aimed at understanding seedling developmental stages and resource mobilisation in Palmyra palm and further to determine the role of cotyledonary sheath (CS) during entire seedling growth. We investigated the developmental changes throughout growth of seedlings by providing different sets of growth conditions. Artificial seed bed made of coconut coir was compared with that of soil conditions to study germination and establishment of seedlings. Phloem loading dye was used to track the flow of nutrients from embryo to cotyledonary sheath. Seed germination in palmyra palm is hypogeal by forming ligular and tubular structures remotely in soil. Eight new organs differentiate from the seed embryo that includes haustorium, ligule, cotyledonary sheath, cataphyll, eophyll, mesocotyle (junction), primary root and mesocotyl roots during seed germination. Among these the first four are temporary organs and they disintegrate once the seedling is well established. The last four organs are responsible for developing acomplete plantlet at later stages. The seed and seedling organs of Palmyra palm have four major storage reserves to support successful germination and firm establishment of seedling. Palmyra has evolved to control the solubilisation, movement and regulation of food among transient seedling organs and carry out translocation of food to the developing and differentiating organs. Seedlings also have developed physiological functions and strategies to mobilise the stored food without losing them at any point of their growth and developmental stages. Water required for seed germination permeates laterally only viacotyledonary sheath which has spongy tissues and lenticels all over. These tissues are the primary mode of water supply as the seedlings lack major root organs in the early stages of development. Flourescent microscopic and anatomical studies were carried out to observe the transport and storage of food substances required during seedling growth. Histochemical studies of seedling organs have revealed the presence of various type of nutrients such as simple sugars, carbohydrates, proteins, amino acids and lipids.

ELECTROPHORETIC SEPARATION OF INDUCIBLE PROTEINS IN MAIZE SEEDLINGS UNDER FUNGICIDE TREATMENT

The aim of the experiment was to study the effect of a various concentrations of a systemic fungicide, metalaxyl on the expression of new proteins during early germination of maize. The study was conducted for 7 days. Our investigation showed the appearance of induced proteins ranging from 100-110kDa, 44kDa, 30kDa and 18-25kDa on different days of observation and different concentration of metalaxyl treatment, which is in line with the ?ndings in various abiotic stresses

Luminosity, temperature, and substrates on the germination of Solanum capsicoides

Solanum capsicoides, commonly known as red soda apple, is a little-studied species with phytotherapeutic characteristics of interest to the pharmaceutical industry. However, little is known about its agronomic characteristics. Therefore, this study aimed to assess the effect of light, different temperatures, and substrates on the germination of S. capsicoides seeds. The study consisted of two experiments: Experiment I - Germination of S. capsicoides seeds under different wavelengths. The seeds were kept on Petri dishes covered with Germitest™ paper moistened with distilled water and exposed to the monochromatic, red, far-red, and dark wavelengths. The experiment was performed in a completely randomized design with four replications of 25 seeds. Experiment II - Germination of S. capsicoides seeds on different substrates and at different temperatures. The seeds were distributed according to six substrates: paper roll, on paper, between sand, on sand, on soil + sand + manure, and between soil + sand + manure. The seeds were also subjected to the constant temperatures of 20, 25, 30, and 35 °C and the alternate temperatures of 20-30, 20-35, 25-30, and 25-35 °C, characterizing a 6 x 8 factorial arrangement. Germination rate, germination speed index, and mean germination time were evaluated. Red soda apple seeds are classified as positive photoblastic, with the highest germination rate occurring at the temperature of 20-35 °C combined with on paper, paper roll, on sand, and between sand substrates.

Seed size and presence of thorns in Caryocar brasiliense on seedling emergence

Commercial propagation of pequizeiro, which stands out among the main native fruits of the Cerrado region for its high economic potential, has been done by seeds, despite their erratic germination. The objective of this study was to evaluate the effect of seed size and presence of thorns on the endocarp on seedling emergence of pequizeiro in the field. Seeds were collected in a pequizeiro commercial area in Canarana-MT, Brazil. Due to availability, 2,353 thorny seeds (3 sizes) and 106 seeds without thorns (medium sized) were tested. Thorny seeds were classified as large ­ longer than 50 mm; medium ­ between 40 and 50 mm; small ­ less than 40 mm. Seedling emergence started after 50 days in all seed categories, extending up to 100 days in the large seeds and to about 1 year after sowing in the small and medium sized ones. Small seeds showed a lower rate and speed of emergence compared to the others. Large seeds showed higher emergence speed compared to the medium ones. The presence of thorns did not affect seedling emergence. These results indicate the advantages of pequizeiro seed classification for sowing purposes.

Evaluation of the phytotoxicity of landfill leachate treated with a Rotating Biological Reactor / Avaliação da fitotoxicidade de lixiviado de aterro tratado com um Reator Biológico Rotativo

ABSTRACT One of the major drawbacks of this final disposal technique is leachate production, which occurs due to the degradation of organic matter and rainwater percolation. The leachate composition is quite varied, with high organic load and various compounds that may be toxic to the environment. To reduce the polluting potential of the leachate, it must be subjected to an appropriate treatment. Biological treatments are widely used in the treatment of leachate, although these technologies have some drawbacks. As an alternative to conventional biological treatments, rotating biological reactors are used. The objective of this study was to determine the efficiency of a pilot scale rotating biological reactors in the removal of biochemical oxygen demand, chemical oxygen demand, and phosphorus from the landfill leachate. Effluent toxicity and rotating biological reactors's ability to remove this toxicity were also verified by calculating the germination index of lettuce (Lactuca sativa) and cucumber (Cucumis sativus) seeds. The reactor showed an average efficiency of 75.99% of biochemical oxygen demand removal, 33.53% of chemical oxygen demand removal, and 16.04% of phosphorus removal. In fact, the cucumber and lettuce seeds proved sensitive to the toxicity of the effluent. For crude leachate, the germination index values of lettuce and cucumber were 13.28 and 49.61%, respectively. In this study, rotating biological reactors obtained a good efficiency in the removal of toxicity from leachate. The germination index of the seeds for the treated effluent was 60.9% for lettuce and 82.49% for cucumber.

RESUMO Diante do crescimento populacional acelerado e do desenvolvimento econômico, grande quantidade de resíduos tem sido gerada nos grandes centros urbanos. Esses resíduos devem ser coletados e dispostos adequadamente, sendo os aterros sanitários os locais mais adequados para a sua disposição. Um dos grandes inconvenientes dessa técnica de disposição final é a produção de um líquido lixiviado, proveniente da degradação da matéria orgânica e da percolação da água da chuva. A composição do lixiviado é bastante variada, com elevada carga orgânica e diversos compostos que podem ser tóxicos ao meio ambiente. Para determinar a toxicidade do lixiviado são amplamente utilizados testes fitotoxicológicos, como o índice de germinação de sementes. Com o intuito de reduzir o potencial poluidor do lixiviado, este deve ser submetido a um tratamento adequado. Tratamentos biológicos são bastante utilizados, mas possuem algumas desvantagens. Como alternativa aos tratamentos biológicos convencionais, surgem os reatores biológicos rotatórios. O objetivo deste estudo foi determinar a eficiência de uns reatores biológicos rotatórios em escala piloto na remoção da demanda bioquímica de oxigênio, da demanda química de oxigênio e de fósforo do lixiviado do aterro. Também foi verificada a toxicidade do efluente e a capacidade dos reatores biológicos rotatórios para remover essa toxicidade por meio do cálculo do índice de germinação de sementes de alface (Lactuca sativa) e pepino (Cucumissativus). O reator utilizado neste trabalho obteve eficiência média de 75,99% de remoção de demanda bioquímica de oxigênio, 33,53% de remoção de demanda química de oxigênio e 16,04% de remoção de fósforo. As sementes de alface e pepino mostraram-se bastantes sensíveis à toxicidade do efluente. Para o lixiviado bruto, os índices de germinação da alface e do pepino foram, respectivamente, 13,28 e 49,61%. Neste estudo, os reatores biológicos rotatórios obtiveram boa eficiência na remoção da toxicidade do lixiviado. O índice de germinação das sementes para o efluente tratado foi de 60,9% para a alface e de 82,49% para o pepino.

Suitability of tetrazolium test for Tamarindus indica L. seeds / Adequação do teste de tetrazolium para sementes de Tamarindus indica L

Seed quality is routinely assessed by direct tests, e.g, the germination test, or indirect tests like the tetrazolium test, which has shoown to be promising in the determine viability and vigor, allowing the diagnosis of the main problems that may affect seed quality, such as mechanic damages, field deterioration and storage. In this respect, this study was conducted to develop a tetrazolium test protocol to evaluate the viability and vigor of Tamarindus indica L. seeds. Before exposing the seeds to the tetrazolium solution, seed preconditioning studies were carried out in which seven soaking times were tested. The soaking time that did not cause damage to the seed embryo and allowed the removal of the seed coat to expose the seed structures to the tetrazolium salt was selected. Then, an experiment was set up in a completely randomized design with a 2x3x3 factorial arrangement involving two seed lots, three soaking times in tetrazolium salt (6, 12 and 16 h) and three salt concentrations (0.075, 0.1 and 0.5%), totaling 18 treatments with four replicates of 25 seeds, evaluated at 40 ºC. For each treatment, the seeds were divided into three classes, namely, viable and vigorous embryos (class 1); viable embryos (class 2) and non-viable or dead embryos (class 3). For a comparison with the tetrazolium test results, the two seed lots were analyzed for water content, germination, emergence, first count, germination speed index, emergence speed index, growth and seedling dry weight. The viability and vigor of T. indica seeds can be evaluated after a soaking period of 48 h and immersion for 6h in tetrazolium salt at the concentration of 0.1%, at 40°C, with provides results similar to conventional seed viability tests. The tetrazolium test proved to be adequate to differentiate T. indica seed lots in terms of viability.(AU)

A qualidade das sementes é avaliada rotineiramente por testes diretos como o de germinação e indiretos como o teste de tetrazólio que tem se mostrado promissor na determinação da viabilidade e vigor possibilitando o diagnóstico dos principais problemas que podem afetar a qualidade das sementes, como danos mecânicos, deterioração de campo e armazenamento. Neste sentido, objetivou-se com este trabalho desenvolver um protocolo do teste de tetrazólio para avaliar a viabilidade e o vigor das sementes de Tamarindus indica L. Antes da exposição das sementes a solução de tetrazólio, foram realizados estudos pre-condicionamento das sementes utilizando sete períodos de embebição sendo selecionado o tempo que não causou danos ao embrião da semente e permitiu a remoção do tegumento para exposição das estruturas da semente ao sal de tetrazólio. Em seguida, o experimento foi instalado em delineamento inteiramente casualizado em esquema fatorial 2x3x3, sendo dois lotes de sementes, três tempos de embebição em sal (6; 12 e 16 horas) e três concentrações de sal de tetrazólio (0,075; 0,1 e 0,5%), totalizando dezoito tratamentos com quatro repetições de 25 sementes, avaliados a 40 ºC. Para cada tratamento as sementes foram divididas em três classes: embriões viáveis e vigorosos (classe 1); embriões viáveis (classe 2) e embriões inviáveis ou mortos (classe 3). Para comparar com o resultado do teste de tetrazólio os dois lotes de sementes foram submetidos a análise de teor de água, germinação, emergência, primeira contagem e índice de velocidade de germinação e emergência, crescimento e massa seca de plântulas. A viabilidade e o vigor das sementes de T. indica podem ser avaliados após um período de 48h de embebição e 6h de imersão na concentração de 0,1% de sal de tetrazólio a 40 ºC, com resultados semelhantes aos testes convencionais de determinação da viabilidade das sementes. O teste de tetrazólio mostrou-se adequado para diferenciar a viabilidade em relação aos lotes de sementes de T. indica.(AU)

Seed mass maturity in the terrestrial bromeliad Hechtia perotensis (Bromeliaceae), endemic to Mexico / Madurez de las semillas en la bromelia terrestre Hechtia perotensis (Bromeliaceae), endémica de México

Abstract Introduction: H. perotensis is a plant with a high potential for ecological restoration because it yields thousands of seeds and grows under low levels of rain, poor soils and contrasting temperatures. However, little is known of the seed mass maturity (high seed germination, low seed fresh weight and low seed moisture content) in this species. Objective: Assess seed germination in the laboratory of H. perotensis during seed development and along the floral stalk (infructescence) in two sites one in rocky location and another near a lake. The hypothesis was that there is a time after flowering in which seeds have highest germination and fresh weight and that the apical, centre and base of the infructescence are different in seed germination and fresh weight in both sites. Methods: Capsules were collected in two sites one in rocky land (Frijol Colorado, Perote, Veracruz) and another near one lake (Alchichica, Puebla), in the months of August, September and November 2016 and January 2017. A repeated measure design (RMD) was used to analyze the effects of infructescence section on seed weight, moisture content and seed germination (41, 87, 152 and 215 days after flowering). Each evaluation time comprised five replicates, each one with 15 seeds. Results: Difference in seed germination, seed weight and moisture content between sections of the infructescence was not significant. However, significant differences were found not only between first and last sample dates, but mainly between first and second dates. Eighty-seven days after flower pollination seed moisture content was lower than 20 % and up to 80 % of seed germinated in both sites of sampling. Conclusions: In this study it was found that the moisture content of H. perotensis seed can be used as an indicator of the physiological maturity of the seed and it is also related to germination of the seed.

Resumen Introducción: H. perotensis es una planta con un alto potencial para la restauración ecológica debido a que produce miles de semillas y crece con niveles bajos de lluvia, suelos pobres y temperaturas contrastantes. Sin embargo, el desarrollo de la madurez de las semillas (germinación alta, baja humedad y bajo peso fresco de semilla) en esta especie es muy poco conocido. Objetivo: Evaluar la germinación en el laboratorio de las semillas de H. perotensis, durante el desarrollo de la semilla y a lo largo del tallo floral (infrutescencia) en dos sitios; uno en un área rocosa y otro cerca de un lago. La hipótesis fue que hay un tiempo después de la floración, en el que las semillas presentan la mayor germinación y menor peso fresco, y que las secciones apical, central y base de la infrutescencia, la germinación y el peso fresco de la semilla son diferentes en los dos sitios. Métodos: se recolectaron las cápsulas en dos sitios uno en un área rocosa (Frijol Colorado, Perote, Veracruz) y otro cerca de un lago (Alchichica, Puebla), en los meses de Agosto, Septiembre y Noviembre de 2016 y Enero de 2017. Se usó un diseño de medidas repetidas (DMR) para analizar los efectos de la sección de la infrutescencia sobre el peso fresco de la semilla, el contenido de humedad y la germinación de la semilla (41, 87, 152 y 215 días, después de la floración). Cada tiempo de evaluación comprendió cinco réplicas y 15 semillas. Resultados: La diferencia en la germinación de la semilla, el peso de la semilla y el contenido de humedad entre las secciones de la infrutescencia no fue significativa. Sin embargo, se encontraron diferencias significativas entre las fechas de muestreo inicial y final, pero sobre todo entre la primera y la segunda fecha. Asimismo, ochenta y siete días después de la polinización de las flores, el contenido de humedad de las semillas fue inferior al 20 % y superior al 80 % en la germinación de las semillas en ambos sitios de muestreo. Conclusiones: en este estudio se encontró que el contenido de humedad de la semilla de H. perotensis, puede usarse como un indicador de la madurez fisiológica de la semilla y también está relacionado con la germinación de la semilla.

Las cabras como dispersores de semillas: aportes y limitaciones para la regeneración del bosque tropical estacionalmente seco de Ecuador / Goats as seed dispersers: contributions and limitations for the regeneration of the seasonally dry tropical forest in Ecuador

Resumen Introducción: Los bosques tropicales estacionalmente secos están sujetos a procesos de degradación crónica que ponen en riesgo su diversidad. La ganadería intensiva dentro de los remanentes de bosque se plantea como una de las principales causas de estos procesos de degradación. Sin embargo, el ganado también podría estar cumpliendo el rol de dispersor de semillas para algunas especies. Objetivo: Evaluar el rol de las cabras en la dispersión de semillas de especies leñosas y las posibles consecuencias de su comportamiento alimenticio sobre la estructura de la vegetación. Métodos: Entre diciembre 2016 y junio 2017 se recolectaron fecas de corrales (N = 38) y parcelas de vegetación (N = 42) de tres localidades de bosque seco en el Suroccidente de Ecuador. Todas las semillas encontradas en las fecas (N = 13 326) se registraron e identificaron taxonómicamente. Para evaluar el efecto de la ingestión de semillas sobre la germinación, se estableció un experimento de siembra de semillas extraídas de fecas procedentes de corrales y semillas recolectadas directamente de la planta. Resultados: Las cabras dispersaron 10 especies leñosas, de las cuales al menos el 50 % son leguminosas. Las semillas de Acacia macracantha representaron más del 70 % de semillas presentes en fecas de cabra. El paso de semillas por el tracto digestivo de las cabras mejoró significativamente el porcentaje y velocidad de germinación en Albizia multiflora, Piscidia carthagenensis y Ziziphus thyrsiflora, mientras que en Choroleucon mangense y Prosopis juliflora no se obtuvo germinación. No encontramos una correlación entre la riqueza de árboles establecidos y el número de especies encontradas en las fecas (χ2 = -0.23, P = 0.53). La composición de especies de semillas dispersadas en las fecas no mostró una dependencia de la localidad, a pesar de que la composición de la vegetación establecida cambia entre localidades. La abundancia de semillas en las fecas no mostró relación con la abundancia de árboles en la vegetación establecida. Conclusiones: Las cabras pueden suplir en cierta medida el rol de otros ungulados localmente extintos, mejorando la germinación de especies de leguminosas con testa dura. Sin embargo, su comportamiento alimenticio y la alta dominancia en la dispersión de ciertas especies puede tener importantes efectos en la estructura de la comunidad.

Abstract Introduction: Seasonally dry tropical forests are subject to chronic degradation processes, which has reduced the populations of some important animal dispersers. Intensive livestock farming within the forest remnants is considered one of the main causes of these degradation processes. However, domestic ungulates could also be fulfilling the role of seed dispersal for some wild species. Objective: To evaluate the role of goats as seed dispersers of woody species and the possible consequences of their feeding behavior on the vegetation structure. Methods: Between December 2016 and June 2017, we collected goat feces from pens (N = 38) and vegetation plots (N = 42) from three locations of dry forest in the Southwestern Ecuador. All the seeds found in the feces (N = 13 326) were recorded and taxonomically identified. To evaluate the effect of gut passage on seed germination, we sowed the seeds found in the goat feces from the pens and seeds collected directly from parent plants. Results: Goats dispersed seeds from ten species, of which at least 50 % are legumes. Acacia macracantha seeds represented ca. 70 % of seeds present in goat feces. The passage of seeds through the digestive tract of goats significantly improved the percentage and speed of germination in Albizia multiflora, Piscidia carthagenensis and Ziziphus thyrsiflora, while in Choroleucon mangense and Prosopis juliflora no germination was registered. We did not find a correlation between the richness of established trees and the number of species found in the goat feces (χ2 = -0.23, P = 0.53). The composition of dispersed seed species in the feces did not show a dependency on the locality, although the composition of the established vegetation changes between localities. The frequency of seeds in the feces did not show a relationship with the tree abundance in the established vegetation. Conclusions: Goats can play, at least partially, the role of locally extinct wild ungulates, improving the germination of legume species with hard coat. However, this positive effect can be blurred by their feeding behavior and high preference for particular species, which can modify the dominance of some species, and result in changes in the composition and structure of the vegetation.

Crop management and its effects on weed occurrence

The objective was to evaluate weed phytosociology and similarities between crop management systems in the Chapadões region. The experiment was conducted at in agricultural area located in the municipality of Chapadão do Sul, MS, during the 2016/17 harvest. Three crop managements strategies were used: (1) cotton/soybean/Urochloa, (2) millet/soybean/millet and (3) millet/soybean/crotalaria. A phytosociological survey of weeds was carried out during soybean cultivation and cover crops growth, in succession. The evaluation area for each management strategy was 0.5 ha. Soybean surveys were carried out in October and January, while the cover crop surveys were performed in February and May. The relative frequency (RF), relative density (RD), relative abundance (AR), and relative importance (RI) of weeds, Venn diagram, and Jaccard and Sorenson similarity indices were evaluated. The management area represented by the cotton/soybean/Urochloa rotation had fewer weed species than others. The species Cenchrus echinatus, Digitaria insularis, Digitaria sanguinalis, Eleusine indica and Commelina benghalensis had the highest phytosociological indeces among the monocotyledons. Attention is required for managing the dicotyledons Amaranthus deflexus, Conyza canadensis and Senna obtusifolia despite their low indices because of herbicide resistant cases. The highest indeces of similarity were found between managements areas 2 and 3, which did not rely on cotton cultivation prior to soybeans.

Storage of corn seeds infested by weevil

Weevil is one of the main pests of stored corn. The adults feed on intact and broken seeds or bran, while the larvae develop by feeding on the inside of the seeds, resulting in a reduction in weight and physiological quality. The objective of this study was to evaluate the effects of Sitophilus zeamais infestation on corn seeds during storage. Corn seeds were infested using insect densities: 0, 36, 72, 108, and 144 individuals per 240 g of seeds. The infested seeds were placed in individualized plastic containers and covered with voile fabric. The seeds were stored for 60 days under laboratory conditions with an average temperature of around 27 °C and relative humidity of 85%. Initially and every 15 days of storage for 60 days, the seeds were evaluated using the parameters: number of S. zeamais individuals, total number of infested seeds, seed weight loss, germination test and seedling emergence in the tray. The population increase of S. zeamais causes a decrease on the physical and physiological quality of stored corn seeds, but the deterioration rate does not increase in the same proportion as the weevil population growth rate.