Antifungal activity of phytotherapeutic preparation of Baccharis species from argentine Puna against clinically relevant fungi

Ethnopharmacological relevance: B. boliviensis and B. tola are used in traditional medicine in the Argentine Puna to treat skin and soft tissue infections and inflammatory processes in humans and animals. Aim of the study: To assess the potential of phytotherapeutic preparations of Baccharis species as antifungal agents against clinically relevant fungi and to determine the chemical composition of the extracts. Material and methods: Phytotherapeutic preparations of B. boliviensis and B. tola collected in Argentine Puna were evaluated as an antifungal agent against clinically relevant fungi (yeast, non-dermatophytes, and dermatophytes) isolated of patients from a local Hospital, and reference strains, using macrodilution and microdilution assays. The bioactivity was supported by UHPLC-OT-MS metabolome fingerprinting. Results: The results revealed that the plant preparations were active against most of evaluated fungal strains; B. boliviensis was more active than B. tola. Dermatophyte fungi strains were the most sensitive isolates. The phytotherapeutic preparation showed Minimal Inhibitory Concentration (MIC) values between 25 and 400 μg GAE/mL and Minimum Fungicidal Concentration (MFC) values between 50 and 400 μg GAE/mL. Regarding the phytochemical analysis, total phenolic and total flavonoid contents of hydroalcoholic preparation of B. boliviensis were greater than those of the B. tola extract. Both Baccharis species showed similar chromatographic patterns, fifty-two compounds were identified based on UHPLC-OT-MS including several terpenoids, flavonoids and phenolic acids that have been identified in this two endemic South American Baccharis species for the first time. Several identified compounds present antifungal properties, the presence of these compounds support the bioactivity of the Baccharis extracts. Conclusions: In this work the traditional use of both Baccharis species as an antimicrobial against commercial products resistant fungal strains was validate, principally against dermatophytes fungi such as T. rubrum, T. mentagrophytes, M. canis, and M. gypseum. These results indicate that the hydroalcoholic preparations could be used for the treatment of fungal infectious.Fil: Carrizo, Silvana Lorena. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; ArgentinaFil: Zampini, Iris Catiana. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Sayago, Jorge Esteban. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Simirgiotis, Mario Juan. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Austral de Chile; ChileFil: Bórquez, Jorge. Universidad de Antofagasta. Facultad de Ciencias Básica. Departamento de Química. Laboratorio de Productos Naturales; ChileFil: Cuello, Ana Soledad. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Isla, Maria Ines. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentin

Aplicación de bacterias lácticas en el desarrollo de alimentos novedosos a base de granos andinos

Actualmente existe una revalorización de los granos andinos (quínoa y amaranto) por sus propiedades nutricionales, funcionales y libres de gluten. Las bacterias lácticas (BL) son ampliamente utilizadas como fermentos en una gran variedad de sustratos. Ciertas BL tienen capacidad de sintetizar vitaminas del grupo B, incrementando el valor nutricional del alimento. La quínoa y el amaranto contienen fitato, compuesto antinutricional, que forma complejos insolubles con oligoelementos afectando su biodisponibilidad. Existen antecedentes de que ciertas BL tienen actividad fitasa capaz de degradar fitato.En base a estos antecedentes, el Objetivo General de esta Tesis fue determinar la biodiversidad de BL de quínoa y amaranto del NOA y diseñar alimentos con mayor valor nutricional y funcional empleando BL autóctonas. La microbiota láctica evaluada en granos y masas ácidas de quínoa y amaranto permitió identificar 73 cepas pertenecientes a diferentes géneros y especies: Lactobacillus (L.) plantarum, L. rhamnosus, L. sakei, Pediococcus (P.) pentosaceus, Enterococcus (E.) casseliflavus, E. mundtii, E. hermaniensis, E. hirae, E. gallinarum, Enterococcus sp., Leuconostoc (Lc.) mesenteroides, E. durans. De acuerdo a la producción de riboflavina (B2) y folatos (B9), capacidad de hidrolizar fitato, degradar almidón, acidificación y producción de exopolisacaridos, se seleccionaron 11 cepas aisladas de quínoa y 7 de amaranto para estudios in situ (masa ácida de quínoa). A partir de estos resultados, se diseñaron cultivos mixtos que fueron evaluados determinándose las combinaciones adecuadas que produzcan mayor concentración de vitaminas y menor concentración de fitato residual. Los resultados llevaron a seleccionar el cultivo mixto (C.M) formado por L. plantarum CRL 2107 + L. plantarum CRL 1964 (1:1). La masa fermentada por CM presentó la mayor concentración de B2 y B9 (5,1 ± 0,4 y 1,6 ± 0,2 µg/g masa, respectivamente) y menor concentración de fitato residual (17,4 ± 1,3 mg/g masa). A fin de evaluar la eficiencia biológica de B9 y B2 producidas por CM en prevenir la deficiencia nutricional en ambas vitaminas, se realizaron ensayos in vivo. Los resultados revelaron que los animales alimentados con la pasta fermentada con el C.M (ACM) tuvieron mayor peso, mayor concentración de B9 en sangre y en órganos y mayor concentración de minerales, comparados con el grupo deficiente (AGD) y/o con el grupo control (AGC). La evaluación histológica del intestino evidenció que los AGD presentaban atrofia de las vellosidades intestinales, mientras que los ACM mostraron una tendencia a la normalización de la forma y del tamaño de las vellosidades. Respecto al estado de B2, los AGD mostraron un valor de EGRAC mayor a 1,3, indicando una deficiencia en B2, mientras que los ACM mostraron valores de EGRAC menores a 1,3, demostrando que la pasta de quínoa fermentada con BL fue capaz de prevenir la deficiencia de B2. De esta manera, se concluye que este trabajo de Tesis puso en evidencia el valor funcional de BL autóctonas y plantea nuevas perspectivas de aplicación de BL con capacidad de degradar fitatos y de producir vitaminas B2 y B9 para el diseño de alimentos bio-enriquecidos con funcionalidad comprobada.There is currently a revaluation of Andean grains (quinoa and amaranth) because of their nutritional, functional and gluten-free properties. Lactic acid bacteria (LAB) are widely used as ferments on a wide variety of substrates. Some LAB have the ability to synthesize B group vitamins, increasing the nutritional value of foods. Quinoa and amaranth contain phytate, an antinutritional compound, which forms insoluble complexes with trace elements affecting their bioavailability. There is evidence that certain LAB have phytase activity capable of degrading phytate. Based on this background, the General Objective of this thesis was to determine the biodiversity of LAB from NOA quinoa and amaranth and to design foods with greater nutritional and functional value using autochthonous LAB. The lactic microbiota was evaluated in grains and quinoa and amaranth sourdoughs: 73 strains were identified belonging to different genera and species: Lactobacillus (L.) plantarum, L. rhamnosus, L. sakei, Pediococcus (P.) pentosaceus, Enterococcus (E.) casseliflavus, E. mundtii, E. hermaniensis, E. hirae, E. gallinarum, Enterococcus sp., Leuconostoc (Lc.) mesenteroides, E. durans. According to the production of riboflavin (B2) and folates (B9), ability to hydrolyze phytate, degrade starch, acidification and production of exopolysaccharides, 11 strains from quinoa and 7 from amaranth were selected for in situ studies (sourdough of quinoa). From these results, mixed cultures were designed using suitable combinations that produce higher concentration of vitamins and decrease the concentration of residual phytate. The results showed that the best mixed cultures (MC) were L. plantarum CRL 2107 + L. plantarum CRL 1964 (1: 1). The dough fermented by MC presented the highest concentration of B2 and B9 (5,1 ± 0,4 y 1,6 ± 0,2 µg / g mass, respectively) and lower concentration of residual phytate (17,4 ± 1,3 mg / g mass). In order to evaluate the biological efficiency of B9 and B2 produced by MC in preventing nutritional deficiency in both vitamins, in vivo trials were performed. The results showed that the animals fed with the MC-fermented paste (PG) had higher live weights, higher concentration of B9 in blood and in organs and higher concentrations of minerals compared to the deficient group (DG) and / or group control (CG). The histological evaluation of the intestine showed that DG presented atrophy of the intestinal villi, whereas the PG group showed normal histology of the intestinal villi. Regarding the state of B2, the DG showed a value of EGRAC greater than 1.3, indicating a deficiency in B2, whereas the PG showed values of EGRAC lower than 1.3, demonstrating that the quinoa paste fermented with LAB was able to prevent B2 deficiency. Thus, it can be concluded that this thesis work showed the functional value of autochthonous LAB and poses new perspectives of LAB application with ability to degrade phytates and to produce vitamins B2 and B9 for the design of bio-enriched foods with proven functionality.Fil: Carrizo, Silvana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Quinoa pasta fermented with lactic acid bacteria prevents nutritional deficiencies in mice

In recent years, quinoa (Chenopodium quinoa Willd), an ancestral crop of the Andean region of South America, has gained worldwide attention due to its high nutritional value. This grain is a good source of several vitamins and minerals; however, their bioavailability is decreased by the presence of antinutritional factors such as phytic acid. These compounds can be reduced using lactic acid bacteria (LAB), that have a GRAS (Generally Recognized as Safe) status and have traditionally been associated with food fermentation due to their biosynthetic capacity and metabolic versatility. The objective of this study was to evaluate the effectiveness of a pasta made with quinoa sourdough fermented by L. plantarum strains producing vitamins B2 and B9 and phytase to prevent vitamins and minerals deficiency using an in vivo mouse model. The results showed that the pasta fermented with the mixed culture containing L. plantarum CRL 2107 + L. plantarum CRL 1964 present increased B2 and B9 levels in mice blood. Likewise, higher concentrations of P, Ca+2, Fe+2, Mg+2 (18.75, 10.70, 0.37, 4.85 mg/dL, respectively) were determined with respect to the deficient group (DG) (9.85, 9.90, 0.26, 3.34 mg/dL, respectively). Hematological studies showed an increase in hemoglobin (14.4 ± 0.6 g/dL), and hematocrit (Htc, 47.0 ± 0.6%) values, compared to the DG (Hb: 12.6 ± 0.5 g/dL, Hto: 39.9 ± 1.1%). Furthermore, histological evaluations of the intestines showed an increase of the small intestine villi length in this latter group. The results allow us to conclude that bio-enrichment of quinoa pasta using LAB could be a novel strategy to increase vitamin and minerals bioavailability in cereal/pseudocereal – derived foods.Fil: Carrizo, Silvana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: de Moreno, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Rollan, Graciela Celestina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Lactic Acid Bacteria from Andean Grain Amaranth: A Source of Vitamins and Functional Value Enzymes

Amaranth is a rediscovered pseudocereal with high nutritional properties. Lactic acid fermentation can increase the functional and nutritional value of foods. The aims of this study were to isolate and evaluate the functionality of lactic acid bacteria (LAB) from amaranth. LAB strains (n = 29) isolated from amaranth sourdough and grains included Lactobacillus (L.) plantarum (n = 8), L. rhamnosus (n = 6), Enterococcus (E.) mundtii (n = 4), E. hermanniensis (n = 3), E. durans (n = 1), Enterococcus sp. (n = 1), Leuconostoc (Lc.) mesenteroides (n = 3), and Lc. mesenteroides subsp. mesenteroides (n = 3). Only 21% of the strains showed the ability to synthesize capsular exopolysaccharides or display ropiness and only 8 strains showed amylolytic activity. L. plantarum CRL 2106 and E. durans CRL 2122 showed the highest phytase activity, which is of importance for mineral bioavailability. L. plantarum CRL 2106 and CRL 2107 and Lc. mesenteroides subsp. mesenteroides CRL 2131 synthesized the highest concentrations of B2 and B9 vitamin (140-250 ng/mL). This study demonstrates the potential of LAB to improve the nutritional and functional values of pseudocereal-derived foods.Fil: Carrizo, Silvana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Montes de Oca, Cecilia Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Hebert, Elvira Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Saavedra, Maria Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Rollan, Graciela Celestina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad del Norte Santo Tomás de Aquino; Argentin

Ancestral Andean grain quinoa as source of lactic acid bacteria capable to degrade phytate and produce B-group vitamins

The lactic acid bacteria (LAB) microbiota of quinoa grains (QG) and spontaneous sourdough (QSS) was evaluated. Different strains of Lactobacillus (L.) plantarum (7), L. rhamnosus (5), L. sakei (1), Pediococcus (Ped.) pentosaceus (9), Leuconostoc (Leuc.) mesenteroides (1), Enterococcus (E.) casseliflavus (2), E. mundtii (3), E. hirae (1), E. gallinarum (12), Enterococcus sp. (1), and E. hermanniensis (2) were isolated, identified and characterized. Only four strains isolated from QSS and eight strains isolated from QG showed amylolytic activity. L. plantarum CRL 1973 and CRL 1970, L. rhamnosus CRL 1972 and L. sakei CRL 1978 produced elevated concentrations of folate with strain CRL 1973 producing the highest concentration (143 ± 6 ng/ml). L. rhamnosus, isolated from QSS, was the LAB species that produced the most elevated concentrations of total riboflavin (> 270 ng/ml) with strain CRL 1963 producing the highest amounts (360 ± 10 ng/ml). Phytase activity, evaluated in forty-four LAB strains from quinoa, was predominantly detected in L. rhamnosus and Enterococci strains with the highest activities observed in E. mundtii CRL 2007 (957 ± 25 U/ml) followed by E. casseliflavus CRL 1988 (684 ± 38 U/ml), Leuc. mesenteroides CRL 2012 (617 ± 38 U/ml) and L. rhamnosus CRL 1983 (606 ± 79 U/ml). In conclusion, this study shows that a diverse LAB microbiota is present in quinoa with important properties; these microorganisms could be used as potential starter cultures to increase the nutritional and functional properties of Andean grains based foods.Fil: Carrizo, Silvana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Montes de Oca, Cecilia Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Laiño, Jonathan Emiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Suárez, Nadia Elina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Rollan, Graciela Celestina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad del Norte Santo Tomás de Aquino; Argentin