Mario Onofre Cortez-Rocha

Anthracnose is a disease caused by phytopathogenic fungi such as Colletotrichum siamense that attacks plants and fruits causing great postharvest losses. Different alternatives for the control of this fungus have been studied. In the present study, we evaluated the in vitro antifungal activity of the methanolic extracts of Baccharis glutinosa (ExB) and Jacquinia macrocarpa (ExJ) individually, as well as in combination with chitosan (CS), along with their toxicity in different models. Using the radial growth technique, it was observed that the mycelial development of C. siamense was altered and reduced during exposure to the different treatments evaluated during the first hours of incubation, indicating a fungistatic effect. While the cell viability, by colorimetric assay using the XTT salt, showed alteration since the chitosan reduced proliferation by 50%, while the plant extracts and their mixtures with chitosan reduced approximately 40% indicating cell damage, which was confirmed by fluorescence microscopy. In addition, toxicity tests demonstrated that the J. macrocarpa extract significantly affected the germination percentage of Lactuca sativa seeds, whereas radicle length was reduced in all treatments except for chitosan. The larval survival test for Artemia salina with the extracts indicated their potential toxicity by causing up to 60% mortality. The results indicate that ExB and ExJ mixed with CS are a good option for controlling C. siamense; however, at the concentrations used, they exhibit a toxic effect on the evaluated models.

Anthracnose is a disease caused by phytopathogenic fungi such as Colletotrichum siamense that attacks plants and fruits causing great postharvest losses. Different alternatives for the control of this fungus have been studied, and in the present study we evaluated the in vitro antifungal activity of chitosan mixed with methanolic extracts of Baccharis glutinosa and Jacquinia macrocarpa as well as toxicity in different models. Using the radial growth technique, it was observed that the mycelial development of C. siamense was altered and reduced during exposure to the different treatments evaluated during the first hours of incubation, indicating a fungistatic effect. While the cell viability, by colorimetric assay using the XTT salt, showed alteration since the chitosan reduced proliferation by 50 %, while the plant extracts and their mixtures with chitosan reduced approximately 40 % indicating cell damage, which was confirmed by fluorescence microscopy. In addition, toxicity tests showed that the extract of J. macrocapra affected the germination percentage of Lactuca sativa seeds while radicle length was reduced in all treatments, except with chitosan alone. The larval survival test for Artemia salina with the extracts indicated their potential toxicity by causing up to 60% mortality. The results indicate that the extracts of B. glutinosa and J. macrocarpa mixed with chitosan are a good option for the control of C. siamense, but at the concentrations used they show a toxic effect in the models evaluated.

El género Enterococcus puede ser encontrado en agua, suelo y en general en todo ambiente, son parte de la microbiota intestinal de animales, incluyendo a humanos. La prevalencia en alimentos se debe principalmente a la resistencia que presentan a las condiciones adversas del ambiente. Setenta y tres aislados de vísceras de pollo fueron confirmados como pertenecientes al género a través de pruebas bioquímicas y fermentación de carbohidratos. La caracterización bioquímica indicó que la especie predominante fue E. casseliflavus. La resistencia de los aislados a antimicrobianos fue notoria para amikacina (42 %), kanamicina (38 %), estreptomicina (55 %), and eritromicina (33 %). El análisis obtenido por ERIC-PCR mostró una alta variabilidad genética entre los aislados. Así también, los aislados mostraron relación entre los perfiles obtenidos con la susceptibilidad a antimicrobianos y por ERIC-PCR. Estos resultados muestran la multi-resistencia de Enterococcus en vísceras de pollo expedidos comercialmente, con una alta variabilidad genética. Además, estos Enterococcus pueden ser potencialmente patógenos en individuos como una bacteria nosocomial y además transferir la resistencia en especies que son estrictamente patógenas y causan enfermedades al humano.

The fungus Aspergillus parasiticus is a contaminant in agricultural crops and its eradication involves the indiscriminate use of harmful synthetic pesticides. In the search for antifungal agents of natural origin, chitosan (Q) and capsaicin (C) are coupled in the form of nanoparticles (Np), which can possess a direct application under specific conditions. Due to their small size, Np can cross through the cell wall, taking the cells into a pro-oxidant environment known as “oxidative stress”, which presents when the reactive oxygen species (ROS) surpass the number of antioxidants in the cell. In the present investigation, nanoparticles of chitosan (Np Q) and nanoparticles of chitosan-capsaicin (Np QC) with an average diameter of 44.8 ± 20.6 nm and 111.1 ± 14.1 nm, respectively, were synthesized, and there was a zeta potential of + 25.6 ± 0.7 mV and + 26.8 ± 6.1 mV, respectively. The effect of the concentration of Np Q (A, B, C, and D), of Np QC (A, B, C, and D), and capsaicin in a solution (control) was evaluated on the viability of the spores, the accumulation of intracellular ROS, and the morphometric changes of A. parasiticus. Acute toxicity of the Np was determined utilizing bioassays with Artemia salina, and acute phytotoxicity was evaluated in lettuce seeds (Lactuca sativa). According to ROS results, capsaicin (control) did not induce oxidative stress in the cell; otherwise, it was observed to have an elevated (p < 0.05) accumulation of ROS when the concentration of Np Q increased. For both, Np Q and Np QC, an inverse physiological pattern relating spore viability and ROS accumulation in the fungus was found; the viability of spores decreased as the ROS accumulation increased. The spore viability of A. parasiticus diminished upon increasing the concentration of chitosan (0.3–0.4 mg/mL) in the Np, while the intracellular accumulation of ROS increased proportionally to the concentration of the nanomaterials in the treatments of Np Q and Np QC. On the other hand, Np QC presented a lower (p < 0.05) toxicological effect in comparison with Np Q, which indicates that the incorporation of bioactive compounds, such as capsaicin, into nanoparticles of chitosan is a strategy that permits the reduction of the toxicity associated with nanostructured materials.

Sorghum’s biological potential can be exploited to formulate healthy foods. The objective of this work was to optimize the germination process of sorghum to maximize the content of free phenolic compounds and antioxidant activity. According to the response surface methodology, the maximum content of total phenols and ferulic acid was obtained at 35°C and 28 h of germination, while the optimal antioxidant activity was obtained at 33.6°C and 28 h and 27.4°C 28 h measured as DPPH and TEAC respectively. The porridges obtained from germinated sorghum flours showed a higher content of total phenols and antioxidant activity compared to non-germinated ones. Porridges formulated with germinated sorghum flour showed an increase of 12% as compared with porridge formulated with flour non-germinated sorghum. Ferulic acid showed a significant reduction in both evaluated porridges.

In this study, cinnamon (Cinnamomum zeylanicum), thyme (Thymus vulgaris) and Schinus molle essential oils (at 125, 250 and 500 µg/mL) were encapsulated in chitosan nanoparticles by ionotropic gelation. Their antimicrobial activity against Staphylococcus aureus ATCC 25923, Enterococcus sp., Escherichia coli ATCC 25922, Klebsiella pneumoniae (BLEE +) and Pseudomona aeruginosa carbapenemase producer was evaluated. The obtained nanoparticles exhibited a regular distribution and spherical shape with size range of 19.7 to 361 nm as observed by using a laser light scattering equipment. It was found that the studied chitosan nanoparticles with essential oils showed high antimicrobial effect than the chitosan and essential oils alone. Enterococcus sp. was the most sensitive bacteria and S. aureus the most resistant. Chitosan nanoparticles and chitosan nanoparticles with cinnamon and thyme essential oils were capable to inhibit growth of Enterococcus sp. and K. pneumoniae. This study demonstrated that chitosan nanoparticles with essential oils were effective against some foodborne pathogens.

This study aimed to determine the ability of Fusarium verticillioides in developing mechanisms to counteract the antifungal effect of a fraction from Jacquinia macrocarpa plant extract (JmAF), as well as the morphological and physiological changes that occur during its exposure. The fungus was exposed to JmAF during consecutive periods. A culture sample was taken weekly to determine radial growth, spore germination and size, and fungal β-1,3-glucanase activity. The results showed that, in the beginning, the radial growth decreased by 85.8%, and spore germination was delayed. As the exposure continued, the fungus showed a recovery, to some extent, in its original characteristics. However, the radial growth of the fungus continued to be inhibited (42.9%) throughout the experiment (7 weeks). The β-1,3-glucanase activity also was inhibited by 36.4% during the first week of exposure to JmAF. However, the activity was recovered after 7 weeks of exposure.

Heavy metals are toxic especially when they are introduced into the environment due to anthropogenic activities such as metallurgy, mining, and tanning. Removing these pollutants has become a worldwide concern since they cannot be degraded into nontoxic forms causing extended effects in the ecosystems. The use of an Aspergillus australensis was evaluated in order to remove Cu²⁺ from simulated wastewater. The fungus was isolated from river sludges contaminated with heavy metals and was first evaluated for the determination of Cu²⁺ tolerance levels. Microscopic fluorescence analysis was carried out to determine the effect of Cu²⁺ presence on the viability, cellular components, polyhydroxyalkanoates production, and oxidative stress of the fungus, as a response to the stress caused by exposure to metal. In order to achieve copper removal, the A. australensis biomass was produced using batch cultures, and the mycelium was immobilized on a textile media in order to compare the copper-removal efficiency of live or dead biomass. The optimal values of pH and temperature for biomass production were established by using a surface response analysis. Live immobilized biomass was capable of removing Cu²⁺ from 1.54 ± 0.19 to 2.66 ± 0.26 mg of copper/ g of dry biomass, while values of 1.93 ± 0.03 to 2.36 ± 0.29 mg of copper/g of dry biomass were observed when dead biomass was used. As was expected, copper removal using biomass varied depending on the pH and temperature used.

Aspergillus and Fusarium are the fungi genera most frequently isolated from cereal grains and other commodities. They are capable of producing mycotoxins, which can affect the human and animal health. Synthetic fungicides have been used to control these fungi, nevertheless, they have acquired resistance and other alternatives are necessary since they now need higher amounts. Therefore, the aim of this study was to evaluate the efficacy of cinnamon, clove and thyme essential oils (EOs) alone and encapsulated in chitosan on the radial growth, spore germination and mycotoxin production by Fusarium verticillioides and Aspergillus parasiticus. The composition of the EOs was determined by gas chromatography mass spectrometry (GC-MS). EOs inhibited radial growth and spore germination of both fungal species better than when they were encapsulated, plus, they reduced mycotoxin production. The major components were eugenol in cinnamon and clove EO (70 and 63%, respectively) and 2-methyl-5-(1-methyethyl)-phenol (46.2%) in thyme EO. The microparticles with clove and thyme EO showed good surface charges, higher than +30 mV and their average size for the three types of microparticles was about 750 nm. Our findings suggest that EOs both alone and encapsulated in chitosan have a fungistatic effect on Fusarium verticillioides and Aspergillus parasiticus.

The indiscriminate use of synthetic fungicides has caused several problems to the environment, which place human and animal health at risk. Due to this fact, the search for natural alternatives to control phytopathogenic fungi growth has increased. This study aims to evaluate the antifungal activity of two essential oils (EOs) and three major components of EOs on the radial growth and spore germination of Fusarium verticillioides and Alternaria tenuissima. Minimum and half‐maximal inhibitory concentrations (CMI and CI₅₀) at 96 h for each treatment were calculated. Lemongrass EO and citral caused the highest inhibition for A. tenuissima (CMI of 1000 μl/l and CI₅₀ of 10 μl/l). For F. verticillioides, the most effective component was geraniol (CMI and CI₅₀ of 1000 and 250 μl/l, respectively). Spore germination rate was delayed by the EOs and major components tested. The use of EOs was effective to control these two fungal species in their different grow stages.