Advances in understanding the enzymatic potential and production of ochratoxin A of filamentous fungi isolated from cocoa fermented beans.

In this work, we determined the degree of filamentous fungi contamination in cocoa fermented seeds and the fungal potential to produce enzymes that could contribute to the cocoa quality. Internal transcribed spacer amplicon sequencing (ITS) showed 11 different species with Byssochlamys spectabilis, Aspergillus niger, Aspergillus flavus, Aspergillus carbonarius, Circinella muscae and Penicillium citrinum as the most abundant species. Our results evidenced intra and inter-species differences in the enzymes production. Cellulases, ß-glucosidase, ornithine decarboxylase and phenylalanine decarboxylase were the most diffused enzymes expressed in the 53 strains here studied. Moreover, A. niger (6/12), A. carbonarius (2/3), and P. citrinum (3/3) showed high pectinolytic activity. Remarkable was the amino decarboxylase activity of P. citrinum, and A. flavus strains. For the first time we reported the presence of B. spectabilis in cocoa fermented beans, which could play an important role in the biogenic amines formation. In addition, we explored the capability of the Aspergillus section Nigri strains, to produce ochratoxin A (OTA) in a cacao model system (CPMS) and in malt extract medium (MEL). We observed that CPMS, but not MEL, stimulated the OTA production in 6 out 15 strains of Aspergillus section Nigri, reaching values ranging between 1.70 and 4995 µg OTA kg-1 dry mycelium.

Passion fruit by-products (Passiflora edulis f. flavicarpa) as a potential fat replacer in ice cream.

In recent years, people have become interested in consuming low-fat foods as this reduces the risk of obesity and cardiovascular disease. For this reason, this study optimized the incorporation of passion fruit epicarp (PFE) as a partial fat substitute in dairy ice cream. For this purpose, a central composite rotational design 22 was performed and 5 response variables were optimized, of which 4 variables were maximized (percentage overrun, hardness, adhesiveness, and color coordinates L*, a*, b*), while the melting rate was minimized as a function of two factors (% PFE and % fat). The stability of three types of ice cream was then evaluated: experimental ice cream (3% milk fat and 0.97% PFE), commercial ice cream (4% milk fat and 2% vegetable fat) and control ice cream (3% milk fat without PFE added), according to their physicochemical and sensory properties during 57 days of storage (-22 °C, 85% relative humidity). The results showed that it was possible to reduce the milk fat by up to 25% by adding PFE (0.97%), which significantly reduced the melting rate and resulted in higher hardness (N), adhesiveness and increased yellowness compared to the commercial samples. The optical microscopy analysis also showed that PFE keeps the air bubbles immobile due to the increased viscosity in the serum phase during the 57-day storage.

Cocoa Shell Infusion: A Promising Application for Added-Value Beverages Based on Cocoa's Production Coproducts.

The cocoa shell (CS) is being incorporated into different food products due to its recognized content of bioactive compounds. In the case of cocoa shell infusions (CSI), the bioactive compounds that manage to be transferred to the infusion have yet to be clearly known, i.e., what is really available to the consumer. In this study, CS was obtained from toasted Colombian Criollo cocoa beans. Three particle sizes (A: >710 µm; B: >425 and <710 µm; C: <425 µm) were evaluated in the CSI, which was traditionally prepared by adding CS to hot water (1%). The decrease in particle size increased the antioxidant capacity (DPPH and ABTS) and the total phenolic compounds. A significant effect (p < 0.05) both of the particle size and of the temperature of tasting was found on some sensory attributes: greater bitterness, acidity, and astringency were due to the greater presence of epicatechin, melanoidins, and proanthocyanidins in the smaller particle sizes. The analysis of the volatile organic compounds showed that the CSI aroma was characterized by the presence of nonanal, 2-nonanone, tetramethylpyrazine, α-limonene, and linalool, which present few variations among the particle sizes. Moreover, analysis of biogenic amines, ochratoxin A, and microbial load showed that CSI is not a risk to public health. Reducing particle size becomes an important step to valorize the functional properties of CS and increase the quality of CSI.

Effect of membrane depolarization against Aspergillus niger GM31 resistant by ultra nanoclusters characterized by Ag2+ and Ag3+ oxidation state.

To date, the impossibility of treating resistant forms of bacteria and fungi (AMR) with traditional drugs is a cause for global alarm. We have made the green synthesis of Argirium silver ultra nanoclusters (Argirium-SUNCs) very effective against resistant bacteria (< 1 ppm) and mature biofilm (0.6 ppm). In vitro and preclinical tests indicate that SUNCs are approximately 10 times less toxic in human cells than bacteria. Unique chemical-physical characteristics such as particle size < 2 nm, a core composed of Ag0, and a shell of Ag +, Ag2+ , Ag3+ never observed before in stable form in ultra pure water, explain their remarkable redox properties Otto Cars (Lancet Glob. Health 9:6, 2021). Here we show that Argirium-SUNCs have strong antimicrobial properties also against resistant Aspergillus niger GM31 mycelia and spore inactivation (0.6 ppm). The membrane depolarization is a primary target leading to cell death as already observed in bacteria. Being effective against both bacteria and fungi Argirium-SUNCs represent a completely different tool for the treatment of infectious diseases.

Influence of Atmospheric Cold Plasma Exposure on Naturally Present Fungal Spores and Physicochemical Characteristics of Sundried Tomatoes (Solanum lycopersicum L.).

This research aimed to evaluate the impact of atmospheric cold plasma (ACP) treatment on the fungal spores naturally present in sundried tomatoes, as well as their influence on the physico-chemical properties and antioxidant activity. ACP was performed with a Surface Dielectric Barrier Discharge (SDBD), applying 6 kV at 23 kHz and exposure times up to 30 min. The results showed a significant reduction of mesophilic aerobic bacteria population and of filamentous fungi after the longer ACP exposure. In particular, the effect of the treatment was assessed on Aspergillus rugulovalvus (as sensible strain) and Aspergillus niger (as resistant strain). The germination of the spores was observed to be reliant on the species, with nearly 88% and 32% of non-germinated spores for A. rugulovalvus and A. niger, respectively. Fluorescence probes revealed that ACP affects spore viability promoting strong damage to the wall and cellular membrane. For the first time, the sporicidal effect of ACP against A. rugulovalvus is reported. Physicochemical parameters of sundried tomatoes such as pH and water activity (aw) were not affected by the ACP treatment; on the contrary, the antioxidant activity was not affected while the lycopene content was significantly increased with the increase in ACP exposure time (p ≤ 0.05) probably due to increased extractability.

The Role of Fungi in the Cocoa Production Chain and the Challenge of Climate Change.

BACKGROUND: The role of fungi in cocoa crops is mainly associated with plant diseases and contamination of harvest with unwanted metabolites such as mycotoxins that can reach the final consumer. However, in recent years there has been interest in discovering other existing interactions in the environment that may be beneficial, such as antagonism, commensalism, and the production of specific enzymes, among others. Scope and approach: This review summarizes the different fungi species involved in cocoa production and the cocoa supply chain. In particular, it examines the presence of fungal species during cultivation, harvest, fermentation, drying, and storage, emphasizing the factors that possibly influence their prevalence in the different stages of production and the health risks associated with the production of mycotoxins in the light of recent literature. Key findings and conclusion: Fungi associated with the cocoa production chain have many different roles. They have evolved in a varied range of ecosystems in close association with plants and various habitats, affecting nearly all the cocoa chain steps. Reports of the isolation of 60 genera of fungi were found, of which only 19 were involved in several stages. Although endophytic fungi can help control some diseases caused by pathogenic fungi, climate change, with increased rain and temperatures, together with intensified exchanges, can favour most of these fungal infections, and the presence of highly aggressive new fungal genotypes increasing the concern of mycotoxin production. For this reason, mitigation strategies need to be determined to prevent the spread of disease-causing fungi and preserve beneficial ones.

The membrane depolarization and increase intracellular calcium level produced by silver nanoclusters are responsible for bacterial death.

This work highlights how our silver ultra nanoclusters (ARGIRIUM-SUNc) hand-made synthesized, are very useful as a bactericide and anti-biofilm agent. The Argirium-SUNc effective antibacterial concentrations are very low (< 1 ppm) as compared to the corresponding values reported in the literature. Different bacterial defense mechanisms are observed dependent on ARGIRIUM-SUNc concentrations. Biochemical investigations (volatilome) have been performed to understand the pathways involved in cell death. By using fluorescence techniques and cell viability measurements we show, for the first time, that membrane depolarization and calcium intracellular level are both primary events in bacteria death. The ARGIRIUM-SUNc determined eradication of different biofilm at a concentration as low as 0.6 ppm. This suggests that the effect of the nanoparticles follows a common mechanism in different bacteria. It is highly probable that the chemical constitution of the crosslinks could be a key target in the disrupting mechanism of our nanoparticles. Since the biofilms and their constituents are essential for bacterial survival in contact with humans, the silver nanoparticles represent a logical target for new antibacterial treatments.

Cacao Pod Husk Flour as an Ingredient for Reformulating Frankfurters: Effects on Quality Properties.

The cocoa pod husk is considered a source of dietary fiber with a high content of water-soluble pectins, bioactive compounds which should be viewed as a by-product with the potential to be incorporated into food. This study aimed to investigate the effect of adding different cocoa pod husk flour (CPHF) levels as a starch replacement for reformulating frankfurters. Results showed that the addition of 1.5 and 3.0% pod husk proportionally increased the frankfurter's fiber content by 0.49 ± 0.08 and 0.96 ± 0.19 g/100 g, which is acceptable for a product that does not contain fiber. Textural properties and sensory characteristics were affected when substituting the starch with CPHF, either totally or partially, although these samples had higher water content, hardness, and adhesiveness while springiness decreased. Non-adverse effects of nitrite on polyphenolic compounds content were evidenced in samples enriched with CPHF. The incorporation of CPHF did not significantly affect the color parameters (ΔE < 3). Finally, the panelists indicated a sensation of the unsalted sausage, suggesting that CPHF may have natural mucoadhesion properties. In conclusion, in formulated meat products such as sausages, plant co-products such as cacao pod husks could be a valid new ingredient to improve technological parameters, functional characteristics, and stability.

Exploring the Capability of Yeasts Isolated from Colombian Fermented Cocoa Beans to Form and Degrade Biogenic Amines in a Lab-Scale Model System for Cocoa Fermentation.

Yeast starters for cocoa fermentation are usually tested according to their enzymatic activities in terms of mucilage degradation and flavor improvement, disregarding their influence on the production or elimination of toxic compounds as biogenic amines (BAs), important for human health. In this work, we tested 145 strains belonging to 12 different yeast species and isolated from the Colombian fermented cocoa beans (CB) for their capability of producing BAs in vitro. Sixty-five strains were able to decarboxylate at least one of the amino acids tested. Pichia kudriavzevii ECA33 (Pk) and Saccharomyces cerevisiae 4 (Sc) were selected to evaluate their potential to modulate BAs, organic acids, and volatile organic compounds (VOCs) accumulation during a simulated cocoa fermentation. The growth of Sc or Pk in the presence of CB caused a significant reduction (p < 0.05) of 2-phenylethylamine (84% and 37%) and cadaverine (58% and 51%), and a significant increase of tryptamine and putrescine with a strong influence of temperature in BA formation and degradation. In addition, our findings pointed out that Pk induced a major production of fatty acid- and amino acid-derived VOCs, while Sc induced more VOCs derived from fatty acids metabolism. Our results suggest the importance of considering BA production in the choice of yeast starters for cocoa fermentation.

Evaluación del potencial de uso de epicarpio de maracuyá deshidratado ( Passiflora edulis f. flavicarpa O. Deg.) en la formulación de yogurt / Evaluation of the potential use of passion fruit dehydrated epicarp ( Passiflora edulis f. flavicarpa O. Deg.) in the formulation of yogurt

RESUMEN La fibra dietética ayuda a reducir el riesgo de padecer algunas enfermedades crónicas, a pesar de esto, su ingesta mundial es menor que los niveles recomendados. Este hecho motiva a la industria alimentaria a incorporar fibra dietética, obtenida de fuentes vegetales en productos alimenticios, como el yogurt. El presente estudio tuvo como objetivo evaluar las principales características de calidad de diferentes formulaciones de yogurt, en función de la incorporación de epicarpio de maracuyá deshidratado (EMD), con alto contenido de fibra dietaria (71,4 ± 0,04%), en concentración de 0 y 0,5% y el efecto que ejerce la grasa láctea presente en la leche cruda de vaca, en concentración de 0, 0,5 y 3,0%. Algunas propiedades fisicoquímicas, tales como pH, acidez titulable (%), viscosidad (cP) y parámetros de color por método CIELab fueron evaluadas en seis formulaciones diferentes. De acuerdo con los resultados obtenidos, se realizó un análisis sensorial y se evaluó la estabilidad de dos formulaciones de yogurt, durante un periodo de almacenamiento de 29 días, a 4°C. La incorporación de EMD contribuyó significativamente al incremento de acidez y la viscosidad en el producto. El EMD favoreció la estabilidad del gel y permitió obtener resultados aceptables en atributos sensoriales hasta el día 15 de almacenamiento. La incorporación de EMD en la elaboración de yogurt permitió enriquecer el valor, debido al aporte de fibra dietaria, la cual, puede actuar como sustituto funcional de la grasa láctea.

ABSTRACT Dietary fiber helps to reduce the risk of suffering from some chronic diseases. Despite this, the global average intake of this component is still lower compared to the recommended levels. This fact has motivated to the food industry to incorporate dietary fiber obtained from vegetable sources in food products. The aim of the present study was the evaluation of the main quality characteristics of different yogurt formulations, as the function of the incorporation of dehydrated passion fruit epicarp with a high dietary fiber content (71,4 ± 0,04%), and concentrations of 0 - 0,5% and the effect of dairy fat present in raw cow milk in concentrations of 0, 0,5 and 3,0%. Some physical-chemical properties such as pH, acidity (% of lactic acid), viscosity (cP) and color parameters by use of CIELab system method were evaluated in 6 different formulations. According to the results, a sensorial analysis was conducted and the stability of two yogurt formulations was determined during a storage period of 29 days at 4°C.The incorporation of EMD significantly contributed to the increase of the acidity and viscosity of the product. EMD favored the stability of yogurt gel and allowed to obtain acceptable results in sensorial attributes until day 15 of storage. The incorporation of EMD in the elaboration of yogurt, allowed the enrichment of the nutritional value due to the contribution of dietary fiber, which can act as a functional substitute of dairy fat.