The role of food preservatives in shaping metabolic profile and chemical sensitivity of fungi - an extensive study on crucial mycological food contaminants from the genus Neosartorya (Aspergillus spp.).

Food preservatives are crucial in fruit production, but fungal resistance is a challenge. The main objective was to compare the sensitivity of Neosartorya spp. isolates to preservatives used in food security applications and to assess the role of metabolic properties in shaping Neosartorya spp. resistance. Sodium metabisulfite, potassium sorbate, sodium bisulfite and sorbic acid showed inhibitory effects, with sodium metabisulfite the most effective. Tested metabolic profiles included fungal growth intensity and utilization of amines and amides, amino acids, polymers, carbohydrates and carboxylic acids. Significant decreases in the utilization of all tested organic compound guilds were observed after fungal exposure to food preservatives compared to the control. Although the current investigation was limited in the number of predominately carbohydrate substrates and the breadth of metabolic responses, extensive sensitivity panels are logical step in establishing a course of action against spoilage agents in food production being important approach for innovative food chemistry.

Comprehensive antifungal investigation of natural plant extracts against Neosartorya spp. (Aspergillus spp.) of agriculturally significant microbiological contaminants and shaping their metabolic profile.

Fungi belonging to the genus Neosartorya (teleomorph of Aspergillus spp.) are of great concern in the production and storage of berries and fruit-based products, mainly due to the production of thermoresistant ascospores that cause food spoilage and possible secretion of mycotoxins. We initially tested the antifungal effect of six natural extracts against 20 isolates of Neosartorya spp. using a traditional inhibition test on Petri dishes. Tested isolates did not respond uniformly, creating 5 groups of descending sensitivity. Ten isolates best representing of the established sensitivity clusters were chosen for further investigation using a Biolog™ MT2 microplate assay with the same 6 natural extracts. Additionally, to test for metabolic profile changes, we used a Biolog™ FF microplate assay after pre-incubation with marigold extract. All natural extracts had an inhibitory effect on Neosartorya spp. growth and impacted its metabolism. Lavender and tea tree oil extracts at a concentration of 1000 µg mL-1 presented the strongest antifungal effect during the inhibition test, however all extracts exhibited inhibitory properties at even the lowest dose (5 µg mL-1). The fungal stress response in the presence of marigold extract was characterized by a decrease of amino acids and carbohydrates consumption and an uptake of carboxylic acids on the FF microplates, where the 10 studied isolates also presented differences in their innate resilience, creating 3 distinctive sensitivity groups of high, average and low sensitivity. The results confirm that natural plant extracts and essential oils inhibit and alter the growth and metabolism of Neosartorya spp. suggesting a possible future use in sustainable agriculture as an alternative to chemical fungicides used in traditional crop protection.

The Antifungal Effect of Pyroligneous Acid on the Phytopathogenic Fungus Botrytis cinerea.

In recent years, climate change has intensified harsh periods of rain alternating with periods of drought, leading to an increase in the presence of phytopathogenic fungi. In this study, we want to analyse the antifungal properties of pyroligneous acid against the fungal phytopathogen Botrytis cinerea. Through the inhibition test, we observed that the application of different dilutions of pyroligneous acid rarefied the growth of the fungal mycelium. Furthermore, we have seen through the metabolic profile that B. cinerea is not able to use pyroligneous acid as a resource or even grow in close contact with this resource. Moreover, we observed that the pre-incubation of the fungus in pyroligneous acid leads to a reduction in biomass production. These results give us hope for the possible use of this natural substance as a possible substance to protect plantations from pathogen attacks.

Soil-Borne Neosartorya spp.: A Heat-Resistant Fungal Threat to Horticulture and Food Production-An Important Component of the Root-Associated Microbial Community.

Soil-borne Neosartorya spp. are the highly resilient sexual reproductive stage (teleomorph) of Aspergillus spp. Fungi of this genus are relevant components of root-associated microbial community, but they can also excrete mycotoxins and exhibit great resistance to high temperatures. Their ascospores easily transfer between soil and crops; thus, Neosartorya poses a danger to horticulture and food production, especially to the postharvest quality of fruits and vegetables. The spores are known to cause spoilage, mainly in raw fruit produce, juices, and pulps, despite undergoing pasteurization. However, these fungi can also participate in carbon transformation and sequestration, as well as plant protection in drought conditions. Many species have been identified and included in the genus, and yet some of them create taxonomical controversy due to their high similarity. This also contributes to Neosartorya spp. being easily mistaken for its anamorph, resulting in uncertain data within many studies. The review discusses also the factors shaping Neosartorya spp.'s resistance to temperature, preservatives, chemicals, and natural plant extracts, as well as presenting novel solutions to problems created by its resilient nature.

Evaluation of Nanocomposite Made of Polylactic Acid and Nanocellulose from Carrot Pomace Modified with Silver Nanoparticles.

In this research, it was proposed to use carrot cellulose nanofibrils (CCNF) isolated from carrot pomace modified with silver nanoparticles (AgNPs) as a filler of polylactic acid (PLA) composites matrix. The new procedure was based on two steps: first, the preparation of nanocellulose modified with metal nanoparticles, and then the combination with PLA. Two concentrations-0.25 mM and 2 mM-of AgNO3 were used to modify CCNF. Then, PLA was mixed with the filler (CCNF/AgNPs) in two proportions 99:1 and 96:4. The influence of CCNF/AgNPs on mechanical, hydrophilic, thermal, and antibacterial properties of obtained nanocomposites was evaluated. The greatest improvement of mechanical properties was observed for composite containing CCNF with 2 mM of AgNPs, which obtained the lowest Young modulus and highest strain at break. The degradation temperature was lower for PLA with CCNF/AgNPs, but crystallization temperature wasn't influenced. The addition of CCNF/AgNPs also increased hydrophilicity. The transmission rates of oxygen, nitrogen, and carbon dioxide also increased after the addition of CCNF/AgNPs to PLA. The antibacterial function against Escherichia coli and Bacillus cereus was obtained after the addition of AgNPs but only at the contact surface with the material made, suggesting the lack of migration of nanoparticles from the composite.

Lab to Field Assessment of the Ecotoxicological Impact of Chlorpyrifos, Isoproturon, or Tebuconazole on the Diversity and Composition of the Soil Bacterial Community.

Pesticides are intentionally applied to agricultural fields for crop protection. They can harm non-target organisms such as soil microorganisms involved in important ecosystem functions with impacts at the global scale. Within the frame of the pesticide registration process, the ecotoxicological impact of pesticides on soil microorganisms is still based on carbon and nitrogen mineralization tests, despite the availability of more extensive approaches analyzing the abundance, activity or diversity of soil microorganisms. In this study, we used a high-density DNA microarray (PhyloChip) and 16S rDNA amplicon next-generation sequencing (NGS) to analyze the impact of the organophosphate insecticide chlorpyrifos (CHL), the phenyl-urea herbicide isoproturon (IPU), or the triazole fungicide tebuconazole (TCZ) on the diversity and composition of the soil bacterial community. To our knowledge, it is the first time that the combination of these approaches are applied to assess the impact of these three pesticides in a lab-to-field experimental design. The PhyloChip analysis revealed that although no significant changes in the composition of the bacterial community were observed in soil microcosms exposed to the pesticides, significant differences in detected operational taxonomic units (OTUs) were observed in the field experiment between pesticide treatments and control for all three tested pesticides after 70 days of exposure. NGS revealed that the bacterial diversity and composition varied over time. This trend was more marked in the microcosm than in the field study. Only slight but significant transient effects of CHL or TCZ were observed in the microcosm and the field study, respectively. IPU was not found to significantly modify the soil bacterial diversity or composition. Our results are in accordance with conclusions of the Environmental Food Safety Authority (EFSA), which concluded that these three pesticides may have a low risk toward soil microorganisms.

Intraspecific functional and genetic diversity of Petriella setifera.

The aim of the study was an analysis of the intraspecific genetic and functional diversity of the new isolated fungal strains of P. setifera. This is the first report concerning the genetic and metabolic diversity of Petriella setifera strains isolated from industrial compost and the first description of a protocol for AFLP fingerprinting analysis optimised for these fungal species. The results showed a significant degree of variability among the isolates, which was demonstrated by the clearly subdivision of all the isolates into two clusters with 51% and 62% similarity, respectively. For the metabolic diversity, the BIOLOG system was used and this analysis revealed clearly different patterns of carbon substrates utilization between the isolates resulting in a clear separation of the five isolates into three clusters with 0%, 42% and 54% of similarity, respectively. These results suggest that genetic diversity does not always match the level of functional diversity, which may be useful in discovering the importance of this fungus to ecosystem functioning. The results indicated that P. setifera strains were able to degrade substrates produced in the degradation of hemicellulose (D-Arabinose, L-Arabinose, D-Glucuronic Acid, Xylitol, γ-Amino-Butyric Acid, D-Mannose, D-Xylose and L-Rhamnose), cellulose (α-D-Glucose and D-Cellobiose) and the synthesis of lignin (Quinic Acid) at a high level, showing their importance in ecosystem services as a decomposer of carbon compounds and as organisms, which make a significant contribution to carbon cycling in the ecosystem.The results showed for the first time that the use of molecular biology techniques (such as AFLP and BIOLOG analyses) may allow for the identification of intraspecific diversity of as yet poorly investigated fungal species with favourable consequences for our understanding their ecosystem function.

Metabolic and Genetic Properties of Petriella setifera Precultured on Waste.

Although fungi that belong to Petriella genus are considered to be favorable agents in the process of microbial decomposition or as plant endophytes, they may simultaneously become plant pests. Hence, nutrition factors are supposed to play an important role. Therefore, it was hypothesized that Petriella setifera compost isolates, precultured on three different waste-based media containing oak sawdust, beet pulp (BP) and wheat bran (WB) will subsequently reveal different metabolic properties and shifts in genetic fingerprinting. In fact, the aim was to measure the influence of selected waste on the properties of P. setifera. The metabolic potential was evaluated by the ability of five P. setifera strains to decompose oak sawdust, BP and WB following the MT2 plate® method and the catabolic abilities of the fungus to utilize the carbon compounds located on filamentous fungi (FF) plates®. Genetic diversity was evaluated using Amplified Fragment Length Polymorphism analysis performed both on DNA sequences and on transcript-derived fragments. P. setifera isolates were found to be more suitable for decomposing waste materials rich in protein, N, P, K and easily accessible sugars (as found in WB and BP), than those rich in lignocellulose (oak sawdust). Surprisingly, among the different waste media, lignocellulose-rich sawdust-based culture chiefly triggered changes in the metabolic and genetic features of P. setifera. Most particularly, it contributed to improvements in the ability of the fungus to utilize waste-substrates in MT2 plate® and two times increase the ability to catabolize carbon compounds located in FF plates®. Expressive metabolic properties resulting from being grown in sawdust-based substrate were in accordance with differing genotype profiles but not transcriptome. Intraspecific differences among P. setifera isolates are described.