Search Results (38)

The disposal of olive mill wastewater (OMWW) poses significant environmental challenges due to its high content of phytotoxic and pollutant compounds. This study aims to explore the chemical composition of OMWW derived from various olive varieties (Buža, Buža puntoža, Istarska bjelica, Leccino, and Rosinjola) and assess its antifungal potential against phytopathogenic fungi from the Botryosphaeriaceae family. OMWW samples were analyzed for their physicochemical properties, phenolic composition via LC-MS/MS, and antifungal activity against Botryosphaeria dothidea (Moug. ex Fr.) Ces. & De Not., Diplodia mutila (Fr.) Fr., D. seriata De Not., Dothiorella iberica A.J.L. Phillips, J. Luque & A. Alves, Do. sarmentorum (Fr.) A.J.L. Phillips, Alves & Luque, and Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. Antifungal efficacy was tested at varying concentrations, alongside the phenolic compounds hydroxytyrosol and vanillic acid. Antifungal activity varied across fungal species and OMWW concentrations. Lower OMWW concentrations inhibited mycelial growth in some pathogens, while higher concentrations often had a stimulatory effect. Among the OMWW treatments, Leccino and Buža showed the most significant antifungal activity against species from the Botryosphaeriaceae family. The results demonstrated significant variability in OMWW composition, with Istarska bjelica exhibiting the highest concentrations of phenolic compounds, sugars, dry matter, and carbon and nitrogen content. The results also highlight the impact of acidification on the phenolic profile of OMWW. Treatment with HCl significantly altered the concentration of individual phenolic compounds, either enhancing their release or contributing to their degradation. Among the two compounds, vanillic acid showed greater efficacy than hydroxytyrosol. In addition, microorganisms isolated from OMWW, including Bacillus velezensis Ruiz-Garcia et al., Rhodotorula mucilaginosa (A. Jörg.) F.C. Harrison, Nakazawaea molendiniolei (N. Cadez, B. Turchetti & G. Peter) C. P. Kurtzman & C. J. Robnett, and Penicillium crustosum Thom, demonstrated antagonistic potential against fungal pathogens, with B. velezensis showing the strongest inhibitory effect. The greatest antagonistic effect against fungi was observed with the species Do. Iberica. The findings highlight the potential of OMWW as a sustainable alternative to chemical fungicides, simultaneously contributing to the management of waste and protection of plants through circular economy principles. Full article

The present study aimed to identify the active chemical compounds, mainly phenolic acids, of Champia parvula and Moringa oleifera, evaluate the pharmacokinetic properties of their primary compounds, and assess a novel method for the biocontrol of blue mold by evaluating the antifungal activity of both extracts. Gas chromatography (GC) and high-performance liquid chromatography (HPLC) were utilized to identify the active chemical compounds, mainly phenolic acids. GC illustrated the presence of long-chain aliphatic fatty acids like eicosanoic acid with the formation of oct-1-en-3-ol compounds attached. Catechin was the main bioactive component among the several bioactive compounds identified by HPLC analysis, exhibiting favorable pharmacokinetic behavior, including good absorption, distribution, and metabolic stability. According to the findings, both extracts had antifungal activity, but C. parvula extract (100 mg/mL) exhibited the strongest in vitro and in vivo antifungal activity, with the highest percentages of inhibition (disk diffusion method) against Penicillium expansum, Penicillium crustosum, and Talaromyces atroroseus, ranging between 62.67 and 100%. C. parvula extract (100 mg/mL) could fully inhibit the pathogenicity and aggressiveness of the five tested strains in apple fruits (in vivo). In conclusion, the extract from C. parvula and M. oleifera shows potential antifungal properties and a high phytochemical content. Full article

Fresh wet vermicelli is highly susceptible to microbial contamination during storage as a result of its high moisture content and rich nutrients, which leads to spoilage and deterioration. In addition to exerting a great impact on the quality of the product, this results in significant economic losses and potential food safety risks. This work aimed to identify spoilage microorganisms via traditional culturing methods and molecular biology techniques. The effects of environmental factors such as temperature and pH on the growth and development of the dominant spoilage fungi were investigated, and the inhibitory effects of both chemical (potassium sorbate) and natural antimicrobial agents (chitooligosaccharides, chitosan, tea polyphenols, citric acid, and ε-polylysine hydrochloride) were evaluated. The results indicated that Penicillium crustosum was the major spoilage microorganism in fresh wet vermicelli, whose optimal growth temperature and pH was 28 °C and 7, respectively. While conidial germination began at 7 h, hyphal formation was only observed after 12 h. Moreover, the findings suggest that both natural and chemical antimicrobial agents can effectively inhibit the growth of P. crustosum, with ε-polylysine hydrochloride being the strongest antimicrobial agent. Overall, the findings of this study provide a scientific foundation for improving the preservation of fresh wet vermicelli, which is of great significance for extending its shelf life and enhancing food safety. Full article

Apple blue mold, caused by the Penicillium species, is a significant postharvest disease, leading to food loss and impacting food safety due to mycotoxin contamination. This study aimed to identify the Penicillium species associated with apple blue mold in Serbia, assess their pathogenicity, and evaluate their patulin production potential. A total of 70 Penicillium isolates were collected from symptomatic apple fruit and identified as P. expansum (92.9%), P. crustosum (4.3%), P. solitum (1.4%), and P. chrysogenum (1.4%). The pathogenicity assay revealed P. expansum strains as the most virulent. Molecular detection of msas gene and HPLC analysis confirmed patulin production exclusively in P. expansum isolates. Principal Component Analysis (PCA) grouped P. expansum strains in two distinctive clusters, while P. crustosum strains clustered separately with P. solitum and P. chrysogenum, yet in distinct positions. This is the first report of P. solitum and P. chrysogenum as causal agents of apple blue mold in Serbia. The results of the study provide insights that might be useful in the development of effective control strategies for apple blue mold, ensuring consumption of healthy and safe apple fruit and apple-based products. Full article

According to the scientific information reviewed, cheese is highly susceptible to contamination by mycotoxin-producing fungi, primarily species from the genera Aspergillus (A. niger, A. flavus) and Penicillium (P. commune, P. solitum, P. palitans, and P. crustosum). Studies on various types of cheese made from cow’s milk report an average concentration of Aflatoxin M1 (AFM₁) at 13,000 ng kg⁻¹, which is alarming since the regulatory limits for AFM₁ in cheese range from 250 to 500 ng kg⁻¹. For instance, limits set by Codex Alimentarius, the European Commission (EC), Turkey, and Iran are 250 ng kg⁻¹. In the Netherlands, the limit is 200 ng kg⁻¹, and in Italy, it is 450 ng kg⁻¹. However, the concentration of mycotoxins frequently exceeds these regulatory limits, including critical mycotoxins such as ochratoxin A, citrinin, and cyclopiazonic acid, which pose significant global health concerns. Therefore, this study aims to review the mycobiota responsible for producing key mycotoxins in cheese and to assess the influence of physicochemical factors on fungal growth and mycotoxin production. By incorporating control strategies such as hygiene practices, pasteurization, and the use of preservatives, this study seeks to improve methodologies in the cheese production chain and mitigate contamination by fungi and mycotoxins. Full article

This work describes the immobilization and the characterization of purified Penicillium crustosum Thom P22 lipase (PCrL) using adsorption, encapsulation, and adsorption–encapsulation approaches. The maximum activity of the immobilized PCrL on CaCO₃ microspheres and sodium alginate beads was shifted from 37 to 45 °C, compared with that of the free enzyme. When sodium alginate was coupled with zeolite or chitosan, the immobilization yield reached 100% and the immobilized PCrL showed improved stability over a wide temperature range, retaining all of its initial activity after a one-hour incubation at 60 °C. The immobilization of PCrL significantly improves its catalytic performance in organic solvents, its pH tolerance value, and its thermal stability. Interestingly, 95% and almost 50% of PCrL’s initial activity was retained after 6 and 12 cycles, respectively. The characteristics of all PCrL forms were analyzed by X-ray diffraction and scanning electron microscopy combined with energy dispersive spectroscopy. The maximum conversion efficiency of oleic acid and methanol to methyl esters (biodiesel), by PCrL immobilized on CaCO₃, was 65% after a 12 h incubation at 40 °C, while free PCrL generated only 30% conversion, under the same conditions. Full article

Difenoconazole (DIF), a demethylation inhibitor fungicide, was registered in 2016 for the control of postharvest diseases of pome fruits. In this study, 162 isolates from P. expansum (n = 31) and 13 other “non-expansum” Penicillium spp., i.e., P. solitum (n = 52), P. roqueforti (n = 32), P. commune (n = 15), P. paneum (n = 9), P. psychrosexuale (n = 8), P. crustosum (n = 5), P. carneum (n = 3), P. palitans (n = 2), along with one isolate each of P. citrinum, P. griseofulvum, P. raistrickii, P. ribium, and P. viridicatum, were collected from multiple packinghouses in the U.S. Pacific Northwest. In vitro sensitivity assays showed similar sensitivities of spores and mycelia across species with the mean EC₅₀ values ranging from 0.01 for P. psychrosexuale (n = 8) to 1.33 μg mL⁻¹ for P. palitans (n = 2), whereas the mean EC_50s were 0.03, 0.12, 0.19, and 0.51 μg mL⁻¹ for P. expansum (n = 31), P. paneum (n = 9), P. solitum (n = 52), and P. crustosum (n = 5), respectively. The recommended rate of DIF controlled P. expansum and P. roqueforti isolates but not all isolates of four other Penicillium spp. on Fuji apples after five months at 1.5 °C. The mixture of DIF + fludioxonil (FDL) (Academy^TM) controlled all the dual-sensitive isolates (DIF^SFDL^S) and DIF single-resistant (DIF^R) isolates among the six species tested but not the FDL^R and dual DIF^RFDL^R isolates. Notable polymorphism was detected in the CYP51 gene of the “non-expansum” species with four mutations located at four residues. Although the isolates analyzed in this study had not previously been exposed to DIF, the findings indicate variable sensitivity levels among the Penicillium spp. Full article

Efficient RNA isolation from filamentous fungi is crucial for gene expression studies, but it poses significant technical challenges due to the robust cell walls and susceptibility of RNA to degradation by ribonucleases. This study presents the effectiveness of two RNA isolation protocols for four species of filamentous fungi: Penicillium crustosum, Penicillium rubens, Penicillium griseofulvum, and Aspergillus fumigatus. Both protocols utilized Fenzol Plus for cell lysis but varied in the mechanical disruption methods: bead-beating versus manual vortexing. The results show that the bead-beater method (Protocol 1) yielded significantly higher RNA quantities, with better purity and integrity, as demonstrated by higher A260/A280 and A260/A230 ratios. RNA concentrations ranged from 30 to 96 µg/g of dry biomass in Penicillium species and up to 52 µg/g in A. fumigatus. The use of chloroform in Protocol 1 also enhanced RNA purity, effectively separating contaminants such as DNA, proteins, and polysaccharides. This optimized protocol is highly efficient and can be applied in routine laboratories handling large numbers of fungal samples, making it a robust method for downstream applications such as cDNA synthesis and transcriptome analysis. Full article

Five unreported ketone compounds—penicrustones A–E—were isolated from the solid fermentation of the endophytic fungus Penicillium crustosum. Their structures were elucidated on the basis of extensive spectroscopic analysis. Their absolute configurations were determined via ECD calculations and single-crystal X-Ray crystallography. All compounds were evaluated for their antimicrobial and antitumor activities. Compounds 4 and 5 showed moderate inhibitory effects on Micrococcus luteus, with MIC values of 12.5 and 25.0 μg/mL, respectively. In addition to this, compound 4 also showed cytotoxicity on tumor cell lines KTC-1 and Hela, with IC₅₀ values of 4.28 and 4.64 μg/mL, respectively. Full article

The American cranberry is a perennial North American fruit plant that is grown successfully on commercial plantations in Poland. The purpose of this study was to recognize filamentous fungi that colonize roots, leaves, and fruits without visible disease symptoms. Pure fungal cultures were isolated from disinfected plant fragments in agar media and identified by sequencing common taxonomic DNA markers such as the ITS region, the TEF-1α, or RPB2 genes. Of the 141 isolates studied, 59% were identified as closely related to soil saprotrophs. They were classified primarily as showing the greatest similarity to type strains of Trichoderma amoenum, Trichoderma dorothopsis, Paraphaeosphaeria sporulosa, and Penicillium murcianum. Additionally, isolates that are most similar to strains of Penicillium crustosum, Aspergillus flavus, and Aspergillus versicolor that produced mycotoxins were detected. The fungi identified as closest to Alternaria geophila, Alternaria senecionicola, Paraphoma radicina, Pestalotiopsis unicolor, Pestalotiopsis scoparia, and Neopestalotiopsis spp., whose hosts are plants other than American cranberry, represented 33.81% of the isolates tested. Only 7.2% of the isolates corresponded to the species of Physalospora vaccinia, Diaporthe vaccinii, and Diaporthe eres, known cranberry pathogens. The results of this study can be used to identify latent plant infection and potential disease risks. Full article

One previously undescribed alkaloid, named penifuranone A (1), and three known compounds (2–4) were isolated from the mangrove endophytic fungus Penicillium crustosum SCNU-F0006. The structure of the new alkaloid (1) was elucidated based on extensive spectroscopic data analysis and single-crystal X-ray diffraction analysis. Four natural isolates and one new synthetic derivative of penifuranone A, compound 1a, were screened for their antimicrobial, antioxidant, and anti-inflammatory activities. Bioassays revealed that penifuranone A (1) exhibited strong anti-inflammatory activity in vitro by inhibiting nitric oxide (NO) production in lipopolysaccharide-activated RAW264.7 cells with an IC₅₀ value of 42.2 μM. The docking study revealed that compound 1 exhibited an ideal fit within the active site of the murine inducible nitric oxide synthase (iNOS), establishing characteristic hydrogen bonds. Full article

We investigated the diversity and occurrence of wood wasps in Lithuania and determined communities of associated fungi. Trapping of wood wasps resulted in three different species, including Sirex juvencus, Urocerus gigas, and Tremex fuscicornis. Fungal culturing from adult females of T. fuscicornis mainly resulted in fungi from the genera Penicillium and Trichoderma. High-throughput sequencing of ITS2 rDNA resulted in 59,797 high-quality fungal sequences, representing 127 fungal OTUs. There were 93 fungal OTUs detected in U. gigas, 66 in S. juvencus, and 10 in T. fuscicornis. The most common fungi were Fusarium sporotrichioides (63.1% of all fungal sequences), Amylostereum chailletii (14.9%), Penicillium crustosum (7.8%), Microascus sp. 2261_4 (5.0%), and Pithoascus ater (2.1%). Among these, only A. chailletii was found in all three insect species with the highest relative abundance in U. gigas (15.2%), followed by S. juvencus (7.7%), and the lowest in T. fuscicornis (0.3%) (p < 0.0003). Correspondence analysis of fungal communities showed a distant placement of different species of wood wasps, indicating that fungal communities in each of these were largely different. In conclusion, the study showed that the economically important tree pathogen A. chailletii was among the most common fungal OTUs associated with siricid wood wasps. Full article

Cellulase and xylanase have been widely studied for bioconversion processes and applied in various industries. The high cost of these enzymes remains to be the major bottleneck for large-scale commercial application of lignocellulosic biorefinery. The use of agroindustrial residues and weeds as fermentation substrates is an important strategy to increase cellulolytic enzymes production and reduce costs. Penicillium crustosum was newly isolated and selected to study its enzyme production during solid-state fermentation (SSF). Natural and pretreated water hyacinth (WH) biomass was used as support, substrate and inducer of cellulases and xylanases. Thermochemical pretreatments of WH biomass at 121 °C and sulfuric acid at three concentrations (0.2, 0.6 and 1 M) were assayed. The pretreatments of WH biomass released mono- and oligo-saccharides that favored fungal growth and enzymes production on SSF. WH is a cost-effective substrate-support and inducer, which to be used as a solid medium, was impregnated with a saline solution, containing only (NH₄)₂SO₄, KH₂PO₄ y MgCl₂. Maximum cellulases (carboxymethylcellulase (CMCase)) and xylanases productions of P. crustosum cultured on SSF were reached using the WH pretreated biomass with H₂SO₄ 0.6 M and 121 °C. The simultaneous CMCase and xylanases production reached (647.51 and 4257.35 U/g dry WH, respectively) are among the highest values ever reported. Full article

β-N-acetylhexosaminidases have great potential in applied biocatalysis owing to their ability to act on a wide range of natural and modified substrates. In this work, β-N-acetylhexosaminidases from four Penicillium crustosum strains (PcHex) were studied. The production strains showed the highest enzymatic activity in the culture medium after 11–14 days of cultivation. The specific activity of the isolated and purified PcHex of hydrolysis of 4-nitrophenyl-N-acetyl-β-D-galactopyranoside was 15–20 U/mg protein. All isolated β-N-acetylhexosaminidases showed similar pH–activity profiles, with the optimum pH being 4.0–5.0 and the optimum temperature being 40 °C–50 °C. Apart from standard substrates, two synthetic substrates (5-bromo-4-chloro-3-indolyl-N-acetyl-β-D-galactoside and 2-chloro-4-nitrophenyl-β-D-galactopyranoside) were tested and successfully hydrolyzed using β-N-acetylhexosaminidase preparations. Maximum activity toward the fluorogenic substrate 4-methylumbelliferyl-N-acetyl-β-D-galactopyranoside (4MU-β-GalNAc) was observed for enzyme preparations from PcHex1 (15.38 U/mg protein) isolated from swab samples of books in the Slovak National Museum in Martin (Slovak Republic) and stored in our laboratory. The same enzyme preparation was used for the selective hydrolysis of β-anomer of 4MU-GalNAc from an anomeric mixture of 4MU-α/β-GalNAc. Thus, pure α-anomer (with the total yield being 90%) was separated from the mixture, which suggests the application potential of these enzymes. Full article

This study investigated 65 (35 in summer and 30 in winter) smallholder dairy cattle feeds from Free State and Limpopo provinces in South Africa from 2018 to 2019 for fungal contamination and assessed the impacts of seasonal variation on fungal contamination levels, isolation frequency, and diversity. Samples were examined for fungal contamination using macro- and microscopic approaches, and their identities were confirmed by molecular means. A total of 217 fungal isolates from 14 genera, including Aspergillus, Fusarium, and Penicillium, were recovered from feeds from both seasons. The most prevalent fungal species recovered were A. fumigatus and P. crustosum. Mycological analyses showed that 97% of samples were contaminated with one or more fungal isolates, with the summer fungal mean level (6.1 × 10³ to 3.0 × 10⁶ CFU/g) higher than that of feeds sampled during winter (mean level: 1.1 × 10³ to 4.1 × 10⁵ CFU/g). Independent sample t-test revealed that the isolation frequencies of the genera Aspergillus and Fusarium were significantly (p ≤ 0.05) higher in summer than winter, while Penicillium prevalence in both seasons was not statistically (p > 0.05) different. Furthermore, the Shannon–Weiner diversity index (H′) revealed a higher fungal diversity in summer (H′ = 2.8) than in winter (H′ = 2.1). This study on fungal contamination could be used for future fungal control and mycotoxin risk management in South Africa. Full article