This study investigated the bioactive composition and antifungal efficacy of neem seed extracts (NSE) prepared using acetone, ethyl acetate, chloroform, methanol, and ethanol against Aspergillus parasiticus and Aspergillus flavus. Gas chromatography-mass spectrometry (GC-MS) analysis revealed solvent-specific compounds profiles: ethanol extract contained 35 bioactive compounds (mainly esters and fatty acids), dominated by n-hexadecanoic acid (28.65%); methanol extract had 29 diketopiperazines, with 3,6-bis(2-methylpropyl)-2,5-piperazinedione (45.45%) as major; chloroform, ethyl acetate, and acetone extracts featured long-chain esters, hydrocarbons, and 2-heptadecanone (33.15%), respectively. All extracts inhibited fungal growth at 500mg/mL, whereas ethanol was most effective at 31.25mg/mL, reducing mycelial biomass by 45%. Methanol and ethanol extracts (250mg/mL) suppressed aflatoxin (AF) (AFB1, AFB2) production by >90%. Six-month feed trials confirmed neem seed powder (NSP) as a potent biopreservative, achieving complete AF inhibition at 30% concentration. These findings highlight neem's potential as a natural antifungal and anti-aflatoxigenic agent.

Inhibition of Aspergillus parasiticus and detoxification of aflatoxin derivatives in tomato paste by adding freeze-dried postbiotic from Lactiplantibacillus plantarum

Synthesis, characterization, antifungal and SERS analysis of mycosynthesized silver nanoparticles against Aspergillus parasiticus

High-protein beverages are valued for their health benefits, but they face threats from toxigenic fungi and mycotoxins This study developed a novel, functional high-protein beverage (HPB) by synergistically combining whey protein concentrate (WPC), fermented by specific bacteria, with Physalis peruviana (golden berry) powder (at a 1:5; w/w). Three bacterial strains—Lactobacillus plantarum (BS-2), L. pentosus (BS-1), and L. paracasei (BS-3)—were involved for the fermentation, individually and in combinations. Fortification with golden berry significantly enhanced the phytochemical profile, increasing phenolic content (22.97 ± 0.56 mg GAE/g), flavonoids (97.15 ± 2.58 mg QE/g), and antioxidant capacity compared to the control. The BS-2 strain exhibited the strongest antifungal activity, inhibiting fungal growth by 53.8 ± 0.41% and completely suppressing the aflatoxin B2 production by Aspergillus parasiticus. Rheological analysis showed that BS-2 and the three-strain mixture significantly increased viscosity (p < 0.05), enhancing beverage stability. Sensory evaluation (n = 40) revealed that the BS-2 fermented Physalis-HPB was the most preferred, earning the highest overall score. During 21-day shelf-life tests under fungal challenge, beverages fermented with BS-2 and mixed strains maintained the lowest fungal CFU counts at 4 °C, demonstrating superior microbial stability. These findings show that combining P. peruviana with probiotic fermentation enhances the functional, sensory, and safety qualities of HPBs, suggesting a promising bio-based approach to reduce fungal spoilage and mycotoxin hazards in dairy beverages.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-24160-y.

Aflatoxin contamination is a serious threat affecting agricultural commodities in storage conditions all over the world. In current investigation work, the combination efficacy of natural fumigants such as terpinen-4-ol: cuminaldehyde and terpinen-4-ol: linalool on growth of the aflatoxigenic fungi Aspergillus flavus and Aspergillus parasiticus are assessed. Chemical characterisation using GC-MS of natural fumigant, the A9 combination showed signature peaks of terpinen-4-ol (39%) and cuminaldehyde (54.59%) which effectively prevented the fungal growth and aflatoxin production in both Aspergillus species. Dose-dependent assay indicated a significant antifungal activity at 7.5 and 10 μL with a complete reduction of aflatoxins. Further, A9 combination treatment showed a decrease of ergosterol biosynthesis, plasma membrane damage and oxidative stress in fungal mycelium. Thereafter, in sorghum and pearl millet the fumigation with A9 combination effectively inhibited fungal proliferation and reduced aflatoxins. Conclusively, the study emphasises the use of natural biofumigant combinations in mitigating the aflatoxin contamination.

Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic compound produced by certain fungi (e.g., Aspergillus flavus and Aspergillus parasiticus). Rapid and ultra-sensitive detection methods for AFB1 in various commodities are in high demand. This study aimed to enhance the sensitivity of a competitive lateral flow immunoassay (LFIA) for AFB1 detection by leveraging a previously developed experimental design strategy, named 4S. This approach comprises four phases—START, SHIFT, SHARPEN, and STOP—and involves the analysis of two reference conditions: NEG (0 ng/mL AFB1) and POS (1 ng/mL AFB1). By generating and overlaying response surfaces, regions of optimal NEG signal and POS/NEG signal ratio (IC%) were identified. Four variables were optimized: two related to the labeled antibody (its concentration and antibody-to-label ratio) and two to the competitor antigen (its concentration and hapten-to-protein ratio). An initial design defined the parameter space, while three subsequent designs did not yield further improvements in sensitivity. A strong anti-correlation was observed between the IC% and competitor parameters. The optimized LFIA-1 exhibited enhanced sensitivity, achieving a limit of detection of 0.027 ng/mL compared to 0.1 ng/mL for the original device. Additionally, the amount of expensive antibody required for device fabrication was reduced by around a factor of four.

Aflatoxins (AFs) are highly toxic and carcinogenic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, posing significant risks to food safety and public health. These mycotoxins commonly contaminate oilseeds such as peanuts, sunflower, cotton and maize seeds, particularly under warm and humid conditions. Controlling and reducing aflatoxin levels in these products is critical to ensuring food safety and complying with international regulatory standards. This review provides a comprehensive overview of current strategies for aflatoxin degradation in oilseeds, with a particular emphasis on physical and microbial approaches. Physical methods including thermal treatment, ammoniation, irradiation and advanced radiation techniques have been explored for their effectiveness in reducing aflatoxin levels. Microbial strategies involving specific bacteria, fungi and their enzymatic systems offer environmentally friendly and biologically based solutions for detoxification. The review critically examines the efficacy, mechanisms of action and limitations of each approach, highlighting the key factors that influence their success in practical applications. Moreover, it discusses the advantages and challenges associated with integrating these methods into existing food processing systems, considering factors such as cost-effectiveness, preservation of nutritional quality, regulatory acceptance and scalability. Finally, the review identifies key areas for future research, emphasizing the need to develop more efficient, sustainable and industrially viable technologies for large-scale aflatoxin decontamination in oilseeds. These advancements are essential for enhancing global food safety, protecting public health and supporting international trade.

This study presents a comprehensive investigation of the essential oil obtained from Satureja candidissima (Munby), with a particular focus on its biological activities. The chemical composition of the oil was determined using gas chromatography (GC) and gas chromatography–mass spectrometry (GC-MS), which led to the identification of 37 compounds accounting for 93.9% of the total oil. The principal constituents were pulegone (32.1%), menthone (23.8%) and neo-menthol (20.2%). The antioxidant activity was assessed using DPPH free radical scavenging and β-carotene–linoleic acid bleaching assays, which indicated low activity in both cases. Antifungal efficacy, determined through disc diffusion and agar dilution methods, demonstrated significant inhibition, particularly against Aspergillus parasiticus and A. niger, which exhibited the lowest minimum inhibitory concentration (MIC) values of 0.250 and 0.5 mg.mL-1, respectively. In vivo anti-inflammatory activity was evaluated in a mice model using carrageenan-induced paw edema. At a dose of 250 mg.kg-1, the essential oil produced moderate anti-inflammatory effects, resulting in 36.31% inhibition of edema. Antinociceptive activity, assessed via the acetic acid-induced writhing test, showed a significant reduction in pain behavior at both 100 and 250 mg.kg1 doses, with effects comparable to the standard analgesic, paracetamol. These findings highlight the potential of S. candidissima essential oil as a promising source of natural antifungal and analgesic agents, with complementary anti- inflammatory properties likely attributed to its phytochemical profile.

Background and Aim: Various feeds and food grains are contaminated with mycotoxins produced by certain fungi along with other secondary metabolites due to long term storage, storage condition, or improper handling during harvest. The economic effects attributed to mycotoxin infections are widely felt in all sectors of the production and consumption of grain products. They are considered a major factor in the spoilage of food stuffs, leading to great economic loss and have implications for food safety and public health. The study aims to isolate and characterize Aflatoxin producing fungi associated with the spoilage of some stored grains within Bauchi metropolis. Place and Duration of Study: This study was carried out in Abubakar Tafawa University (ATBU), Bauchi, Nigeria, in a period duration of August 2024 to January 2025. Methodology: Grain samples were obtained from various stores in markets and transported to the laboratory. Grains samples were aseptically inoculated in a set of three Petri dishes each containing of Potato Dextrose Agar (PDA) and incubated for 5 days at 30°C. Pure culture was obtained from identified isolate after Identification of the isolates morphologically and phenotypically. Pure isolates were subjected to Polymerase Chain Reaction (PCR) by extracting the DNA following the Bioneers kit manufacturers manual and gel electrophoresis was ran on the pure DNA extracted. Aflatoxin test was done on samples using Thin Layer Chromatography (TLC). Grain samples were ground into fine powder using mortar and pestle. Extraction was done on the samples and 10µl of the extract was added to the TLC plate 1cm from the bottom edge and allowed to dry. The TLC plate was placed in the developing chamber with the solvent at the bottom, allowing the solvent to rise up by capillary action. When solvent reached the top of the plate, the plate was allowed to air-dry completely. Sulfuric acid spray was used to visualize the separated compounds and plate was observed using spectrophotometer. Results: The findings revealed a diverse array of Aspergillus species capable of producing aflatoxins, notably Aspergillus flavus and Aspergillus parasiticus. Aspergillus was identified as the predominant fungal species, accounting for 51.47% of isolates, followed by Rhizopus (22.06%), Penicillium (19.12%), and Fusarium (7.35%). Significant aflatoxin contamination was observed across various grain types, with millet and groundnut exhibiting high contamination rates and concentrations. Molecular analysis further confirmed the presence and co-expression of aflR and aflS genes in Aspergillus flavus isolates, indicating strong aflatoxigenic potential. Conclusion: This study highlights the prevalence and some of the genetic diversity of the aflatoxin producing fungi focussing on the aflR and aflS gene which is one of the genes that plays a crucial role in the biosynthesis of aflatoxins. The study emphasizes the need for targeted interventions to mitigate aflatoxin contamination and ensure food safety. The prevalence of aflatoxin producing fungi in this study was 65% based on the TLC result showing that 65 samples have exceeded the NAFDAC permissible limit. Aspergillus flavus and Aspergillus parasiticus were the primary aflatoxin producers observed among the fungal isolates. The relationship between the grain type and aflatoxin contamination is highly significant indicating that the effect of grain type on aflatoxin production is consistent independent of storage conditions.

Abstract In sub-Saharan African countries, the maize crop is a major staple food crop, providing up to 70% of the population’s total caloric intake. Aflatoxins, toxic secondary metabolites primarily produced by Aspergillus flavus and Aspergillus parasiticus , pose significant threats to food safety, public health, and agricultural economies in sub-Saharan Africa (SSA), where maize is a staple crop. This review synthesizes current knowledge on the mechanisms of aflatoxin production and its management within maize cropping systems of SSA. The occurrence and severity of aflatoxin contamination are influenced by multiple factors, including high temperatures, drought stress, insect damage, poor post-harvest handling, and inadequate storage conditions. Maize has been linked to being contaminated by roughly eighteen (18) different forms of aflatoxins, which are severely poisonous, and contribute to public health issues. The review explores the biological and environmental triggers of aflatoxin biosynthesis, highlighting molecular pathways and fungal-host interactions. Additionally, it evaluates integrated management strategies encompassing host resistance, good agricultural practices, biocontrol agents (such as Aflasafe ), proper harvesting, drying, and storage techniques. Socio-economic and institutional barriers to effective aflatoxin control are also discussed, along with policy and research recommendations. The review also emphasizes on the necessity to apply novel and existing techniques to prevent aflatoxin. The study featured the need for a multidisciplinary and region-specific approach to sustainably mitigate aflatoxin risks in SSA. Best bet recommendations are provided given different levels of scenarios at the farmer, farm plot, maize farming systems, and eventually the nodes across the entire maize value chain.

Aflatoxins, which are potentially genotoxic and carcinogenic substances, are mainly produced by the Aspergillus section Flavi, including Aspergillus flavus and A. parasiticus. Current Aspergillus spp. detection is often based on molecular methods, such as real-time PCR and loop-mediated isothermal amplification (LAMP), targeting genes of the aflatoxin biosynthetic cluster. In this study, we developed a Lab-on-a-Chip (LoC) method based on real-time PCR and on LAMP for the specific detection of aflatoxigenic strains of A. flavus and A. parasiticus from infected hazelnuts. LoC-LAMP and LoC-real-time PCR assays were tested in terms of specificity, sensitivity, speed, and repeatability. The microfluidic chip allowed quick, specific, sensitive, simple, automatized, cheap, and user-friendly detection of aflatoxigenic strains of A. flavus and A. parasiticus. The LoC-LAMP showed a limit of detection (LOD) of 10 fg of DNA, while the LoC-real-time PCR showed a LOD of 10 pg of DNA. Achieving comparable sensitivity to that of LAMP and real-time PCR techniques, both LoC methods developed in this work offer the advantages of automation, minimal sample requirements, reagent requirements, and cost-effectiveness. Overall, the developed methods open the perspective for alternative monitoring of aflatoxigenic fungi in the agri-food industry.

Aflatoxin B1 (AFB1) is a mycotoxin mainly produced by Aspergillus flavus and Aspergillus parasiticus. The genotoxic capacity of AFB1 has been demonstrated in a range of genotoxicity assays. The aim of the present study was to evaluate the cytotoxic and genotoxic effects of AFB1 and fumonisin B1 (FB1) mixtures on hepatocellular carcinoma cell line (HepG2), as well as the inflammatory effects on human neutrophils of AFB1 at environmentally relevant concentrations. The cytotoxicity was evaluated using the MTT assay. The cytostatic and genotoxic effects were evaluated using the micronucleus technique, and the inflammatory effect of AFB1 was determined by fluorescence microscopy to evaluate neutrophil extracellular traps (NETs). Additionally, we have explored the response of the system in a coexposure treatment with FB1. Significant increases in micronucleus frequency, apoptosis, and necrosis were observed after treatment with 10-μM AFB1 or 10-μM AFB1 + 50-μM FB1. FB1 did not have a synergistic effect with AFB1. A relative impairment of the genetic material reflected by nucleoplasmic bridges, nuclear budding, and donut-shaped nuclei was observed. On the other hand, the assessment of the inflammatory response showed that AFB1 induces the formation of extracellular traps, apparently dose dependent.

ABSTRACTThis study investigated the use of Rosmarinus officinalis (rosemary) essential oil in a microemulsion (ME) formulation for the treatment of ocular fungal keratitis, a serious infection that can lead to blindness. The ME was characterized in terms of pH, stability, homogeneity, osmolarity, and other physicochemical properties. The Hen's Egg Test‐Chorioallantoic Membrane assay was used to assess ocular irritancy, and antifungal efficacy was evaluated using the minimum inhibitory concentration method. The oil extraction yielded 1.2%, with eucalyptol (37.89%) as the major compound. The formulation showed a suitable pH (6.96), particle size of 11.35 nm, good stability, and was classified as non‐irritant with an irritation score = 1.5. It exhibited strong antifungal activity against Candida parapsilosis (96.1%), C. krusei (100%), C. albicans (99.1%), Fusarium graminearum (90.5%), and Aspergillus parasiticus (79.5%). The results highlight the potential of rosemary essential oil as a base for developing eye drops for the treatment of fungal keratitis.

Aspergillus parasiticus is a fungus recognized for producing highly carcinogenic mycotoxins. In this study, we collected 38 isolates of A. parasiticus from fields in South Georgia. We performed whole genome re-sequencing and developed 38 draft genome assemblies of A. parasiticus. The average genome size was 38.7 Mb, with larger genomes (~40 Mb) found in peanut fields in Turner County. Scaffold N50 was recorded highest for isolates collected from the corn fields of Tifton. The average BUSCO completeness score for these assemblies was 99.1%. The genome sequences generated for these 38 isolates will serve as a valuable genomic resource for the community working on aflatoxin mitigation strategies in crops.

ABSTRACTMycotoxins in food and feed are a significant health risk, even more so than pesticides and synthetic waste. These toxic secondary metabolites are produced by various fungal species, particularly after fungal colonization of crops. Aflatoxins produced mainly by Aspergillus flavus and Aspergillus parasiticus are among the most concerning mycotoxins. These fungi can colonize a range of crops, including maize and wheat, and produce aflatoxins both in the field and during post‐harvest. Aflatoxin B1 (AFB1) is the most toxic and carcinogenic, with demonstrated genotoxic, immunosuppressive, teratogenic, and hepatotoxic effects. Aflatoxins are stable in food and feed and can persist in the food chain, potentially appearing in milk as AFM1. Due to their toxicity, aflatoxins are strictly regulated globally, including in the European Union under Commission Regulation 2023/915. Climate change is increasing the frequency and concentration of mycotoxins in crops. The current control methods, including antifungals and synthetic chemicals, are ineffective and harmful, leading to the need for “greener” solutions. Recent research suggests that mushroom metabolites, particularly polysaccharides from species like Pleurotus eryngii, have potential in inhibiting aflatoxin synthesis. This study explores the effects of mycelial culture filtrates and aqueous extracts from two varieties of Tunisian Pleurotus eryngii on the growth and aflatoxin production of Aspergillus flavus.

The current study was aimed to assess the chemical compositions of Melia azedarach Linn (Chinaberry) leaves aqueous extracts. Additionally, the extracts were also tested to investigate its antifungal potentials against Aspergillus flavus and Aspergillus parasiticus. Leaf extract of M. azedarach was obtained by maceration technique, subsequently analyzed using UV-Visible Spectrophotometer, Fourier Transform Infrared (FTIR) and Gas Chromatography-Mass Spectrometry (GC-MS). The total phenolic and flavonoids contents were; 67.5 ± 0.4 mg GAE/g DW and 12.7 ± 0.2 mg QE /g DW respectively. The presence phytochemicals were confirmed from various functional groups recorded in FT-IR spectra. The results were further validated through GC-MS analysis where a total of 18 compounds were identified with seven major compounds; namely 1-Butanol, 3-methyl-, acetate (11.53%), followed by coumaran (10.04%), (R, S)-2-propyl-5-oxohexanal (7.07%), 10-octadecenoic acid, methyl ester (5.16%) and 5,7-Octadien-2-one, 3-acetyl (3.06%). The extract exhibited antifungal activities against two major aflatoxin-producing fungi, A. flavus and A. parasiticus. The aqueous extract (31.25 to 500 mg mL-1) was active to inhibit the spore germination, mycelial growth, biomass production and aflatoxin biosynthesis. Spore germination was significantly reduced, with maximum inhibition of 83% against A. flavus and 85% against A. parasiticus at 500 mg mL-1. Mycelial growth and fungal biomass were markedly declined with increasing trend in extract concentration. The recorded biomass inhibition was 73.2% and 76.9% respectively against A. flavus and A. parasiticus. The extract also significantly suppressed the aflatoxin production in the selected fungal strains at higher concentrations, exceeded from 75% with respect to aflatoxins B₁, B₂, G₁, and G₂. The findings suggest that M. azedarach leaves extract is a valuable source of bioactive compounds possessing strong antifungal and anti-aflatoxigenic properties and could be considered as a promising natural alternative for controlling aflatoxins contamination in agricultural food sectors.

Aspergillus species are found all over the globe and may grow on a broad range of surfaces. Many species may cause food and spice degradation, as well as the development of aflatoxins, which is a public health concern in Nigeria and other tropical regions across the globe. Four species of Aspergillus were identified in four culinary spices regularly eaten in Adamawa state’s Mubi north local government area. Spice samples included turmeric, garlic, ginger, clove, and nutmeg. The agar plate technique was utilised for inoculation and identification of Aspergillus species. Spice samples were inoculated on ampicillin-modified potato dextrose agar and cultured for seven days at 28ºC to isolate mixed fungus colonies. The following aspergillus species were isolated in spices: garlic (Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, and Aspergillus parasiticus), nutmeg (Aspergillus fumigatus and Aspergillus parasiticus), ginger, and clove (Aspergillus niger and Aspergillus flavus), and tumerric (all species). The technique used for the testing of aflatoxins is ELISA process, therefore the total aflatoxins discovered are turmeric 0.13, clove 0.052498, ginger 0.052962, garlic 0.059992, and nutmeg 0.035. The obtained results show that the analysed spices eaten in Mubi North Local Government in Adamawa State provide low or no public danger. Given the significant impact of the toxins, there is a need for enhanced spice handling, preservation, and storage to reduce fungal infection. Keywords: Spices, Aflatoxins, Aspergillus species and Fungi

ABSTRACTConversion of agro‐waste into carbon dots (CDs) transforms biomass into sustainable nanomaterials, enabling advanced applications in packaging and ensuring food safety. Bambara groundnut pericarp powder (BGPP) was hydrothermally processed at 200°C for varying durations (3–12 h) to synthesize BGP‐CDs. BGP‐CDs exhibited differences in size (1.34–15.02 nm), shape, and chemical composition, as characterized by transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) analyses. All samples exhibited excellent optical properties, including photoluminescence and UV barrier capability. Notably, BGP‐CDs/3 (synthesized for 3 h) at 100 µg/mL demonstrated the highest UV‐blocking efficiency against UV‐A (35.62%) and UV‐B (92.64%) lights. BGP‐CDs synthesized for 12 h exhibited remarkable antimicrobial activities against selected species (Escherichia coli, Listeria monocytogenes, Aspergillus flavus, and Aspergillus parasiticus) due to their nanoscale size; however, their reduced size negatively impacted human cell viability. Antioxidant activity was highest in BGP‐CDs synthesized for 6 h (BGP‐CDs/6), as determined by different assays. Incorporating BGP‐CDs/6 (4%, w/w) into gelatin/polylactic acid (PLA) bilayer film enhanced UV and water barrier properties, antioxidant capacity, and antibacterial efficacy compared to the control film. Additionally, Asian seabass slices (AS‐S) packaged in pouches made from active bilayer film had the maintained color with negligible pH change. Microbial growth and lipid oxidation in AS‐S were retarded as indicated by the lower rise in total viable count (TVC) and psychrophilic bacterial count (PBC) as well as thiobarbituric acid reactive substances (TBARS) values over 12 days of storage at 4°C. Thus, the developed multifunctional packaging film offers an innovative strategy that could impede the quality loss of perishable fish slices.

Aflatoxins, toxic secondary metabolites produced primarily by Aspergillus flavus and Aspergillus parasiticus, pose a significant global health concern due to their frequent presence in crops, food, and feed-especially under climate change conditions. This review addresses the growing threat of aflatoxins by analyzing recent advances in detection and mitigation. A comprehensive literature review was conducted, focusing on bioremediation, physical and chemical detoxification, and fungal growth inhibition strategies. The occurrence of aflatoxins in water systems was also examined, along with current detection techniques, removal processes, and regulatory frameworks. Emerging technologies such as molecular diagnostics, immunoassays, biosensors, and chromatographic methods are discussed for their potential to improve monitoring and control. Key findings highlight the increasing efficacy of integrative approaches combining biological and technological solutions and the potential of AI-based tools and portable devices for on-site detection. Intelligent packaging and transgenic crops are also explored for their role in minimizing contamination at the source. Overall, this review emphasizes the importance of continued interdisciplinary research and the development of sustainable, adaptive strategies to mitigate aflatoxin risks, thereby supporting food safety and public health in the face of environmental challenges.

Given fungi's critical role in public health and their impact during pandemics such as COVID-19, precise identification and classification are essential. Additionally, fungi hold significant value in medical and economic applications. For this work, fungi were isolated from various fruit. The fungi were initially identified based on their morphological characteristics using microscopic techniques. To achieve a comprehensive characterization, the eight fungal species were analyzed using rapid and cost-effective spectroscopic techniques, including attenuated total reflectance Fourier transform infrared spectroscopy (ATR FT-IR), Raman spectroscopy (RS), and ultraviolet-visible spectroscopy (UV-Vis). Fungal samples were used in the powder form, generating distinct spectral fingerprints in the biochemical region specific to components such as proteins, lipids, polysaccharides, carbohydrates, and nucleic acids. Results demonstrated the efficacy of these spectroscopic approaches for rapid and accurate identification, enabling discrimination between fungal species and reliable classification at the genus level. The results showed the species were identified as Aspergillus parasiticus, Phytophthora spp., Chaetomium globosum, Penicillium digitatum, Penicillium sp., Penicillium italicum, Rhizoctonia solani, and Myrothecium roridum. This highlights the potential of these techniques as efficient tools for fungi identification.