Revelation of antifungal susceptibility of botanicals and genetic diversity of aflatoxin-producing Aspergillus flavus in chili (Capsicum annuum)

Mst. Sabiha Sultana, Nusrat Jahan, Shamim Ahmed Kamal Uddin Khan, S.M. Abdullah Al Mamun, Md. Mustafizur Rahman

Abstract


Aflatoxins associated with Aspergillus flavus pose a significant risk to food safety and public health. This study determined the minimum inhibitory concentration (MIC) of five medicinal plant extracts against A. flavus. Molecular identification, performed by amplifying the ITS rDNA region, confirmed the isolate as A. flavus (99.80% similarity to the GenBank sequence OP480002.1). The antifungal activities of botanical extracts (Piper betle, Centella asiatica, Azadirachta indica, Mentha spicata, and Ocimum tenuiflorum) at different concentrations (600 mg/ml, 300 mg/ml, and 100 mg/ml) were evaluated using disc diffusion and broth dilution methods, with Fluconazole as a reference. Except for M. spicata, all extracts exhibited inhibitory activity, with A. indica showing the highest inhibition (19.83 mm at 600 mg/ml), surpassing Fluconazole (15.67 mm). The MIC values were lowest for Fluconazole (4.68 mg/ml), followed by A. indica (9.375 mg/ml). RAPD-PCR analysis revealed significant genetic diversity among isolates, with 12 primers generating 50 polymorphic bands. Genetic similarity ranged from 73% to 99.2%, with the highest diversity observed between isolates AS3 and AS4. These findings highlight the potential of these medicinal plant extracts as natural alternatives to synthetic fungicides at specific concentrations for reducing aflatoxin contamination in agricultural products. Furthermore, the observed genetic diversity may help in assessing the aflatoxin production capacity of A. flavus in different chili cultivars.

Keywords


Aspergillus flavus, Medicinal plant extracts, Genetic diversity, Therapeutic value, MIC

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References


Adebiyi, J.A., Kayitesi, E., Adebo, O.A., Changwa, R., Njobeh, P.B., 2019. Food fermentation and mycotoxin detoxification: An African perspective. Food Control 106, 106731.

Basak, S., Guha, P., 2017. Betel leaf (Piper betle L.) essential oil microemulsion: Characterization and antifungal activity on growth, and apparent lag time of Aspergillus flavus in tomato paste. LWT 75, 616-623.

Behiry, S.I., Hamad, N.A., Alotibi, F.O., Al-Askar, A.A., 2022. Antifungal and antiaflatoxigenic activities of different plant extracts against Aspergillus flavus. Sustainability 14(19), 12908.

Bharathi, T., Kolanjinathan, K., Saranraj, P., 2014. Antimicrobial activity of solvent extracts of Ocimum tenuiflorum, Azadirachta indica, and Phyllanthus amarus against clinical Pathogens. Global Journal of Pharmacology 8(3), 294-305.

Burdon, J., Silk, J., 1997. Sources and patterns of diversity in plant-pathogenic fungi. Phytopathology 87(7), 664-669.

Chongo, G., Gossen, B.D., Buchwaldt, L., Adhikari, T., Rimmer, S., 2004. Genetic diversity of Ascochyta rabiei in Canada. Plant disease 88(1), 4-10.

Chukwudi, H.C., and Ezeabara, C.A., 2018. Phytochemical screening and in vitro antimicrobial activities of Mimosa invisa Mart. leaves and stems. Bioscience Horizons: The International Journal of Student Research, 11, hzy019.

Di Ciaccio, L., Catalano, A., López, P., Salvat, A., 2020. In vitro antifungal activity of Peltophorum dubium (Spreng.) taub. extracts against Aspergillus flavus. Plants 9(4), 438.

FAOSTAT., 2017. FAO, available online: http://www. fao. org/faostat/en/# data. QC (accessed on January 2018).

Hossain, M.N., Talukder, A., Afroze, F., Rahim, M.M., Begum, S., Haque, M.Z., Ahmed, M. M., 2018. Identification of aflatoxigenic fungi and detection of their aflatoxin in red chili (Capsicum annuum) samples using direct cultural method and HPLC. Advances in Microbiology 8(1), 42-53.

Hudzicki, J., 2009. Kirby-Bauer disk diffusion susceptibility test protocol. American Society for Microbiology 15, 55-63.

Jannat, M., Masud, M., Nusrat, M., Bashar, S., Mita, M.M., Hossain, M.I., and Islam, M.R., 2022. Aflatoxins and fumonisins contamination of maize in Bangladesh: an emerging threat for safe food and food security. maize genetic resources: Breeding Strategies and Recent Advances 18-69.

Kavitha, K., Vijaya, N., Krishnaveni, A., Arthanareeswari, M., Rajendran, S., Al-Hashem, A., Subramania, A., 2020. Nanomaterials for antifungal applications. In Nanotoxicity. Elsevier 385-398.

Kennedy, Z.J., Paranitharan, V., Karthikeyan, S., 2022. Mycotic contamination and aflatoxin potential of molds in Capsicum annum (chili), and chili powder commercialized in south Indian markets. Toxicon 210, 109-114.

Khan, M.A., Asghar, M.A., Iqbal, J., Ahmed, A., Shamsuddin, Z.A., 2014. Aflatoxins contamination and prevention in red chillies (Capsicum annuum L.) in Pakistan. Food Additives & Contaminants: Part B 7(1), 1-6.

Maida, C.M., Milici, M.E., Trovato, L., Oliveri, S., Amodio, E., Spreghini, E., and Barchiesi, F., 2008. Evaluation of the disk diffusion method compared to the microdilution method in susceptibility testing of anidulafungin against filamentous fungi. Journal of Clinical Microbiology 46(12), 4071-4074.

Makhuvele, R., Naidu, K., Gbashi, S., Thipe, V.C., Adebo, O.A., Njobeh, P.B 2020. The use of plant extracts and their phytochemicals for control of toxigenic fungi and mycotoxins. Heliyon 6(10).

Margaret, A., Yolanda, H., 2013. Antifungal activity of neem leaf ethanol extract on Aspergillus flavus. Universa Medicina 32(2), 80-85.

McDonald, B.A., McDermott, J.M., 1993. Population genetics of plant pathogenic fungi. Bioscience 43(5), 311-319.

Mesterhazy, A., 2024. Food safety aspects of breeding maize to multi-resistance against the major (Fusarium graminearum, F. verticillioides, Aspergillus flavus) and minor toxigenic fungi (Fusarium spp.) as well as to toxin accumulation, trends, and solutions-a review. Journal of Fungi 10(1), 40.

Müller, S., Schlöder, F., Stutzki, J., Winnewisser, G., 2005. The cologne database for molecular spectroscopy, CDMS: a useful tool for astronomers and spectroscopists. Journal of Molecular Structure 742(1-3), 215-227.

Njoki, L.M., Okoth, S.A., Wachira, P.M., 2017. Effects of medicinal plant extracts and photosensitization on aflatoxin producing Aspergillus flavus (Raper and Fennell). International Journal of Microbiology 2017, 1-9.

Okayo, R.O., Andika, D.O., Dida, M.M., K’Otuto, G.O., Gichimu, B.M., 2020. Morphological and molecular characterization of toxigenic Aspergillus flavus from groundnut kernels in Kenya. International Journal of Microbiology 2020, 1-10.

Prakash, B., Kumar, A., Singh, P.P., Songachan, L.S., 2020. Antimicrobial and antioxidant properties of phytochemicals: Current status and future perspective. Functional and Preservative Properties Of Phytochemicals 1-45.

Saitou, N., Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4(4), 406-425.

Shahi, S.K., Patra, M., Shukla, A.C. Dikshit, A., 2003. Use of essential oil as botanical-pesticide against post harvest spoilage in Malus pumilo fruit. BioControl 48, 223-232.

Shrivastava, D.K., Swarnkar, K., 2014. Antifungal activity of leaf extract of neem (Azadirachta indica Linn). International Journal of Current Microbiology and Applied Sciences 3(5), 305-308.

Singh, P., Cotty, P.J., 2019. Characterization of Aspergilli from dried red chilies (Capsicum spp.): insights into the etiology of aflatoxin contamination. International Journal of Food Microbiology 289, 145-153.

Sneath, P.H., Sokal, R.R., 1973. Numerical Taxonomy: The Principles and Practice of Numerical Classification. WF Freeman & Co., San Francisco, 573 p.

Szabo, B., Toth, B., Toth Toldine, E., Varga, M., Kovacs, N., Varga, J., Mesterházy, A., 2018. A new concept to secure food safety standards against Fusarium species and Aspergillus flavus and their toxins in maize. Toxins 10(9), 372.

Varga, J., Tóth, B., Kocsubé, S., Farkas, B., Szakács, G., Téren, J., & Kozakiewicz, Z., 2005. Evolutionary relationships among Aspergillus terreus isolates and their relatives. Antonie Van Leeuwenhoek, 88, 141-150.




DOI: https://doi.org/10.33804/pp.009.01.5475

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