Screening of Aflatoxin Production by Aspergillus flavus Isolates from Petroleum-contaminated Soil
Fungi are eukaryotic, heterogeneous, unicellular to filamentous, spore-bearing, and chemoorganotrophic organisms which lack chlorophyll. This present study was carried out to isolate and identify fungi from petroleum-contaminated soil. Several fungal genera which included Rhizopus spp., Mucor spp., Penicillium spp., Rhizoctonia spp., Aspergillus spp., Alternaria spp., and Cladosporiumspp. were isolated using potatoes dextrose agar, Czapek-Dox Agar, and Aspergillus flavus Differentiation Agar culture media that comparable with co-amoxiclav (1g) and chloramphenicol to prevent the growth of any bacteria. The direct plate and serial dilution agar plate methods were used for the isolation of fungi. Based on results, Aspergillusand Mucor spp. were the most predominant genera and had the highest number of colonies in the soil samples. In this investigation, seven out of 27 soil samples were morphologically (macroscopically and microscopically) identified, such as A. flavus. Aflatoxigenicity of A. flavus was detected using characteristics in Aspergillus differentiation agar and colony fluorescence on exposure to ultraviolet light. Moreover, molecular approaches were used for the detection of aflatoxigenic of the A. flavus isolates. Three structural (aflD, aflO, and aflP) and one regulatory (aflR) gene of the aflatoxin gene cluster of A. flavus were targeted for amplification by the polymerase chain reaction method. The aflatoxigenic of all six A. flavus isolates was detected molecularly which contained two structural (aflD, aflP)genes out of three structural genes, while there was no specific amplification of the aflO gene in the fourth, fifth, and sixth A. flavus which issimilar to the aflR gene in the first and second A. flavus.
Abdel-Hadi, A., 2011. Molecular Ecology of Aspergillus Section Flavi Species: Approaches to Understanding the Role of Aflatoxin Genes in Aflatoxin Biosynthesis.
Abdel-Hadi, A., Carter, D., and Magan, N., 2011. Discrimination between aflatoxigenic and non-aflatoxigenic Aspergillus section Flavi strains from Egyptian peanuts using molecular and analytical techniques. World Mycotoxin Journal, 4(1), pp.69-77. DOI: https://doi.org/10.3920/WMJ2010.1223
Agarwal, P., 2017. Isolation and characterization of Tyrosinase (a carbon trapping enzyme) producing microorganisms, in the agricultural soil of Western Uttar Pradesh and the study of enzymatic activity of Tyrosinase produced. Biochemistry and Molecular Biology Letters, 3(1), p.105.
Almoammar, H., Bahkali, A.H., and Abd-Elsalam, K.A., 2013. A polyphasic method for the identification of aflatoxigenic ‘Aspergillus’ species isolated from Camel feeds. Australian Journal of Crop Science, 7(11), p.1707.
Blakely, J.K., Neher, D.A. and Spongberg, A.L., 2002. Soil invertebrate and microbial communities, and decomposition as indicators of polycyclic aromatic hydrocarbon contamination. Applied Soil Ecology, 21(1), p.71-88. DOI: https://doi.org/10.1016/S0929-1393(02)00023-9
Carbone, I., Jakobek, J.L., Ramirez‐Prado, J.H., and Horn, B.W., 2007. Recombination, balancing selection and adaptive evolution in the aflatoxin gene cluster of Aspergillus parasiticus. Molecular Ecology, 16(20), pp.4401-4417. DOI: https://doi.org/10.1111/j.1365-294X.2007.03464.x
Criseo, G., Bagnara, A., and Bisignano, G., 2001. Differentiation of aflatoxin‐ producing and non-producing strains of Aspergillus flavus group. Letters in Applied Microbiology, 33(4), pp.291-295. DOI: https://doi.org/10.1046/j.1472-765X.2001.00998.x
Degola, F., Berni, E., Dall’Asta, C., Spotti, E., Marchelli, R., Ferrero, I., and Restivo, F.M., 2007. Amultiplex RT‐PCR approach to detect aflatoxigenic strains of Aspergillus flavus. Journal of Applied Microbiology, 103(2), pp.409-417. DOI: https://doi.org/10.1111/j.1365-2672.2006.03256.x
Dheeb, B.I., Ismail, E.N., AL-mishhadani, I.I.H., Majeed, S.M., and Majeed, D.M., 2014. Astudy of the expression of aflatoxin B1 regulatory gene in clinical and environmental Aspergillus flavus using real-time PCR. International Journal of Sciences: Basic and Applied Research, 17(1), pp.417-427.
Erami, M., Hashemi, S.J., Pourbakhsh, S.A., Shahsavandi, S., Mohammadi, S., Shoushtari, A., and Jahan, S.Z., 2007. Application of PCR on detection of aflatoxinogenic fungi; short communication. Archives of Razi Institute, 62, pp.95-100.
Farrag, A.A., Shehata, R.M., EL-Sheikh, H.H., Abo-Dahab, N.F., and Ali, A.M., 2017. Diversity and biotechnological applications of some fungi that isolated from unusual soil samples in Egypt. Journal of Ecology of Health and Environment, 5(1), pp.23-33. DOI: https://doi.org/10.18576/jehe/050104
Gallo, A., Stea, G., Battilani, P., Logrieco, A.F., and Perrone, G., 2012. Molecular characterization of an Aspergillus flavus population isolated from maize during the first outbreak of aflatoxin contamination in Italy. Phytopathologia Mediterranea, 51, pp.198-206.
Giusiano, G.E., Piontelli, E., Fernández, M.S., Mangiaterra, M.L., Cattana, M.E., Kocsubé, S., and Varga, J., 2017. Biodiversity of species of Aspergillus section Fumigati in semi-desert soils in Argentina. Revista Argentina de microbiologia, 49(3), pp.247-254. DOI: https://doi.org/10.1016/j.ram.2017.02.002
Hanson, J.R., 2008. Chemistry of Fungi. Royal Society of Chemistry, United Kingdom. DOI: https://doi.org/10.1039/9781847558329
Harris, S.D., 2008. Branching of fungal hyphae: Regulation, mechanisms and comparison with other branching systems. Mycologia, 100(6), pp.823-832. DOI: https://doi.org/10.3852/08-177
Hedayati, M.T., Pasqualotto, A.C., Warn, P.A., Bowyer, P., and Denning, D.W., 2007. Aspergillus flavus: Human pathogen, allergen and mycotoxin producer. Microbiology, 153(6), pp.1677-1692. DOI: https://doi.org/10.1099/mic.0.2007/007641-0
Houshyarfard, M., Rouhani, H., Falahati-Rastegar, M., Malekzadeh-Shafaroudi,S., Mehdikhani-Moghaddam, E., and Chang, P.K., 2014. Gene deletion patterns in non-aflatoxigenic isolates of Aspergillus flavus. Mycologia Iranica, 1(2), pp.87-97.
Hussain, A., Afzal, A., Irfan, M., and Malik, K.A., 2015. Molecular detection of aflatoxin-producing strains of Aspergillus flavus from peanut (Arachis hypogaea). Turkish Journal of Agriculture-Food Science and Technology, 3(5), pp.335-341. DOI: https://doi.org/10.24925/turjaf.v3i5.335-341.123
Jasuja, N.D., Saxena, R., and Joshi, S.C., 2013. Isolation and identification of microorganisms from playhouse agriculture soil of Rajasthan. African Journal of Microbiology Research, 7(41), pp.4886-4891. DOI: https://doi.org/10.5897/AJMR2012.2413
Khalid, S., Hussain, N., and Imran, M., 2018. Detection of aflatoxigenicity of Aspergillus flavus, based on potential gene marker, from food and feed samples. Journal of Food Safety, 38, p.e12448. DOI: https://doi.org/10.1111/jfs.12448
Kvenvolden, K.A., and Cooper, C.K., 2003. Natural seepage of crude oil into the marine environment. Geo-Marine Letters, 23(3-4), pp.140-146. DOI: https://doi.org/10.1007/s00367-003-0135-0
Machida, M., and Gomi, K., Eds., 2010. Aspergillus: Molecular Biology and Genomics. United Kingdom: Horizon Scientific Press.
Martins, M.L., Martins, H.M., and Bernardo, F., 2001. Aflatoxins in spices marketed in Portugal. Food Additives and Contaminants, 18(4), pp.315-319. DOI: https://doi.org/10.1080/02652030120041
Nair, S.C., Bhagobaty, R.K., Nampoothiri, K., Kalaigandhi, V., and Menon, K.R.K., 2014. Detection of Aflatoxin production by fungi in spice samples using HPLC and direct visual cultural methods. Innovative Romanian Food Biotechnology, 14, pp.1-12.
Raja, M., Praveena, G., and William, S.J., 2017. Isolation and identification of fungi from soil in Loyola College campus, Chennai, India. International Journal of Current Microbiology and Applied Sciences, 6(2), pp.1789-1795. DOI: https://doi.org/10.20546/ijcmas.2017.602.200
Ramírez-Camejo, L.A., Zuluaga-Montero, A., Lázaro-Escudero, M., Hernández Kendall, V., and Bayman, P., 2012. Phylogeography of the cosmopolitan fungus Aspergillus flavus: Is everything everywhere? Fungal Biology, 116(3), pp.452-463. DOI: https://doi.org/10.1016/j.funbio.2012.01.006
Rodrigues, P., Soares, C., Kozakiewicz, Z., Paterson, R., Lima, N., and Venâncio,A., 2007. Identification and characterization of Aspergillus flavus and aflatoxins. In: Communicating Current Research and Educational Topics and Trends in Applied Microbiology. Formatex, Mumbai.
Rohilla, S.K., and Salar, R.K., 2012. Isolation and characterization of various fungal strains from agricultural soil contaminated with pesticides. Research Journal of Recent Sciences ISSN, 2277, p.2502.
Scherm, B., Palomba, M., Serra, D., Marcello, A., and Migheli, Q., 2005. Detection of transcripts of the aflatoxin genes aflD, aflO, and aflP by reverse transcription-polymerase chain reaction allows differentiation of aflatoxin producing and non-producing isolates of Aspergillus flavus and Aspergillus parasiticus. International Journal of Food Microbiology, 98(2), pp.201-210. DOI: https://doi.org/10.1016/j.ijfoodmicro.2004.06.004
Soni, S.K., 2007. Microbes: A Source of Energy for the 21st Century. New India Publishing, New Delhi. DOI: https://doi.org/10.59317/9788194281542
Temu, G.E., 2016. Molecular Identification of Aspergillus strains and quick detection of aflatoxin from selected common spices in Tanzania. Journal of Scientific Research and Reports, 10, pp.1-8. DOI: https://doi.org/10.9734/JSRR/2016/26102
Yazdani, D., Zainal, A.M., Tan, Y.H., and Kamaruzaman, S., 2010. Evaluation of the detection techniques of toxigenic Aspergillus isolates. African Journal of Biotechnology, 9(45), pp.7654-7659.
Yin, Y.N., Yan, L.Y., Jiang, J.H., and Ma, Z.H., 2008. Biological control of aflatoxin contamination of crops. Journal of Zhejiang University Science B, 9(10), pp.787-792 DOI: https://doi.org/10.1631/jzus.B0860003
Copyright (c) 2023 Sawan M. Mirkhan, Taha J. Omar
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors who choose to publish their work with Aro agree to the following terms:
Authors retain the copyright to their work and grant the journal the right of first publication. The work is simultaneously licensed under a Creative Commons Attribution License [CC BY-NC-SA 4.0]. This license allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
Authors have the freedom to enter into separate agreements for the non-exclusive distribution of the journal's published version of the work. This includes options such as posting it to an institutional repository or publishing it in a book, as long as proper acknowledgement is given to its initial publication in this journal.
Authors are encouraged to share and post their work online, including in institutional repositories or on their personal websites, both prior to and during the submission process. This practice can lead to productive exchanges and increase the visibility and citation of the published work.
By agreeing to these terms, authors acknowledge the importance of open access and the benefits it brings to the scholarly community.