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Abstract
Some secondary metabolites produced by fungi are carcinogenic, hepatotoxic, and/or cause birth defects in humans and animals. We developed and optimised bio-analytical tools for detection of metabolites, aflatoxins and evaluated the effectiveness of the methods in co-infected maize tissues. Isolate KSM012 (atoxigenic) demonstrated no peaks and no blue fluorescence on HPLC and TLC plates respectively confirming non-toxicity. AFB1 and AFB2 were produced by Isolate KSM015 in addition to AFG1 and AFG2, which is an indication of possible SBG morphotype. The limits of quantification and detection ranged from 0.02 to 35.81 µg/mL and 0.01–6.8 µg/mL, respectively. The best mass spectrum with lowest noise was obtained at 100% ACN and sterile water spiked with 0.1% formic acid at a flow rate of 0.3 mL/min. The positive ion mode with electrospray ionisation application exhibited better fragmentation for mycotoxins. In total 17 metabolites were detected by targeted and formula mass. KDVI maize line exhibited high fungal colonisation in comparison to GAF4 at equal co-infection ratio 50:50. AFB1 and AFG2 were remarkably higher in GAF4 in comparison to sensitive KDV1 (p ˂ 0.05). The detection limits, linearity and sensitivity showed the method developed was suitable for the determination of mycotoxin in comparisons to the guide-lines of European Commission 657/EC 2002.
References
Atehnkeng J, Donner M, Ojiambo PS, Ikotun B, Augusto J, Cotty PJ, Bandyopadhyay R. 2016. Environmental dis-tribution and genetic diversity of vegetative compatibil-ity groups determine biocontrol strategies to mitigate aflatoxin contamination of maize by a spergillus flavus. Microb Biotechnol. 9(1):75–88. doi:10.1111/1751-7915.12324.
Bandyopadhyay R, Ortega-Beltran A, Akande A, Mutegi C, Atehnkeng J, Kaptoge L, Senghor AL, Adhikari BN, Cotty PJ. 2016. Biological control of aflatoxins in Africa: current status and potential challenges in the face of climate change. World Mycotoxin J. 9(5):771–789. doi:10.3920/WMJ2016.2130.
Bertuzzi T, Rastelli S, Mulazzi A, Pietri A. 2012. Evaluation and improvement of extraction methods for the analysis of aflatox-ins B1, B2, G1 and G2 from naturally contaminated maize. Food Anal Methods. 5(3):512–519. doi:10.1007/s12161-011-9274-5.
De LF, Andrew A, Esther S, Jenny B, Hejazi F, Hayes S, Morrison IEG, Prior BA, McGenity TJ, Rangel DEN, et al. 2015. Concomitant osmotic and chaotropicity - induced stres-ses in Aspergillus wentii : compatible solutes determine the biotic window. Curr Genet. 61(3):457–477. doi:10.1007/ s00294-015-0496-8.
[EC] European Commision 2002 Commission Decision No 657/ EC. August 17. Commission decision of 12 August 2002 implementing council directive 96/23/EC concerning the per-formance of analytical methods and the interpretation of results (notified under document number C (2002) 3044)-(Text with EEA relevance) (2002/657/EC). Off J Eur Communities. 29.
[EC] European Commission. 2011. Method validation & quality control procedures for pesticide residue analysis in food and feed. Doc No SANCO/12495/2011. 2011;41. Filtenborg O, Frisvad JC, Svendsen JA. 1983. Simple screening method for moulds producing intracellular mycotoxins in pure cultures. Appl Environ Microbiol. 45(2):581–585.
doi:10.1128/AEM.45.2.581-585.1983.
Frisvad JC, Smedsgaard J, Larsen TO, Samson RA. 2004. Mycotoxins, drugs and other extrolites produced by species in Penicillium subgenus Penicillium. Stud Mycol. 49:201–241.
Gallo A, Masoero F, Bertuzzi T, Piva G, Pietri A. 2010. Effect of the inclusion of adsorbents on aflatoxin B 1quantification in animal feedstuffs. Food Addit Contam - Part A. 27
(1):54–63. doi:10.1080/02652030903207219.
Hayat A, Paniel N, Rhouati A, Marty JL, Barthelmebs L. 2012. Recent advances in ochratoxin A-producing fungi detection based on PCR methods and ochratoxin A analysis in food matrices. Food Control. 26:401–415. doi:http://dx. doi.10.1016/j.foodcont.2012.01.060
Helrich K. 1990. AOAC: official methods of analysis (Volume 1). 15th ed. Arlington (VA): Contam Agric Chem;
p. 1298.
Herzallah SM. 2009. Determination of aflatoxins in eggs, milk, meat and meat products using HPLC fluorescent and UV detectors. Food Chem. 114(3):1141–1146. doi:10.1016/j. foodchem.2008.10.077.
Horn BW, Greene RL. 1995. Vegetative compatibility within populations of Aspergillus flavus, A. parasiticus, and
A. tamarii from a peanut field. Mycologia. 87(3):324–332. doi:10.1080/00275514.1995.12026537.
Hruska Z, Rajasekaran K, Yao H, Kincaid R, Darlington D, Brown RL, Bhatnagar D, Cleveland TE. 2014. Co-inoculation of aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus to study fungal invasion, colonization, and competition in maize kernels. Front Microbiol. 5:122. doi:10.3389/fmicb.2014.00122.
Huang C, Jha A, Sweany R, DeRobertis C, Damann KE. 2011. Intraspecific aflatoxin inhibition in Aspergillus flavus is thigmoregulated, independent of vegetative compatibility group and is strain dependent. PLoS One. 6(8):e23470. doi:10.1371/journal.pone.0023470
Jaime-Garcia R, Cotty PJ. 2006. Spatial distribution of Aspergillus flavus and its toxigenic strains on commercial cottonseed from south Texas and its relationship to afla-toxin contamination. Plant Pathol. 55(3):358–366. doi:10.1111/j.1365-3059.2006.01358.x.
Kok A. 1986. Effects of degradation of visual stimuli on com-ponents of the event-related potential (ERP) in go/no go reaction tasks. Biol Psychol. 23(1):21–38. doi:10.1016/0301-0511(86)90087-6.
Li P, Zhang Q, Zhang D, Guan D, Liu X, Fang S, Wang X, Zhang W 2011. Aflatoxin measurement and analysis. Chapter in ‘aflatoxins - detection, measurement and con-trol’, Irineo Torres-Pacheco, intechOpen. https://www.inte chopen.com/books/aflatoxins-detection-measurement-and
-control/aflatoxin-measurement-and-analysis
Li P, Zhang Q, Zhang W. 2009. Immunoassays for aflatoxins. Trends Anal Chem. 28(9):1115–1126. doi:10.1016/j. trac.2009.07.003.
Malachová A, Sulyok M, Beltrán E, Berthiller F, Krska R. 2014. Optimization and validation of a quantitative liquid chro-matography–tandem mass spectrometric method covering 295 bacterial and fungal metabolites including all regulated mycotoxins in four model food matrices. J Chromatogr A. 1362:145–156. doi:10.1016/j.chroma.2014.08.037.
Medina A, Rodríguez A, Sultan Y, Magan N. 2015. Climate change factors and Aspergillus flavus: effects on gene expression, growth and aflatoxin production. World Mycotoxin J. 8(2):171–179. doi:10.3920/WMJ2014.1726.
Mehl HL, Cotty PJ. 2010. Variation in competitive ability among isolates of Aspergillus flavus from different vegetative
compatibility groups during maize infection. Phytopathology®. 100(2):150–159. doi:10.1094/PHYTO-100-
2-0150.
Mehl HL, Cotty PJ. 2011. Influence of the host contact sequence on the outcome of competition among Aspergillus flavus isolates during host tissue invasion. Appl Environ Microbiol. 77(5):1691–1697. doi:10.1128/AEM.02240-10.
Mehl HL, Cotty PJ. 2013. Influence of plant host species on intraspecific competition during infection by Aspergillus flavus. Plant Pathol. 62(6):1310–1318. doi:10.1111/ppa.12038. Mehl HL, Jaime R, Callicott KA, Probst C, Garber NP, Ortega-Beltran A, Grubisha LC, Cotty PJ. 2012. Aspergillus flavus diversity on crops and in the environment can be exploited to reduce aflatoxin exposure and improve health. Ann NY Acad
Sci. 1273(1):7–17. doi:10.1111/j.1749-6632.2012.06800.x.
Mitema A, Okoth S, Rafudeen MS. 2018. Vegetative compat-ibility and phenotypic characterization as a means of deter-mining genetic diversity of Aspergillus flavus isolates. Fungal Biol. 122(4):203–213. doi:10.1016/j.funbio.2017.11.005.
Mitema A, Okoth S, Rafudeen MS. 2019. The development of a qPCR assay to measure aspergillus flavus biomass in maize and the use of a biocontrol strategy to limit aflatoxin production. Toxins. 11(3):179. doi:10.3390/toxins11030179. Möller TE, Nyberg M. 2004. Efficiency of different extraction solvent mixtures used in analyses of aflatoxins from a certified peanut meal reference material. Food Addit Contam. 21
(8):781–785. doi:10.1080/02652030412331282394.
Newman D, Cragg G, Snader K. 2003. Natural products as sources of new drugs over the period 1981-2002. J Nat Prod. 66(7):1022–1037. doi:10.1021/np030096l.
Nielsen K. 2003. Fungal metabolite screening: database of 474 mycotoxins and fungal metabolites for dereplication by standardised liquid chromatography–UV–mass spectrome-try methodology. J Chromatogr A. 1002(1–2):111–136.
Nielsen KF, Smedsgaard J. 2003. Fungal metabolite screening: database of 474 mycotoxins and fungal metabolites for dereplication by standardised liquid chromatography-UV-mass spectrometry methodology. J Chromatogr A. 1002(-1–2):111–136. doi:10.1016/S0021-9673(03)00490-4.
Papp E, Klara H-Otta GZ, Mincsovics E. 2002. Liquid chro-matographic determination of aflatoxins. Microchem J. 73 (1–2):39–46. doi:10.1016/S0026-265X(02)00048-6.
Payne GA, Yu J. 2010. Ecology, development and gene regula-tion in Aspergillus flavus. Mol Biol Genomics. 8:157–171. Pramanik BN, Mirza UA, Ing YH, Liu Y-H, Bartner PL,
Weber PC, Bose AK. 2002. Microwave-enhanced enzyme reaction for protein mapping by mass spectrometry : A new approach to protein digestion in minutes. Protein Sci. 11 (11):2676–2687. doi:10.1110/ps.0213702.
Raeder U, Broda P. 1985. Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol. 1(1):17–20. doi:10.1111/j.1472-765X.1985.tb01479.x.
Songsermsakul PER-F. 2008. A review of recent trends in appli-cations of liquid chromatography-mass spectrometry for determination of mycotoxins liquid chromatography-mass spectrometry for. J Liq Chromatogr Relat Technol. 6076(11–-12):1641–1686. doi:10.1080/10826070802126395.
Stevenson A, Cray JA, Williams JP, Santos R, Sahay R, Neuenkirchen N, McClure CD, Grant IR, Houghton J, Quinn JP, et al. 2015. Is there a common water-activity limit for the three domains of life? Isme J. 9(6):1333–1351. doi:10.1038/ismej.2014.219.
Sulyok M, Krska R, Schuhmacher R. 2007. A liquid chromato-graphy/tandem mass spectrometric multi-mycotoxin method for the quantification of 87 analytes and its application to semi-quantitative screening of moldy food samples. Anal Bioanal Chem. 389(5):1505–1523. doi:10.1007/s00216-007-1542-2.
Taylor SL, King JW, Richard JL, Greer JI. 1993. Analytical-scale supercritical fluid extraction of aflatoxin B1 from field-inoculated corn. J Agric Food Chem. 41(6):911. doi:10.1021/jf00030a014.
Trucksess MW, Nesheim S, Eppley RM. 1984. Thin layer chromatographic determination of deoxynivalenol in wheat and corn. J Assoc Off Anal Chem. 67(4):4244.
Villas-bo SG, Smedsgaard J, Nielsen J. 2005. Mass spectro-metry in metabolome analysis. Mass Spectrom Rev. 24 (5):613–646. doi:10.1002/mas.20032.
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