Colonization of grapes berries by Alternaria sp. and their ability to produce mycotoxins
Keywords:Alternaria, altenuene, alternariol, alternariol monomethylether, grapes
Our research focused on identify the Alternaria species from grapes (surface sterilized berries and non-surface sterilized berries) of Slovak origin and characterize their toxinogenic potential in in vitro conditions. We analyzed 47 samples of grapes, harvested in years 2011, 2012 and 2013 from various wine-growing regions. For the isolation of species, the method of direct plating berries and surface-sterilized berries (using 1 % freshly pre-pared chlorine) on DRBC (Dichloran Rose Bengal Chloramphenicol agar) was used. For each analysis was used 50 berries. Only undamaged berries have been used for analysis. The cultivation was carried at 25 ±1°C, for 5 to 7 days in dark. After incubation, the colonies of Alternaria were transferred on PCA - potato-carrot agar and CYA - Czapek-yeast extract agar and cultured for 7 days at room temperature and natural light. A total 4 species-groups of the genus Alternaria were isolated from grapes berries: Alternaria alternata (1369 isolates), Alternaria arborescens (734 isolates), Alternaria infectoria (143 isolates), and Alternaria tenuissima (3579 isolates). According to European Union legislation mycotoxins produced by species genus Alternaria are not monitored in foods and food commodities. Mycotoxins such as alternariol and alternariol monomethylether are mutagenic and genotoxic in various in vitro systems. Selected strains were tested for production of altenuene, alternariol monomethylether and alternariol. In neither case of A. infectoria species-group isolates was confirmed the production of tested mycotoxins in in vitro conditions by TLC method. The ability to produce altenuene, alternariol monomethylether and alternariol in in vitro conditions was detected in isolates of Alternaria alternata, Alternaria arborescens and Alternaria tenuissima species-groups. Isolates of Alternaria alternata species-group (44 tested isolates) were able to produce altenuene (24 isolates), alternariol monomethyleter (42 isolates) and alternariol (43 isolates). Only one isolate did not produce any mycotoxins. Isolates of Alternaria arborescens species-group (38 tested isolates) were able to produce altenuene (24 isolates), alternariol monomethyleter (33 isolates) and alternariol (36 isolates). Only two isolates did not produce any mycotoxins. Isolates of Alternaria tenuissima species-group (87 tested isolates) were able to produce altenuene (42 isolates), alternariol monomethyleter (41 isolates) and alternariol (73 isolates). Thirteen isolates did not produce any mycotoxins.
Andersen, B., Kroger, E., Roberts, R. G. 2001. Chemical and morphological segregation of Alternaria alternata, A. gaisen and A. longipes. Mycological Research, vol. 105, no. 3, p. 291-299. https://doi.org/10.1017/s0953756201003446
Andersen, B., Kroger, E., Roberts, R. G. 2002. Chemical and morphological segregation of Alternaria arborescens, A. infectoria and A. tenuissima species-groups. Mycological Research, vol. 106, no. 2, p. 170-182. https://doi.org/10.1017/s0953756201005263
Andersen, B., Nielsen, K. F., Fernández Pinto, V., Patriarca, A. 2015. Characterization of Alternaria strains from Argentinean blueberry, tomato, walnuts and wheat. International Journal of Food Microbiology, vol. 196, p. 1-10. https://doi.org/10.1016/j.ijfoodmicro.2014.11.029
Barata, A., Malfeito-Ferreira, M., Loureiro, V. 2012. The microbial ecology of wine grape berries. International Journal of Food Microbiology, vol. 153, no. 3, p. 243-259. https://doi.org/10.1016/j.ijfoodmicro.2011.11.025
Bau, M., Bragulat, M. R., Abarca, M. L., Minguez, S., Cabañes, F. J. 2005. Ochratoxigenic species from Spanish wine grapes. International Journal of Food Microbiology, vol. 98, no. 2, p. 125-130. https://doi.org/10.1016/j.ijfoodmicro.2004.05.015
Chunmei, J., Junling, S., Qi´an, H., Yanlin, L. 2013. Occurrence of toxin-producing fungi in intact and rotten table and wine grapes and related influencing factors. Food Control, vol. 31, no. 1, p. 5-13. https://doi.org/10.1016/j.foodcont.2012.09.015
Diguta, C. F., Vincent, B., Guilloux-Benatier, M., Alexandre, H., Rousseaux, S. 2011. PCR ITS-RFLP: A useful method for identifyting filamentous fungi isolates on grapes. Food Microbiology, vol. 28, no. 6, p. 1145-1154. https://doi.org/10.1016/j.fm.2011.03.006
EFSA CONTAM Panel (EFSA Panel on Contaminants in the Food Chain), 2011. Scientific Opinion on the risks for animal and public health related to the presence of Alternaria toxins in feed and food. EFSA Journal 2011, vol. 9, no. 10, 97 p. https://doi.org/10.2903/j.efsa.2011.2407
Gautam, A., Sharma, S., Bhadauria, R. 2009. Detection of toxigenic fungi and mycotoxins in medicinally important powdered herbal drugs. The Internet Journal of Microbiology, vol. 7, no. 2, p. 1-8. https://doi.org/10.5580/104b
González, H. H. L., Pacin, A., Resnik, S. L., Martinez, E. J. 1996. Deoxynivalenol and contaminant mycoflora in freshly harvested Argentinean wheat in 1993. Mycopathologia, vol. 135, no. 2, p. 129-134. https://doi.org/10.1007/bf00436463
Labuda, R., Tančinová, D. 2006. Fungi recovered from slovakian poultry feed mixtures and their toxinogenity. Annals of Agricultural and Environmental Medicine, vol. 13, no. 2, 193-200. PMid:17195991
Labuda, R., Piovarčiová, Z., Tančinová, D. 2008. Sensitivity of the Alternaria infectoria species-group to cycloheximide. Letters in Applied Microbiology, vol. 46, no. 6, p. 673-675. https://doi.org/10.1111/j.1472-765x.2008.02370.x
Logrieco, A., Moretti, A., Solfrizzo, M. 2009. Alternaria toxins and plant diseases: an overview of origin, occurrence and risks. World Mycotoxin Journal, vol. 2, no. 2, p. 129-140. https://doi.org/10.3920/wmj2009.1145
López, P., Venema, D., de Rijk, T., de Kok, A., Scholten, J. M., Mol, H. G. J., de Nijs, M. 2016. Occurrence of Alternaria toxins in food products in the Netherlands. Food Control, vol. 60, p. 196-204. https://doi.org/10.1016/j.foodcont.2015.07.032
Lorenzini, M., Zapparoli, G. 2014. Characterization and pathogenicity of Alternaria spp. strains associated with grape bunch rot during post-harvest withering. International Journal of Food Microbiology, vol. 186, p. 1-5. https://doi.org/10.1016/j.ijfoodmicro.2014.06.008
Magnoli, C., Violante, M., Combina, M., Palacio, G., Dalcero, A. 2003. Mycoflora and ochratoxin-producing strains of Aspergillus section Nigri in wine grapes in Argentina. Letters in Applied Microbiology, vol. 37, no. 2, p. 179-184. https://doi.org/10.1046/j.1472-765x.2003.01376.x
Marin, S., Ramos, A. J., Cano-Sancho, G., Sanchis, V. 2013. Mycotoxins: Occurrence, toxicology, and exposure assessment. Food and Chemical Toxicology, vol. 60, p. 218-237. https://doi.org/10.1016/j.fct.2013.07.047
Mašková, Z., Tančinová, D., Barboráková, Z., Felšöciová, S., Císarová, M. 2012. Comparison of occurrence and toxinogenity of Alternaria spp. isolated from samples of conventional and new crossbred wheat of slovak origin. Journal of Microbiology, Biotechnology and Food Sciences, vol. 1, p. 552-562.
Mašková, Z., Tančinová, D., Rybárik, Ľ., Felšöciová, S., Kačinová, J. 2013. Colonization of grape berries by representatives of the genus Alternaria and their occurence in the stum. Journal of Microbiology, Biotechnology and Food Sciences, vol. 2, p. 1754-1764.
Ostrý, V., Škarková, J., Procházková, I., Kubátová, A., Malíř, F., Ruprich, J. 2007. Mycobiota of Czech wine grapes and occurrence of ochratoxin A and Alternaria mycotoxins in fresh grape juice, must and wine. Czech Mycology, vol. 59, p. 241-254.
Ostrý, V. 2008. Alternaria mycotoxins: an overview of chemical characterization, producers, toxicity, analysis and occurrence in foodstuffs. World Mycotoxin Journal, vol. 1, no. 2, p. 175-188. https://doi.org/10.3920/wmj2008.x013
Piovarčiová, Z., Labuda, R., Tančinová, D. 2007. Výskyt Alternaria spp. na zrnách potravinárskej pšenice dopestovanej na Slovensku v sezóne 2006. In Proceeding from workshop MICROMYCO 2007. České Budějovice : Department of soil biology, Czech Republic, p. 124-132.
Prendes, L. P., Merín, M. G., Andreoni, M. A., Ramirez, M., Morata de Ambrosini, V. I. 2015. Mycobiota and toxicogenic Alternaria spp. strains in Malbec wine grapes from DOC San Rafael, Mendoza, Argentina. Food Control, vol. 57, p. 122-128. https://doi.org/10.1016/j.foodcont.2015.03.041
Polizzotto, R., Andresen, B., Martini, M., Grisan, S., Assante, G., Musetti, R. 2012. A polyphasic approach for the characterization of endophytic Alternaria strains isolated from grapevines. Journal of Microbiological Methods, vol. 88, no. 1, p. 162-171. https://doi.org/10.1016/j.mimet.2011.11.009
Robiglio, A. L, Lopez, S. E. 1995. Mycotoxin production by Alternaria alternata strains isolated from Red Delicious apples in Argentina. International Journal of Food Microbiology, vol. 24, no. 3, p. 413-417. https://doi.org/10.1016/0168-1605(94)00035-5
Rotem, J. 1994. The genus Alternaria: biology, epidemiology and pathogenicity. St. Paul, MN: The American Phytopathological Society, 326 p.
Rousseaux, S., Diguta, C. F., Radoï-Matei. 2014. Non-Botrytis grape-rotting fungi responsible for earthy and moldy off-flavors and mycotoxins. Food Microbiology, vol. 38, p. 104-121. https://doi.org/10.1016/j.fm.2013.08.013
Serra, R., Braga, A., Venâncio, A. 2005. Mycotoxin-producing and other fungi isolated from grapes for wine production, with particular emphasis on ochratoxin A. Research in Microbiology, vol. 156, no. 4, p. 515-521. https://doi.org/10.1016/j.resmic.2004.12.005
Serra, R., Lourenço, A., Alípio, P., Venâncio, A. 2006. Influence of the region of origin on the mycobiota of grapes with emphasis on Aspergillus and Penicillium species. Mycological research, vol. 110, no. 8, p. 971-978. https://doi.org/10.1016/j.mycres.2006.05.010
Samson, R. A., Hoekstra, E. S., Frisvad, J. C., Filtenborg, O. 2002. Introduction to food- and airborne fungi. Utrecht: Centraalbureau voor Schimmelcultures, 2002. 389 p. ISBN 90-70351-42-0. https://doi.org/10.5580/104b
Simmons, E. G. 1994. Alternaria themes and variations (106-111). Mycotaxon, 1994, vol. 50, p. 409-427.
Simmons, E. G. 2007. Alternaria, An Identification Manual. Utrecht : CBS Fungal Biodiversity Centre, 2007. 775 p. ISBN 978-90-70351-68-7.
Simmons, E. G., Roberts, R. G. 1993. Alternaria themes and variations. In Mycotaxon, vol. 73, p. 109-140.
Tančinová, D., Rybárik, Ľ., Mašková, Z., Felšöciová, S., Císarová, M. 2015. Endogenal colonization of grapes berries. Journal of Microbiology, Biotechnology and Food Sciences vol. 4, sp. no. 1, p. 69-73. https://doi.org/10.15414/jmbfs.2015.4.special1.69-73
Tournas, V. H., Katsoudas, E. 2005. Mould and yeast flora in fresh berries, grapes and citrus fruits. International Journal of Food Microbiology, vol. 105, no. 1, p. 11-17. https://doi.org/10.1016/j.ijfoodmicro.2005.05.002
Woundenberg, J. H. C., Seidl, M. F., Groenewald, J. Z., de Vries, M., Stielow, J. B., Thomma, B. P. H. J., Crous, P. W. 2015. Alternaria section Alternaria: Species, formae speciales or pathotypes? Studies in Mycology, vol 82, p. 1-21. https://doi.org/10.1016/j.simyco.2015.07.001
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