PLANT BIOLOGY
ANIMAL BIOLOGY
SUBSCRIPTION
E-SUBSCRIPTION
 
MAP
MAIN PAGE

 

 

 

 

Kononenko G.P., Piryazeva E.A., Burkin A.A. Toxin-producing small-spore Alternaria species from oat grain contaminated with alternariol. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2023, Vol. 58, № 3, p. 567-575.
(how to cite this article)

doi: 10.15389/agrobiology.2023.3.567eng

UDC: 636.085.19:636.086.1/.3:632.4

 

TOXIN-PRODUCING SMALL-SPORE Alternaria SPECIES FROM OAT GRAIN CONTAMINATED WITH ALTERNARIOL

G.P. Kononenko✉, E.A. Piryazeva, A.A. Burkin

All-Russian Research Institute of Sanitary, Hygiene and Ecology — Branch of FSC Skryabin and Kovalenko All-Russian Research Institute of Experimental Veterinary RAS, 5/1, Zvenigorodskoe sh., Moscow, 123022 Russia, e-mail
kononenkogp@mail.ru (✉ corresponding author), piryazeva01@yandex.ru, aaburkin@mail.ru

ORCID:
Kononenko G.P. orcid.org/0000-0002-9144-615X
Burkin A.A. orcid.org/0000-0002-5674-2818
Piryazeva E.A. orcid.org/0000-0001-5443-3213

Final revision received March 18, 2023
Accepted April 07, 2023

For many years, the problem of grain infestation with toxin-forming fungi Alternaria has been under the close attention (S.M. Tralamazza et al., 2018). Extensive studies have been carried out on wheat (M.T. Amatulli et al., 2013; M.E. Müller, U. Korn, 2013) and barley (V. Sanchis et al., 1993; T.T.T. Nguyen et al., 2018). The grain of oats has been studied much less, and it is still unclear which species of fungi of this genus and to what extent are responsible for the accumulation of the toxin alternariol (AOL). In our country, data have been obtained on the infection with Alternaria fungi of oat grains from a number of regions (O.P. Gavrilova et al., 2016; Yu.I. Vargach et al., 2019), as well as grain samples of varieties and lines from the VIR collection detected in field tests (A.S. Orina et al., 2017), however, the frequency of occurrence of the toxin was estimated for several regional lots in total (A.A. Burkin et al., 2015; G.P. Kononenko et al., 2020). In this study, it was established for the first time that the species A. tenuissima (Nees et T. Nees:Fries) Wiltshire, which is known as an active AOL producer, and to a lesser extent representatives of A. arborescens E.G. Simmons and the ‘A. infectoria’ complex can participate in the contamination of oat grains. The aim of the work was to study the species affiliation and toxin-forming ability of Alternaria fungi isolated from oat grain with natural AOL contamination. The object of the study was a sample obtained in October 2020 from an agricultural enterprise of the Moscow Province (Odintsovo District) containing AOL in the amount of 630 ppb. Isolation of pure fungal cultures was carried out after surface sterilization of grain and its sowing on Chapek-Dox agar containing bile and antibiotics. Color, structure, and growth rate of colonies were described on yeast extract sucrose agar (YES) and potato-carrot (PCA) on day 8. To assess their toxin formation, oat grain and a panel of four mycological media were used — PCA, malt extract agar (MEA), hay infusion agar (HAY) and an analog of vegetable agar (V-8). After cultivation (7 days, 25 °C, without lighting) and extraction of biomass samples with a mixture of acetonitrile and water in a volume ratio of 84:16, AOL was determined by ELISA test (A.A. Burkin, G.P. Kononenko, 2011) with a detection limit of 0.01 μg/g. In the subepidermal mycobiota of the studied sample, representatives of the genus Alternaria were predominant, the degree of infection was 36.0 %, and they were accompanied by fungi Fusarium spp. (14.7 %) and Epicoccum spp. (2.7 %). After carrying out mycological procedures for the isolation and identification of Alternaria cultures, they were assigned to the species A. tenuissima (Nees et T. Nees:Fries) Wiltshire (7 strains), A. arborescens E.G. Simmons (2 strains) and to the species complex ‘A. infectoria' (4 isolates). On the grain substrate, all strains of A. tenuissima, A. arborescens and three isolates of ‘A. infectoria’ produced AOL in amounts of 370, 5 and 0.8 μg/g. When testing cultures under the same conditions on agar media, the intensity of AOL accumulation in A. tenuissima was highest on HAY and MEA (56 and 23 μg/g), in A. arborescens and ‘A. infectoria’ — on PCA and MEA. Taking this into account, commercial substrate MEA and oat grains are recommended for in vitro evaluation of the biosynthetic potential of fungi and their involvement in contamination of AOL grains in an expanded format. The cultural and morphological features of several cultures of A. arborescens and ‘A. infectoria’ and their ability to AOL biosynthesis are discussed in a comparative aspect.

Keywords: oat grain, Alternaria tenuissima, A. arborescens, ‘A. infectoria’, alternariol, ELISA.

 

REFERENCES

  1. Amatulli M.T., Fanelli F., Moretti A., Mule G., Logrieco A.F. Alternaria species and mycotoxins associated to black point of cereals. Mycotoxins, 2013, 63(1): 39-46.
  2. Müller M.E., Korn U. Alternaria mycotoxins in wheat — a 10 years survey in the Northeast of Germany. Food Control, 2013, 34(1): 191-197 CrossRef
  3. Tralamazza S.M., Piacentini K.C., Iwase C.H.T., De Oliveira Rocha L. Toxigenic Alternaria species: impact in cereals worldwide. Current Opinion in Food Science, 2018, 23: 57-63 CrossRef
  4. Sanchis V., Sanclemente A., Usall J., Viñas I. Incidence of mycotoxigenic Alternaria alternata and Aspergillus flavus in barley. Journal of Food Protection, 1993, 56(3): 246-248 CrossRef
  5. Nguyen T.T.T., Kim J., Jeon S.J., Lee C.W., Magan N., Lee H.B. Mycotoxin production of Alternaria strains isolated from Korean barley grains determined by LC-MS/MS. International Journal of Food Microbiology, 2018, 268: 44-52 CrossRef
  6. Fodder oats: a world overview. J.M. Suttie, S.G. Reynolds (eds.). Plant Production and Protection Series No. 33. FAO, Rome, 2004.
  7. Sacchi C., González H.H.L., Broggi L.E., Pacin A., Resnik S.L., Cano G., Taglieri D. Fungal contamination and mycotoxin natural occurrence in oats for race horses feeding in Argentina. Animal Feed Science and Technology, 2009, 152(3-4): 330-335 CrossRef
  8. EFSA (European Food Safety Authority). Scientific opinion on the risks for animal and public health related to the presence of Alternaria toxins in feed and food. EFSA Journal, 2011, 9(10): 2407-2505 CrossRef
  9. Lee H.B., Patriarca A., Magan N. Alternaria in food: ecophysiology, mycotoxin production and Toxicology. Mycobiology, 2015, 43(2): 93-106 CrossRef
  10. Gruber-Schley S., Thalmann A. The occurrence of Alternaria spp. and their toxins in grain and possible connections with illness in farm animals. Landwirtschaftliche Forschung, 1988, 41(1-2): 11-29.
  11. Logrieco A., Bottalico A., Solfrizzo M., Mule G. Incidence of Alternaria species in grains from Mediterranean countries and their ability to produce mycotoxins. Mycologia, 1990, 82(4): 501-505 CrossRef
  12. Häggblom P., Stepinska A., Solyakov A. Alternaria mycotoxins in Swedish feed grain. Proc. 29th Mycotoxin-Workshop. Gesellschaft für Mykotoxin Forschung, Stuttgart-Fellbachm, 2007: 35.
  13. Uhlig S., Sundstøl Eriksen G., Skow Hofgaard I., Krska R., Beltrán E., Sulyok M. Faces of changing climate: semi-quantitative multi-mycotoxin analysis of grain grown in exceptional climatic conditions in Norway. Toxins, 2013, 5(10): 1682-1697 CrossRef
  14. Tittlemier S.A., Blagden R., Chan J., Roscoe M., Pleskach K. A multi-year survey of mycotoxins and ergosterop in Canadian oats. Mycotoxin Research, 2020, 36: 103-114 CrossRef
  15. De Colli L., De Ruyck K., Abdallah M.F., Finnan J., Mullins E., Kildea S., Spink J., Elliott Ch., Danaher M. Natural co-occurrence of multiple mycotoxins in unprocessed oats grown in Ireland with various production systems. Toxins, 2021, 13(3): 188 CrossRef
  16. Babič J., Tavčar-Kalcher G., Celar F.A., Kos K., Knific T., Jakovac-Strajn B. Occurrence of Alternaria and other toxins in cereal grains intended for animal feeding collected in Slovenia: A three-year study. Toxins, 2021, 13(5): 304 CrossRef
  17. Gavrilova O.P., Gannibal F.B., Gagkaeva T.Yu. Fusarium and Alternaria fungi in grain of oats grown in the North-Western Russia regarding cultivar specificity. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2016, 51(1): 111-118 CrossRef
  18. Vargach Yu.I., Golovin S.E., Loskutov I.G. Trudy po prikladnoy botanike, genetike i selektsii, 2019, 180(3): 96-105 CrossRef (in Russ.).
  19. Orina A.S., Gavrilova O.P., Gagkaeva T.Yu., Loskutov I.G. Symbiotic relationships between aggressive Fusarium and Alternaria fungi colonizing oat grain. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2017, 52(5): 986-994 CrossRef
  20. Gavrilova O.P., Gagkaeva T.Yu., Orina A.S., Markova A.S., Kabashov A.D., Loskutov I.G. Trudy po prikladnoy botanike, genetike i selektsii, 2020, 181(2): 134-144 CrossRef (in Russ.).
  21. Burkin A.A., Kononenko G.P., Gavrilova O.P., Gagkaeva T.Yu. Sovremennaya mikologiya v Rossii, 2015, 5(5): 221-223 (in Russ.).
  22. Kononenko G.P., Burkin A.A., Zotova E.V. Veterinariya segodnya, 2020, 2(33): 139-145 CrossRef (in Russ.).
  23. Orina A.S., Gavrilova O.P., Gagkaeva T.Yu., Gannibal F.B. Mikologiya i fitopatologiya, 2020, 54(5): 365-377 CrossRef (in Russ.).
  24. Orina A.S., Gavrilova O.P., Gagkaeva T.Yu., Gogina N.N. Vestnik zashchity rasteniy, 2021, 104(3): 153-162 CrossRef (in Russ.).
  25. Metodicheskie rekomendatsii po vydeleniyu i kolichestvennomu uchetu mikroskopicheskikh gribov v zerne [Methodological recommendations for the isolation and quantitative accounting of microscopic fungi in grain]. Moscow, 2006 (in Russ.).
  26. Simmons E.G. Alternaria. An identification manual. Utrecht, CBS Fungal Biodiversity Centre, 2007.
  27. Gannibal F.B. Monitoring al’ternariozov sel’skokhozyaystvennykh kul’tur i identifikatsiya gribov roda Alternaria. Metodicheskoeposobie[Monitoring of Alternariosis of agricultural crops and identification of fungi of the genus Alternaria. Methodical manual]. St. Petersburg, 2011 (in Russ.).
  28. Kononenko G.P., Piryazeva E.A., Burkin A.A. Production of alternariol in the populations of grain feed-associated small spore Alternaria species. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2020, 55(3): 628-637 CrossRef
  29. Introduction to food- and airborne fungi. R.A. Samson, E.S. Hoekstra, J.C. Frisvad, O. Filtenborg (eds.). CBS, Utrecht, 2000.
  30. Burkin A.A., Kononenko G.P. Prikladnaya biokhimiya i mikrobiologiya, 2011, 47(1): 79-83 (in Russ.).
  31. Gagkaeva T.Yu., Gavrilova O.P., Orina A.S., Kazartsev I.A., Gannibal F.B. Mikologiya i fitopatologiya, 2017, 51(5): 292-298 (in Russ.).
  32. Piryazeva E.A., Kononenko G.P. Sovremennaya mikologiya v Rossii, 2017, 7: 175-177 (in Russ.).
  33. Ustyuzhanina M.I., Burkin A.A., Kononenko G.P., Piryazeva E.A., Zotova E.V. Alternative assay media for alternariol production by Alternaria species. Proc. VIII Int. Conf. on Environmental, Industrial and Applied Microbiology — BioMicroWorld2018 «Global progress in applied microbiology: a multidisciplinary approach». A. Méndez-Vilas (ed.). Badajoz, Formatex Research Center, 2018: 1-5.
  34. Kononenko G.P., Ustyuzhanina M.I., Orina A.S. Multi-substrate screening the ability to produce alternariol among Alternaria arborescens strains. Journal of Veterinary Science & Technology, 2019, 10: 41-42.
  35. Zwickel T., Kahr S.M., Rychlik M., Müller E.H. Chemotaxonomy of mycotoxigenic small-spored Alternaria fungi – Do multitoxin mixtures act as an indicator for species differentiation? Frontiers in Microbiology, 2018, 9: 1368 CrossRef
  36. Ramires F.A., Masiello M., Somma S., Villani A., Susca A., Logrieco A.F., Luz C., Meca G., Moretti A. Phylogeny and mycotoxin characterization of Alternaria species isolated from wheat grown in Tuscany, Italy. Toxins, 2018, 10(11): 472 CrossRef
  37. Oviedo M.S., Sturm M.E., Reynoso M.M., Chulze S.N., Ramirez M.L. Toxigenic profile and AFLP variability of Alternaria alternata and Alternaria infectoria occurring on wheat. Brazilian Journal of Microbiology, 2013, 44(2): 447-455 CrossRef
  38. Kosiak B., Torp M., Skjerve E., Andersen B. Alternaria and Fusarium in Norwegian grains of reduced quality — a matched pair sample study. International Journal of Food Microbiology, 2004, 93(1): 51-62 CrossRef
  39. Kelman M.J., Renaut J.B., Seifert K.A., Mack J., Yeung K. K.-C., Sumareh M.W. Chemotaxonomic profiling of Canadian Alternaria populations using high-resolution mass-spectrometry. Metabolites, 2020, 10(6): 238 CrossRef
  40. Patriarca A., da Cruz Cabral L., Pavicich M.A., Nielsen K.F., Andersen B. Secondary metabolite profiles of small-spored Alternaria support the new phylogenetic organization of the genus. International Journal of Food Microbiology, 2019, 291: 135-143 CrossRef

 

back

 


CONTENTS

 

 

Full article PDF (Rus)

Full article PDF (Eng)