Significant occurrence of aflatoxin M1 and M2 in milk, 2012

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Eszterágpuszta : GOSZ maize test, ear rot data of

across eight locations, natural infection,.

1, Szabolcs Lehoczki-Krsjak 1, Éva Toldi 1,

Szeged, Hungary Agricultural University, Dept

conference, April 13, Szeged, Hungary 1 Infected

7 9 Infected ears % Why the problem is important?

99 3 3 Relation of incidence and severity in the

verticillioides Significant occurrence of

1 Eszterágpuszta : GOSZ maize test, ear rot data of maize hybrids GOSZ maize ear rot tests, means across eight locations, natural infection,. Aspergillus flavus as a new resistance problem in maize (Zea mays L.) Ákos Mesterházy 1 Mónika Varga 1, Beáta TóthT 1, Szabolcs Lehoczki-Krsjak 1, Éva Toldi 1, Ferenc Bagi 1 Cereal Research non-profit Ltd. Szeged, Hungary Agricultural University, Dept Plant Pathology, Novi Sad, Serbia ToxFreeFeed conference, April 13, Szeged, Hungary 1 Infected ears as % S e v e r i ty x % 3 3 y =.x +.9 R =.77 3 y =.37x -.97 R =.73 Maize hybrid registration Tests, Székkutas, Infected ears % Why the problem is important? Infected ears as % y =.771x +.3 R = Relation of incidence and severity in the variety registration trial of NÉBIH,, n=1, means for 7 locations y =.3777x R = Infected ears % F. verticillioides F. verticillioides Significant occurrence of aflatoxin M1 and M in milk, F. verticillioides F. graminearum 3 A. flavus F. verticillioides, Aspergillus flavus, Penicillium sp. A. flavus F. verticilliodes

Epidemyiology At female flowering warm and humid weather prefers all. Thereafter: F. graminearum warm and rainy August and September F. verticillioides warm and dry wearter Aspergillus spp.

7 The TOXIN triangle Mesterházy Toxigenic species, isolate Environment Toxin Resistance level Toxins F.. verticillioides, F. proliferatum,, F.

. s, B1, B, G1, G Trait Aspergillus flavus infection and aflatoxin production in maize hybrids, Fert. felület let Genotype Check Asp1 Asp Asp3 H1 1.3 1. 1. 1.9 % H 1. 1. 1.3 1. mg/kg 1. 1.3 1. 1. H..99.

2 Epidemyiology At female flowering warm and humid weather prefers all. Thereafter: F. graminearum warm and rainy August and September F. verticillioides warm and dry wearter Aspergillus spp.. Hot and dry weather Irrigation at flowering prefers higher incidence, severity and toxin contamination. 7 The TOXIN triangle Mesterházy Toxigenic species, isolate Environment Toxin Resistance level Toxins F.. verticillioides, F. proliferatum,, F. moniliforme: : Fumonisins B1, B, B3, B, B, total number above /: : DON (3-ADON, -ADON, 3-DON 3 glükozid) és s Zearalenon (α-( és β- zearalenol) Aspergillus spp.. s, B1, B, G1, G Trait Aspergillus flavus infection and aflatoxin production in maize hybrids, Fert. felület let Genotype Check Asp1 Asp Asp3 H % H mg/kg H H The situation is more complex than in wheat, more toxins and more complicated genetics in host. 9 Human limit. mg/kg, feedstuffs:. mg/kg Maize samples of the same hybrids in -, their aflatoxin B1 and fumonisin B1+B content Artificial inoculation 11 Mycotoxins B1 (µg/kg) FB1+FB (mg/kg) B1 (µg/kg) FB1+FB (mg/kg) B1 (µg/kg) FB1+FB (mg/kg) Positive samples (%) Maximum value of positive samples n.d. 7, n.d.,,, n.d.,1 n.d. 3,7, n.d.,9 n.d. 1,,9,7 n.d. 1,1 n.d., 3,1 1, 11

MKK maize resistance test against four toxigenic ear rot pathogens, severity, R MS Resistance types at toothpick method : R= resistant MS =medium susceptible 3 MKK MKK13 MKK7 MKK MKK1 MKK19 MKK3 MKK3

1***.71*** Kontr.739***.37***.99*** Asp. term.7.**.7**.3*.9***.7***.99***.***.* 13 ***P =.1, ** P =.

3 3 P99 Dunasil DKC9 DKC9 DKC9 Somacorn NKThermo Árpád Kansas Phileaxx Amanita Esflato DKC13 PR37F73 Dascipio DKC37 Mv1 DKC7 DKC79 Mv DKC7 Mv37 Essensor DKC311 DKC Alexxandra RH9111 Finkas Surreal Mv

3 MKK maize resistance test against four toxigenic ear rot pathogens, severity, R MS Resistance types at toothpick method : R= resistant MS =medium susceptible 3 MKK MKK13 MKK7 MKK MKK1 MKK19 MKK3 MKK3 MKK11 MKK1 MKK Correlations MKK MKK17 MKK MKK MKK3 MKK MKK7 MKK3 MKK1 MKK MKK MKK3 MKK37 MKK MKK31 MKK MKK MKK MKK3 MKK9 MKK MKK3 MKK33 MKK1 MKK1 MKK9 Kontr Asp S S= susceptible Trait Kontr Asp.79***.1***.71*** Kontr.739***.37***.99*** Asp. term.7.**.7**.3*.9***.7***.99***.***.* 13 ***P =.1, ** P =.1 1 Resistance of maize hybrids against Fusarium spp with silk channel inoculation, Mycored maize ear rot resistance test with toothpick method against Fusarium spp. 3 3 P99 Dunasil DKC9 DKC9 DKC9 Somacorn NKThermo Árpád Kansas Phileaxx Amanita Esflato DKC13 PR37F73 Dascipio DKC37 Mv1 DKC7 DKC79 Mv DKC7 Mv37 Essensor DKC311 DKC Alexxandra RH9111 Finkas Surreal Mv Superbia Sufavor PR37N1 Lucius DKC99 Cobalt Kontr. Kozak SBL1 SzegediTC9 SBL GK Boglár SBL SBL Szegedi 39 SBL1 SzegediTC37 Bella Szegedi 3 SBL13 SBL Narew ISH33 Szegedi 7 SBL1 Kenéz SBL17 Szegedi 33 SBL3 Kisérleti SBL19 ISH3 SBL SBL Beym SBL7 Szegedi 1 SBL9 Kisérleti Sze SBL1 ISH3v Csanád Szegedi SBL11 Szegedi 37 Sarolta SzegediTC13 SBL Szegedi 33 SzegediSC SBL ISH SzegediTC F.gram F.culm F.vert Term. Kontr Kontr All significant at P =.1 % term..***.11***.799*** Natural.*** ***.17***.9***.733***.3*** ***P =.1 1 Mycored maize ear rot resistance test, silk channel method, Szeged, Hungarian maize club resistance test against toxigenic ear rot pathogens in maize, silk channel method, SBL7 Szegedi 3 SBL1 Szegedi 7 SBL17 SBL Szegedi 1 SzegediTC37 SBL SBL3 ISH3 Kenéz SBL19 SBL13 Szegedi 33 SBL Szegedi 33 SBL9 Bella SBL1 GK Boglár SBL SBL Szegedi Kozak SBL SzegediTC9 SzegediTC SBL1 Narew Szegedi 37 SzegediTC13 SBL Szegedi 39 ISH33 SBL1 Kisérleti SzegediSC ISH Beym SBL Sarolta Csanád SBL11 Kisérleti Sze ISH3v term. F.vert F.gram F.culm Kontr. Check.7 Correaalations Control Kontr Contr All significant at P =.1 % 17 All significant at P =.1 %-on 1

Resistance of Hungarian and Serbian maize

toothpick inoculation, means of isolates,

Check 9 7 SZ-M SZ-M1 SZ-M3 SZ-M SZ-M SZ-M9 SZ-M7

Kontroll. -.7.7***.*.97 -. Kontroll.191** -. -.9.3**.

SZ-M SZ-M13 SZ-M SZ-M1 SZ-M SZ-M SZ-M11 SZ-M1

Natural hybrids to toxigenic fungi, means for

NS3 NS3 GKBoglár Sze39 SzeTC37 NS3 NS Sze3 NS NS

SzeTC Correlations Afl Kontr.7***.33.919 Afl..19.3 Kontr.

4 Resistance of Hungarian and Serbian maize hybrids against toxigenic fungi following toothpick inoculation, means of isolates, Szeged, Resistance of Hungarian-Serbian maize hybrids to toxigenic fungi according to isolates, ear coverage (%), Szeged, Afl Check 9 7 SZ-M SZ-M1 SZ-M3 SZ-M SZ-M SZ-M9 SZ-M7 SZ-M17 SZ-M19 SZ-M SZ-M1 Összefüggések Afl Kontroll ***.* Kontroll.191** **.1.7***.933*** *** P =.1, ** P =.1, * P =. SZ-M SZ-M13 SZ-M SZ-M1 SZ-M SZ-M SZ-M11 SZ-M1 SZ-M A1 A Natural Resistance of Hungarian and Serbian maize hybrids to toxigenic fungi, means for Szeged and Novi Sad,, coverage (%) 3 3 Afl Kontr NS3 NS3 GKBoglár Sze39 SzeTC37 NS3 NS Sze3 NS NS NS1 GKT Sze33 Kenéz Sarolta NS NS31 Sze1 NS3 SzeTC Correlations Afl Kontr.7*** Afl Kontr ***.9***.93*** *** P =.1 1 3

5 SZM Very strong natural overinfection after Fusarium and without it ert Term. ert A. flavus MKK7 MKK13 Natural MKK1 7 MKK9 ert Term 3 9

6 What should be done? To increase the comatibility of maize production, keeping the competitivity in food and feed safety as in profitability also in epidemic years. Both low toxin contamination and high yielding are priorities. In maize no applicable fungicide technology exists, therefore the role of resistance is larger Resistance research and breeding Very limited informations are at hand for important breeding questions like how resistance to different toxigenic fungi correlates. Very limited genetic information is present, much less than in wheat. We need new resistance sources to detect first in own material, and also in others. The financial background should be secured. 3. Variety registration The artificial inoculations, separately for the most important toxigenic fungi should be introduced into the registrtation, keeping the evaluation of natural infection. This work should be supplemented with toxin analyses, as the whole trade and industry is based on toxin limits, therefore the registration cannot avoid it, even it is not cheap.. It is highéy y important to screen the registered hybdids as this is the most effective mean to increase feed safety within -3 3 years. 3. Fusarium toxin survey For aflatoxins a large number of data were produced, they should be summarized to receive exact information on the severity of the epidemic. This should be made also for the most important Fusarium Integrated control No means are successful enough, the whole production from the hybrid choice to the feeding must be done under toxicological control and methods should be developed that allow a much more effective grain and silage preservation and quality. What to do with the infected parties? Detoxication, until now no effective method, only toxin binders are in use for aflatoxins, for the others the situation is better. Colour selection under UV light in labs. Development of industrial optic technology as for wheat happened Detoxification with strong oxidative agents, research is necessary, no developed method available Target: Grain and silage 3 3

7 Summary There are highly significant resistance differences between maize e hybrids to toxigenic pathogens. Significant infection severity differences were recorded against Aspergillus flavus at artificial anad natural infection. From the resistance side, the A. flavus epidemic was not a big surprise. It seems that the genetic background to F. graminearum/f. culmorum should not be identical with the resistance to F. verticillioides / A. flavus group. This maked breeding more complicated. For this reason only the breeding for complex resistance is meaningful where all four pathogens are considered. As there are numerous hybrids with this resistance structure, the t project can be managed. Acknowledgements The Authors thank the support of projects FP7 MycoRed, A ToxFreeFeed HUSRB/// HU/SR, OTKA grants K and K77 As the toxin level is decisive in the measuring resistance to toxigenic pathogens, the toxin content should be measured. It is important to introduce artificial inoculation methods into the breedin against F. graminearum, F. culmorum, F. verticillioides and A. flavus. The evaluation of the natural infections should be kept Thank you for your attention 39

mycotoxin-contaminated contaminated food or feed

mycotoxin-contaminated contaminated food or feed Poisons in our Food By Melvin Newman, Ph.D. Professor Entomology and Plant Pathology Foods that may contain toxins Introduction Fungi: produce an array of chemical products called secondary metabolites.