MYCOTOXIN DETECTION. Toksiner på dansk Per Laustsen

Transkript

1 MYCOTOXIN DETECTION Toksiner på dansk Per Laustsen

2 Welcome Established in 1980 by Dr. Pearse Lyons

3 Top 5 animal health company The Alltech Advantage Only company that is: - Privately-held - Non-pharmaceutical Growing consistently over 36 years Moving towards a $4bn company

4 Our Core is FERMENTATION OPTIONAL HEADER Yeast: BIO-MOS, INTEGRAL A +, NUPRO YEA-SACC, MYCOSORB A + LIQUI-PLEX AGRO-MOS Bacteria: ALL-LAC, LACTO-SACC SOIL-SET Enzymes: ALLZYME SSF ALLZYME VEGPRO YEAST Algae: ALL G-RICH Beer & Spirits: KENTUCKY ALE Range including KENTUCKY BOURBON BARREL ALE LYONS SPIRITS including TOWN BRANCH BOURBON, PEARSE LYONS RESERVE

5 Alltech: Growth through acquisition

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7 Main moulds and mycotoxins Field Origin Storage Origin Fusarium Deoxynivalenol Zearalenone T-2 Toxin Fumonisin Moniliformin Nivalenol Fusaric Acid Aspergillus Aflatoxin Ochratoxin Sterigmatocystin Fumitremorgens Fumigaclavines Fumitoxins Gliotoxin Penicillium Ochratoxin PR Toxin Patulin Penicillic Acid Citrinin Penitrem Cyclopiazonic acid

8 Tjekliste Reduceret foderudnyttelse Reduceret libido Opkast Aborter Dårlig immunitet Uensartet huld Pludselige dødsfald Reduceret tilvækst Uforklarlig ændring i fertilitet Færre levendefødte Ringere malkning Øget sygdomsfrekvens Rektal eller vaginal prolaps Hvis du kan svare JA til en af ovennævnte påstande kan mykotoxiner være forklaringen.

9 Ochratoxin

10 Zearalenone

11 IT ENDS-UP BEING A BIASED DECISION Aflatoxins Aflatoxicol Aflatoxin B1,B2,B2α Aflatoxin G1,G2,G2α Aflatoxin M1 Austocystin A Averufin 5-Methoxysterigmatocystin Dihydroxysterigmatocystin Methoxysterigmatocystin Sterigmatocystin Norsolorinic acid Parasiticol Trichothecenes Nivalenol Fusarenone X Deoxynivalenol 3-Acetyldeoxynivalenol 15-O-Acetyl-4-DON Scirpentriol 15-Acetoxyscirpenol Diacetoxyscirpenol 3α -DAS Neosolaniol T-2 Triol HT-2 Toxin T-2 Toxin Iso-T-2 toxin Acetyl-T-2 toxin Trichodermin Trichodermol 7-a-Hydroxytrichodermol Verrucarol 4,15-Diacetylverrucarol Trichothecin Trichothecolone Macrocyclic trichothecens Isosatratoxin F Roridin A,E,H,L-2 Satratoxin G,H Trichoverrin A,B Trichoverrol A,B Verrucarin A,J Ochratoxins Ochratoxin A,B,α Ochratoxin A,B,α-ethyl ester Ochratoxin A,B,α-methyl ester Sphinganin myotoxins AAL toxin TA1,TA2 AAL toxin TB1,TB2 Fumonisin B1,B2,B3,B29 Cytochalasins Chaetoglobosin A,C Cytochalasin A,B,C,D,E,H,J Dihydrocytochalasin B Dihydrocytohalasin B γ lactone Zeralenons α,β-zearalanol Zearalanone α,β-zearalenol Zearalenone Roquefortines, ergot amines and alkaloids Agroclavine-I Auranthine Aurantiamine (alkaloid 302) Aurantioclavine Chanoclavine-I Costaclavine Cyclopenin Cyclopenol Cyclopeptin Dihydroergotamin Elymoclavine Epoxyagroclavine-I Ergocristine Ergometrin Ergotamin Fumigaclavine B,C Marcfortine A,B Meleagrin Oxalin Pyroclavine Roquefortine A,B,C,D Rugulosuvine Rugulovasine A,B Secoclavine α-ergocryptin C Atranone & precursors Atranone A,B,E,F,H,J 6-Hydroxydolaballa-3,7,12- -trien-14-one 3,4-Epoxy-6-hydroxy- dolabella- -7,12-diene-one Tremorgens Fumitremorgen B Lolitrem B Lolitriol Territrem A,B,C Penitrem A Penitremone A 19-Acetoxypaxillin Deoxypaxillin Paxillin a-paxitriol b-paxitriol C Janthitrem B Peptides Alamethicin α-amanitin Antimycin A1,A2,A3,A4,A5 Apicidin Cyclo(Phe-Ser) Cyclosporin Distamycin A Enniatin A1,A2,B,B1 Malformin A1,A2,B1,B2,C Phalloidin Valinomycin Others 2-(4-Hydroxyazobenzene)- benzoic acid 2,4,6-Trihydroxytoluene 2,5-Dihydroxybenzoic acid 3,5-Dihydrotoluene 3,7-Dimethyl-8-hydroxy-6- methoxy-isochroman 3-Methylsalicylic acid 5-Methylbenzene-1,2,3 triol 6-Methylcitreoisocumarin Abscisic acid 2-Acetyl-4(3H)-quinazolone α-acetyl-g-methyltetronic acid Aflatrem Alizarin Altenuene Alternariol Alternariol-methyl ether Altersolanol A Altertoxin I Amphotericin B Anacine Anhydrofusarubin p-anisaldehyde Antibiotic Y Arabenoic acid (Verrucolone) Aurofusarine Ascochitine Aspergillic acid Aspergillic acid-polymer Asperthecin Asteltoxin Asterric acid Austamide Austdiol Baccatin III Barnol Beauvericin 1,3-Benzendiol Benzoic acid Bikaverin Bis-dethio-bis(methylthio)- gliotoxin Bostrycin Bostrycoidin Brassicasterol Brefeldin A Brevianamide A Butenolide Byssochlamic acid Caffeic acid Caffeine Calphostin C Canadensolide Candidusin C Canescin Carlosic acid Carolic acid Catenarin Chaetocin Chetomin Chromanol 1, 2, 3 Chrysazin Chrysogine Chrysophanol Cinnamic acid Circumdatin A, B, C Citreoisocumarin Citreomontanin Citreoviridin A, X Citrinin Citromycetin Cladofulvin Cladosporin Clerocidin Communesin A, B Compactin Culmorin Curvularin Cyclochlorotine Cyclopaldic acid Cyclopiazonic acid Cynodontin Daldinin D Bis-dechlorogeodin Dechlorogriseofulvin Dehydrocarolic acid Dehydrocurvularin Deoxybostrycin Dermoglaucin Des-acetylpebrolide Desertorin A,B, C Des-methoxyviridiol Desmosterol Dethiosecoemestrin Diaporthin Diaportin acid Diaportinol Dichlorodiaportin cis-dihydrofusarubin trans-dihydrofusarubin Dihydrojasmonic acid 29,39-Dihydrosorbicillin Dihydroxyaflavinine 3,5-Dimethyl-6- hydroxyphthalide Dimethylphthalate Dipicolinic acid Dithiosilvatin Duclauxin Dustanin Echinulin Emestrin Emestrin B Emindole DA,DB Emodin Epicorazine A Epi-dechlorogriseofulvin Epoxysuccinic acid Equisetin Ergosterol Erythroglaucin Ethisolide Expansolide Ferulic acid Flavoglaucin Formyl-xanthocillin X Frequentin Fructigenine Fucosterol Fulvic acid Fumagillin Fumagillol Fumiguinazoline F Fusaproliferin Fusaric acid Fusarin C Fusarochromanone C Fusarubin Fusicoccin Fusidic acid Fusoxysporone Gallic acid Gentisylalcohol Gibberellic acid Gibepyrone A,F Gliotoxin Gliovirin Griseofulvin Griseophenone C Helminthosporin Helvolic acid Histamin 59-Hydroxyasperentin p-hydroxybenzoic acid p-hydroxycinnamic acid 5-Hydroxymaltol 4-Hydroxymellein ω-hydroxypachybasin p-hydroxyphenylacetic acid Indolacetic acid Insulicolide A,B Insulicolide B Iodinine Islandicin Isoemodin Isomarticin Italicic acid Italicic acid-methyl ester Italinic acid Jasmonic acid Jasmonic acid-methyl ester Javanicin Kojic acid Kotanin Lactone 1,2 Lambertellin Lanosterol Ascorbic acid Leucyltryptophanyldiketopi perazine Lichexanthone L-Phenylalanin L-tyrosin Luteoskyrin Macrosporin Maculosin Marticin 3-Methoxyviridicatin 6-Methylsalicylic acid Mevinolin Mitomycin C Mitorubrin Mitorubrinic acid Mitorubrinol Mitorubrinol acetate C Mizoribine Mollisin Moniliformin Monorden Mycophenolic acid Myriocin Nectriafurone Nidulin Nigragillin 3-Nitropropionic acid Nominine Norjavanicin Norlichexanthone Nortryptoquivaline Oleic acid Oosporein Ophiobolin G Orsellinic acid Oxalic acid Pachybasic acid Pachybasin Palitantin Paracelsin Paspaline Paspalinin Penicillic acid Penicillin G Phaseic acid Phoenicin Phomarin Physcion PI-3 PR-1635 Preechinulin PR-imine Propyl-3,4,5- trihydroxybenzoate PR Toxin Puberuline Puberulonic acid Purpurin Purpurogenone Pyrogallol 2- Pyruvoylaminobenzamide Questin Questinol Radicinin Ravenelin Riboflavin Roseopurpurin Rubratoxin A,B Rugulosin Salicin Salicyclic acid Sclerotigenin Scytalidine Scytalone Secalonic acid D Shikimic acid Skyrin SMTP-6,-7,-8 Solaniol Soranjidiol Sorbicillin Spinulosin Staplabin Steckiin Stemphone Stipitatic acid Sulochrin Taxol Tentoxin Tenuazonic acid Terrein Terrestric acid Terretonin Thiamin, hydrochloride Toluhydroquinone Tomatine Torreyol 3,4,5-Trihydroxy-7- methoxy-2- methylanthraquinone Trisdechloronornidulin Trypacidin Tryptophan Tryprostatin B Tyramine Usnic acid Vermiculin Verrucine A,B Verrucofortine Verrucosidin Verrucosine Verruculogen Verruculotoxin 10-Epi-verruculotoxin Vertinolide Violaceic acid Viomellein Vioxanthin Viridamine Viridicatic acid Viridicatin Viridicatumtoxin Viridiol Viriditoxin Visoltricin Walleminol Walleminone Wortmannin Xanthocillin X Xanthomegnin

12 INCREASE ANALYTICAL PRECISION / MULTI-DETECTION Qualitative test Kit Semi-Quantitative test Kit FIA RIA ELISA Nano-Biosensors GC-UV HPLC-UV HPLC-PDA HPLC-ECD GC-MS LC-MS 1.00e6 9.00e5 LC/LC-MS/MS >300 pesticides 8.00e5 7.00e5 Intensity, cps 6.00e5 5.00e5 4.00e5 3.00e5 2.00e5 1.00e5 Yes No Time, min

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15 RISK ANALYSIS Aflatoxin (B1) Aflatoxins (B1+B2+G1+G2) Ochratoxins (A+B) Type B Trichothecenes* Type A Trichothecenes** Fumonisins (B1+B2+B3) Zearalenone Group Fusaric Acid Other Penicillium Mycotoxins*** Other Aspergillus Mycotoxins**** Ergot Toxins REQ (Risk Equivalent Quantity)

16 INTERPRETATION: RISK EVALUATION TEF/TEQ WHAT ARE TEF AND TEQ? Toxic Equivalency compare to a single reference: Aflatoxin B1 TEF: TOXIC EQUIVALENT FACTORS Poultry Swine Dairy Mycotoxins Groups High risk (ppb) TEFs High risk (ppb) TEFs High risk (ppb) TEFs Aflatoxins B Total Aflatoxins Ochratoxins Other Aspergillus Toxins Trichothecenes B Trichothecenes A Fumonisins Zearalenones Penicillium Toxins Ergot Alkaloids TEQ: TOXIC EQUIVALENT QUANTITIES Evaluate the total toxicity given mycotoxin mixture, based on amount and TEF: TEQ total = C tox 1 TEF tox 1 + C tox 2 TEF tox C tox n TEF tox n

17 What is ISO/IEC 17025: International Standard designed to recognize the competence of Testing Laboratories. This accreditation is accepted worldwide by Governments, businesses and laboratories. 2. Objective third-party assessment of the managerial and technical requirements of the lab ensuring the accuracy and impartiality of analytical results. 3. Technical competence of personnel and availability of technical resources. This ensures personnel are qualified for their positions and are provided with proper training, instrumentation and environment to perform their duties. 4. Quality system that continually improves ability to consistently produce valid test results. Includes review and updating of work instructions and forms. Includes an established process for corrective and preventive actions within the lab. 5. Ensures confidentiality and impartiality of analytical results. Parent company has no influence on results.

18 Maksimal sikker lagringstid (døgn) m.h.t. tilvækst af lagersvampe Antal døgn ved afgrødens vandindhold % Temperatur ºC ,5 1,7 1, , ,5

19 tørring Luften (evt. opvarmet) anvendes for at transportere vandet bort Vandindhold % Mængde vand (ton) at tørre bort ved forskellige mængder korn (ton): ,7 18,6 37,2 55, ,9 55,8 83,7

20 Ligevægtstabel mellem relativ luftfugtighed og vandindhold i korn og frøafgrøder Relativ luftfugtighed i % Byg 10,5 11,1 11,8 12,6 13,4 14,3 15,2 16,3 17, ,9 Hvede 10,6 11,3 12,3 13,1 13,5 14,3 15,2 16,3 17,4 19,1 20,5 Havre 9,7 10,3 11,3 11,9 12,5 13,3 14,2 15,4 16,7 18,5 21 Rug 10,3 10,9 11,6 12,4 13,2 13,9 14, , ,5 Raps 5,7 6,1 6,6 7 7, ,3 12,7 14,3 Ærter 10, ,9 12,7 13,5 14, , Rajgræs 8,6 9,3 10, , ,2 15,6 17,4 19,8 23,3 Opvarmning af luften med 1 C = 4% reduktion af relativ luftfugtighed Plantørring: max. 6ºC opvarmning af udeluften inkl. blæserens egenopvarmning (ca. 1ºC)

21 Betydning af varmetilsætning 750 ton hvede, 6,5 meter lagringshøjde i silo med omrører Nedtørring % giver 35 ton vand at tørre bort 22 kw blæser og Jumbo 200 olieovn 18 C og 70% luftfugtighed Betyder følgende: Tørretid uden varme, ca. 30 døgn Tørretid med varme, ca. 6 døgn Samme omkostning til energi, men meget kortere tørretid!

22 Storage Contamination Score 1 clean silo with no accumulated crusts on its walls. Score 2 Silo with crusts accumulated on its walls. Score 3 Many old crusts On the walls of the silo.

23 Hvad gør man hvis man har problemer? Destruktion af råvaren Fortynde den kontaminerede råvare Brug af mykotoxinbinder

24 Stabilitet ved ændringer af ph Effekt af ph på MTB-100 s absorbtion af aflatoxin B1 % Absorberet Devegowda et al., 1994

25 Mycosorb binder IKKE Mineraler og Vitaminer Vitamin levels Vitamin A (IU/g) Vitamin E (mg/g) Negative Control Control + Mycosorb Contaminated feed Contaminated feed + Mycosorb Treatments Meissonnier et al., 2008

26 Evne til at binde en bred vifte af mykotoxiner MYCOTOXIN FORSKER ÅR Aflatoxins Stanley and Co-workers 2004 Aflatoxins Diaz and Whitlow 2004 Zearalenone Yiannikouris and Co-workers 2004 T-2 toxin Manoj and Devegowda 1999 T-2 toxin Dvorska and Surai 2001 DAS Pavicic and Spring 2001 Ergot alkaloids Merrill and Co-workers 2007 KOMBINATIONER AF MYKOTOXINER Aflatoxin, ochratoxin Raju and Devegowda 2000 & T-2 toxin DON and Zearalenone Swamy and Smith 2002 DON and Zearalenone Chowdhury and Smith 2005 DON and Zearalenone Raymond, Smith and Swamy 2003 DON and Zearalenone Diaz and Smith 2006 DON and Zearalenone Korosteleva and Smith, DON and Zearalenone Girish and Smith,

27 Effektivt ved lav iblanding Clay A Mycosorb Mahesh & Devegowda, 1996

28 Peer-reviewed in vivo data

29 Krav til absorbent Stabilitet Ikke binde mineralet/vitaminer Dokumenteret effekt Hurtig binding In vivo data

30 dålig opbevaring

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