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1 ซ ต าเทคโนโลย แก ป ญหาสารพ ษเช อรา

2 ซ ต าเทคโนโลย แก ป ญหาสารพ ษเช อรา

3

4 Mycotoxin ความท าทาย

5 Key Raw material : Mycotoxin contamination Aflatoxin Zearalenone Fumonisin T2 Toxin Soy Bean Meal Corn Baked Beans Fishmeal Broken Rice Casava Rice Bran DDGS ท มา : คณะส ตวแพทยศาสตร มหาว ทยาล ยเกษตรศาสตร ว ทยาเขตก าแพงแสน

6 Mycotoxin : Challenge

7 Production of mycotoxins Harvest Fusarium Claviceps Mycotoxins Aspergillus Penecillium Field Time Storage 7 7

8 Mycotoxin Binder

9 Mycotoxin Management 真 菌 毒 素 管 理 方 案 DO NOT Feed Contaminated raw material DILUTION 稀 释 AMMONIZATION 氨 化 处 理 MYCOTOXIN BINDER 霉 菌 毒 素 吸 附 剂 BIOTRANSFORMATION 生 物 转 化 SUPPORTIVE TREATMENT 补 充 治 疗

10 Available Ingredient to Control Mycotoxin Yeast Cellwall Clay Activated Charcoal Synthetic Enzyme Herb Quick response ** **** **** **** ** Polar Mycotoxin *** **** **** **** Nonpolar Mycotoxin ** *** **** **** Zearalenone ** *** *** *** Fumonisin * ** ** Ochratoxin ** * ph Supportive **** * Immunity *** * Palatability *** ** Liver Function ** Combine Nutrient *** **** Easy denature *** ** 没 有 单 一 的 解 决 方 案!! 10

11 Effects of mycotoxins on animals Poultry Aflatoxin Growth reduction Reduced feed efficiency Immunosupression Anemia Reduced antibiotic effectiveness Anorexia Liver damage Reduced egg production Increased mortality Reduced skin pigmentation Lethargy Reduced ability to metabolize fat, starch, and protein Increased blood clotting time Zearalenone Increased mortality Increased susceptibility to disease Increased blood clotting time Ochratoxin Nephrotoxicity Reduced feed consumption Pale beaks Weakness and lameness Ataxia Tremors Diarrhea Death T-2 Toxin Digestive disorders Increased mortality Bloody diarrhea Reduced egg production Intestinal hemorrhages Increased susceptibility to disease Deoxynivalenol Reduced reproductive performance Increased mortality Reduced feed efficiency Diarrhea Reduced growth Reduced egg production Neural disturbances Increased susceptibility to disease Increased blood clotting time Source: CAST, Dairlyland labs

12 Effects of mycotoxins on animals Swine Aflatoxin Liver damage Hydrolyzation into M1 and is eliminated in milk ( piglets) Immunosupression Acute signs: anorexia, depression, ataxia, epitasis Chronic signs: reduced feed efficiency, reduced milk production, icterus, decreased appetite Deoxynivalenol Reduced feed consumption / feed refusal Vomiting Ergot alkaloids Reduced growth Vertigo, staggers, convulsions, temporary posterior paralysis Decreased peripheral blood supply Reduced reproductive efficiency of sows Eventual death T-2 Toxin Immunosupression Frequent defecation Vomiting Weight loss Feed refusal Source: CAST, University of Minnesota Ochratoxin Reduced feed efficiency Growth reduction Liver and kidney damage Increased mortality Zearalenone Hyperestrogenism Vulvovaginitis Vaginal or rectal prolapses in pre-pubertal gilts Enlarged uterus, swollen or twisted uterus, shrunken ovaries In boars, testes atrophy, enlarged mammary glands, decreased fertility

13 Effects of mycotoxins on animals Ruminants Aflatoxin Reduced feed efficiency Growth reduction Hydrolyzation into M1 and is eliminated in milk ( humans) Tachycardia Tachypnoea Liver damage Death T-2 Toxin Feed refusal Reduced production Gastroenteritis and intestinal hemorrhages Reduced immunological response in calves Abortions Death Deoxynivalenol Reduced feed consumption / feed refusal Vomiting Reduced milk production Increased count of somatic cells in milk Reduced reproductive efficiency Esporidesmine Eczema facial Zearalenone Hyperestrogenism Vulvovaginitis Vaginal secretion Abortions Reduced feed consumption Reduced milk production Reduced reproductive efficiency Source: NCSU, CAST

14 Aflatoxins fatty liver kidney disorders leg and bone problems pigmentation problems (carcasses, egg yolk) reduced hatchability of eggs, smaller eggs, eggshell quality affected coccidiosis vaccine failure reduced immunity, lower resistance to diseases, bacteria, viruses reduced performances (weight gain, FCR, mortality)

15 Ochratoxin increased mortality reduced growth and FCR feed refusal diarrhoea, tremors and other neural malfunctions (high doses). reduced egg production weaker eggshell eggs with blood and meat spots.

16 Trichothecenes (ex T-2) lesions at the edges of the beaks lesions in the intestine. decreased feed intake, weight loss anomalous feathering in chicks drop in egg production eggs with weaker shells

17 Maximum levels of mycotoxin recommended in poultry feeds (Dr. Jowaman Khajarern Thailand) Mycotoxin (ppb) Animal AF ZEN T2 OC FMN DON Duckling < <100 <300 <300 Starter Broiler < <300 <300 Grower Broiler < <200 <500 <300 Layer/Breeder < <100 <300 <300 AF = Aflatoxin ZEN = Zearalenone T2 = T-2 toxin OC = Ochratoxin FMN = Fumonisins DON = Deoxynivalenol

18 Immune Suppressive Aflatoxin Vomitoxin, T - 2 toxin Ochratoxin Fumonisins

19 研 究 和 开 发 選 擇 材 料 與 納 米 結 構 特 點

20 How do mycotoxins act? Impact to production Health,Immunity 2.Absorp Mycotoxin at intestine and transform at liver x x x x x x 1.Mycotoxin contaminate in feed Mycotoxin destroy target organ and be deactivated by liver kidney

21 Our Learning of Mycotoxin 1. Polar Mycotoxin can absorped by Clay and Yeast Cellwall: Aflatoxin 2. Non Polar Mycotoxin,have some surface charge can absorped by clay and Yeast Cellwall but need to create environment : Zeralenone 3. Non Polar Mycotoxin,No surface charge need to change Mycotoxin s structure for more polarity and synergistic effect in nano level (Nanocomposit action): Fumonisin /DON /Ochatoxin 4. We have to absorp Mycotoxin As much as Posible before get into liver 5. Liver and Kidney be Performed for Eliminate Mycotoxin 6. Mycotoxin have > 200 type and combination effect need to manage as Broad Spectrum effect 24

22 Zetatechnology L Tonic We develop Nanocomposite by enhance the absorption performance and support Enzyme s Immobilization support Biotransformer action Totally, we are the first Nanocomposite plus Biotansformer in Mycotoxin binding action Our products are Theurapeutic Program planning for Feed Industry (Prevent +Treatment before lose) 25

23 Zetatechnology L Tonic 26

24 Select Raw Material : Characterization STP (cc/g) Zeta Relative Pressure (p/p0) Surface Area Specification of Zeta L-Tonic Product Characteristic of Surface area BET Surface area 面 积 [m 2 /g] Average pore size 小 孔 尺 寸 (Nanometer) Total No.Pore 小 孔 总 数 (cc/g) Type of pore IUPAC Zeta L-Tonic Mesopore

25 Mode of Action of Zeta technology 1 Attraction 2 Absorption Polar mycotoxins (Aflatoxin, CPA, Ergot) Ca ++ Na O + O-CH O 3 O O O ß-ceto-lactone of Aflatoxin Non-polar mycotoxins (DON, FB1, OTA, T-2, ZEA) Adsorbent Pore system: Macro Meso Micro Submicro Liquid phase

26 Zetatechnology L Tonic 29

27 Plant Protease Mechanism Plant protease mechanisms, Ochratoxin A can be cleaved by enzymatically (a). Enzymes that catalyze the hydrolytic cleavage of Ochratoxin A are called plant protease. In the active sites of this plant protease, the eponymous residue is usually paired with a proton-withdrawing group to promote nucleophilic attack on the Ochratoxin A. Close-up on the action, three amino acids located near the active site (Asn175, His259, and Cys25) are labeled (b). Finally, products are released, and enzyme is free to bind other Ochratoxin A molecules (c). 30

28 Select material by Characteristic s Nano structure Charateristic % Absorption Normal Clay Modified Clay Zetatechnology Zetatechnology +Biotransformer Surface Area m2/g pore diametre nm Total pore cm3/g Aflatoxin* Zearalenone* Fumonisin* Ochartoxin** Dosage per ton (KG) * Ku Vet lab ** Central lab

29 Select & Develop Raw Material Nano structure Pore 小孔 Big Pore 大孔 Clays HSCAS Modified Clays Pore size 孔尺寸 Pore volume 孔总数 Zeta potential Nano composite

30 Select & Develop Raw Material Nano structure Pore Big Pore Clays HSCAS Modified Clays Pore size 孔尺寸 Pore volume 孔总数 Zeta potential Nano composite

31 Select & Develop Raw Material Nano structure Nanocomposite testing : Normal Clay 粘 土 Nanocompozite Modified Clay 改 性 粘 土 Nanocompozite +Biotransformer

32 2.Enzymatic Detoxification & Biotransformer

33 Enzymatic Biotransformation 酵 素 生 物 转 化 It is the detoxification of Mycotoxins through Biotransformation by enzyme produce by microorganism. The mycotoxin molecule is breakdown into more simple molecule 简 单 的 分 子 and less toxic as compare to the original form. The key to Enzymatic Biotransformation is the right amount of enzyme unit present 毒 毒 性 减 少 Ezymatic Biotransformation

34 Enzymatic Detoxification 酵 素 生 物 转 化 Enzyme Dosage 用 量 Zearalenone 玉 米 赤 霉 烯 酮 毒 素 % Bio transformation 生 物 转 化 率 Fumonisin B1 伏 马 菌 素 Ochratoxin A 赭 曲 霉 毒 素 Mycopase Mycopase 50 mg/10 ml 100 mg/10 ml nd. nd. nd. nd. nd %

35 Biotransformer & Nanocomposite Performance Zeta L-Tonic Product Dosage 用 量 Zearalenone 玉 米 赤 霉 烯 酮 毒 素 % Absorption 吸 附 率 Fumonisin B1 伏 马 菌 素 Ochratoxin A 赭 曲 霉 毒 素 Zeta L-Tonic 100 mg/10 ml 82.7% 83.5% 90.1%

36 Our Learning of Mycotoxin 40

37 ซ ต าเทคโนโลย : นว ตกรรมสารด ดจ บช วพ ษเช อรา

38 Zeta CARE in support Environment Herbal Technology Support Liver Function 1-3,1-6 Betaglucan Immuno-stimulation Feed Intake Structural Absorption Organic Glycine Chelate Functional Supplement Zn Mn Cu Zeta Oxidant Diatomateus Earth Natural Essential oil

39 Zeta CARE : Glycine Chelate Zn Mn Cu Zn Chelate Zn, Increased Mn, Cu absorbtion of Ca Egg shell quality Immune enhancer Pigment of egg shell Mn Chelate Improved fertility and bone structure Cu Chelate Pigment of egg shell Immune enhancer

40 Key Success of Zeta technology 1 Composition 1 Nanocomposite 2 BioTransformer 3 Support Liver Kidney Function 2 Mode of Action 1Fast absorb before transform in liver 2 Bio Transformer 3 Support Liver Kidney Function 3 Solve Key Mycotoxin Laboratory & Field result Fast Action 快 操 作

41 KLEAN Greentech Mycotoxin Nano Composite Zeta-CARE Y Y Y Y Y Y Y Y Y Y Y Y Y YY Y YY Y YY Y YY Y YY Y YY Y YY Y YY Y YY Y YY Y Y Y Y Y Y Y Y Y Y 45

42 Why Zeta-Technology Work!! Zeta Composite Bio Zn, Mn, Cu Transformer 1-3,1-6 Beta glucan Zeta Care % Absorption 吸 附 率 Zearalenone %* Fumonisin %* Ochatoxin %** Aflatoxin %* * Veterinary Sci Faculty,Kasetsart University,2014 * * Central lab,2014 Source: Central lab ศ นย ห องปฏ บ ต การกลาง,ส งหาคม 2556

43 Mycotoxin Binding CapacityAnalysis 黴 菌 毒 素 的 吸 附 能 力 Veterinary Sci Faculty,Kasetsart University,2014

44 Comparison of different Mycotoxin Binder on Zearalenone & Fumonisin 不 同 黴 菌 毒 素 吸 附 劑 的 比 較 Absorption ph 6 (%) Dose kg/ton 用 量 Zearalenone 200 ppb Fumonisin B ppb Comp Ags Comp MF Comp MT Zeta ZAD Zeta TOF Zeta L Tonic Zeta Plus Zeta Central lab,2014

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46 ADD VALUE SERVICE 额 外 服 务 原 料 检 测 找 到 快 解 决 方 案

47 Raw Material Risk Low Risk Moderate Risk High Risk 1. % Mycotoxin Positive Less than 25 % positive % More than 75 % ) 2. Mycotoxin toxic Level Less 100 % % more than150 % 3.% CV Sampling Random less 10 % % More than 35 % 4. How long of Stock Less than 7 day 7-10 ว น More than 7-10 day (ท มา : Cargill Animal Nutrition, Romer labs )

48 Theurapeutic Program Planning: KGT Tools Therapeutic Program Planning TPP 與 KGT 的 工 具 治 療 規 劃 方 案 Normal Loss before TPP AVERAGE DAILY GAIN Infection TPP Profit from TPP Treatment Profit from Treatment Death TIME IN DAYS

49 Dosage of Zeta Products Preventive dose: 0.5 kg/t Standard dosage: 1.0 kg/t In case of serious problems: Small Animal : up to 2.0 kg Breeder : up to 1.5 kg

50 Thank you for your Supports National Innovation Award 2014 Economic Zetatechnology : Innovative Mycotoxin Binding Innovator : Dr Kitti Supchukun Dr Kittiwut Kasemwong

51 ผลงานว จ ยและพ ฒนาของ ศน. ได ร บรางว ลในระด บชาต และนานาชาต National Innovation Award 2014 Honorable Mention in Economics Award Seoul International Invention Fair 2014 Silver Prize VIII International Warsaw Invention Show 2014 Special Award ดร. ก ตต ว ฒ เกษมวงศ ห วหน าห องปฏ บ ต การนาโนเพ อผล ตภ ณฑ อาหารและการเกษตร ร วมก บ บร ษ ท คล นกร นเทค จ าก ด ศ กษาค นคว ว จ ยและพ ฒนาผล ตภ ณฑ นว ตกรรมด ด า จ บสารพ ษจากเช อราปนเป อนในว ตถ ด บอาหารส ตว (Mycotoxin binder) ภายใต ช อทาง การค า ZETATECHNOLOGY ผล ตภ ณฑ ด งกล าว ได ร บรางว ลนว ตกรรมแห งชาต ป ๒๕๕๗ รางว ลชมเชยด านเศรษฐก จ นอกจากน นย งได ร บรางว ลในเวท การประกวด ผลงานว จ ยและส งประด ษฐ ระด บนานาชาต ได แก รางว ลพ เศษจากงาน International Warsaw Invention Show 2014 (IWIS 2014) โดยสมาคมน กประด ษฐ ประเทศ โปแลนด เม อว นท ๑๔-๑๖ ต.ค. ๒๕๕๗ ณ กร งวอร ซอประเทศโปแลนด และรางว ลเหร ยญ เง นจากงาน Seoul International Invention Fair (SIIF2014) โดยส าน กงาน ทร พย ส นทางป ญญาแห งสาธารณร ฐเกาหล (KIPO) ณ กร งโซลสาธารณร ฐเกาหล ใต เม อ ว นท ๒๗-๓๐ พ.ย. ๒๕๕๗

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