CAPTEX T2

1. CAPTEX T2

2. Effect of the different mycotoxins in on the different farmed animals • SPECIES • Poultry • Broilers • Layers • Breeder • Sheep • Goats • Fish • Shrimp Mycotoxins • Aflatoxin (AFB) • Fumonisin (FB) • Ochratoxin (OTA) • T-2 Toxin (T-2) • Zearalenone (ZEN) • Vomitoxin or Deoxynivalenol (DON)

3. MYCOTOXIN CONCENTRATIONS IN COMPLETE FEED IN EUROPE (318 samples from 10 countries, 2012)

4. How does CAPTEX T2 deactivates mycotoxins (mode of action) 3 different strategies to counteract a broad spectrum of mycotoxins • Preventing further mold growth • Adsorption by minerals • Biotransformation by enzymatic activity

5. 1. Preventing further mold growth Sodium Propionate • acts as a mold inhibitor to prevent mold growth and proliferation • minimizes risk of having mycotoxin-producing molds further proliferating in feed. • has no activity against mycotoxins, but, always, when you ascertain mycotoxins presence, there is still a residual presence of molds.

6. 1. Preventing further mold growth Minimum inhibitory concentration of propionic acid (g/kg diet) on bacteria and molds

7. 2. Adsorption by minerals CAPTEX T2 contains 2 main aluminosilicates • Calcium Bentonite (montmorillonite) • Zeolite (clinoptilolite)

8. 2. ABsorption by minerals Bentonite / Montmorillonite • originally created from the breakdown (weathering) of volcanic ash. • is a silicate with a layered polar crystalline microstructure which adsorbs organic substances either on its external surfaces or within its interlaminar spaces. • Used to bind mainly Aflatoxin.

9. 2. ABsorption by minerals Molecular structuresof Bentonite (Montmorillonite) 2:1 clay structure: one octahedral sheet sandwiched between two tetrahedral sheets The mycotoxins that are bound by the montmorillonite are those that can physically enter into the interlayer space. The width of the interlayer space is 0.25 to 0.7 nanometers in the dry state and 1 nanometer in the hydrated state. Aflatoxins and ochratoxins can enter into this space

10. 2. ABsorption by minerals Zeolites / Clinoptilolite • is of (sea or lake) sedimentation origin with a unique, complex crystalline structure. • honeycomb (tetrahedral) framework of cavities and channels act like cages, trapping mycotoxins. • Used to deactivate Zearalenone, Ochratoxin and Fumonisin.

11. 2. ABsorption by minerals molecular structuresof Clinoptilolite 3-dimentional crystalline Structure with an 8-ring and 10-ring channels has a cage-like structure, with pores and channels running through the crystal. The cage and surrounding mineral carries a net negative charge, making it one of the few negatively charged minerals found in nature

12. 2. ABsorption by minerals molecular structuresof Bentonite(Clinoptilolite) Because of its cage-like structure and negative charge, clinoptilolite has the ability to draw and trap within and on itself positively charged toxic particles that fit into the pores and channels of the cage It acts as molecular sieves and absorb substances of a low-molecular compounds (mycotoxins) Used to adsorb Zearalenone, Ochratoxin and Fumonisin, Deoxynivalenol

13. 2. ABsorption by minerals CAPTEX • Belongs to a new generation of mycotoxindeactivators, which are classified as highly purified and activated clays. • The minerals undergo a special process that involves; • Physical treatment(Micronisation) • Chemical treatment which greatly increases its adsorbent efficiency

14. 2. ABsorption by minerals Physical Treatment and active surface of CAPTEX T2 • The high affinity of binding is due to the fact that the product is modified by the micronisation process to be extremely fine, 150,000 particles/gram. • This provides for a larger surface area that increases the possibility of interacting with mycotoxins. • Though extremely fine, caution is given not to have any particles size of less than 5 microns in order to avoid dustiness as well as inhalation issues with the people that are handling the product.

15. 2. ABsorption by minerals Chemical Treatment and CEC ofCAPTEX T2 • CEC (Cationic Exchange Capacity) is important as it explains the water absorption capacity of the product. • The lower the CEC the better it is, as its water absorption capacity decreases. In the other hand, if CEC is below of 35 then it starts losing its affinity to mycotoxins. • CEC over 100, means that the product has high in water absorption capacity and consequently some nutrients can be trapped. • CAPTEX T2 has a CEC of approximately 55. IDEAL!!

16. 2. ABsorption by minerals CAPTEX T2 doesnotbindeithernutrient or drugs!Maximum size of Captex T2 holesis 50 microns

17. 2. ABsorption by minerals Physical & Chemical Treatment These treatments enable the minerals to form a stable irreversible complex that immobilizes the target mycotoxins in the gastrointestinal tract of animals and by reducing their bio-availability, they are prevented from being absorbed through the gut and into the blood circulation, so thus eliminated through faeces. 

18. 3. Biotransformation • is the enzymatic degradation of mycotoxins that leads to non-toxic metabolites. In this case, Chitinase is this hydrolytic enzyme capable of deactivating mycotoxins by degrading their molecules. • Chitinase is incorporated into yeast cells by our patented process and becomes active only at intestinal level when yeast cells are lyzed by the intestinal enzymes and cell content is released. • Chitinase is able to form non toxic de-epoxy metabolite by removing oxygen from the epoxide group of the trichothecenemycotoxin. • This action mimics the detoxifying process carried out by carboxylesterase (a microsomal enzyme from liver) that selectively hydrolyses the C-4 acetyl group of T-2 toxin.

19. 3. Biotransformation Molecule of T-2 and what happens to it after Biotransformation by Chitinase

20. 3. Biotransformation Biotransformationof T-2 toxin This action mimics the detoxifying process carried out by carboxylesterase (a microsomal enzyme from liver) that selectively hydrolyses the C-4 acetyl group of T-2 toxin to yield HT-2 toxin

21. CAPTEX T2 Not all glucomannans are the same! • Prior to their inclusion into CAPTEX-T2, Doxal’s Esterified Glucomannans are treated by another Chitinase, an enzyme which is able to reduce, dramatically, their chitin content. • Chitin content is supposed to increase the alkali insolubility of β-glucans and to decrease the cell wall flexibility, so that the toxin molecule has a restricted access to the complexing chemical sites.

22. Glucans and Chitincontents of varioussources of Saccharomycescerevisiae Vopato I. Bizzini B. -1998 Italian Project M.S.T./09/96

23. CAPTEX T2 TRIALS Toxins binding capacity of three feed additives in vitro • Each mycotoxin was solved at the level of 50 µg into 200 ml of methanol, and kept under gentle stirring throughout the test period, in a 250 ml volumetric flask. • 100 mcg and 250 mcg aliquots of each Clay, Bentonite and CAPTEX-T2 were solved into the flasks, and kept under gentle stirring for 20 minutes; one flask of each mycotoxins was left as blank. • After 20 minutes, small aliquots were collected from each volumetric flask and then assayed by High Performance Liquid Chromathography.

24. CAPTEX T2 TRIALS Toxins binding capacity of three feed additives in vitro

25. CAPTEX T2 TRIALS Toxins binding capacity of three feed additives in vitro

26. CAPTEX T2 TRIALS Toxins binding capacity of three feed additives in vitro

27. CAPTEX T2 TRIALS Toxins binding capacity of three feed additives in vitro

28. CAPTEX T2 TRIALS TOXINS BINDING CAPACITY OF CAPTEX on T-2 toxin IN VITRO

29. CAPTEX T2 TRIALS Trial in vivoPoultryfield - Results

30. Thanks for the kindattention