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)
FEED AF
FUMONISIN
OTA T2
DON
ZEA
% positive samples 84 66 65 39 57 37
Av. Concentration (ppb) 18
106 21
111 76

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
CAPTEX T2 combines in one product 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.
Sodium Propionate (the sodium salt of propionic acid) acts as a mold inhibitor to prevent mold growth and proliferation
Thereby minimizing the risk of having mycotoxin-producing molds that further proliferate in the feed.
Sodium Propionate 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
Bacteria
Propionic
Salmonella typhimurium
1.5
Pseudomonas aeroginosa
2.0
Escherichia coli
Staphylococcus aureus
2.5
Listeria monocytogenes
Campylobcter jejuni
Clostridum botulinum
Clostridum perfringes
Molds
Propionic
Aspergillus Niger
2.5
Penicillum expansum
1.3
Fusarium nivale
Cladosporium sp.

7. 2. ABsorption by minerals
Through a number of screening studies the best aluminosilicate adsorbents with regard to mycotoxin deactivation and safe application where selected.
A premium blend of minerals was designed that as part of CAPTEX T2 product guarantees maximum, pH-independent activity at an inclusion rate as low as 1 kg/ton without removing essential nutrients from the diet.
Through a number of screening studies the best aluminosilicate adsorbents with regard to mycotoxin deactivation and safe application where selected.
A premium blend of minerals was designed that as part of CAPTEX T2 product guarantees maximum, pH-independent activity at an inclusion rate as low as 1 kg/ton without removing essential nutrients from the diet.

8. 2. Adsorption by minerals
CAPTEX T2 contains 2 main aluminosilicates
Calcium Bentonite (montmorillonite)
Zeolite (clinoptilolite)
Through a number of screening studies the best aluminosilicate adsorbents with regard to mycotoxin deactivation and safe application where selected.
A premium blend of minerals was designed that as part of CAPTEX T2 product guarantees maximum, pH-independent activity at an inclusion rate as low as 1 kg/ton without removing essential nutrients from the diet.

9. Bentonite / Montmorillonite
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.

10. Molecular structures of Bentonite (Montmorillonite)
2. ABsorption by minerals
Molecular structures of 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
Vimonses, V., Lei, S., Jin, B., Chow, C., Saint, C. (2009) Kinetic study and equilibrium isotherm analysis of Congo Red adsorption by clay materials. Chemical production Journal, Volume 148, Issues 2–3, 15, Pages 354–364

11. Zeolites / Clinoptilolite
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.

12. molecular structures of Clinoptilolite
2. ABsorption by minerals
molecular structures of 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
Manley and Holmes, 1989, Minerals with natural advantage, New Scientist, March 25, 1989, pp

13. molecular structures of Bentonite (Clinoptilolite)
2. ABsorption by minerals
molecular structures of 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
Manley and Holmes, 1989, Minerals with natural advantage, New Scientist, March 25, 1989, pp

14. 2. ABsorption by minerals
CAPTEX
Belongs to a new generation of mycotoxin deactivators, 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

15. Physical Treatment and active surface of CAPTEX T2
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.

16. Chemical Treatment and CEC of CAPTEX T2
2. ABsorption by minerals
Chemical Treatment and CEC of CAPTEX 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!!

17. 2. ABsorption by minerals
CAPTEX T2 does not bind either nutrient or drugs! Maximum size of Captex T2 holes is 50 microns
Name of compound
Minimum
particle size
(microns)
VITAMIN A
350
VITAMIN D3
200
VITAMIN B12
120
FOLIC ACID
IRON SULFATE
250
ZINC SULFATE
160
TYLOSINE
150
AMOXYCILLIN
Name of mycotoxins
Maximum
particle size
(microns)
AFLATOXIN 1
ZEARALENONE
OCHRATOXIN
FUMONISIN T2
VOMITOXIN

18. Physical & Chemical Treatment
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. 

19. 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 trichothecene mycotoxin.
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.
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 the yeast cell by the patented process and becomes active only at intestinal level when the yeast cell is lyzed by the intestinal enzymes and the cell content is released.
Chitinase is able to form non toxic de-epoxy metabolite by removing oxygen from the epoxide group of the trichothecene mycotoxin.
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.

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

21. Biotransformation of 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
CAPTEX reduces the toxicity of T-2 toxin by been able to form de-epoxy metabolites (which are essentially non toxic compounds), by removing oxygen from the epoxide group of the trichothecene mycotoxin. This action seems to mimic 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.

22. Not all glucomannans are the same!
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.
Our product contains Glucomannans, in an esterified form. Glucomannans are known to be able to protect the intestinal mucosa from toxins adhesion. But it is very important that the natural content of chitin, in glucomannans, be the minimum possible, in order to increase the capacity of the substance to be “flexible”.

23. Vopato I. Bizzini B. -1998 Italian Project M.S.T./09/96
Glucans and Chitin contents of various sources of Saccharomyces cerevisiae
Yeast Sources
Total Glucans (%)
(1.3)-glucans (%)
(1.3)-glucans (%)
Chitin (%)
Saccharomyces cerevisiae 1
13.2
7.2
6.0
9.0
Saccharomyces cerevisiae 2
14.4
7.4
7.0
9.2
Saccharomyces cerevisiae 3
13.5
7.3
6.2
9.1
Saccharomyces cerevisiae 4
13.9
7.6
6.3
10.0
Doxal’s
S.C. 2234
18.2
10.6
2.7
Doxal’s Yeast source for Glucomannans has a very high content of glucans, specially beta-Glucans, and an extremely low level of Chitin.
Vopato I. Bizzini B Italian Project M.S.T./09/96

24. Toxins binding capacity of three feed additives in vitro
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.
A test, carried out in vitro, comparing the capacity of three compounds to bind various mycotoxins, under same conditions.
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. Seven replicates for each mycotoxin were prepared.
100 µg and 250mcg 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, Column Kromasyl, fluorescence detector, excitation 274 nm, flow rate 1 mL/min, emission 440 nm.

25. Toxins binding capacity of three feed additives in vitro
CAPTEX T2 TRIALS
Toxins binding capacity of three feed additives in vitro
AFLATOXIN
CLAY AT 100 MCG
7.40
MCG
CLAY AT 250 MCG
2.40
BENTONITE AT 100 MCG
BENTONITE AT 250 MCG
1.00
CAPTEX-T2 AT 100 MCG
4.05
CAPTEX-T2 AT 250 MCG
0.00
BLANK
49.90
These results indicate that, out of 50 mcg of toxin, Clay at first dosage has been able to bind most of it, as only 7.40 mcg, at last, were found not bound to it.
These results tend to demonstrate also that there is a dose-depending response. When Aflatoxin is the only contaminant, also very cheap and common minerals, like clay and/or bentonite are effective in binding the toxin.
Anyhow Captex T-2, as 250mcg, was the only one able to bind 100% of the toxin.

26. Toxins binding capacity of three feed additives in vitro
CAPTEX T2 TRIALS
Toxins binding capacity of three feed additives in vitro
ZEARALENONE
CLAY AT 100 MCG
42.60
MCG
CLAY AT 250 MCG
36.60
BENTONITE AT 100 MCG
27.70
BENTONITE AT 250 MCG
20.00
CAPTEX-T2 AT 100 MCG
15.10
CAPTEX-T2 AT 250 MCG
9.90
BLANK
49.70
When the contaminating toxin is Zearalenone, administration of Clay is performing very poorly. Bentonite has a limited effect. Captex-T2 is very effective. Again, there is a dose-depending response (higher dose, better results).

27. Toxins binding capacity of three feed additives in vitro
CAPTEX T2 TRIALS
Toxins binding capacity of three feed additives in vitro
OCHRATOXIN
CLAY AT 100 MCG
6.90
MCG
CLAY AT 250 MCG
3.10
BENTONITE AT 100 MCG
7.00
BENTONITE AT 250 MCG
2.95
CAPTEX-T2 AT 100 MCG
3.60
CAPTEX-T2 AT 250 MCG
0.20
BLANK
50.10

28. Toxins binding capacity of three feed additives in vitro
CAPTEX T2 TRIALS
Toxins binding capacity of three feed additives in vitro
FUMONISIN
CLAY AT 100 MCG
47.48
MCG
CLAY AT 250 MCG
46.10
BENTONITE AT 100 MCG
44.90
BENTONITE AT 250 MCG
42.50
CAPTEX-T2 AT 100 MCG
27.40
CAPTEX-T2 AT 250 MCG
15.05
BLANK
49.30
In case of Fumonisin, both Clay and Bentonite are ineffective.

29. TOXINS BINDING CAPACITY OF CAPTEX on T-2 toxin IN VITRO
CAPTEX T2 TRIALS
TOXINS BINDING CAPACITY OF CAPTEX on T-2 toxin IN VITRO
T-2 TOXIN
Activated carbon AT 100 MCG
45.63
MCG
Activated carbon AT 250 MCG
42.14
CAPTEX-T2 AT 100 MCG
22.42
CAPTEX-T2 AT 250 MCG
15.21
BLANK
49.20
In case of Fumonisin, both Clay and Bentonite are ineffective.

30. Trial in vivo Poultry field - Results
CAPTEX T2 TRIALS
Trial in vivo Poultry field - Results
Feed
consumption
Body
weight
mortality
Plasmatic
calcium
g/day g %
mM/L
group A
24,7
344
1,66
2,180 ± 0,16
group B
16,4
265,4
8,33
1,64 ± 0,66
group C
23,1
326,4
2,09 ± 0,22
A total of 540 day-old chicks (Ross)(Gallus gallus) was randomly divided into three groups, of 180 birds each.
Standard feed and water were provided ad libitum, for 20 consecutive days.
Group A was fed standard blank feed (Control Positive).
Group B was fed standard feed, contaminated by 3 ppm Aflatoxin (Control Negative)
Group C was fed standard feed, contaminated by 3 ppm Aflatoxin and fortified by 0.3% CAPTEX (Test).
The Aflatoxin used in this test was supplied by Bidifarm - Italy, and was produced by Aspergillus parasiticus.
Body weights and feed consumption were recorded every seven days.
Mortality was recorded daily.
We may conclude that:
the addition of 3 ppm Aflatoxin in feed for day-old chicks, fed continuously for 20 consecutive days, causes lower Feed Intake, lower Growth, increases Mortality and moreover it decreases Calcium Plasmatic levels.
The addition of Captex to feed for day-old chicks is very effective in significantly reducing the toxicity effects caused by 3 ppm Aflatoxin fed continuously for 20 days.

31. Thanks for the kind attention