Repox50
A novel, natural polyphenol partial
replacement for vitamin E in feeding diets
Repox50
Presentation overview
 The problem
– Oxidative stress
 The solution
– Antioxidants
 Our recommended solution
– Repox50
Repox50
 The problem
– Oxidative stress
 The solution
– Antioxidants
 Our recommended solution
– Repox50
Repox50
Reactive oxygen species

Reactive oxygen species (ROS) are chemically-reactive molecules
containing oxygen. Examples include oxygen ions and peroxides.

Increased ROS can result in significant damage to cell structures. This
cumulates into a situation known as oxidative stress.

Antioxidants are used to counteract the effect of these increased ROS
levels often in combination with the addition of extra minerals and vitamins
to the diet.
Repox50
Some causes of oxidative stress?
Poor
nutrition
package
Poor living
conditions
Infections
Oxidative Stress
Repox50
Problems resulting from oxidative stress

Growth inhibition

Muscle degeneration, leading to inferior meat quality

Suppression of the immune system response

Loss of flavour, texture, appearance and in some cases nutritional value of
the meat
Repox50
 The problem
– Oxidative stress
 The solution
– Antioxidants
 Our recommended solution
– Repox50
Repox50
What are antioxidants?

Antioxidants are phytochemicals, vitamins and other nutrients that protect
our cells from damage caused by free radicals.

In vitro en in vivo studies have shown that antioxidants help prevent the free
radical damage that is associated with disease.

Antioxidants have the ability to neutralize free radicals without becoming a
free radicals themselves, however, when the antioxidant neutralises a free
radical it becomes inactive. Therefore we need a continuous supply of
antioxidants.
Repox50
Antioxidants in action
Repox50
Some commonly used antioxidants

Vitamin E
– A fat-soluble antioxidant that stops the production of reactive oxygen species
formed when fat undergoes oxidation.

Vitamin C
– Vitamin C or L-ascorbic acid or L-ascorbate is an essential
nutrient for humans and certain other animal species, in which it functions as
a vitamin.

Selenium
–
Selenium is necessary for cellular function in most, if not all, animals, forming the
active centre of the enzymes glutathione peroxidase and thioredoxin reductase.
Repox50
 The problem
– Oxidative stress
 The solution
– Antioxidants
 Our recommended solution
– Repox50
Repox50
Repox50 the natural choice

Super concentrated polyphenols extracted from specifically selected
botanical sources.

Works in synergy with Vitamin E, able to replace up to 50% resulting the
same or superior protective effect, thus creating more cost effective
inclusion into in poultry diets.

Sustainably produced from natural renewable resources.

Safe and easy to handle.
Repox50
Repox50 Heat Stability
OVERVIEW
Two representative samples of Repox50 were received from Phytologica Ltd,
batch number LAB/PHY01. This sample was separated into 2 samples for test
purposes. Sample 1 and Sample 2.
• Sample 1 was subjected to an increase in temperature for 60 minutes in
an autoclave at a temperature of 120°C.
• Sample 2 was untreated.
• Sample 1 and Sample 2 were analysed for polyphenols using a cyclic
voltammetry technique (CV) used routinely in the Citrox QC laboratory.
Repox50
Repox50 Heat Stability
LAB/PHY01 - Sample 1 (120°C)
MondaydEC1514W.efwdat : Page 21
40
i 1 ( A)
20
0
-20
-40
-1.2
-0.8
-0.4
0
E (V)
0.4
Mode: Cyclic, Initial E: -1200mV, Final E: -1200mV, Rate: 80mV/ s, Step W: 25ms, Upper E: 1200mV, Lower E: -1200mV, Cycles: 5 (Mon, Dec 15, 2008, 4:00:59 PM)
Some data points in the recording are out of range. This may affect the results.
0.8
1.2
Repox50
Repox50 Heat Stability
LAB/PHY02 - Sample 2 (untreated)
MondaydEC1514WJE.efwdat : Page 26
40
i 1 ( A)
20
0
-20
-40
-1.2
-0.8
-0.4
0
E (V)
0.4
Mode: Cyclic, Initial E: -1200mV, Final E: -1200mV, Rate: 80mV/ s, Step W: 25ms, Upper E: 1200mV, Lower E: -1200mV, Cycles: 5 (Mon, Dec 15, 2008, 4:08:13 PM)
Some data points in the recording are out of range. This may affect the results.
0.8
1.2
Repox50
Repox50 Heat Stability
Conclusion
• “there were no discernable differences in the voltammograms which can
be thought of as “fingerprints” of the polyphenol activity”
Testing carried out by Dr. David J. Lax BSc MRSC PhD Chemical Research Services
Repox50
Some scientific evidence

Sparing effect of procyanidins from Vitis vinifera on vitamin E: in vitro studies
–

Select flavonoids and whole juice from purple grapes inhibit platelet function and
enhance nitric oxide release
–

Carini M, Maffei Facino R, Aldini G, CalloniI MT, Bombardelli E, Morazzoni P, 1998 : The protection of
polyunsaturated fatty acids in micellar systems against UVB-induced photo-oxidation by procyanidins from
Vitis vinifera L., and the protective synergy with vitamin E. International J. of Cosmetic Science, Volume 20
Issue 4 Page 203-215,August 1998.
Freedman JE, Parker C 3rd, Li L, Perlman JA, Frei B, Ivanov V, Deak LR, Iafrati MD, Folts JD. , 2001. Select
flavonoids and whole juice from purple grapes inhibit platelet function and enhance nitric oxide release.
Circulation. 2001 Jun 12;103(23):2792-8.
Catechins and procyanidins in Mediterranean diets
–
AUGER Cyril, AL-AWWADI Najim, BORNET Aurélie, ROUANET Jean-Max, GASC Francis, CROS Gerard,
TEISSEDRE Pierre-Louis : Catechins and procyanidins in Mediterranean diets Food research International
2004, vol. 37, no3, pages. 233-245 [13 page(s) (article)] (1 p.1/4)
Repox50
In vitro antioxidant trials
Trial Objective
To measure the antioxidant activity at 0%, 25%, and 50% of the vitamin E substitued by Repox50 and
compare with control.

Growth Control: No antioxidant added

100% Vitamin E: 0% substitution
Increase of antioxidant
activity compared to no

75% Vitamin E: 25% substition
antioxidant addition

50% Vitamin E: 50% substitution
Conclusion

100% Vitamin E antioxidant activity
is 250% higher than negative control

Replacement of 25% and 50% of
Vitamin E by Repox50 the antioxidant
activity is still identical to 100% Vitamin E
33.00%
33.00%
34.00%
Repox50
33.00%
34.00%
Poultry Applications
33.00%
Repox50
Poultry diet trials
Effect of Repox50 and Vitamin E on Growth Performance, Nutrient Digestibility
in Chickens

120 one-day-old male broiler chicks were obtained from a commercial hatchery and housed in an
environmentally controlled room over a period of 3 weeks. All diets were formulated to meet or
exceed the minimum NRC (1994) requirements for broiler chickens.

At the end of the experimental period, birds were weighed, and feed consumption was recorded

The addition of increasing concentration of Repox50 in the chicken diets did not impair growth
performance (BW, feed consumption, and feed efficiency) compared with the birds fed the non
supplemented and supplemented vitamin E diets

Antioxidant activity in vitamin E and Repox50 diets exhibited significantly higher scavenging free
radical capacity than controls.

Similarly, the birds fed vitamin E and Repox50 diets exhibited significantly higher scavenging
free radical capacity in excreta than those fed control diets.
Repox50
BANGKOK ANIMAL RESEARCH CENTER CO., LTD.
HEAD OFFICE
487/1 SI AYUTTHAYA ROAD, KHWAENG THANON PHAYATHAI, KHET
RATCHATHEWI,
BANGKOK 10400, THAILAND
Evaluation of Repox50 in Broilers under
research station circumstances
Trial No. AB11238B
Repox50
Trial Detail
Objective
This trial was conducted to evaluate the effect of replacing vitamin E by Repox
50 on performance of broilers.
Materials and methods
The trial was conducted at BARC research facility during July 6 – August 10,
2011. Six hundred newly hatched male broiler chicks of commercial strain (AA
Plus) were randomly allocated to 3 treatments with 8 replications in a
randomized complete block design experiment, using 25 chicks per
experimental unit. Practical corn-soybean diet with 100 g/t added vitamin E 50
was used as control diet and two levels of replacing of vitamin E 50 by Repox50.
Treatment diets were presented as below.
Diet 1 = Control diet: practical diet with 100 g/tone added Vitamin E 50
Diet 2 = Replacing 25% of added Vitamin E of diet 1 by Repox 50
Diet 3 = Replacing 50% of added Vitamin E of diet 1 by Repox 50
Repox50
Composition of starter and grower diets
Code
B103-1
B422
B417-1
B202
B601-2
B602-2
B603
B908
B703
B502
B501
B503
B605
B606
B804
B910
Ingredient
Corn 7.35%
Soybean meal (DH) 49.0%
Full fat soybean 36.0%
Soybean oil
MCP
Lime stone 39% Ca
Salt
Pellet binder
Vitamin/mineral permix (100g/t of vitamin E 50)
DL-Methionine
L-Lysine HCl
L-Threonine
Sodium bicarbonate
Choline chloride 60%
Toxin binder
Coccidiostat (Salinomycin 12%)
TOTAL
COST
Price
(Baht / kg)
11.50
14.07
19.17
40.00
18.50
1.40
4.00
70.00
55.00
150.00
72.00
78.00
15.00
64.00
200.00
250.00
Baht / kg
Starter
Grower
0-18 d
Control diet
18-35 d
Control diet
49.09
31.56
10.00
4.82
1.78
1.06
0.33
0.30
0.20
0.27
0.16
0.03
0.20
0.10
0.05
0.05
57.67
25.94
8.00
4.67
1.44
0.90
0.23
0.30
0.20
0.18
0.07
0.20
0.10
0.05
0.05
100.00
15.47
100.00
14.93
Repox50
Nutrients of starter and grower diets
NUTRIENT
1
2
3
9
11
12
13
17
18
19
20
21
22
23
24
25
26
46
47
48
49
50
52
53
55
56
57
58
59
66
67
68
69
78
79
80
Weight
Dry matter
ME. For poultry
Crude protein
Crude fat
Linoleic acid
Crude fiber
Dig.Lys (Poultry)
Dig.Met (Poultry)
Dig.Cys (Poultry)
Dig.M+C (Poultry)
Dig.Thr (Poultry)
Dig.Trp (Poultry)
Dig.Arg (Poultry)
Dig.Val (Poultry)
Dig.Ile (Poultry)
Dig.Leu (Poultry)
Lysine
Arginine
Methionine
Met + Cys
Cystine
Threonine
Tryptophan
Histidine
Isoleucine
Leucine
Valine
Phenylalanin
Calcium
Phosphorus-total
Phosphorus-avail
Non phytate
Sodium
Chloride
Salt
Unit
Kg
%
Kcal/kg
Kcal/kg
Kcal/kg
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
%
1
87.85
3050
23.0
9.05
4.29
3.22
1.24
0.58
0.31
0.89
0.77
0.20
1.42
0.96
0.83
1.69
1.41
1.59
0.62
1.00
0.38
0.92
0.27
0.62
1.02
1.89
1.11
1.07
0.86
0.75
0.43
0.50
0.20
0.23
0.36
1
87.62
3100
20.0
8.73
4.16
3.01
1.00
0.46
0.29
0.75
0.65
0.17
1.22
0.84
0.72
1.55
1.14
1.36
0.50
0.84
0.35
0.78
0.23
0.55
0.88
1.71
0.97
0.94
0.72
0.66
0.36
0.42
0.16
0.18
0.27
Repox50
Statistical Analysis
Feed intake, body weight gain, feed conversion ratio, liability, FRAP value and
TBA value were determined and were subjected to ANOVA as RCBD. Significant
differences among treatment groups were detected by LSD tests (least square
deviation).
Repox50
Results
Table1 Evaluation of Repox 50 in broilers1 under research station
circumstances (0-18 days of age).
Treatment
Final
Body
Feed
Feed
Feed
intake
conversion
conversion
ratio2
ratio3
Repox 50
body
weight
weight
With Vit E 50
(replacing of
weight
gain
gain
100%
Vit E 50)
(g)
(g)
(g)
(g)
1
Control diet
-
44
838
793
957
1.207
1.210
99.5
2
As diet 1
25 %
44
835
790
956
1.210
1.217
99.0
3
As diet 1
50 %
44
833
789
964
1.222
1.228
99.0
P-value
0.7026
0.6850
0.4195
0.1181
0.1213
0.8815
Pooled SEM
3.576
3.564
4.448
0.005
0.006
0.809
C.V.%
1.21
1.27
1.31
1.15
1.33
2.31
Male broilers (Arbor Acres Plus).
Feed conversion ratio corrected for mortality and culls.
3 Feed conversion ratio uncorrected for mortality and culls.
2
Livability
Diet
Group
1
Initial
(%)
Repox50
Table 2 Evaluation of Repox 50 in broilers1 under research station
circumstances (18-35 days of age).
Treatment
Final
Body
Feed
Feed
Feed
intake
conversion
conversion
ratio2
ratio3
Repox 50
body
weight
weight
With Vit E 50
(replacing of
weight
gain
gain
100%
Vit E 50)
(g)
(g)
(g)
(g)
1
Control diet
-
44
838
793
957
1.207
1.210
99.5
2
As diet 1
25 %
44
835
790
956
1.210
1.217
99.0
3
As diet 1
50 %
44
833
789
964
1.222
1.228
99.0
P-value
0.7026
0.6850
0.4195
0.1181
0.1213
0.8815
Pooled SEM
3.576
3.564
4.448
0.005
0.006
0.809
C.V.%
1.21
1.27
1.31
1.15
1.33
2.31
Male broilers (Arbor Acres Plus).
Feed conversion ratio corrected for mortality and culls.
3 Feed conversion ratio uncorrected for mortality and culls.
2
Livability
Diet
Group
1
Initial
(%)
Repox50
Table 3 Evaluation of Repox 50 in broilers1 under research station
circumstances (0-35 days of age).
Treatment
Group
Initial
Final
Body
Feed
Feed
Feed
intake
conversion
conversion
ratio2
ratio3
Diet
Repox 50
body
weight
weight
With Vit E 50
(replacing of
weight
gain
gain
100%
Vit E 50)
(g)
(g)
(g)
(g)
Control diet
-
44
2565
2521
3798
1.507
1.524
98.0
2
As diet 1
25 %
44
2571
2527
3826
1.515
1.521
98.5
3
As diet 1
50 %
44
2580
2535
3831
1.511
1.526
98.0
P-value
0.7960
0.7926
0.4966
0.5555
0.9726
0.9641
Pooled SEM
15.253
15.243
20.676
0.005
0.015
1.508
1.68
1.71
1.53
0.91
2.70
4.34
Male broilers (Arbor Acres Plus).
Feed conversion ratio corrected for mortality and culls.
3 Feed conversion ratio uncorrected for mortality and culls.
2
(%)
1
C.V.%
1
Livability
Repox50
Table 4 Evaluation of Repox 50 on fat oxidation ability in broilers1 under
research station circumstances.
Treatment
Group
Body
Feed
Feed
Feed
intake
conversion
conversion
ratio2
ratio3
Repox 50
body
weight
weight
With Vit E 50
(replacing of
weight
gain
gain
100%
Vit E 50)
(g)
(g)
(g)
(g)
Livability
(%)
1
Control diet
-
44
2565
2521
3798
1.507
1.524
98.0
2
As diet 1
25 %
44
2571
2527
3826
1.515
1.521
98.5
3
As diet 1
50 %
44
2580
2535
3831
1.511
1.526
98.0
P-value
0.7960
0.7926
0.4966
0.5555
0.9726
0.9641
Pooled SEM
15.253
15.243
20.676
0.005
0.015
1.508
1.68
1.71
1.53
0.91
2.70
4.34
Male broilers (Arbor Acres Plus).
Thiobarbituric acid
3 Ferric reducing antioxidant power
2
Final
Diet
C.V.%
1
Initial
Repox50
Fish Applications
Repox50
Repox50 will enhance colour and shelf life via replacing a portion of
Vitamin E within the diet
Repox50
Polyphenols can improve body color and meat brilliance of cultured fishes,
preventing decrease in astaxanthin levels in the fish.
Polyphenols added by weight from 0.004% 40ppm to 0.04% 400ppm.
Sakiura US 6,509,030, B2
Cultured fish carotenoid and polyphenol added feed for improving fish body color tone and fish meat brilliance
Repox50
Meat Applications
Repox50
Some scientific evidence

Ahn, J., Grün, I.U., Fernando, L.N. 2002. Antioxidant properties of natural plant extracts containing
polyphenolic compounds in cooked ground beef. J. Food Sci. 67(4), 1364-1369.

Ahn, J., Grün, I.U., Mustapha, A. 2007. Effects of plant extracts on microbial growth, color change,
and lipid oxidation in cooked beef. Food Micro. 24, 7-14

Bekhit, A.E.D., Geesink, G.H., Ilian, M.A., Morton, J.D., Bickerstaffe, R. 2003. The effects of
natural antioxidants on oxidative processes and metmyoglobin reducing activity in beef patties.
Food Chem. 81(2), 175-187

Brannan, R.G., Mah, E. 2007. Grapeseed extract inhibits lipid oxidation in muscle from different
species during refrigerated and frozen storage and oxidation catalyzed by peroxynitrite and
iron/ascorbate in a pyrogallol red meat model system. Meat Science. 77(4), 540-546.

Jayaprakasha, G.K., Singh, R.P., Sakariah, K.K. 2001. Antioxidant activity of grape seed (vitis
vinifera) extracts on peroxidation models in vitro. Food Chem. 73(3), 285-290.
Repox50
Factors affecting meat colour
•
•
•
Vitamin E feeding of cattle
– Prevents oxidation; retards conversion of myoglobin to metmyoglobin
Bacteria
– Produce metmyoglobin, choleglobin, and sulfmyoglobin pigments
Curing
– Nitrosylhemochromogen is the stable cured meat pigment
Repox50
Repox50 improves meat colour
Repox50
Factors affecting meat colour
Exposure to long-term or short term stress
• Effects glycogen content of muscle and ultimate pH of muscle
• Long Term Stress: DFD (dark cutter)Transport, Hunger, Fear, Aggression
• Ultimate pH above 5.9 (beef), 6.5 (pork)
• Short Term Stress: PSE Usually only problematic in pork
• Ultimate pH below 5, generally problem can be overcome with
enhancement
Thank you for your time
Any questions?
Repox50