H. Farkas, Jog Raj, J. Bosnjak-NeumĂźller & Marko Vasiljevic PATENT CO, Serbia
in vitro & in vivo STUDIES
on
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Global use of feed materials in the production of animal feed is increasing the risk of chemical and microbiological contaminants in food-producing animals. The feed can be contaminated with microorganisms, mycotoxins, animal by-products, organic pollutants and toxic metals. Mycotoxin contamination is now recognized as a global issue in agricultural production, especially in the livestock sector.
HMore than 300 mycotoxins are known so far but only a small number are relevant to the feed
MYCOTOXINS
industry. The most important
In this article we are going to
mycotoxins are Aflatoxins produced
Mycotoxins are secondary
by Aspergillus, Ochratoxins produced
metabolites that are produced by
by Penicillium and Fumonisin,
moulds stressed by environmental
Zearalenone and Deoxynivalenol
and management factors, such as
produced by Fusarium sp.
focus on analysis of mycotoxins, in vitro and in vivo studies carried on MINAZEL PLUS, obtained by organic modification of the surface of clinoptilolite, known for its capacity of adsorbing polar and non-polar mycotoxins.
extreme temperatures, drought, flooding and harvesting techniques.
In the following trials, MINAZEL PLUS, which is referred to as MP, was used.
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In vitro testing using Adsorption & Desorption method LESS POL AR MYCOTOXIN ADSORPTION MINAZEL PLUS is a result of organic modification of clinoptilolite surface, well known for adsorbing polar and non-polar mycotoxins. By firmly attaching long-chain organic cations on surface of the mineral, the new active centres for adsorbing less polar mycotoxins become available.
pH
ADSORPTION/DESORPTION
The adsorbet MP is highly effective across a wide spectrum pH range as it absorbs a greater percentage of mycotoxins in low pH gastric juices and does not desorb mycotoxins in the presence of intestinal juices where pH is close to neutral.
The analysis carried out in our laboratory using LC-MS/MS (Combined analysis with Liquid Chromatography and Mass Spectrometry) and have revealed that MP can adsorb all major mycotoxins more than 80% with very low desorption.
Once adsorbed to the clinoptilolite, mycotoxins are not desorbed through the intestinal tract.
The adsorbent MP can adsorb in the in vitro studies:
100% of AFB1 88% of Zearalenone 91% of Ochratoxin A 98% of FB1
Adsorption(%) Aflatoxin B1
The desorption rate was:
Ochratoxin A
Desorption(%)
0% for AFB1 2,3% for Zearalenone
100%
91%
1,0% for Ochratoxin A 3,9% for FB1
Aflatoxin B1
0%
1% Ochratoxin A
Zearalenone
88%
Fuminosin B1
98,7%
2,3% Zearalenone
3,9% Fuminosin B1
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SPEED OF ADSORPTION Exposure to mycotoxins is mostly by ingestion. Only the
The adsorbent MP can adsorb
fraction of an orally ingested mycotoxin, in a certain food
more than 78% of AFB1, FB1,
matrix, that finally reaches the systemic circulation and is
ZON and OTA within five minutes
distributed throughout the entire body, exert its toxic effect.
of contact with mycotoxins.
The bioaccessibility of the mycotoxins corresponds with the number mycotoxins present in the alimentary bolus and that are available for absorption in the small intestine. Certain mycotoxins are absorbed faster after oral intake. After 30 min they can be found in the blood, and after 60 min in the liver.
Speed of adsorption (LC-MS/MS)
Aflatoxin B1
90.2%
5 min
Zearalenone
99.6%
30 min
100%
60 min
Ochratoxin A
79.5%
5 min
78.5%
5 min
83%
93.2%
30 min
60 min
Fumonisin B1
86.3%
87.4%
79.7%
86.6%
87.3%
30 min
60 min
5 min
30 min
60 min
Time
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In vivo studies AIM The aim of this study was to
LIVE WEIGHT (G)
investigate the efficacy of the mycotoxin binder in the prevention
1st day
14th day
28th day
49th day
of the detrimental effects of
T-I
15020±52,07
13565±62,06
13190±62.85
11320±64,04
Ochratoxin A (OTA) in laying hens.
T-II
14968±34,20
13905±41,62
14150±54.17
14435±29.07
T-III
14805±76,95
14550±67,68
14770±35.98
13665±34,05
MATERIAL & METHODS Animals 48 laying hens (27-week-old)
T-IV
14865±59,49
15005±44,70
15155±34.76
15340±50,56
Positive Control
14970±38.32
b
15350±29.27
15840±49,90
15975±42,70
Negative Control
15025±40,34
15055±47.97
15260±35,30
15565±36,10
were used in this study.
Table 1. Body weight of laying hens during the trial (g).
Statistically significant results
TREATMENTS & DIETS The hens were randomly divided
RESULTS
to six equal groups and were fed during 7 weeks with the following diets: Laying hens fed with diets contaminated with OTA, and Negative Control
supplemented with 0,2% of MP, achieved better production
Standard diet without mycotoxins
results compared to the results recorded in the hens that were given food contaminated with OTA without binder.
Treatment I Standard diet + 1 mg/kg OTA
The supplementation of MP in the standard diet, by itself resulted in the higher body
Treatment II
weight, number of eggs
Standard diet + 0,25 mg/kg OTA
and improved feed
Treatment III Standard diet + 0,25mg/kg OTA + 0,2% MP
conversion ratio than in the control group of hens (fed with
Treatment IV
standard diet
Standard diet +
without any
1 mg/kg OTA + 0,2% MP
addition).
Positive Control Standard diet without mycotoxins + 0,2% MP
PARAMETERS Influence of OTA on production parameters (body weight, feed consumption, number and mass of eggs) was monitored.
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During the experiment, starting from the second week, the BW of hens was significantly lower in T-I group in relation to all other experimental and control groups.
Also, a group of hens which received just the adsorbent MP in commercial diet, had significantly higher BW compared to the hens in T-I, T-II and T-III group (Table 1).
Compared to the all experimental and control group, feed consumption was significantly lower in the hens that received diet with 1 mg/kg of OTA.
Also, the hens of T-II group, which are given diet with 0,25 mg/kg OTA, had periodically lower BW in comparison with the hens in T-III, T-IV group, as well as in comparison with hens in the MP and Positive Control groups.
Table 2. Feed consumption (g) These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008) Statistically significant results
CONSUMPTION (G)
1st week
2nd week
3rd week
4th week
5th week
6th week
7th week
T-I
655±5,31,01
675±72,17
724,3±59,3
794,3±73,0
841,7±69,51
879,3±15,12
815,0±53,62
T-II
913,6±14,06
921,4±24,62 946,4±6,26
944,3±5,34
943,6±14,35
938,6±28,97
951,4±5,56
T-III
932,1±9,51
950,7±3,45
951,4±6,90
895,7±109,4 868,6±56,91
898,6±31,19
925,0±22,73
T-IV
945,7±4,49
950,7±5,34
952,1±4,88
944,3±14,56 954,3±4,49
944,3±17,18
952,1±10,75
PC
951,4±7,48
954,3±5,34
956,4±3,78
960,0±0,0
959,3±1,89
956,4±4,75
958,6±2,44
NC
951,4±10,69
949,3±6,07
955,0±7,07
957,9±5,66
958,6±2,44
956,4±2,44
957,9±2,67
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Nº EGGS Number of eggs in T-I group during the 2nd and 3rd weeks of the study, was significantly lower than the number of eggs in the other experimental and control groups. This trend continued until the end of the experiment.
It is interesting that a group of hens which received 1 mg/kg OTA + 0,2% MP (T-III), produced a significantly higher number of eggs in relation to T-I group. This result indicates the achievement of the protective effect of the mycotoxin binder against OTA in the feed of hens.
1st week
2nd week
3rd week
4th week
5th week
6th week
7th week
T-I
52±0,14
40±0.42
39,0±0,20
47±1,38
46±0,53
45±0,97
38±0,53
T-II
52±0,37
53±0,53
55,0±0,14
55±0,38
55±0,14
54±70,48
54±0,18
T-III
54±0,28
54±0,48
55,0±0,14
53±0,53
53±0,3
53±0,78
53±0,20
T-IV
54±0,18
55±0.38
54±0,18
54±0,48
55±0,14
56 ± 0,0
53±0,3
PC
52±0.0
56±0,0
56,0±0,0
56±0,0
56±0,0b
56 ± 0,0
56±0,0
NC
51±0.3
52±0,20
55±0,14
56±0,0
55±0,14
54±0,48b
50±0,14
The highest number of eggs produced hens that have received commercial diet containing 0.2% MP.
CONCLUSIONS
The OTA expressed a negative impact on body weight and feed consumption in both the applied concentrations. The addition of MP 0,2% in the feed, improves all production parameters in laying hens, regardless of whether they are fed with or without OTA.
The feed consumption in hens treated with OTA was lower compared to the results in other groups. Adding MP in food contaminated with OTA showed a protective action with respect to harmful effects of mycotoxins on the observed physiological parameters in hens. A clear protective effect against OTA by the adsorbent MP was exhibited on the number of laid eggs.
Table3. Number of eggs Statistically significant results These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008)
The results obtained show that the adsorbent MP is a potent adsorbent, which can reduce the harmful effects of mycotoxins on the health and production parameters in animals.
References upon request
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