In vitro & in vivo studies on MINAZEL PLUS

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.

H. Farkas, Jog Raj, J. Bosnjak-Neumüller & Marko Vasiljevic
PATENT CO, Serbia

Mycotoxins

Mycotoxins are secondary metabolites that are produced by moulds stressed by environmental and management factors, such as extreme temperatures, drought, flooding and harvesting techniques.

More than 300 mycotoxins are known so far but only a small number are relevant to the feed industry. The most important mycotoxins are Aflatoxins produced by Aspergillus, Ochratoxins produced by Penicillium and Fumonisin, Zearalenone and Deoxynivalenol produced by Fusarium sp.
In this article we are going to 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.

In the following trials, MINAZEL PLUS, which is referred to as MP, was used.

Analysis

Test in vitro with ADSORPTION & DESORPTION



LESS POLAR 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.
Once adsorbed to the clinoptilolite, mycotoxins are not desorbed through the intestinal tract.

pH

The adsorbent 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.

ADSORPTION/DESORPTION

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.



Adsorbent(%)



The adsorbent MP can adsorb in the in vitro studies:
  • 100% of AFB1
  • 88% of Zearalenone
  • 91% of Ochratoxin A
  • 98% of FB1


Desorption(%)

The desorption rate was:
  • 0% for AFB1
  • 2,3% for Zearalenone
  • 1,0% for Ocratoxina A
  • 3,9% for FB1


SPEED OF ADSORPTION

Exposure to mycotoxins is mostly by ingestion. Only the fraction of an orally ingested mycotoxin, in a certain food matrix, that finally reaches the systemic circulation and is distributed throughout the entire body, exert its toxic effect.
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)



In vivo studies



AIM
The aim of this study was to investigate the efficacy of the mycotoxin binder in the prevention of the detrimental effects of Ochratoxin A (OTA) in laying hens.


MATERIAL & METHODS

Animals
48 laying hens (27-week-old) were used in this study.

TREATMENTS & DIETS
The hens were randomly divided to six equal groups and were fed during 7 weeks with the following diets:

  • Negative Control - Standard diet without mycotoxins
  • Treatment I - Standard diet + 1 mg/kg OTA
  • Treatment II - Standard diet + 0,25 mg/kg OTA
  • Treatment III - Standard diet + 0,25mg/kg OTA + 0,2% MP
  • Treatment IV - Standard diet + 1 mg/kg OTA + 0,2% MP
  • 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.

Table 1. Body weight of laying hens during the trial (g).
RESULTS
Laying hens fed with diets contaminated with OTA, and supplemented with 0,2% of MP, achieved better production results compared to the results recorded in the hens that were given food contaminated with OTA without binder.
The supplementation of MP in the standard diet, by itself resulted in the higher body weight, number of eggs and improved feed conversion ratio than in the control group of hens (fed with standard diet without any addition).
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) Datos estadísticamente signi cativos

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.
The highest number of eggs produced hens that have received commercial diet containing 0.2% MP.

Table 3.Number of eggs, Statistically significant results. These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008)


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.
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




Micotoxicosis prevention
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