Efficacy of Minazel Plus® in reducing detrimental effect of Ochratoxin A in laying hens
Published: November 3, 2015
By: Nedeljkovic Trailovic Jelena (Asociate Professor, Department of Nutrition and Botany, Faculty of Veterinary Medicine, University of Belgrade, Serbia) and Vasiljevic Marko ( Patent komerc d.o.o, Mišicevo, Serbia).
Summary
The aim of this study was to investigate the efficacy of modified inorganic adsorbent Minazel Plus® (MZ) in the prevention of the detrimental effects of Ochratoxin A (OTA) in laying hens. Fourthly eight (n=48) laying hens (27 week old) were used in this study. Therefore, hens were randomly divided to six equal groups, and were feeding 7 weeks by standard diet in addition to: E-I group - 1 mg/kg OTA; E-II group 0,25 mg/kg OTA; E-III group - 0,25mg/kg OTA + 0,2% of MZ; E-IV group 1 mg/kg OTA + 0,2% of MZ; E-V group 0,2% MZ. The control group of hens was fed only standard diet, without any addition. Influence of OTA on production parameters (body weight, feed consumption, number and mass of eggs), as well as pathohistology changes of kidney tissue were monitored. Laying hens fed with diets contaminated with OTA, and supplemented with 0,2% of MZ, achieved better production results compared to the results recorded in the hens that were given food contaminated with OTA without adsorbent. Interestingly, the supplementation of MZ 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). These finding needs to be further investigated and clarified, but clearly point to the protective potential of MZ against the toxic effects of OTA in laying hens.
a) Minazel Plus®, is a trademark of a modified inorganic adsorbent property of Patent Commerce and in Mexico, of Lapisa, S. A. de C.V.
Introduction
Mycotoxins are secondary fungal metabolites, whose presence in feed and foodstuffs. Ochratoxin A (OTA) is one of the most known mycotoxins with greatest public health and agro-economic significance (Duarte 2011). Whereas, OTA is considered a potent nephrotoxic and hepatotoxic effects (Stormer and Lea, 1995). In commercial birds, consumption of OTA-contaminated diets has been related to clinical signs of toxicosis, involving reduced feed intake, disorder of egg production and egg quality (Prior and Sisodia, 1978, Haazele et al., 1993; Verma at al., 2003). Consumption of OTA by birds may also be associated with the appearance of OTA residues in eggs (PiskorskaPliszczynka and Juszkiewicz 1990). MINAZEL® Plus is adsorbent created as result of ion exchange reaction between inorganic captions on the mineral surface and organic captions. Addition of organic cations is used in order to change mineral surface. Result of this addition is not a simple mixture of mineral and organic phase, but a completely new compound, organic complex. New active centers, which are formed on the mineral surface, ensure efficient binding (over 90%) not only of polar mycotoxins (Aflatoxins, Ergot Alkaloids, etc.), but also of non-polar mycotoxins (Zearalenone, Ohratoxin A, T-2 toxin, etc.)
In the order to investigate the efficacy of a modified inorganic adsorbent MINAZEL® Plus in the prevention of the detrimental effects of OTA in laying hens, the presented study was organized.
Material and methods
Birds and Diets:
Fourthly eight (n=48) laying Lowman Brown hens (27 week old) were used in this study. Average body weight of birds was 1520±87,29 g, while study lasted 49 days. Birds were placed in a light-controlled (16L:8D) and temperature-controlled (22°C) room in the wire cages with unlimited access to drinking water. Hens were randomly divided in to six equal groups, and been fed 7 weeks by standard diet in addition to: E-I group - 1 mg/kg OTA; E-II group 0,25 mg/kg OTA; E-III group - 0,25mg/kg OTA + 0,2% of MZ; EIV group 1 mg/kg OTA + 0,2% of MZ; E-V group 0,2% MZ. The control group of hens (C) was fed only standard diet, without any addition. Hens were fed once a day, while standard diet considered complete mixtures (with or without the above additions), whose raw and chemical composition was according to the NRC recommendations (National Research Council, 1994).
The contaminated diet was prepared using OTA obtained by contamination of corn with Aspergillus spp. OTA was produced using a culture of Aspergillus ochraceus Wilhelm NRRL 263.67 from the Dutch collection. Tested adsorbent - MINAZEL® Plus (MZ) is originated from the zeolite tuff that was modified by the addition of an organic cation.
All procedures were done in accordance to the guidelines of the Ethical Committee of the Faculty of Veterinary Medicine, Belgrade University as well as EU Directive 2010/63/EU for animal experiments.
Hens Production Performances end Egg quality
Eggs were collected daily to measure egg weight, also daily were measured feed consumptions, while body weights were recorded weekly.
Histopathological examinations of kidneys
The samples for histopathological examinations were fixed in a solution of 10% neutral formalin and absolute ethanol and molded by the standard paraffin technique. Tissue samples (5-8 µm) were then stained by the hematoxylin eosin technique as described by (Scheuder and Chalk, 1986).
Statistical analysis
All results were statistically analyzed, for differences between groups, by an analysis of variance (ANOVA). The results were processed by the use of Graph Pad Prism® 5.0 software (Graph Pad Software Inc., San Diego, California, USA). All values are expressed as the mean ± SE.
Results and Discussion
Table 1. Body weight of laying hens during the trail (g).
During the experiment, starting from the second week, the BW of hens was significantly lower in E-I group in relation to all other experimental and control groups. Also, a group of hens which received just the MZ in commercial diet, had significantly higher BW compared to the hens in E-I, E-II and E- III group (Table 1). These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008).
Table 2. Feed consumption (g)
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 E-II group, which are given diet with 0,25 mg/kg OTA, had periodically lower BW in comparison with the hens in E-III, E-IV group, as well as in comparison with hens in MZ and control group. These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008).
Table 3. Number of eggs.
Number of eggs in E-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% MZ (E-III), produced a significantly higher number of eggs in relation to E-I group. This result indicates the achievement of the protective effect of the adsorbent against OTA in the feed of hens. The highest number of eggs produced hens that have received commercial diet containing 0.2% MZ. These results are consistent with the data of Duarte et al. (2011) and Denli et al. (2008).
Table 4. Mass of yolk/hens/group
Weight of yolks (separated from the egg white by a separator) was measured on the technical balance, accuracy of 0.01 g. The presented results indicate that during the 2nd, 3rd and 4th week, the average mass of yolk in laying hens E-I group (1mg/kg OTA) was decrease. However, the average weight of egg yolks in these hens was significantly lower in relation to the results in group E-IV (0.25 mg/kg OTA + 0.2% Minazel plus) and the control group. During the 6th and 7th week of study, statistically significant difference in mean yolks weight appeared between hens in E-I group and all the other experimental and control groups. Probably, as a result of long-term presence of OTA in feed, during the last two weeks of the study took a place significant depletion of nutrients reserves in hen’s bodies. Therefore, significant differences in the mean weight of yolk between E-I and all other experimental and control groups, was recorded. However, among the other experimental and control groups statistically significant difference in weight of yolks was not appeared.
After 7 weeks of study hens were sacrificed and kidney samples were taken for histopathological examination. In hens E-I group (1mg / kg OTA) on histopathologic examination were observed degenerative changes in the tubules (blurry swelling, vacuolar degeneration and hydrops of tubulocytes). Different degree of desquamation of tubulocytes and necrobiotic changes in the cells nuclei was recorded in 8/8 birds figure 1. Tubular atrophy and dilatation of the lumen of tubules was proved in 8/8 hens, while hemorrhages were present in the tissues of the kidney in 5/8 examined birds.
In hens E-II group histopathological examination of kidney tissue was detected different degree of degenerative changes. Degenerative changes in the tubules (blurry swelling, vacuolar degeneration and hydrops of tubulocytes), different degree of desquamation of tubulocytes and necrobiotic changes in the cells nuclei were seen in the tissue of all tested birds (8/8) figure 2. Furthermore, tubular atrophy was present in 6/8 hens and dilatation of the lumen of tubules in 6/8 hens.
Histopathological changes were less expressed in the kidney of hens that were fed diet containing OTA and adsorbent. In hens of E-III group (1 mg/kg OTA + 0,2% MZ), degenerative changes in the tubules (blurry swelling), necrobiotic changes (karyopyknosis) and tubular atrophy were seen in the tissue 6/8 hens figure 3. Dilatation of the tubular lumen was observed in 6/8 birds, while the cuboidal metaplasia of the parietal layer of Bowman's capsule (glomerulopathy) was seen in 2/8 examined hens.
Significantly less pronounced changes were recorded in histopathological examination of the kidney tissue of hens that received a lower dose of OTA and adsorbent, E-IV (E-IV 0,25 mg/kg OTA + 0,2% MZ). Degenerative changes in the tubules (blurry swelling) and tubular atrophy were seen in the tissue of 7/8 and 6/8 examined hens, respectively figure 4. Histopathological examination are consistent with the findings of Duarte et al , 2011 aand Bozzo et al , 2008)
Figure 1: E-I group degenerative changes in the tubules (blurry swelling, vacuolar degeneration and hydrops of tubulocytes 8/8 hans
Figure 2: E-II group Tubular atrophy 6/8 hans
Figure 3: 2: E-III group Tubular atrophy 6/8 hans
Figure IV: E-IV group Tubular atrophy 6/8 hans
Acknowledgement:
This work was supported by grant TR31087 Ministry of Education, Science and Technological Development of The Republic of Serbia.
Conclusions
Based on our results, it is clear that the OTA expressed a negative impact on body weight and feed consumption in both the applied concentrations. On the other hand, addition of MZ 0,2% in the feed, improves all production parameters in laying hens, regardless of whether they are fed with or without OTA. Also, the feed consumption in hens treated with OTA was lower compared to the results in other groups. Adding MZ 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, MZ exhibited on the number of laid eggs, while the same effect was observed in the average weight of yolk. Finally, we especially emphasize, that the addition of the MZ in the hens feed contaminated with OTA, decrease in the intensity and frequency of histopathological changes in the kidneys tissue.
All the results obtained show that Minazel plus is a potent adsorbent, which can reduce the harmful effects of OTA on the health and production parameters in laying hens.
Literature:
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Prior, M. G. and C. S. Sisodia. 1978. Ochratoxicosis in white legorn hens. Poult. Sci. 57,169.
Haazele, F. M., W. Guenter, R. R. Marquardt and Frohlich, A.A: 1993. Beneficial effects of dietary ascorbic acid supplement on hens subjected to ochratoxin A toxicosis under normal and high ambient temperatures. Can. J. Anim. Sci. 73:149-157.
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Bozz, G. Ceci, E. Bonerba, E. Desantis S. an Tantillo, G. 2008. Ochratoxin A in laying hans: High-Performance Liquid Chromatography detection and cytological and histological Analysis of target tissues. J. Appl. Poult. Res. 17. 151-156.
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