Journal of Veterinary Science & Medical Diagnosis ISSN: 2325-9590

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Research Article, J Vet Sci Med Diagn Vol: 6 Issue: 1

Aluminium Silicate Clay as Mycotoxin Adsorbent in Dairy Cattle Feed

Starý J, Čoudková V*, Vrbová A, Svoboda V and Maršálek M
Department of Zootechnical Sciences, University of South Bohemia, Czech Republic
Corresponding author : Mgr. Veronika Čoudková
PhD Student, Department of Zootechnical Sciences, University of South Bohemia, Mažice 33, 391 81 Veselí nad Lužnicí, Czech Republic
Tel: +420721268702
E-mail: [email protected]
Received: December 30, 2016 Accepted: January 18, 2017 Published: January 23, 2017
Citation: Starý J, Čoudková V, Vrbová A, Svoboda V, Maršálek M (2017) Aluminium Silicate Clay as Mycotoxin Adsorbent in Dairy Cattle Feed. J Vet Sci Med Diagn 6:1. doi: 10.4172/2325-9590.1000221


The objective of the current study was to monitor the most common Fusarium mycotoxins Zearalenone (ZEA), T-2 toxin (T-2) and Deoxynivalenol (DON): in the occurence in dairy cows diet and its effect on dairy cows. Livestock health, welfare and productivity may be severely compromised by consumption of DON, T-2 toxin, ZEA and by interactions among these mycotoxins. Hydrated Sodium Calcium Aluminosilicate (HSCAS) helps to prevent the absorption of mycotoxins in the gut to prevent its symptoms. Influence of HSCAS intake to milk yield, components of milk and dairy diseases was evaluated. Research was located in South Bohemia in Central Europe. Four different types of feed which included grain mixes, corn silage, grass silage and hay, were sampled by ELISA. All samples of surveyed types of animal feed were positive for some of the monitored mycotoxins, however any of them didn‘t exeed guide values of EU Comittee recommendation for the content of mycotoxins in livestock feed. Co-occurrence of two mycotoxins was found at 56% of feed samples. It was observed 180 cows with HSCAS intake and 180 cows in the control group. It is concluded the positive impact of HSCAS intake on somatic cells count (SCC) in the milk by significant (p<0.05) reducing their number, on average 33,000 cells/ml. The occurence of diseases on average decreased by 17%, the largest positive effect was observed in the case of metabolic disorders. The results indicate that using of HSCAS has positive effect on milk yield, components of milk and occurence of cattle diseases by reduced impact of long-term exposure to subliminal doses of mycotoxins.

Keywords: Dairy cows; Zearalenone; T-2 toxin; Deoxynivalenol; Hydrated sodium calcium aluminosilicate


Dairy cows; Zearalenone; T-2 toxin; Deoxynivalenol; Hydrated sodium calcium aluminosilicate


The interest in mycotoxins began when aflatoxins were found to be carcinogens and to be widespread in foodstuffs and feedstuffs. Today, mycotoxins and mouldy feedstuffs are known causes of animal disease [1]. Mycotoxins can be formed on crops in the field, during harvest or during storage processing or feeding. Molds are present throughout the environment. The spores are high in the soil and in plant debris and lie ready to infect the growing plant in the field. Field diseases are characterized by yield loss, quality loss and mycotoxin contamination. Mold growth and the production of mycotoxins are usually associated with extremes in weather conditions leading to plant stress or hydration of feedstuffs to poor storage practices, low feedstuff quality and inadequate feeding conditions [2]. Metabolic and reproductive disorders may act indirectly through a subsequent decrease in milk yield and reproductive performance [3]. In cattle, mycotoxin consumption is associated with a decrease in feed intake, weight loss, reduced milk production, lack of response to diet change and terapies [4]. Mycotoxin-contaminated feeds impair farm operations as well as feed production in various ways: mycotoxins are invisible, odourless and cannot be detected by smell or taste, but can reduce performance in animal production significantly. The best control is the prevention of mycotoxins in the field, which is supported by proper crop rotation and fungicide administration at the right time. In the case of toxin manifestation, measures are required that act specifically against certain types and groups of toxins. Adsorptive compounds can be used for reduction of potency of mycotoxins in general. While adsorbents have proved to be efficient against some mycotoxin-induced toxicosis, alternative strategies such as enzymatic or microbial detoxification, have been used recently for counteracting impacts of certain fungal toxins [5]. It is known that some mycotoxins are generally termostable as much as 400°C [6], and thus, can be transfered to food, even after microbial stabilization steps [7]. Mycotoxin levels in corn silage and concentrated feeds were the factors most correlated with mycotoxin concentrations in milk [8]. There is also an indirect risk due to the carry over of toxins and their metabolites to edible animal products such as milk, meat, and eggs [9]. Mycotoxins are metabolized in the liver and the kidneys and also by microorganisms in the digestive tract [10].
Regarding animal feed, five mycotoxins (aflatoxins, DON, ZEA, fumonisins and ochratoxin A) are covered by EU legislation [11]. Transgressions of these limits are rarely observed in official monitoring programs [12]. The most frequently detected mycotoxins in feed for dairy cows are T-2, DON and ZEA. T-2 and DON belonging to nonmacrocyclic trichothecenes have a direct effect on the composition of rumen microflora and after absorption causes immunosuppression and damage many organs, firstly, liver, gastrointestinal tract and reproductive tract [13]. DON has the greatest prevalence in feedstuffs (between 20 and 100%) and is found in forage feeds, in forage corn in particular, and in ingredients for concentrate feeds [2,4,14]. DON is a Fusarium produced mycotoxin that is one of the most commonly detected in feed. It is sometimes called vomitoxin because it was first associated with vomiting in swine. Surveys have shown DON to be a primary mycotoxin associated with swine disorders including feed refusals, diarrhea, emesis, reproductive failure, and deaths [15]. DON and ochratoxin A are examples of mycotoxins that are transformed into less toxic metabolites in the rumen [16]. The impact of DON on dairy cattle is not established, but clinical data show an association between DON contamination of diets and poor performance in dairy herds, but without establishing a cause and effect [15]. DON may have adverse health effects after acute, short-term, or long-term administration. After acute administration, DON produces two characteristic toxicological effects: decrease in feed consumption (anorexia) and emesis (vomiting) [17].
ZEA (previously known as F-2 toxin) was produced by some Fusarium species Fusarium graminearum, Fusarium culmorum, Fusarium cerealis, Fusarium equiseti, Fusarium crookwellense and Fusarium semitectum. These fungi infected contaminants of cereal crops worldwide [18,19]. Toxicity of and its metabolites was related to the chemical structure of the mycotoxins, similar to naturally occurring estrogens [20]. Zearalenone and its metabolites bind to estrogen receptors, causing hyperstrogenic syndrome with all its manifestations [5], edema of the external genitalia, endometrium and mammary gland, ovarian disrupts the cycle and an increase in infectious and non-infectious diseases of the reproductive system and mammary gland [21].
The T-2 is a mycotoxin produced mainly by fungi of the genus Fusarium sporotrichioides [22] and belongs to the group of nonmacrocyclic trichothecenes [23]. Following ingestion, T-2 causes acute and chronic toxicity and induces apoptosis in the immune system and fetal tissues. T-2 is usually metabolized and eliminated after ingestion, yielding more than 20 metabolites [24]. Ruminants are considered to be less sensitive towards mycotoxins than monogastric animals because rumen microbiota has mycotoxin-detoxifying capacities [25]. This is based on the assumption that the rumen flora degrades and inactivates mycotoxins, thus protecting the animal [26]. This is different for dairy cows which combine milk production with pregnancy; this group is the most susceptible to all errors in breeding care, first of all to deficiencies in nutrition. The feed for dairy cows, attention is directed to mycotoxins formed by molds linked to the main component of the diet. Although not forgetting the source of feed in the form of concentrates.
As a prevention against mycotoxicoses several types of products have been used to help prevent the absorption of mycotoxins in the gut to prevent symptoms of mycotoxicoses, including both inorganic and organic-based products [27,28]. In practice is commonly used addition of supplements based on aluminosilicate clay [29], possibly live yeast cultures, or modified yeast walls [30,31]. The role of these binders is to adsorb and reduce the intestinal absorption of mycotoxins to reduce the toxic effects for livestock and the carry-over of toxin compounds to animal products [32]. In last several decades, various binders of different origins have ben investigated for their efficacy and capacity to adsorb mycotoxins [33]. The binding efficacy of mineral adsorbent is related to the structure of both the binders and the mycotoxins. The charge distribution, surface area and pore size of the adsorbents and the charge distribution, polarity and shapeof the mycotoxins singnitificantly contribude to the overall binding compatibility [34-36]. Regarding the applicability of aluminosilicates for the binding of mycotoxins, it can be concluded that they are very effective in preventing aflatoxicosis, but their efficacy against ZEA, ochratoxin, and trichothecenes is limited. In addition to the narrow binding range concerning different mycotoxins, aluminosilicates have the disadvantage of showing high inclusion rates for vitamins and minerals [27].


Farm characteristics
The study was carried out for a period of 31 months from July 2010 to December 2012 in Southern Bohemia. Average milk yield of Vleckviehn (Czech pied cattle) in this region was 6760 kg milk with 4.38% fat and 3.58% protein. Total Mixed Ration (TMR) contained 22 kg of corn silage, 14 kg of haylage, 0.75 kg of barley straw, 8.5 kg of grain mixes for dairy cows, 0.5 kg of hay, 0.15 kg of feeding limestone and 0.5 kg of soya extracted meal. A herd of 180 cows in production was observed, where was added HSCAS (5 kg per tonne of feed per day, via the TMR) to the feed and the control group consisted of 180 cows as well. The average annual temperature in the monitored area was 7.1°C, average annual rainfall was 729 mm per year and altitude ranged from 450 to 520 meters above sea level.
Co-bind AZ was used with HSASC to belongs to the group of highly purified clay, it is the combination of layering illite (K,H3O)(A l,Mg,Fe)2(Si,Al)4O10[(OH)2,(H2O)] and Chloritu (Mg,Fe)6 (Si,Al)4O10 (OH)8. The characteristic structure of the purified aluminosilicate clay allows not only a simple adsorption, but on the basis of layout of electric charges bonds with the polar group of mycotoxins, to a large extent are formed and covalent bonds which make it possible adsorption of molecules of mycotoxins with neutral or weak charge as in case of ZEA. Co-bind is tested in vitro for efficacy to ochratoxin, ZEA and Fumonisins.
Milk sampling and analysis
Milk samples were collected regularly 5-8 times a month with an autosampler in the period from June 2010 to December 2012. A total of 382 samples (191 in each group of cows) were evaluated, where was determined the content of milk fat, milk protein, urea, somatic cell count and average month milk yield (according to EUROPEAN STANDARD EN ISO 707).
Feed sampling – ELISA
Feed samples were collected continuously at monthly intervals directly from the storage area and TMR from the feeding table. There were collected and evaluated 124 samples of feed. Basic 1000 g sample was subsequently adjusted as needed ELISA (NOACK Ridascreen) to 2-5 g, and analyzed by ELISA. There were determined mycotoxins ZEA, DON and T-2. Under EC legislation [11] Member States should ensure that samples are simultaneously analyzed for the presence of DON, ZEA, ochratoxin A, fumonisin B1 + B2 and T-2 and HT-2 toxin, in order to assess the extent of co occurrence.
Health records
Health records were divided into five basic groups according to their economic significance or breeding importace. Diseases of the mammary gland, reproductive disorders, metabolic disorders, disease of the extremities, a group of diseases which are not classified, and which contained less common disorders such as skin problems, eye disorders, general febrile disease without direct relation to the illness of the aforementioned groups. Each occurrence of a new disease was categorized and evaluated monthly.
The Analysis of Variance statistical method was employed for check the existence of statistical differences in milk yeld and occurence of cattle diseases between two groups of dairy cows (180 cows receiving HSASC and 180 cows in the control group). It was performed by the software Statistica 12. The significance level of the test using the standard α = 0.05 cutoff, the null hypothesis has been rejected when p<0 .05 and not rejected when p>0 .05 [37].

Results and Discussion

Occurrence of mycotoxins
All samples of feed were positive for some of the monitored mycotoxins. In the total of 124 samples the percentage of positive samples was at 59%, 80%, 25% for the ZEA, T-2 and DON, respectively. Rate of positive samples of different types of feed are shown in Figure 1. Placinta et al. [30] concluded that on a global scale, cereal grains and animal feed may be subject to multiple contamination with trichothecenes, ZEA and fumonisins, the major mycotoxins of Fusarium fungi. Schllenberger et al. [38] monitored the occurrence and distribution of a spectrum of trichothecene toxins in different parts of maize plants in Germany. The percentage of positive samples was at 37% for the T-2 and 64% for the DON. With regards to the toxicity of maize fractions it should be considered that in plants chopped for silage making, uncontaminated fractions may be mixed with those strongly contaminated with mycotoxins [39], and that trichothecenes in naturally contaminated corn cob mixes and corn silages are still present after ensiling [39,40].
Figure 1: Rate of positive feed samples for occurrence of mycotoxins in monitored types of feed (ZEA = Zearalenone, T-2 = T-2 toxin and DON = Deoxynivalenol).
The spectrum of toxins in each type of feed, their mean, minimum and maximum contents are desribed in Table 1. The observed levels correspond to occurence of appropriate mycotoxins in other areas of Europe [41,42]. The highest levels of mycotoxins were observed in grain mixes (DON 1529 μg/kg feed) and in corn silage (DON 1329 μg/kg feed). Mean content of DON in corn silage was higher by 64 μg/ kg than content, which Driehuis et al. [4] mention. Amount of ZEA in samples of corn silage was lower by 316,4 μg/kg than amount in study of Schollenberger et al. [38]. Eckard et al. [43] found amount of T-2 in corn silage lower by 235 μg/kg. Mean content of ZEA in grain mixes was higher by 41 μg/kg, but DON was lower by 117 μg/ kg in comparison with Zachariasova et al. [44]. Obremski et al. [13] in study from USA demonstrated negative effect of T-2 in content in feed more than 350 ppb: a decrease in feed intake, weight loss of dairy cows and decrease of lactation curve. In this study there were found 17 feed samples (14%) with a T-2 over 350 ppb (μg/kg), of which 10 (8%) were corn silage, 5 (4%) grass silage and in two cases (2%) the grain mexes. None of the positive samples exeeded guide values of EU Comittee recommendation for the content of mycotoxins in livestock feed [11]. Eropean guide values for the content of mycotoxins in feed for livestock are 5,000 ppb (μg/kg) in grain mixes, 12,000 ppb in corn silage and 8,000 ppb in hay/grass silage in case of DON and 500 ppb (g/kg) in grain mixes, 3,000 ppb in corn silage and 2,000 ppb in hay/grass silage in case of ZEA. There are no guide values for T-2 in Europe. Common with other physiologically active compounds, the Fusarium mycotoxins are capable of inducing both acute and chronic effects. The effects observed are often related to dose levels and duration of exposure. It is clear that chronic intake of Fusarium and indeed other mycotoxins by farm livestock is inevitable and there are numerous cases of suspected mycotoxicoses on a worldwide basis [45]. In the long term altered the body’s defenses participate in the higher incidence of disorders of individual organ systems in cattle especially reproductive tract, breast, gastrointestinal and limb. Therefore when such evidence of deterioration in production and the health status of dairy cows appears it is important to think about the risk of mycotoxins and reduce their negative impact [46-48]. In cattle, despite its relative resistance to mycotoxins, there is a danger of longterm effects of subliminal doses. Chronic exposure of farm animals to DON is a continuing hazard in Canada, the USA and continental Europe [45].
Table 1: The levels of mycotoxins in surveyed types of animal feed: grain mixes, corn silage, grass silage and hay (in μg/kg; N = number of positive samples, μ ± SD = mean ± standard deviation).
Several Fusarium mycotoxins may co-occur in a particular feed ingredient or in compound feedingstuffs. In general, combinations of Fusarium mycotoxins result in additive effects, but synergistic and/ or potentiating interactions have been observed and are of greater concern in livestock health and productivity. Additive and synergistic effects between known and unidentified mycotoxins may account for enhanced adverse effects observed on feeding Fusarium-contaminated diets [45]. EU Comittee recommends [11], that member states should ensure that samples are simultaneously analyzed for the presence of deoxynivalenol, zearalenone, ochratoxin A, fumonisin B1 + B2 and T-2 and HT-2 toxin, in order to assess the degree of co-occurrence. In this study co-occurrence of monitored mycotoxins was determinated. All three monitored mycotoxins did not occure togehther in one sample. At 64 % samples, the occurence of two types of mycotoxins was detected (T-2 and ZEA at 57%, T-2 and DON at 5%, ZEA and DON at 2%). Only one type of mycotoxin was found at 36% of samples (T-2 at 18% and DON at 18%). ZEA did not occur independently. Detailed results for particular types of feed are demonstrated in Table 2. According to Streit et al. [12] the frequent detection of mycotoxin co-occurrence even in studies screening for a limited number of analyses underlines the importance of multi-mycotoxin analysis methods. Placinta et al. [30] note that the co-occurrence of individual trichothecenes in cereal grains and animal feed is serious enough that it is not possible yet to quantify the extent of any resulting interactions on animal health and performance.
Table 2: Co-occurrence of mycotoxins in monitored dairy cattle feed (ZEA = Zearalenone, T-2 = T-2 toxin and DON = Deoxynivalenol).
Changes in milk yield
For most of monitored milk characteristics positive impact of HSCAS intake was detected (Table 3). In case of milk yield, its use is demonstrated by an increase in anverage of 78.83 kg of milk per month (2.75%). Yiannikouris and Jouany [9] claim, that mycotoxins fed in amounts up to 300 μg/kg feed have a negative effect on milk production, if the content of mycotoxins in the feed exceeds the limit of 600 μg/kg feed, milk yield decreases. Geiger et al. [49] confirm these values in their study and have not seen an increase in milk production after adding adsorbents to feed. Use of HSCAS was completely without any effect in the case of milk fat amount and only a minimal difference was found in the values of milk proteins (0.02%). Similar conclusions reached also Nageswara Rao and Chopra [50] who carried out their experiment in small ruminants. The content of urea is lower by an average of 15.96 mg/ml (6%).
Table 3: Influence of Hydrated Sodium Calcium Aluminosilicate (HSCAS) adsorbent to milk yield and milk components (N = number of months/milk samples; significance:*p<0.05, - p>0.05).
Only in the case of somatic cells count (SCC) in the milk, significant effect of HSCAS intake was proven. The number of somatic cells in the milk was significanty (p<0.05) reduced, on average by 33,000/ml (13,5%). These results are consistent with the conclusions of Fink-Gremmels [26]. Obremski et al. [13] demonstrated negative effect of T-2 in content of 350 ppb in the feed on lactation curve, by its decrease. Prolonged exposure of DON levels from 2.6 to 6.4 ppm in the feed leads to a reduction in milk yield of 1.35 liters /cow /day [51].
Health disorders
The incidence of health disorders directly related to the levels of mycotoxins. Their direct toxic effect is particularly rare in ruminants, more often leads to impaired general health, decrease of the immune response of orgasnism and increase of pressure on the cow, particularly in the transit period.
Disruption of rumen fermentation by affecting of rumen microorganisms leads to a change in the composition of volatile fatty acids (impact of mycotoxins on in vitro organisms in the rumen content). This results in changes of content of milk fat and milk protein, and in general milk production, chronic stress leads to a decrease in immune mechanisms, higher incidence of mastitis, chronic endometritis and load liver [52]. Feeding by naturally contaminated feed leads to a decrease in serum immunoglobulins levels [53].
The impact of HSCAS intake on the incidence of diseases was studied. The decrease of the amount of newly-morbidities recorded in the period of one month occurred in all studied groups of disorders. A significant (p<0.05) positive effect was proved on the total number of newly reported illnesses per month, the value of which fell on average by 17 %. From the results in Table 4 it is obvious, that the HSCAS intake leads to decrease in the incidence of newly recorded disease by an average of two cases per month. A significant effect of the HSCAS intake on the number of newly reported metabolic disorders was proved, the decline was an average of 50%. This result agrees with Driehuis et al. [4]. The smallest decrease of new monthly reported disorders was observed in the case of diseases of the mammary gland. ZEA causes swelling vulva, vulvovaginitis, swollen mammary, endometrial hyperplasia, ovarian cysts and reduced milk production [21]. According to Gaumy at al. [54] estrogenic effect manifests itself after a minimum of 4-7 day exposure. When feed contains 0.5-1 μg/kg feed, negative effect of ZEA result in changes in reproductive performance [1]. In this study we did not obtain these values of ZEA in samples, however Whitlow et al. [15] highlight the potentiating effect in co-occurrence of DON and ZEA.
Table 4: Influence of Hydrated Sodium Calcium Aluminosilicate (HSCAS) adsorbent to health disorders (N = number of monitored months, mean = mean of number of newly reported diseases per month; significance:* p<0.05, - p>0.05, other diseases = less common diseases).


Analysis of fodder for three most frequently occurring mycotoxins (DON, T-2 and ZEA) showed 100% contamination, although in no feed were found above the limit value. Nevertheless, long-term exposure to low doses of mycotoxins and their interactions can lead to disruption of the health condition of dairy cows and reduce milk production. HSCAS appears to be effective feed supplement for reducing negative effects in the case of low contents of mycotoxins in the feed. This is manifested as a decline in the incidence of metabolic diseases, improving milk yields, with higher milk fat and decrease of the number of SCC.


This work would not have been possible without the financial support of Grant Agency of University of South Bohemia GAJU- 019/2016/Z.


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