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Pii: s0378-4274(01)00360-5

Toxicology Letters 122 (2001) 179 – 188 Alexander Huwig a, Stefan Freimund a, Othmar Ka¨ppeli b, Hans Dutler b a Swiss Federal Institute of Technology, Laboratory of Technical Chemistry, Uni6ersita¨tstrasse 6, 8092 Zurich, Switzerland b ABAC R&D Ltd., Strangenstrasse 1a, 8304 Wallisellen, Switzerland Received 18 December 2000; received in revised form 20 April 2001; accepted 30 April 2001 Abstract
The contamination of animal feed with mycotoxins represents a worldwide problem for farmers. These toxins originate from molds whose growth on living and stored plants is almost unavoidable particularly under moistconditions. Mycotoxin-containing feed can cause serious diseases in farm animals resulting in suffering and evendeath and thus can cause substantial economic losses. The most applied method for protecting animals againstmycotoxicosis is the utilization of adsorbents mixed with the feed which are supposed to bind the mycotoxinsefficiently in the gastro-intestinal tract. Aluminosilicates are the preferred adsorbents, followed by activated charcoaland special polymers. The efficiency of mycotoxin binders, however, differs considerably depending mainly on thechemical structure of both the adsorbent and the toxin. This review describes the most important types of adsorbentsand the respective mechanisms of adsorption. Data of the in vitro and in vivo efficacy of detoxication are given.
2001 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Review; Mycotoxin detoxication; Activated charcoal; Aluminosilicate; Clay; Zeolite 1. Introduction
ins, ochratoxin A, trichothecenes, zearalenone,and fumonisins.
Mycotoxins are produced by several fungi, par- Cereal plants may be contaminated by myco- ticularly by many species of Aspergillus, Fusarium, toxins in two ways. First, there are fungi growing Penicillium, Cla6iceps, and Alternaria. They com- as pathogens on plants; secondly, there are fungi prise a group of several hundreds of chemically growing saprophytically on stored plants. In this different toxic compounds (William, 1989; Moss, context, it has to be considered that not all of 1996; Rotter et al., 1996; Sweeney and Dobson, these fungi form mycotoxins, i.e. the detection of 1998). The most common mycotoxins are aflatox- fungi is not the same as the detection of mycotox-ins because many fungi are not able to producemycotoxins or produce them in different amounts * Corresponding author. Tel.: + 41-1-8391018; fax: + 41-1- depending on the substrate on which they are E-mail address: (S. Freimund).
growing. However, high incidence rates of con- 0378-4274/01/$ - see front matter 2001 Elsevier Science Ireland Ltd. All rights reserved.
PII: S 0 3 7 8 - 4 2 7 4 ( 0 1 ) 0 0 3 6 0 - 5 A. Huwig et al. / Toxicology Letters 122 (2001) 179 – 188 tamination of cereal grains and animal feed have Rood, 2000). These methods include fermentation been reported worldwide (Placinta et al., 1999; procedures with microorganisms. One example is Spahr et al., 1999), so that the contamination of diets by mycotoxins and the carry-over of myco- Fla6obacterium auranticum) to harmless degrada- toxin related compounds through the food chain tion products. The conversions, however, are gen- (Ramos and Hernandez, 1996) have to be accu- erally slow and incomplete (Sweeney and Dobson, rately controlled. Although in terms of acute toxi- 1998; Arici, 1999; Bata and La´sztity, 1999; city even the most poisonous of the mycotoxins is far less toxic than the botulinum toxin (Moss, Chemically, some mycotoxins can be destroyed 1996), the consumption of mycotoxin contami- with calcium hydroxide monoethylamine (Bauer, nated diet may induce acute and long-term 1994), ozone (McKenzie et al., 1997; Lemke et al., chronic effects resulting in a teratogenic, carcino- 1999) or ammonia (Park, 1993). Particularly the genic (mainly for liver and kidney), oestrogenic, ammoniation is an approved procedure for the or immunsuppressive impact not only on animals detoxication of aflatoxin-contaminated feed in but also on man whereas animals usually suffer some U.S. states as well as in Senegal, France, more due to grain of lower quality (D’Mello et and the UK. The average ammoniation costs vary al., 1999; Steyn and Stander, 1999; Casteel and between 5 and 20% of the value of the commodity Rottinghouse, 2000). In addition to the toxic ef- (Coker, 1998). Main drawbacks of this kind of fects, a mycotoxin contaminated diet may lead to other consequences like feed refusal, poor feed against other mycotoxins and the possible deterio- conversion, diminished body weight gain, in- ration of the animals health by excessive residual creased disease incidence due to immune suppres- sion, and interference with reproductive capacities The physical methods are focused on the re- (CAST, 1989; Lindemann et al., 1993; Kubena et moval of mycotoxins by different adsorbents al., 1998a) which are responsible for great eco- added to mycotoxin-contaminated diets (Ramos et al., 1996a) with the hope of being effective in In order to avoid mycotoxicosis, several strate- the gastro-intestinal tract more in a prophylactic gies have been investigated (Doyle et al., 1982; rather than in a therapeutic manner. At present, Park, 1993; Bauer, 1994; Ramos and Hernandez, however, the utilization of mycotoxin-binding ad- 1997) which can be divided into pre- and post- sorbents is the most applied way of protecting harvest technologies and into biological, chemical, animals against the harmful effects of decontami- The best procedure to prevent the effect of mycotoxins is the minimizing of the mycotoxinproduction itself (Miedaner and Reinbrecht, 2. Efficacy of different adsorbents for the binding
1999), e.g. by harvesting the grain at maturity and of mycotoxins
low moisture and storing it at cool and dry condi-tions which is difficult to perform in countries Herein, the adsorbents are discussed particu- with a warm and humid climate. Furthermore, the larly concerning efficacy, specificity, and the growth of fungi and therefore the production of mechanism of the adsorption process. The latter is mycotoxins is limited by the use of propionic acid similar to a chemical reaction and therefore, the or ammonium isobutyrate. Feed additives like release of free energy (DG) is the driving force of antioxidants, sulphur-containing amino acids, vi- every adsorption. The most important feature of tamins, and trace elements can be useful as detox- the adsorption is the physical structure of the adsorbent, i.e. the total charge and charge distri- Biological methods are not yet used in practice bution, the size of the pores and the accessible though the number of corresponding patents in- surface area. On the other hand, the properties of creases continously (Erber, 1996; Duvick and the adsorbate molecules, the mycotoxins, like po- A. Huwig et al. / Toxicology Letters 122 (2001) 179 – 188 larity, solubility, size, shape and — in case of ries one negative charge which has to be compen- ionized compounds — charge distribution and sated by positive charges, usually sodium ions as dissociation constants play a significant role, too.
in zeolite A. Zeolites are similar to molecular Therefore, the efficacy of every adsorption process sieves as well as to ion exchange resins and are has to be investigated in regard to the particular suitable for the distinction of different molecules by size, shape, and charge. HSCAS contain cal-cium ions and protons which are exchanged against the naturally occuring sodium ions. Theyare a type of montmorillonite belonging to phyl- Activated charcoal which is formed by pyrolysis losilicates which are composed of layers of alu- of organic materials is a very porous non-soluble minium and silicon connected in a 1:1 or 2:1 powder with a high surface to mass ratio (500 – 3500 m2/g). Since the 19th century it has been The applicability of aluminosilicates for the ad- used as an antidote against poisoning. Therefore, sorption of mycotoxins (Table 1) has been studied it might also inactivate mycotoxins. In aqueous for more than 20 years (clays: Masimango et al., solution, it can adsorb most of the mycotoxins 1978; zeolites: Mumpton and Fishman, 1977; efficiently (Table 1) whereas different activated HSCAS: Davidson et al., 1987; Ramos and Her- charcoals have less or even no effects against nandez, 1997). Phillips et al. (1988) analyzed the mycotoxicosis (Table 2). This might be due to the in vitro binding capacities of different adsorbents fact that activated charcoal is a relatively unspe- which were representative for the major chemical cific adsorbent and, hence, essential nutrients are classes of aluminas, silicas, and aluminosilicates also adsorbed particularly if their concentrations and selected HSCAS as a possible suitable candi- in the feed are much higher compared to those of date for in vivo trials concerning the prevention of the mycotoxin. In other trials with goats, how- aflatoxicosis in chicken. HSCAS was shown to ever, it was shown that high doses of activated have a high affinity for aflatoxin B forming a charcoal are beneficial in an acute poisoning situ- complex which was stable at temperatures of 25 ation concerning the intake of high amounts of and 37°C, in a pH range of 2 – 10, and in an eluotropic series of solvents. When HSCAS wasadded in a concentration of 0.5% to chicken diets 2.2. Aluminosilicates (zeolites, HSCAS, clays) containing 7.5 mg/kg aflatoxin B , the growth inhibitory effects were significantly decreased. In Most studies related to the alleviation of myco- this study, the adsorption of HSCAS was thought toxicosis by the use of adsorbents are focused on to be chemisorption including the formation of aluminosilicates, mainly zeolites and hydrated strong bonds. Two years later, Phillips et al.
sodium calcium aluminosilicates (HSCAS), and (1990a) interpreted the binding mechanism as the aluminosilicate-containing clays, all consisting of formation of a complex by the b-carbonyl system aluminates, silicates and some interchangeable of the aflatoxin with ‘uncoordinated edge site’ ions, mainly alkali metal and alkaline earth metal aluminium ions. Thus, HSCAS can be used as an ions (Barrer, 1989; Mumpton, 1999). Clay miner- ‘inorganic sponge’ sequestering aflatoxins in the als are primarily layered silicates with the com- gastro-intestinal tract of farm animals. Ramos et mon chemical formula [Si O2−] , e.g. kaolin al., 1996b investigated the adsorption of aflatox- Al (OH) Si O . Zeolites are composed of tetrahe- ins to montmorillonite according to Freundlich drons of SiO and AlO as the two fundamental and Langmuir isotherm calculations. They ob- building blocks with the metal atom at the center tained a better fit of their adsorption data em- of each tetrahedron. The common chemical for- ploying the Freundlich isotherm and suggested therefore the presence of a heterogeneous surface zeolite A {Na [Al Si O ]·27 H O} . While the with different adsorption centers having different SiO -unit is electrically neutral, the AlO -unit car- affinities for the adsorbate or the co-existence of A. Huwig et al. / Toxicology Letters 122 (2001) 179 – 188 different adsorption mechanisms or both. The use of aluminosilicates for the adsorption of other cyltrimethylammonium resulting in an increased mycotoxins was also tested, but with little success hydrophobicity of the clay surface following a (Bauer, 1994; Ramos et al., 1996b; Lemke et al., high affinity to the hydrophobic zearalenone. In 1998) except of a chemically modified montmoril- contrast, a closely related organophilic phyllosili- lonite with a binding capacity for zearalenone of cate showed a significantly lower binding capacity 108 mg/g (Lemke et al., 1998). This clay was (Schall et al., 2000). A surprisingly high binding Table 1In vitro adsorption of mycotoxins by different adsorbents Acti6ated charcoalActivated charcoal Aluminosilicates (Ethacal®, Novasil™, perlite, zeobrite) afl afl, aflatoxin; och, ochratoxin A; zea, zearalenone; tri, trichothecenes; fum, fumonsins; cpa, cyclopiazonic acid; erg, ergotamine.
a Derivatized with long-chain quarternary ammonium residues.
b 40% sterilized yeast, 60% fermentation residua of beer production.
Table 2In vivo adsorption of mycotoxins by different adsorbents Acti6ated charcoalActivated charcoal Decreased excretion of afl M , no protective effects against Significant increase in body weight gains 100% survival of goats given a lethal dose Significant reduction of the och concentration in blood, bile, Growth inhibitory effects on pregnant rats significantly diminished; ability of reproduction warranted Growth inhibitory effects of broiler chickens diminished by 64 No significant effect (broiler chickens); ethacal® alone reduced feed intake and body weight and increased water consumption Growth inhibitory effects on broiler chickens diminished by 38 Reduction of bioavailability of aflatoxins in the liver and blood of chickens in a dose-dependent manner Growth inhibitory effects on chickens diminished by 50–100%; feed conversions improved in a dose-dependent fashion; no fullprotection against liver or spleen weight changes by afl Significant decrease of urinary excretion of afl M in turkey poults when HSCAS simultaneously dosed with afl Growth inhibitory effects on chickens diminished by 55–100% 68% decrease in mortality of growing male turkey poults Growth inhibitory effects on chickens diminished by 39–68% (2.5 mg afl/kg feed) and by 46–88% (5 mg afl/kg feed) Growth inhibitory effects on broiler chicks completely Growth inhibitory effects on pigs diminished by 87–89% Growth inhibitory effects on pigs diminished by 80% Decrease of growth inhibitory effects, protective effects on Growth inhibitory effects on chickens diminished by 85–100% (Ethacal®, NovaSil™,perlite, zeobrite) Growth inhibitory effects on average daily gain of pigs 3-Phase study: nursery, growing, metabolism phase; performance and liver function were enhanced, but not allfunctions restored Growth inhibitory effects on average daily gain of pigs Decreased food consumption of quail chicks diminished by 57%, growth inhibitory effects diminished by 70% Growth inhibitory effects on chickens diminished by 65%, no effect against toxicity of och, little effect against toxicity ofcombined toxins Growth inhibitory effects on broiler chickens diminished by Growth inhibitory effects on turkey poults diminished by 55–100% only for afl, no effect against T-2 induced toxicity Growth inhibitory effects on chickens diminished by 85% (afl), Growth inhibitory effects on young broiler chickens diminished Kubena et al., 1998b by 43%; no significant effect against tri toxicosis Reproductive effect of zea alleviated; protection against increase in gestation length, decrease in litter size andincrease in kit mortality of mink No reduction of the och concentration in blood, bile, No reduction of the och concentration in blood, bile, afl, aflatoxin; och, ochratoxin A; zea, zearalenone; tri, trichothecenes; fum, fumonsins; cpa, cyclopiazonic acid. The efficacy of each adsorbent was estimated by theeffects on, for instance, the animal performance, clinical chemistry parameters, or body weight gain. As far as possible, it was calculated as percentage of the decreaseof growth inhibitory effects.
a 40% sterilized yeast, 60% fermentation residua of beer production.
A. Huwig et al. / Toxicology Letters 122 (2001) 179 – 188 capacity of 290 mg/g for the alkaloid ergotamine (consisting of 40% sterilized yeast and 60% fer- mentation residua of yeasts used for beer produc- tion) is dependent on the pH being at maximum Related to in vivo trials, the amount of an in acidic solutions (at pH 3: 8.6 mg/g, at pH 8: 1.2 adsorbed mycotoxin is difficult to calculate.
mg/g). However, in trials with pigs employing a Therefore, the efficacy of adsorption has to be feed supplement of 5% of yeast, only a slight determined by the animal performance, e.g. body reduction of the ochratoxin A concentration in weight gain, feed intake, mortality, concentrations blood plasma, bile, and tissues was achieved. By of the corresponding mycotoxin in blood, tissues, the use only of yeast cell walls instead of whole and organs. The results from such feeding trials cells, the adsorption of mycotoxins can be en- hanced. The cell walls harboring polysaccharides Regarding the applicability of aluminosilicates (glucan, mannan), proteins, and lipids exhibit nu- for the binding of mycotoxins, it can be concluded merous different and easy accessible adsorption that they are very effective in preventing aflatoxi- centers including different adsorption mecha- cosis, but their efficacy against zearalenone, nisms, e.g. hydrogen bonding, ionic, or hydropho- ochratoxin, and trichothecenes is limited. In addi- bic interaction. Therefore, it was possible to bind tion to the narrow binding range concerning dif- 2.7 mg zearalenone per gram of cell walls. The binding was rapid and reached equilibrium after disadvantage of showing high inclusion rates for only 10 min, which is superior to commercial available clay-based toxin binders (Vo¨lkl andKarlovsky, 1998, 1999).
In another context, it was shown that yeast killer toxins were adsorbed by the polysaccharides and not by the proteins or fatty acids of yeast cell Cholestyramine is an anion exchange resin walls (Radler and Schmitt, 1987) and that this which is used for the binding of bile acids in the adsorption was not unspecific because cellulose gastro-intestinal tract and for the reduction of low and glycogen were not able to bind killer toxins.
density lipoproteins and cholesterol. The in vitrobinding capacity of this resin for ochratoxin Aand zearalenone was 9.6 mg/g (Bauer, 1994) andmore than 0.3 mg/g (Ramos et al., 1996b), respec- 3. Conclusion
tively, but in vivo, cholestyramine had only a verysmall effect on the reduction of the ochratoxin The applicability of different binders for the concentration in blood, bile, and tissues.
adsorption of mycotoxins was first investigated by Another adsorbent is crospovidone (polyvinyl- in vitro experiments demonstrating that most of pyrrolidone), a highly polar amphoteric polymer the mycotoxins were sufficiently bound by at least the in vitro adsorbance of which was measured as one adsorbent (Phillips et al., 1988, 1990b; Bauer, 0.3 mg/g for zearalenone by Ramos et al. (1996b).
1994; Galvano et al., 1997, 1998; Huebner et al., Up to now, this polymer has not been tested in 1999), which was possibly derivatized, e.g. em- ploying cetylpyridinium or hexadecyltrimethylam- polyvinylpyrrolidone recently showed increased values up to 2.1 mg/g (Alegakis et al. 1999).
exhibiting high binding capacities in vitro werefurther tested in lifestock and it was shown that 2.3.2. Yeast and products from yeast some adsorbents are suitable to alleviate the toxic Besides its excellent nutritional value, yeast or effects of specific mycotoxins. The addition of yeast cell walls can also be used as adsorbents for HSCAS for example resulted in almost total pro- mycotoxins (Gru¨nkemeier, 1990; Bauer, 1994).
tection against aflatoxicosis (Kubena et al., 1988; The in vitro adsorption of ochratoxin by yeast Doerr, 1989; Ramos and Hernandez, 1996), but A. Huwig et al. / Toxicology Letters 122 (2001) 179 – 188 its efficacy against zearalenone and ochratoxin cate decrease the bioavailability of aflatoxin in the chicken.
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Microsoft word - assignments_2013.docx

ASSIGNMENT 2 for Marianna Marra The dataset ear.dat is based on 214 children with acute otitis media (OME) who participated in a randomized clinical trial (Mandel, 1982,Pedriatic Infectious Diseases, 1, 310-316). Each child had OME at the beginning of the study in either one (unilateral cases) or both (bilateral cases) ears. Each child was randomly assigned to receive a 14-days cours


TRIAZOL® fluconazol Cápsula 150 mg MODELO DE BULA – RDC 47/2009 I - IDENTIFICAÇÃO DO MEDICAMENTO: Nome comercial: Triazol® Nome genérico: fluconazol APRESENTAÇÕES Triazol® cápsulas de 150 mg em embalagens contendo 1, 2 ou 4 cápsulas. VIA DE ADMINISTRAÇÃO: ORAL USO ADULTO COMPOSIÇÃO Excipientes: lactose, croscarmelose sódic

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