Non-essential and essential trace element concentrations in meat in cattle reared under organic, intensive or conventional production systems

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1 Non-essentil nd essentil trce element concentrtions in met in cttle rered under orgnic, intensive or conventionl production systems Isel Blnco-Penedo, Mrt López Alonso, Mrt Mirnd, Joquin Hernández, José Luis Benedito, Richrd Frederick Shore To cite this version: Isel Blnco-Penedo, Mrt López Alonso, Mrt Mirnd, Joquin Hernández, José Luis Benedito, et l.. Non-essentil nd essentil trce element concentrtions in met in cttle rered under orgnic, intensive or conventionl production systems.,, (), pp.-. <./>. <hl-> HAL Id: hl- Sumitted on Mr HAL is multi-disciplinry open ccess rchive for the deposit nd dissemintion of scientific reserch documents, whether they re pulished or not. The documents my come from teching nd reserch institutions in Frnce or rod, or from pulic or privte reserch centers. L rchive ouverte pluridisciplinire HAL, est destinée u dépôt et à l diffusion de documents scientifiques de niveu recherche, puliés ou non, émnnt des étlissements d enseignement et de recherche frnçis ou étrngers, des lortoires pulics ou privés.

2 Non-essentil nd essentil trce element concentrtions in met in cttle rered under orgnic, intensive or conventionl production systems Journl: Mnuscript ID: TFAC--.R Mnuscript Type: Originl Reserch Pper Dte Sumitted y the Author: -Jun- Complete List of Authors: Blnco-Penedo, Isel; University of Sntigo de Compostel, Animl Pthology Alonso, Mrt; University of Sntigo de Compostel, Animl Pthology Mirnd, Mrt; University of Sntigo de Compostel, Ciencis Clínics Veterinris Hernández, Joquin; University of Sntigo de Compostel, Animl Pthology Benedito, José; University of Sntigo de Compostel, Animl Pthology Shore, Richrd; Lncster Environment Centre, c Centre for Ecology & Hydrology Methods/Techniques: Metls nlysis - ICP, Toxicology - niml study Additives/Contminnts: Toxic elements, Trce elements Food Types: Animl products met

3 Pge of Non-essentil nd essentil trce element concentrtions in met in cttle rered under orgnic, intensive or conventionl production systems I. Blnco-Penedo, M. López-Alonso *, M. Mirnd, J. Hernández, J.L. Benedito, R.F. Shore c Universidde de Sntigo de Compostel, Deprtmento de Ptoloxí Animl, Fcultde de Veterinri, Lugo, Spin. Universidde de Sntigo de Compostel, Ciencis Clínics Veterinris, Fcultde de Veterinri, Lugo, Spin. c Centre for Ecology & Hydrology, Lncster Environment Centre, Lirry Avenue, Bilrigg, Lncster LA AP, UK. Astrct We evluted if differences in non-essentil nd essentil trce element ccumultion in eef-cttle rered under different systems (including orgnic, conventionl nd intensive mngement) were reflected in the met derived from these nimls. Diphrgm muscle from clves from nine frms were nlysed. Muscle cdmium concentrtions were low (< µg/kg wet wt.) nd muscle rsenic, mercury nd led concentrtions were elow limits of detection (<, nd µg/kg respectively) in most (-%) smples; there were no significnt differences etween frms. Essentil trce element concentrtions in muscle were generlly within dequte physiologicl rnges nd, lthough they vried significntly etween frms, this ws not pprently relted to mngement prctices. There were no significnt correltions in element concentrtions etween muscle nd liver or kidney (orgn concentrtions tht etter reflect exposure), except for colt

4 Pge of (positive ssocition) nd zinc (negtive ssocition). Non-essentil nd essentil trce element concentrtions in muscle in our study nimls thus did not generlly reflect differences in exposure. This is prticulrly relevnt for nimls rered in systems (such s orgnic frms) where cttle re exposed to somewht higher levels of non-essentil elements (proly due to soil ingestion when grzing) ut lso cn suffer from minerl deficiencies. Keywords: eef cttle; frming systems; non-essentil elements; essentil trce elements; muscle; nutrition. *Corresponding uthor: M López-Alonso. E-mil: mrt.lopez.lonso@usc.es.

5 Pge of Introduction There hve een vrious food scres over the pst decde tht hs involved met products. These include Slmonell in chicken, ntiiotics in pork, Escherichi coli nd nned growth promoters in eef, nd the BSE crisis (Trrnt, ; Gmelli et l. ). These hve contriuted to growing consumer wreness of food production methods tht do not use chemicls nd tht re more in hrmony with the nturl environment. This, together with promotion of the importnce of helthy food, hs contriuted to the recent rpid development of orgnic frming in the Europen Union (Vrst nd Hovi, ; von Borell nd von Sorensen, ; Willer nd Yussefi, ). Orgnic frming prctices represent n lterntive to the progressive intensifiction tht hs occurred in conventionl niml production. The qulity of orgnic eef is ffected y the production system used, prticulrly with respect to the grzing nd exercise regimes which re integrl components of ny orgnic eef production system (Nielsen nd Thmsorg, ). It is well documented tht grzing cttle involuntrily ingest certin mount of soil (up to %; Thornton nd Arhms, ) which cn led to significnt exposure to non-essentil elements nd other toxic compounds (such s pesticides, microiologicl toxins, nd medicinl products) tht my e present in the soil (Flndysz et l., ; Shrpe nd Livesey, ). This exposure scenrio hs lso een descried for other livestock species, such s orgniclly-rered pigs (Linden et l. ) nd hens (Kijlstr ). However, home-grown feed, nd ccompnying restrictions on minerl supplementtion, cn lso led to dietry deficiencies in orgniclly frmed nimls (Flndysz, ; Vrst nd Hovi,

6 Pge of ). In prticulr, restrictions on feed supplements nd prophylctic prsite control in orgnic eef production cn led to minerl deficiencies (Roderick nd Hovi, ; McNeidhe, ) nd result in poor ody condition nd production. Thus, there re potentil conflicts etween prctices tht re employed to ensure food sfety nd promote more nturl livestock production nd those used to enhnce niml welfre (Vrst nd Hovi, ). In terms of met qulity, different feeding systems hve een studied to ssess product qulity in reltion to post-mortem proteolysis, tenderness, met flvour (Andersen et l. ) nd chemicl residues minly hormones nd ntiiotics (Smith et l. ). Although it hs een well documented tht frm prctices cn significntly ffect ssimiltion y cttle of non-essentil elements in key orgns such s the liver nd kidneys (López-Alonso et l. ), there is no informtion, s fr s we re wre, on how cn such prctices ffect non-essentil nd essentil trce element concentrtions in muscle, the min cttle product eten y people. We hve recently exmined how the ssimiltion of non-essentil nd essentil trce element concentrtions differs in the liver nd kidneys of eef cttle in NW Spin etween frms tht vry in their production systems, prticulrly in the extent to which they grze cttle on psture nd provide dietry concentrtes (Blnco- Penedo et l., ). Clves tht were from frms which lrgely or exclusively grzed livestock on psture nd provided little or no minerl supplementtion hd the highest tissue concentrtions of non-essentil elements. We hypothesise tht this is ecuse psture-grzed cttle ingest soil prticles to

7 Pge of which non-essentil elements dhere. There ws lso evidence of some essentil trce element deficiency in cttle from frms tht use low mounts of concentrtes nd do not provide minerl supplements. The ojective of the current study ws to evlute if differences in non-essentil nd essentil trce element ccumultion y cttle tht re relted to frm production prctices re reflected in met destined for humn consumption. Mteril nd methods Frm selection Frms were selected from the districts of Brll (B), Montederrmo (M) nd Vill (V) in Glici (NW Spin). In ech district, conventionl (C), intensive (I) nd orgnic (O) frm from the sme neighourhood were selected. The frms were similr in most respects other thn their grzing nd supplementry feeding regimes. Detiled informtion including frm size, feeding regime, other husndry nd mngement prctices, nd non-essentil nd essentil trce element concentrtions in soils nd diet (forge nd concentrte feed) hve een summrised elsewhere (Blnco-Penedo et l., ). Smple collection A smple of diphrgm muscle (out g) ws collected t the time of slughter, which ws when nimls were etween nd months old. Smples were

8 Pge of pcked in plstic gs, immeditely plced on ice, trnsported to the lortory, then stored t -ºC until processing. Smple nlysis Approximtely g su-smples were digested in ml of concentrted nitric cid (Suprpur grde, Merck) nd ml of % w/v hydrogen peroxide in microwve digestion system (Milestone, Ethos Plus; Itly). Digested smples were trnsferred to polypropylene smple tues nd diluted to ml with ultr-pure wter. Elements present t very low concentrtions (rsenic (As), cdmium (Cd), chromium (Cr), colt (Co), nickel (Ni), mercury (Hg) nd led (P)) were determined y inductively coupled plsm mss spectrometry (ICP-MS; VGElementl PlsmQud SOption) wheres elements tht occurred t higher concentrtions (copper (Cu), iron (Fe), mngnese (Mn), molydenum (Mo), selenium (Se) nd zinc (Zn)) were determined y inductively coupled plsm tomic emission spectrometry (ICP-OES; Perkin Elmer Optim DV). An nlyticl qulity control progrmme ws pplied throughout the study. Blnks were run longside smples nd concentrtions in smples were lnk corrected s necessry. The limits of detection in the cid digest were clculted s three times the stndrd devition of the regent lnks (Tle ) nd were sed on the men smple weight nlysed. Anlyticl Qulity ssurnce Anlyticl recoveries were determined from Certified Reference Mteril (Pig kidney CRM, R Reference Mterils, Belgium) tht ws nlysed longside

9 Pge of unknowns. There ws generlly good greement etween the mesured nd certified or indictive vlues (Tle ). The CRM ws not certified for Co nd Mo nd nlyticl recoveries were determined for these elements using smples spiked t concentrtion tht gve sornce vlues some - times greter thn the norml levels in muscle. Men recoveries were % nd % respectively. The precision of the nlyticl method, clculted s the reltive stndrd devition (RSD) of Co nd Mo concentrtions in digests of the sme smple, were etween. nd. %. Sttisticl nlysis All sttisticl nlyses were done using the progrm SPSS for Windows (v..). Non-detectle concentrtions were ssigned vlue of hlf the detection limit when clculting men element concentrtions in muscle. Dt were tested using Kolmogorov-Smirnov test nd were generlly not normlly distriuted. They were therefore log-trnsformed efore nlysis, nd verge concentrtions re therefore given s geometric mens. One-wy Anlysis of Vrince followed y Tukey s honest significnt difference (HSD) post-hoc tests were used to test for differences in non-essentil nd essentil trce element concentrtions etween frms. The significnce of correltions etween the levels of non-essentil nd essentil trce elements in muscle nd liver nd kidney in ech frm were clculted using Spermn rnk correltion nlysis. In ll cses sttisticl significnce ws tken to e indicted y p<..

10 Pge of Results nd Discussion Non-essentil element concentrtions in muscle from cttle from different frms re presented in Tle. Overll, concentrtions were low, nd with the exception of Cd, most smples (. % for As nd Hg,.% for P) hd levels which were not detectle. The reminder of smples hd concentrtions close to the detection limit. Cd concentrtions were ll elow µg/kg wet weight. In generl, non-essentil element residues in met in this study were similr to those descried in previous studies in cttle in NW Spin (López-Alonso et l. ; ) nd were within the rnge of those descried in other countries (for review see López-Alonso et l. ). None of the smples nlysed in the current study exceeded the mximum dmissile levels for Cd nd P (. nd. mg/kg fresh weight respectively) estlished y the Europen Commission (). The Europen Commission hs not estlished sttutory limits for As nd Hg ut Hg residues, when detected, were -fold lower thn those llowed in fish (Europen Commission ). No significnt differences were found etween frms in muscle Cd concentrtions (men vlues rnged from. to. µg/kg wet weight) nor in the proportion of smples with detectle residues for the other non-essentil elements (Tle ). There ws no significnt ssocition etween Cd concentrtions in muscle nd those in the liver or kidney (Tle ). This is rodly consistent with findings elsewhere tht suggest tht muscle concentrtions of non-essentil elements, except perhps for As, re not closely relted to the level of exposure, (Vremn et l. ).

11 Pge of Essentil trce element concentrtions in muscle from cttle from different frms re presented in Figure. In generl, nd with the exception of Se, essentil trce element concentrtions in cttle muscle were within the rnges descried y Puls () s dequte nd were similr to those reported for cttle muscle in most other countries (for review see Jorhem et l. ; López-Alonso et l. ). In contrst, Se concentrtions in cttle from ll frms in the current study were ove the norml rnge (.-. mg/kg wet weight; Puls ) nd were higher thn concentrtions reported in cttle elsewhere (Jorhem et l. ). Indeed, most nimls in the present study hd muscle Se concentrtions within the high (.-. mg/kg) or toxic (.-. mg/kg) rnge. However, the rnges for essentil element concentrtions proposed y Puls () re not comprehensive nd do not tke into ccount possile differences etween different types of muscle. Comprison of results from seprte studies superficilly suggest tht essentil trce element concentrtions in muscle vry - fold etween cttle from different countries (Jorhem et l. ; López-Alonso et l. ) even when essentil trce element concentrtions in the liver nd kidneys re comprle nd within the dequte physiologicl rnge. However, differences in muscle element concentrtions more likely reflected differences in the type of muscle nlysed. For exmple, cttle diphrgm hs een found to contin nerly twice the Cu nd Se concentrtion of pectorl muscle (López-Alonso et l. ; López-Alonso unpulished dt), perhps reflecting greter metolic ctivity in the diphrgm. There were sttisticlly significnt differences etween frms for most essentil trce element concentrtions in muscle (Figure ). As with inter-frm differences in

12 Pge of liver nd kidney concentrtions (Blnco-Penedo et l. ), there ws no evidence of ny specific pttern of differences etween regions or different types of frm. Cttle from the orgnic frm in Montederrmo () hd the lowest men Co, Cu, Ni nd Se concentrtions found in our study. This ws the only frm where high proportion of nimls hd heptic Co nd Se concentrtions tht were within the mrginl or deficient rnge, nd (together with the orgnic frm in Brll ()), hd clves with heptic Cu concentrtions tht were elow the rnge considered dequte (Blnco-Penedo et l. ). Clves from the frm lso hd significntly higher Zn muscle concentrtions thn cttle from other frms lthough, surprisingly, clves from the frm hd the lowest men Zn heptic nd renl concentrtions tht we recorded (Blnco-Penedo et l. ). The highest men concentrtions in muscle of most of the essentil trce elements (Cu, Fe, Mn, Mo nd Se) were in cttle from the intensive frm in Vill (), lthough it did not correspond with higher intke of these elements in the diet or higher concentrtions in the liver or kidney (Blnco-Penedo et l. ). When nlysing the reltionship etween essentil trce element concentrtions in muscle nd those in the liver or kidney (dt from Blnco-Penedo et l. ), the only significnt positive ssocition ws for Co; muscle Co concentrtions were positively ssocited with Co concentrtions in oth liver nd kidney (Figure ; Tle ). There ws lso significnt negtive ssocition etween Zn concentrtions in the muscle nd those in the liver nd kidney (Tle ). Tking into ccount tht essentil trce element concentrtions in the liver, nd to lesser extent in the kidney, re the est indictors of the minerl sttus in clves (López-

13 Pge of Alonso et l. ), our results indicte tht, t lest under the conditions of our study, essentil trce element concentrtion in muscle is generlly not indictive of minerl sttus. This is presumly ecuse muscle, unlike the liver nd kidney, does not hve essentil trce element storge cpcity. At dequte dietry intkes, minerl concentrtions in muscle my e most closely relted to protein synthesis nd the predominnt type of metolism in the muscle (Schricker et l. ). Although our results did suggest tht muscle Co might correlte with Co sttus, this hs not een found elsewhere. Vn Ryssen et l. () compred tissue Co concentrtions in cttle given little or no dietry Co supplementtion (Co requirement in cttle is. mg/kg DM; NRC ) with those in cttle given lrge Co supplements ( nd mg/kg DM). The uthors found tht lthough liver nd pncretic Co concentrtions significntly incresed with Co supplementtion (increse from. to. mg/kg dry weight in liver, nd from. to. mg/kg dry weight in the pncres), there ws no concomitnt increse in muscle Co (. vs. mg/kg dry weight). It is possile tht the low dietry levels of Co in cttle in our study were elow those necessry to mximise metolic ctivity (lthough norml or dequte Co concentrtions hve not een estlished for muscle; Puls ) nd so Co concentrtions in the muscle of cttle from frms where Co nutrition ws etter my hve significntly incresed longside concentrtions in the liver nd kidney. The negtive ssocition tht we found etween Zn concentrtions in the muscle nd those in the liver nd kidney (Figure ) ws lso surprising. In previous study

14 Pge of in cttle with dequte Zn sttus in NW Spin (López Alonso ), Zn concentrtions in the liver nd kidney were strongly nd positively correlted (s in this study; R s =., p<.) ut there ws no significnt ssocition etween muscle Zn nd either liver or kidney Zn. In cttle, Zn tissue concentrtions re efficiently regulted y homeosttic mechnisms nd, once optiml physiologicl concentrtions ( mg/kg DM; NRC ) re reched, Zn supplementtion hs no significnt effect on Zn muscle levels (Kessler et l. ). Differences in tissue Zn concentrtions etween cttle receiving dequte Zn dietry concentrtions could e due to fctors such s ge, sex, or production clss (milk or eef cttle) (Puschner et l. ). It is lso possile tht other dietry components my influence the distriution of zinc in the ody. For exmple other elements such s Cu nd Cd hve similr chemicl nd physicl properties to Zn nd compete for metolic inding sites in metllothioneins (López-Alonso et l. ). It is possile tht the negtive ssocition etween Zn concentrtions in the muscle nd liver nd kidney in the current study my e due, t lest in prt, to some of these confounding fctors. Conclusions Our results indicte tht differences in non-essentil nd essentil trce element concentrtions in the liver nd kidney of cttle from different production systems on frms in Glici re not reflected in the non-essentil nd essentil trce element content in the met. This is especilly relevnt for nimls rered in systems (such

15 Pge of s orgnic frms) where wened clves re exclusively rered y eing grzed on psture nd, s result, ccumulte elevted heptic nd renl non-essentil element levels (proly due to soil ingestion) ut lso cn suffer from minerl deficiencies. Acknowledgements This study ws supported y the Xunt de Glici (Spin) (PGIDTRAPR). I. B. P. is recipient of reserch fellowship (AP- ) from the Ministry of Science of Spin. We thnk Nieves Muñoz for sttisticl ssistnce. References Andersen HJ, Oksjerg N, Young JF, Therkildsen M.. Feeding nd met qulity- future pproch. Met Sci. :-. Blnco-Penedo I, Benedito JL, Shore RF, Mirnd M, Cstillo C, López-Alonso M.. Frming fctors ffecting the ccumultion of toxic chemicl elements in clves in NW Spin. Sumitted.. Blnco-Penedo I, Shore RF, Mirnd M, Benedito JL, López-Alonso M.. Fctors ffecting trce element sttus in clves in NW Spin. Livest Sci. (- ):-.

16 Pge of Europen Commission. Commission Regultion EEC/. Setting Mximum Levels for certin contminnts in Foodstuffs. Commission of the Europen Communities, Brussels. Flndysz J.. Some toxic nd essentil trce metls in swine from Northern Polnd. Sci Totl Environ. :-. Flndysz J, Koteck W, Knnn K.. Mercury, led, cdmium, mngnese, copper, iron nd zinc concentrtions in poultry nd sheep from the northern prt of Polnd. Sci Totl Environ. :-. Gmelli D, Nspetti S, Viro D.. Why re consumers uying orgnic met nd milk? A qulittive study of the Itlin mrket. Proceeding of the st SAFO Workshop; - Septemer; Florence, Itly. Jorhem L, Sundström B, Astrnd C, Hegglund G.. The levels of zinc, mngnese, selenium, chromium, nickel, colt, nd luminium in the met, liver nd kidney of Swedish pigs nd cttle. Z Leensmi Unters Forsch. :-. Kessler J, Morel I, Dufey P-A, Gutzwiller A, Stern A, Geyer H.. Effect of orgnic zinc resources on performnce, zinc sttus nd crcss, met nd clw qulity in fttening ulls. Livest Sci. :-. Kijlstr A.. The role of orgnic nd free poultry production systems on the dioxin levels in eggs. Proceedings of the rd SAFO Workshop, Septemer, Flenty. Reding.

17 Pge of Linden A, Andersson K, Oskrsson A.. Cdmium in orgnic nd conventionl pig production. Arch Environ Contm Toxicol. :-. López Alonso M.. Estudio de los principles elementos contminntes en Gndo vcuno de Glici. PhD thesis, [Spin]: University of Sntigo de Compostel. López-Alonso M, Benedito JL, Mirnd M, Cstillo C, Hernández J, Shore RF.. Toxic nd trce elements in liver, kidney nd met from cttle slughtered in Glici (NW Spin). Food Addit Contm. :-. López-Alonso M, Benedito JL, Mirnd M, Cstillo C, Hernández J, Shore RF.. Contriution of cttle products to dietry intke of trce nd toxic elements in Glici, Spin. Food Addit Contm. :-. López-Alonso M, Prieto Montñ F, Mirnd M, Cstillo C, Hernández J, Benedito JL.. Interctions etween toxic (As, Cd, Hg nd P) nd nutritionl essentil (C, Co, Cr, Cu, Fe, Mn, Mo, Ni, Se, Zn) elements in the tissues of cttle from NW Spin. BioMetls :-. McNeidhe FS.. Psture mngement nd composition s mens of minimizing minerl disorders in orgnic livestock. Proceedings of the th NAHWOA Workshop; Novemer; Rodding, Denmrk. NRC (Ntionl Reserch Council) (). Nutrient requirements of Beef Cttle. th revised edn. Ntionl Acdemic Press, Wshington DC.

18 Pge of Nielsen BK, Thmsorg SM.. Welfre, helth nd product qulity in orgnic eef production: A Dnish perspective. Livest Sci. :-. Puls R.. Minerl levels in niml helth. Clerrook: Sherp Interntionl. Puschner B, Choi Y-K, Tegzes JH, Thurmond.. Influence of ge, sex, nd production clss on liver zinc concentrtion in clves. J Vet Dign Invest. :-. Roderick S, Hovi M.. Animl helth nd welfre in orgnic livestock systems: identifiction of constrints nd priorities. A report to MAFF p.. Schricker BR, Miller DD, Stouffer JR.. Content of zinc in selected muscles from eef, pork, nd lm. J Food Sci. :-. Shrpe RT, Livesey CT.. Surveillnce of suspect niml toxicoses with potentil food sfety implictions in Englnd nd Wles etween nd. Vet Rec. :-. Smith GC, Heton KL, Sofos JN, Ttum JD, Aronson MJ, Clyton RP.. Residues of ntiiotics, hormones nd pesticides in conventionl, nturl nd orgnic eef. J Muscle Foods. :-. Trrnt PV.. Some recent dvnces nd future priorities in reserch for the met industry. Met Sci. :S-S. Thornton I, Arhms P.. Soil ingestion mjor pthwy of hevy metls into livestock grzing contminted lnd. Sci Totl Environ. :.

19 Pge of Vrst M, Hovi M. (). Orgnic livestock production nd food qulity: review of current sttus nd future chllenges. Proceedings of the nd SAFO Workshop; Mrch ; Witzenhusen, Germny. Vn Ryssen JBJ, Miller WJ, Gentry RP, Nethery MW.. Effect of dded dietry colt on metolism nd distriution of rdioctive selenium nd stle minerls. J Diry Sci. :-. von Borell E, von Sorensen JT.. Orgnic livestock production in Europe: ims, rules nd trends with specil emphsis on niml helth nd welfre. Livest Sci. :-. Vremn K, vn der Veen NG, vn der Molen EJ, de Ruig WG.. Trnsfer of cdmium, led, mercury nd rsenic from feed into tissues of fttening ulls: chemicl nd pthologicl dt. Neth J Agr Sci. :-. Willer M, Yussefi M.. The World of Orgnic Agriculture: Sttistics nd Emerging Trends. IFOAM, Bonn.

20 Pge of Tle. Results of the nlyticl qulity control progrmme used in the determintion of non-essentil nd essentil trce elements Element Blnk (n=) Detection limit Certified Reference Mteril Men ± SD (µg/l) (µg/g) certified levels men ± SD (mg/kg) nlysed levels men ± SD (mg/kg) As. ±... ±.. ±. Co. ± ±. Cr. ±.. (.-.). ±. Cd. ±... ±.. ±. Cu ±...±.. ±. Fe. ±.. ± ±. Hg. ±... ±.. ±. Mn. ±...±.. ±. Mo. ± ±. Ni. ±.. (.). ±. P. ±... ±.. ±. Se. ±...±.. ±. Zn. ±.. ± ±. numers in prentheses re indictive vlues

21 Pge of Tle. Non-essentil (As, Cd, Hg nd P) element concentrtions in muscle (µg/kg wet weight) in cttle in our study. Arevitions for frms re s follows B: Brll, M: Montederrmo, V: Vill, C: Conventionl, I: Intensive, O: Orgnic Frm As Cd Hg P N (<ld) () () () Geometric men ND. ND. Rnge ND-ND.-. ND-ND ND-. N (<ld) () () () Geometric men.. ND. Rnge ND-..-. ND-ND ND-. N (<ld) () () () Geometric men ND... Rnge ND-ND.-. ND-. ND-. N (<ld) () () () Geometric men ND... Rnge ND-ND.-. ND-. ND-. N (<ld) () () () Geometric men.. ND. Rnge ND-..-. ND-ND ND-. N (<ld) () () () Geometric men.. ND. Rnge ND-..-. ND-ND ND-. N (<ld) () () () Geometric men ND. ND. Rnge ND-ND.-. ND-ND ND-. N (<ld) () () () Geometric men ND... Rnge ND-ND.-. ND-. ND-. N (<ld) () () () Geometric men ND... Rnge ND-ND.-. ND-. ND-. N is the numer of smples nlysed nd numers in prenthesis indictes the numer of smples tht were elow the limit of detection (N<LoD); ND: non detected

22 Pge of Tle. Rnk correltions etween men non-essentil nd essentil trce element concentrtions in muscle nd liver nd kidney in ech frm in cttle in our study. Results re expressed s Spermn rnk correltions coefficient nd proility (*p<., * p<., *** p<.). Correltions hve een not clculted for As, Hg nd P ecuse most smples hd not detectle concentrtions Element (N) Muscle vs. liver Muscle vs. kidney Cd () Co (). **. *** Cr () Cu ().. Fe () Ni ().. Mn ().. Mo (). -. Se (). -. Zn () -. ** -. *

23 Pge of Figure cptions Figure. Br chrt showing essentil trce element concentrtions in muscle (expressed s geometric mens nd geometric stndrd error) in the frms in our study. Arevitions for frms re s follows B: Brll, M: Montederrmo, V: Vill, C: Conventionl, I: Intensive, O: Orgnic. Different letters denote sttisticlly significnt differences etween frms t p<.. Figure. Sctterplot showing the reltionship etween Co concentrtions in the muscle nd liver nd kidney. Figure. Sctterplot showing the reltionship etween Zn concentrtions in the muscle nd liver nd kidney.

24 Figure. Co (µg/kg) Cr (µg/kg)... Cu (mg/kg) Fe (mg/kg) Mn (mg/kg)..... Mo (mg/kg) Ni (µg/kg) Se (mg/kg) Zn (mg/kg) c c c c Co (µg/kg) Cr (µg/kg)... Cu (mg/kg) Fe (mg/kg) Mn (mg/kg)..... Mo (mg/kg) Ni (µg/kg) Se (mg/kg) Zn (mg/kg) c c c c c c c Pge of

25 Pge of Co muscle (µg/kg) Co muscle (µg/kg) Co liver (µg/kg) Co kidney (µg/kg) Figure.

26 Pge of Zn muscle (mg/kg) Zn muscle (mg/kg) Figure. Zn liver (mg/kg) Zn kidney (mg/kg)