Roczniki Naukowe Polskiego Towarzystwa Zootechnicznego, t. 5 (2009), nr 4

The effect of a minerał mixtureon cobalt łevełs in dairy cowsin the region of southern Podłasie

Krzysztof Górski I,Agata Paprocka I,Elżbieta Bombik I,Teresa Bombik", Leon Sa ba

'Unlversity of Podlasie, Department of Reproduction and Animai Hygiene,B. Prusa 14,08-110 Siedlce2University of Life Sciences in Lublin, Department af Animai Hygiene and Environrnent,Akademicka 13,20-950 Lublin

The investigation of cobalt suppły of dairy cows was carried out on four farms A, B, C andD, łocated in the region of southern Podlasie, Cobalt concentrations we re examined in thetrophic system: soi!-płant-animal. Soi! sampłes were collected once from a 0-15 cm łayer (bya soi! sampling stick) at the peak of the vegetation season. Additionally, the pH of theexamined soi! was tested. The feeds were sampłed regułarly during the year. B100d wascollected 4 times: at the beginning of drying-off period (about 60 days before całving), 10-14days before całving, after the first month of łactation and after the second month of łactation.Hair was collected twice: 10-14 days before parturition (I) and after the first month ofłactation (II). It was found that the feeds met the cobalt requirements. Low Co concentrationswe re detected especially in cows' hair. A minerał mixture was composed taking into conside-ration the geobiochemicał conditions of the farm łocation. The influence of the mixture onminerał metabolism was defined on the basis of changes in the hair eontent of Co. Theexperimentał mixture Bovifosfornag'" improved the minerał status of cows on all the exami-ned farms, and resulted in the increase of cobalt concentration in cows' hair.

KEY WORDS: dairy cows / minerał mixture / cobalt / soil / feed

There are a number of mineraIs whieh are believed to be essential for animals. Theyinclude eight mieraelements, that is: iron (Fe), eopper (Cu), manganese (Mn), zine (Zn),eobalt (Co), iodine (I) and selenium (Se). Inadequate supply of animals with mineraIsfrequently leads to disturbances in body funetions and pathologieal eonditions whiehare ealled produetion diseases [18,29,31,37,50]. It is espeeially important to preventeobalt defieieney in ruminants, in partieular high produeing animals whieh, when fedlow-eobalt diets, develop a number of non-specific symptoms [8, 18, 24]. Unlikemonogastric animals, ruminants are more prane to cobalt deficiencies as the rate ofeobalt absorption in the digestive tract is much lower. Moreover, cows secrete substan-

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tial amounts of this element with milko As a result of factors mentioned above, cobaltdeficiency is often determined in dairy cows [24].

In the body of land mammals cobalt is accumulated mainly in flesh organs andmuscles. Cobalt eontent in these tissues ranges between 0.005- 0.5 mg/kg. As a rule,cobalt concentration in body liquids does not exceed 1 ug/I [20]. An increased ruminantdem and for cobalt and considerable sensitivity of ruminants to Co deficiency result fromthe fact that the discussed element is a component of cobalamin (vitarnin B 12) synthe-sized by rumen microorganisms. The ex tent of synthesis mainly depends on Co eontentin feed. Therefore, the deficiency in cobalt is ultimately a deficiency in vitamin BI2[15, 18,20,22,23,24,27,28,46,49].

Cobalamine plays a very important role in the process of red blood celi formation,metaboli sm of nucleic acids and proteins as well as oxidation and reduction reactions(activation of enzymatic processes) [9,20,49]. Insufficient vitamin B 12 level mayaIsoresult in disturbed transformation ofpropionates into succinates in liver, which increasespropionie acid concentration in blood and leads to appetite loss and reduced feed intake.Such a situation is more likely when cows are fed diets of mainly concentrated feeds[18, 27, 48, 49]. Young animals are more sensitive to cobalt deficiency (their vitaminB 12reserves are lower). Moreover, sheep are more sensitive to cobalt deficiency com-pared to cattle [17, 18,21,27,49].

Before typical deficiency symptoms of individual mineraIs occur in ani maI s, theydisplay poorer performance and lower resistance [27, 50]. As a result, a fuli picture ofmineral metaboli sm in ruminants can be attained by applying complex examinations,like clinical studies, evaluation of feeding regimes and conditions of housing, analysesof performance indicators and laboratory analyses [27,41]. Every time a need arises tocompensate mineral deficiencies in ruminants, any such examinations should be prece-ded by studies of biogeochemical conditions in the system: soil-plant-animal. Thecompensation is usually implemented through an application of mineral supplementscontaining mineraIs in the form of carbonates, oxides, chlorides andsulphates [2, 10,17,18,24,41,42,43,44,45].

Diagnosis of disturbances in animaI cobalt metabolism includes analysis of vitaminB 12 concentration in serum, determination of cobalt and vitamin B 12 contents in liverand of methylomalonic acid and cobalt levels in serum. According to some authors, itis advisable to test hair concentration of cobalt [8, 34, 36, 43, 49].

The objectives of the work were: to evaluate the cobalt status of cows on the basi sof soil, feed and tissue analyses on the selected farms, situated in the southern Podlasie;and to determine the feeding value of the mineral mixture Bovifosfomag'P formulatedtalking into account biogeochemical conditions of the area where the animals weremaintained as well as the mineral profile of animaI tissues.

Materiał and methods

A three-year study was performed in the region of southern Podlasie on four dairyfarms. The farms were denoted by the letters A, B, C and D. The animals raised on the

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farms were of Polish Hołstein-Friesian breed, Błack and White variety, aged between4 and 7 with an average milk performance of around 4000 kg. Twełve cows weresełected from each farm. The animałs were clinically heałthy and had similar rearingparameters as well as various physiołogicał statuses. The study was carried out over theperiod of summer and winter feeding. In winter the animałs were housed in barns wherethere were maintained appropriate zootechnicał parameters required for dairy cows [40].Feeding regimes of cows were based on rations inc\uding corn siłage, meadow hay,barley straw and a concentrated mixture. In summer, the rations incłuded green foragefrom meadows and cułtivated fiełds suppłemented by hay and barley straw. Nutritionałrequirements of the cows were established on the basis of Feeding Standards [33].

The study was divided into two stages. During the first one-year stage we evałuatedthe characteristics of minerał metaboli sm in cows. The evałuation took into account thełocał biogeochemicał conditions and was based on examination of cobałt eontent in thesoił, feeds, serum and hair. Twełve animałs were sełected from each farm (6 łactatingand 6 dry cows). In the second stage, the bałanced minerał mixture Bovifosfornag'f wasintroduced to the diet in the amount ot' 150 g per cow per day (Tab. l and 2). Over theperiod ot' 14 days, the cows from the experimentał groups (D) were offered the mixturetogether with the concentrated feed in the morning to łet the animałs get used to thenew regime. The controł cows (K) did not receive an addition of the minerał mixture.The effect of minerał suppłement on cobałt status in dairy cows' hair was determineafter two years of supplementation.

Table 1 - Tabela 1Raw material composition (macroelements) of the experimental mixture Bovifosfornag'"Skład surowcowy (makroelementy) mieszanki Bovifosfornag'" w wersji użytej w doświadczeniu

Specification Pure ingredient eontent

Wyszczególnienie Zawartość czystego składnika

(g) Ca P Mg Na

(g) (g) (g) (g)

Ca(H2P04h 350 60 95(monocalcium phosphate)(fosforan jednowapniowy)Ca,(PO.h 100 39 20(tricalcium phosphate)(fosforan trójwapniowy)MgO 175 105(magnesium oxide)(tlenek magnezu)CaCO, 200 80(ground limestone)(kreda pastewna)NaCI 175 70(forage salt)(sól pastewna)Total - Razem 1000 179 115 105 70

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Table 2 - Tabela 2Raw material composition (microelements) of the experimental mixture Bovifosfornag'"Skład surowcowy (mikroelementy) mieszanki Bovifosfornag'" w wersji użytej w doświadczeniu

Specification Pure ingredient eontentWyszczególnienie Zawartość czystego skladnika

(g) Zn Cu Fe Mn Se I Co(g) (g) (g) (mg) (mg) (mg) (mg)

ZnSo47H20 22.000 5.0(zine sulfate)(siarczan cynku)CuS045H20 4.000 1.0(cupric sulfate)(siarczan miedzi)FeS047H20 5.000 1.2(ferrous sulfate)(siarczan żelaza)MnCOJ 0.020 10.0(manganese carbonate)(węglan manganu)Na2Se04 0.050 200(sodium selenate)(selenian sodu)KI 0.040 30.0(potassium iodide)(jodek potasu)CoS047H20 0.015 3.0(cobalt sulfate)(siarczan kobaltu)Total - Razem 31.125 5.0 1.0 1.2 10.0 20.0 30.0 3.0

Soi! sampies were collected once from pastures and cultivated fields at the peak ofthe vegetation season by means of a soi I sampling stick from the surface layer (0-15cm) following the current recommendations [4]. Cobalt eontent was determined in thesampies by soi! mineralization in a mixture of concentrated acids HN03 + HCI04. Thedetermination was performed using the atomie absorption spectrophotometer UNICAM939. Moreover, we determined soil pH [35J. Feed sampies were collected in all theperiods when the feeds were introduced as diet components. Following drying andgrinding, the sampies were chemically analyzed (wet materialization) as outlined byOstrowska et al. [35]. Co eontent in feed was determined by atomie absorption spect-rometry AAS by means of the apparatus UNICAM 939. During each period, blood wassampled four times from the external jugular vein of selected cows. The sampling dateswere as follows: the beginning of drying off, that is, day 60 (date I), 10-14 days priorto calving (date II), after the first month of lactation (date III) and after two months oflactation (date IV). The blood was centrifuged to obtain serum which was chilled at-20oC and then successively analyzed. As recommended by Brochart [7J, sampies weretaken of white hair (which had just grown back following shaving) from the back. Therewere two sampling dates: I 00-14 days prior to calving) and II (after the first week oflactation). The hair was cleansed and degreased using detergents and alcohol. Dried and

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weighed hair sam pies were placed in Teflon® containers which were filled withspect-rally clean, concentrated nitric acid and hydro gen peroxide (Merck, Germany). FolIo-wing microwave mineralization, the sampies were subjected to spectrometric analysis.Co eontent in serum and hair was determined by the lCP (Inductively Coupled Plasma)method using the plasma spectrometer lCP Philips SC-PU 7000 attached to the Ul tra-sonic-Nebulizer. The detailed methodology is presented in the works by Bodak andDobrzański [5] as well as Górecka [11]. The study results were statistically analyzedby means of Excel 2007 and Statistica 97, version 5. Differences between group meanswere investigated using t-Student test.

Results and discussion

Cobalt-rich plants, which grow mainlyon soils with adequate cobalt concentrations,are the main source of this element for cattle. Soi l eontent of cobalt ranges from 0.1 to100 mg/kg d.m. and depends on many factors. The lowest cobalt concentration is insandy podzolic soils and the highest in brown loamy and sil tYloam alluvial soils as wellas rendzinas. Manganese oxides in the soil stimulate cobalt binding as a result of whichit becomes less available for plants. The more acid soils get, the faster is the process ofbinding [20, 47]. Cobalt plays an important role in the processes of biological methy-lation because it is a component of methyl vitamin B 12. Thus, it is crucial that cobaltis present in the soil [20]. Nitrogen and phosphorus fertilization is a major factordecreasing cobalt availability for plants [l].

Soils containing under 0.2 mg/kg d.m. Co are assumed to be low in cobalt. Soilconcentrations of cobalt in the soils examined were adequate for plants (Tab. 3). The

Table 3 - Tabela 3Soil pH and the cobalt levels of grassland (farm A, B, C, D)Zestawienie wyników badań odczynu i poziomu kobaltu w glebach użytków zielonych (ferma A, B, C, D)

Farm Soil type

Ferma Typ glebyNumber

of bulk sampIesLiczba próbek

zbiorczych

pH Comglkg dry matter

mg/kg suchej masyx Sd

A podzolic from loamy sandbielicowa wytworzonaz piasków gliniastych

10 6.0 1.13 0.30

B podzolic from loamy sandbielicowa wytworzonaz piasków gliniastych

10 6.0 1.07 0.57

C podzolic from sandbielicowa wytworzonaz piasków luźnych

la 6.1 1.01 0,42

D podzolic from sandbielicowa wytworzonaz piasków luźnych

10 6.1 0.88 0.18

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values of concentration feli within the limits reported for Polish soils by other authors.Increased cobalt concentrations in surface soil layers may be indicative of an influenceof anthropogenic factors (e.g. industrial pollution) [20].

Plants concentration of cobalt is positively correlated with the Co soi l concentra-tion. While plant cobalt deficiencies are infrequent, because this element is easilyassimilated by plants; there have been recorded instances of Co toxicity in plants withcobalt eontent of over 60 mg/kg d.m. Manganese and iron are known to be antagonistsof cobalt. Mn and Fe immobilize cobalt during geochemical processes. Cobalt is of greatimportance to legumes as it is a component of porphyrins which are the building blocksof cobalamin coenzyme participating in processes of nitrogen fixation from the air [20].The appropriate cobalt supply to animals depends to a great extent on availability ofcobalt in plants. The feed eontent of cobalt which satisfies ruminant requirementsshould be 0.1 mg/kg d.m. [14, 16,20,24,27,32,33]. In turn, the cobalt dos e which istoxic for cattle is over l mg Co/kg body weight/day [18, 31]. The feed concentrationsof cobalt recorded in the present study posed no threat to the animals as they indicatedneither cobalt deficiency nor excess (Tab. 4). Obvious symptoms of cobalt deficiencyoccur in ruminants when the feed eontent of this element is under 0.05 mg/kg d.m. [17].

Maize silageKiszonka z kukurydzyPasture green forageZielonka pastwiskowaConcentrated mixtureMieszanka treściwaMedow haySiano łąkoweBarley strawSloma jęczmienna

Number Co (mg/kg dry matter)of bulk sampIes Co (mg/kg suchej masy)

Liczba próbekA B C D

zbiorczych

10 x 0.25 0.33 0.38 0.41Sd 0.13 0.12 0.24 0.15

10 x 0.19 0.21 0.24 0.20Sd 0.11 0.10 0.12 0.11

10 x 0.17 0.18 0.21 0.19Sd 0.08 0.08 0.09 0.09

10 x 0.16 0.19 0.17 0.18Sd 0.07 0.10 0.08 0.08

10 x 0.14 0.16 0.11 0.12Sd 0.01 0.02 0.03 0.02

Table 4 - Tabela 4Comparison of the level of cobalt (mg/kg d.m.) in the feeds for cattle on farrns A, B, C, DZestawienie poziomu kobaltu (mg/kg s.m.) w paszach dla bydła z ferm A, B, C, D

Type of feedsRodzaj badanej paszy

The dairy cows' mineral status can be monitored on the basis of serum and hairconcentrations of elements [3]. Mineral analysis of serum to determine the cobaltconcentration yielded average levels of this element ranging from 0.58 to 0.87 urnol/l(Fig. l).

In comparison with the levels reported by other workers [30, 45], in the presentstudy there were recorded higher cobalt concentrations in cows maintained in the region

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0.90

0,80

0.10.• 0,60E:g. 0,.50o§. 0,40

030

0.20

0.10

0,00dried oH - zasuszone

of southern Podlasie, However, they did not reach the adeguate level which, accordingto Puls [38], should exceed 1.0 umol/l. Hence the results may indicate that in this casethere occurred antagonistic response of cobalt to other microelements (e.g. copper andzinc) whose levels in the region of the southern Podlasie had been determined in theprevious work [13]. In the present study, there were observed statistical differencesbetween cobalt concentrations in lactation and dry cows belonging to herds B (P~O.Ol),C (P~0.01) and D (P~0,05),

AB • Iólctatlng - lakŁujące

Cfarms - fermy D

Fig. l. Mean Co levels in the blood serum of cows according to their physiological status (A, B -statistically significant differences at PSO.OI; a, b - statistically significant differences at PSO.OS)Rys. l. Średni poziom Co w surowicy krwi krów z uwzględnieniem grupy fizjologicznej (A, B - różniceistotne statystycznie przy PSO,Ol; a, b - różnice istotne statystycznie przy PSO,OS)

Many authors state that hair is a useful type of material for analysis because it canbe easily collected and used to evaluate properties of mineral metaboli sm in ruminants[6,26,41]. It is generally believed that, due to regular accumulation of minerais in hair,determination of hair levels of individual minerais yields data on the mineral eoncent-rations in animai bodies [3, 8]. Some workers add that hair mineral composition isa good reflection of deficiency or excess of an element if it results from a long-terminfluence of a factor (e.g. environmental contamination with the element, low feedeontent of minerais) [26, 37,39], The average cobalt levels in the hair of dairy cows inthe present study ranged between 0.13 and 0.19 mg/kg d.m. (Fig. 2). The levels are lowcompared with the values reported by other authors [37,42,45].

The decreased cobalt \evels in the cows coupled with increased calcium concentra-tions (calcium is a heavy Co antagonist) in the study region [12] necessitated supple-mentation of cattle feeding rations with Co salt to improve mineral metaboli sm anddetermine an effect of supplements on cobalt levels in animai tissues [29,42].

Due to low hair levels of Co and the long experimental period, it was useful to testthe hair samp\ed from the animals. Because of economical reasons, the mineral mixture,formulated following recommendations, was supplemented with cobalt sulfate, although

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D •• lactating . laktulące

Fig. 2. Mean Co levels in the hair of cows according to their physiological statusRys. 2. Średni poziom Co w sierści krów z uwzględnieniem grupy fizjologicznej

carbonates and oxides are believed to be most valuable [19, 25]. There was found a verysmali and slow effect of Bovifosfornag'P on hair concentration of cobalt. The mineralsupplement increased Co levels in the hair of experimental animals (Fig. 3). Statisticallysignificant differences were found in farm A (l st sampling) and C (2nd sampling).Similar statistical tendencies were reported by other authors [6, 43].

0.35 ~.35: __ ~------------ _

O:W V-...---.. O~.L -_0.25 t O;25~ .17 _ _

~E t/" --;;;-,; 0.20 l 0.17 ----u:rr __~Jt V --- 0,1V:;--"t;:o~O.15 1//' 0.12·

E E 0.10 r/.»:

0,05

~, '. .

0.00 I -:-r:--r K~' ---:-.-l-----!~.I Farm A- Ferma A . D

Farm B - Ferma BFarm C - Ferma C

Fig. 3. Mean Co le vel s in the hair of control and experimental cows according to the date of sampling(K - control group, D - experimental group; I, II - sampling dates; a, b - statistically significantdifferences at P~O.OS)Rys. 3. Średni poziom Co w sierści krów, z uwzględnieniem grupy kontrolnej i doświadczalnej orazkolejności pobrania (K - grupa kontrolna, D - grupa doświadczalna; l, II - pobrania; a, b - różnice istotnestatystycznie przy P~O,OS)

128

The present study showed that cobalt levels in the soils of southern Podlasie werewithin the limits of background values for these kinds of soils in Poland. Feed eoncen-tration of cobalt varied according to feed type and could not result in Co deficiency orexces s in the animals examined. Tissue analysis indicated reduced cobalt levels in cows'hair. Lower Co concentration in the body might result frorn an influence of other feedmineral components, in particular Ca, Cu and Zn. In order to supplement the cobaltdeficiencies and improve metaboli sm of macroelements and microelements, we·appliedthe mineral mixture Bovifosfornag'P which was formulated foIIowing recommendationsand monitoring studies. The mixture positively influenced mineral metabolism of cows,which was reflected in increased cobalt levels in hair.

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Krzysztof Górski, Agata Paprocka, Elżbieta Bombik,Teresa Bombik, Leon Saba

Wpływ dodatku mieszanki mineralnej na poziom kobaltuu krów mlecznych z regionu południowego Podlasia

Streszczenie

W ciągu trzech lat przeprowadzono na terenie południowego Podlasia badania mające na celuocenę stanu zaopatrzenia krów w kobalt na podstawie analizy gleb, pasz i tkanek organizmu(surowica, sierść). Następnie u krów mlecznych z czterech ferm (A, B, C, D) określono przydat-ność żywieniową mieszanki mineralnej Bovifosfornag'", której skład uwzględniał warunki biogeo-chemiczne panujące w miejscu ich bytowania oraz profil pierwiastkowy tkanek zwierzęcych.Próbki gleb pobierano jednorazowo z pastwisk i pól uprawnych w szczycie sezonu wegetacyjnegoza pomocą laski gleboznawczej. Zawartość kobaltu oznaczono metodą ASA. W próbkach określo-no również odczyn (pH) badanych gleb. Próbki pasz do oznaczeń pobierano we wszystkichokresach wprowadzania ich do dawek pokarmowych. Zawartość Co w paszach oznaczono metodąASA. Krew od wybranych krów pobierano czterokrotnie z żyły powierzchniowej szyi w następu-jących terminach: około 60 dnia przed porodem, 10-14 dni przed porodem, po pierwszym miesiąculaktacji i po dwóch miesiącach laktacji. Sierść pobierano z części grzbietowej dwukrotnie: przydrugim i trzecim pobraniu krwi. Zawartość Co w surowicy i włosach oznaczono metodą ICP.Stwierdzono, że poziom kobaltu w glebach z rejonu południowego Podlasia mieścił się w grani-cach wartości rejestrowanych dla tego typu gleb w Polsce. Koncentracja kobaltu w paszach byłazmienna w zależności od ich rodzaju i nie stanowiła zagrożenia nadmiaru bądź niedoboru Cou zwierząt w analizowanych fermach. Na podstawie badań tkanek organizmu wykazano obniżonepoziomy kobaltu w sierści krów. Zastosowana mieszanka Bovifosfomag®, jako uzupełnieniestwierdzonych niedoborów, wywarła korzystny wpływ na metabolizm mineralny u krów, co wy-rażało się zwiększonymi poziomami kobaltu w sierści.

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