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Effect of iron supplementation on plasma levels of vitamins A,
E and C in piglets
a ,
*
c b ,1 c¨
Florian J. Schweigert
, Herbert Gurtler , Anita Baumane
, Margit Wahren ,
cMonika Leo
a
¨
Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114 –116, D-14558 Bergholz-Rehbrucke, Germany b
Department of Veterinary Physiology, Veterinary Faculty, University of Leipzig, Semmelweisstrasse 4, D-04103 Leipzig, Germany c
Department of Veterinary Physiological Chemistry, Veterinary Faculty, University of Leipzig, Semmelweisstrasse 4, D-04103 Leipzig,
Germany
Received 23 September 1997; received in revised form 4 September 1998; accepted 1 June 1999
Abstract
The objective of this study was to investigate the effects of two different commercially available iron–dextran preparations for intramuscular injection of piglets on haematological parameters including haemoglobin, haematocrit and mean corpuscular haemoglobin concentration (MCHC), as well as on plasma levels of vitamins A, E and C. Piglets were either control animals or administered two different iron–dextran preparations (200 mg iron) at day 3 and blood samples were taken by puncture of the cava vein at 3 (before injection), 10, 17 and 24 days after birth. Both iron preparations were efficient to prevent anaemia from day 10 onwards (P,0.001) with no differences between the iron preparations used. While no significant differences were observed for the plasma level of vitamin E, the administration of iron caused a more pronounced decrease in plasma vitamin C than in control animals with significant differences observed already at day 10 (P,0.001). Furthermore, the administration of iron prevented the decrease in the concentration of retinol in plasma as observed in control piglets, resulting in higher plasma retinol levels at day 24 (P,0.01). This would indicate an improvement of the vitamin A status by stabilisation of plasma retinol levels in piglets after birth. Because of the overall role of vitamin A in growth and tissue integrity, this could be of importance for the well-being of the developing piglet. 2000 Elsevier Science B.V. All rights reserved.
Keywords: Pig; Iron; Vitamin A; Vitamin E; Vitamin C
1. Introduction
Iron has important functions in the body as a component of haemoglobin and numerous other iron-*Corresponding author. Tel.: 0149-33200-88527 / 528; fax: containing proteins. Increased incidence of infectious
149-33200-88573.
diseases associated with iron deficiency may be E-mail address: fjschwei@rz.uni-potsdam.de (F.J. Schweigert)
1 attributed to an impairment of activities of enzymes Present address: Department of Animal Nutrition, Latvia
Uni-versity of Agriculture, Liela Strasse 2, Jelgava, LV-3001, Latvia. containing iron in cells of the immune system (Beard
et al., 1996). However, apart from being essential to Within one litter, the maximal possible number of cell metabolism, iron can also promote the damage piglets were equally distributed among the control of lipids and membranes by the generation of free and treatment groups. During the experiment, piglets radicals (Herbert et al., 1996). Since iron deficiency remained with their dam. On the third day after birth, and iron overload result in major disturbances, the the piglets of the two experimental treatments were body iron balance is tightly regulated (Bothwell, injected intramuscularly with 1 ml of the iron– 1995). In piglets, iron deficiency is a major problem dextran preparations containing 200 mg / ml of iron because body stores at birth are small (Ekman and [Ursoferran 200 (iron (III)-hydroxide–dextran–hep-Jwanska, 1966) and sow milk contains insufficient tone acid), Serum-Werk Bernburg, Germany – group amounts of iron compared to the need of piglets U (n521) or Myofer 200 (iron (III)-hydroxide–
¨
during the first weeks of life (Kirchgessner et al., dextran), Hoechst Veterinar GmbH, Germany – 1982). Therefore, it is necessary to supply iron to group M (n521)]. Control piglets (group C; n521) piglets orally or parenterally shortly after birth in were injected with 1 ml of physiological saline. order to maintain proper development (Furugouri, Blood samples were obtained on days 3 (before
¨
1972; Gurtler et al., 1979). injection at this day), 10, 17 and 24 after birth from Iron is known to interact with numerous other the cava vein. The haematocrit value was measured dietary components (Lynch, 1997). In this context, by microhaematocrit capillaries, and the concentra-the interaction of iron as a generator of free radicals tion of haemoglobin was measured with the cyan-with the antioxidative vitamins E and C has been methaemoglobin method according to van Kampen studied (Tollerz and Lannek, 1964; Herbert et al., and Zijlstra (1961). The mean corpuscular haemo-1996; Berger et al., 1997). Additionally, the inter- globin concentration (MCHC) was calculated from action of iron and vitamin A has elicited consider- haemoglobin and the haematocrit. Vitamins A and E able interest because the deficiencies of both micro- were extracted from plasma after deproteinisation nutrients are two of the most prevalent nutritional with an equivalent volume of ethanol twice into three problems of humans in developing countries (Bloem, volumes of n-hexane (0.05% BHT, butylated hy-1995). Numerous studies address the effect of vita- droxytoluene). The combined organic extracts were min A on iron metabolism but very little is known dried under nitrogen flow and redissolved in metha-with regard to the effect of iron on vitamin A nol–ethanol (80:20; v / v). Vitamins A (retinol) and E metabolism (Bloem, 1995). (a-tocopherol) were separated and quantified using a The objective of this field study was to evaluate reversed-phase high-performance liquid chromatog-the effect of two different commercially available, raphy (HPLC) system (Waters, Eschborn, Germany) injectable iron–dextran preparations on haematologi- on an RP-18 column (5 mm, 12534 mm; Grom, cal parameters, retinol and antioxidative vitamins E Germany) with methanol as a solvent at a flow-rate and C in plasma, because differences in composition of 1 ml (1–3 min), 1.5 ml (3–7 min) and 2 ml (7–19 might not only affect availability (haematological min) (Schweigert, 1990). A fluorescence spec-parameters), but oxidative properties (vitamins) as trophotometer was used for the quantification of well. vitamin E (excitation 295 nm; emission 330 nm). Identification and quantification of vitamins were obtained by comparison of retention time and peak
2. Materials and methods areas (at 325 nm for vitamin A) with external
standards (Serva, Heidelberg, Germany). Vitamin C The effects of iron supplementation on was determined in the plasma after deproteinisation haematological parameters and on plasma levels of with 0.6 M HClO using dinitrophenylhydrazine as4 vitamins A (retinol), E and C were investigated in reagent (Albanese et al., 1975). All solvents or nursing piglets (German Landrace3German chemicals used were of high purity commercial
´
Edelschwein3Pietrain) from day 3 to 24 under field grade (Merck, Darmstadt, Germany).
groups for vitamin C at the beginning of the experi- administration of iron was an effective way to ment, changes in response variables in plasma were prevent anaemia in piglets during the first weeks of partitioned using General Linear Model (GLM) life. This is in accordance to numerous previous
¨
procedure of SAS (SAS, 1988) for repeated mea- studies in piglets (Furugouri, 1972; Gurtler et al., surement design. When a significant difference was 1979).
found a LSD post-hoc test was used to determine the
cause of the significant difference. Between-subject 3.2. Vitamins in blood plasma factors were transformed into contrast (repeated) for
analysis. The effects of iron supplementation on vitamin A, E and C concentrations in plasma are presented in Table 2. No differences between the two groups of 3. Results and discussion piglets treated with different forms of iron were observed. The higher values for vitamin E on day 3 3.1. Haematological parameters post-partum compared to adult animals may be attributed to the uptake of colostrum and milk rich in Data on haematological parameters (Table 1) this vitamin (Schweigert et al., 1991; Hidiroglou et showed that from day 10 onwards the piglets of the al., 1993; Mahan, 1994). Plasma values of vitamin C untreated control group had lower haemoglobin (P, in general and the observed decrease over time 0.001) and haematocrit values (P,0.001) than iron- correspond to published data (Yen and Pond, 1981, treated animals. All untreated animals developed a 1983). The decrease of vitamin A level with age in hypochromic anaemia. No differences in haemoglo- control animals (P,0.01) and the more pronounced bin, haematocrit and MCHC between the two groups decrease in vitamins E and C concentrations in all of treated animals were found. The intramuscular animals (P,0.01), irrespectively of treatment, may
Table 1
Effect of intramuscular iron administration on haematological parameters of piglets (mean6SD and number of samples)
Group Day
Table 2
Effect of intramuscular iron administration on plasma levels of vitamins A, E and C in piglets (mean6SD and number of samples)
Group Day
Values in the same column with different superscripts are different (a, b P,0.05; c, d P,0.01; e, f P,0.001). Values in the same row with capital superscripts differ from the previous ones (A P,0.05; B P,0.01; C P,0.001).
1
Numbers of samples correspond to those given for retinol.
oxidant. The iron-dependent reduction of H O2 2 supporting normal haematopoiesis as well as the generates hydroxyl radicals (Herbert et al., 1996). immune system the observed effect of iron on This increase in free radicals promote the damage of vitamin A levels might be of additional benefit for lipids and therefore cell membranes, and influences the growing pig, by possibly reducing infectious the status of antioxidative vitamins and carotenoids diseases as one of the major cause of morbidity and in plasma and body tissues (Whittaker et al., 1996; mortality in commercial animal production.
Parkkila et al., 1996). The lower levels of vitamin C and to a lesser extend of vitamin E in iron-treated
piglets might be a consequence of iron-associated Acknowledgements mechanism that lead to a disappearance of both
antioxidative vitamins from plasma. Similar effects A.B. received a grant from the Saxonian Ministry have been observed in pigs, where a parenteral of Science and Art and the Hanns-Seidel-Stiftung administration of iron results in lower plasma vita- e.V., Germany.
min E concentrations compared to untreated animals (Loudenslager et al., 1986). In addition, vitamin E
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