Bioavailability of two organic forms of zinc in comparison to zinc sulphate for weaning pigs fed a diet composed mainly of wheat,barley and soybean meal

This study was performed to compare the bioavailability of two organic zinc compounds, a zinc glycinate complex and a zinc amino acid chelate with that of zinc sulphate in growing pigs fed a basal diet composed mainly of wheat, barley and soybean meal. The experiment included 96 pigs with an average body weight of 8 kg, allotted to ten groups of nine to ten pigs each. The first group received the basal diet, containing 42 mg of native zinc per kg, without zinc supplementation over a period of five weeks. The other nine groups received the basal diet supplemented with 15, 30 or 50 mg of zinc/kg as zinc sulphate, zinc glycinate or the zinc amino acid chelate. Pigs fed the unsupplemented diet had a lower growth performance (body weight gain, feed conversion ratio) than the other nine groups. Supplementation of 15 mg zinc/kg diet (irrespective of zinc form) was sufficient to yield optimum growth performance. Plasma zinc concentration and activity of alkaline phosphatase were rising with increasing zinc supplementation levels up toa maximum reached at a supplementary level of 30 or 50 mg/kg diet for activity of alkaline phosphatase and plasma zinc concentration, respectively. The response of those parameters to zinc supplementation did, however, not differ between thethree zinc compounds considered. The apparent digestibility of zinc from the diet was also not different for the three zinc compounds. In conclusion, these findings show that the bioavailability of the two organic zinc compounds did not differ from that of zinc sulphate in growing pigs fed a diet with wheat, barley and soybean meal as major components.     

Bioavailability of two organic forms of zinc in comparison to zinc sulphate for weaning pigs fed a diet composed mainly of wheat, barley and soybean meal

This study was performed to compare the bioavailability of two organic zinc compounds, a zinc glycinate complex and a zinc amino acid chelate with that of zinc sulphate in growing pigs fed a basal diet composed mainly of wheat, barley and soybean meal. The experiment included 96 pigs with an average body weight of 8 kg, allotted to ten groups of nine to ten pigs each. The first group received the basal diet, containing 42 mg of native zinc per kg, without zinc supplementation over a period of five weeks. The other nine groups received the basal diet supplemented with 15, 30 or 50 mg of zinc/kg as zinc sulphate, zinc glycinate or the zinc amino acid chelate. Pigs fed the unsupplemented diet had a lower growth performance (body weight gain, feed conversion ratio) than the other nine groups. Supplementation of 15 mg zinc/kg diet (irrespective of zinc form) was sufficient to yield optimum growth performance. Plasma zinc concentration and activity of alkaline phosphatase were rising with increasing zinc supplementation levels up to a maximum reached at a supplementary level of 30 or 50 mg/kg diet for activity of alkaline phosphatase and plasma zinc concentration, respectively. The response of those parameters to zinc supplementation did, however, not differ between the three zinc compounds considered. The apparent digestibility of zinc from the diet was also not different for the three zinc compounds. In conclusion, these findings show that the bioavailability of the two organic zinc compounds did not differ from that of zinc sulphate in growing pigs fed a diet with wheat, barley and soybean meal as major components.     

Granulation mechanism of a UASB reactor supplemented with iron

The aim of this paper was to propose a granulation mechanism in order to interpret all the experimental observations that arose during experiments on two UASB reactors, where one was supplemented with ferrous iron at a dose of 0.01 g Fe(2+) per g COD feed. This supplementation with ferrous iron allowed COD removal of more than 98% at a loading rate of 9 g COD/L per day, which was 24% higher than for the reactor not receiving the ferrous iron. Moreover, in the Fe(2+)-dosed reactor, a higher increase of the granule diameter was observed. Indeed, the granule diameter in the Fe(2+)-dosed reactor at the end of the experiments was 56% greater than that of the control reactor. This mechanism describes the course of anaerobic granule growth. The formation of the inorganic precipitate of ferrous sulphide constitutes the inert nuclei around which the biomass is attached. This initiates the formation of new granules.     

Bioavailability study of dried microencapsulated ferrous sulfate — SFE 171 — by means of the prophylactic-preventive method

Microencapsulated ferrous sulfate with soy lecithin (SFE-171) has been used as an iron source for the fortification of milk and dairy products. With the purpose to extend the use of this agent to other kind of foods or even to pharmaceutical preparations for oral administration, the SFE-171 was turned into a fluid powder (SFE-171-P) by means of vacuum drying. The iron bioavailability (BioFe) of SFE-171-P was evaluated in this work by means of the prophylactic-preventive method in rats, using ferrous sulfate as reference standard. Both iron sources were separately added to a basal diet of low iron content in a concentration of 10 mg iron/kg diet. Two groups of 10 weaned rats 25 days old received the fortified diets during 28 days, while a third group of the same size received the basal diet without iron additions. The weights and haemoglobin concentrations (HbC) of every animal were determined before and after the treatment, thus allowing the calculation of the mass of iron incorporated into haemoglobin (HbFe) during this period. The BioFe of the iron sources were obtained as the percentage ratio between the HbFe and the mass of iron consumed by each animal. The results were also given as Relative Biological Value (RBV), which relates the BioFe of the studied source with that of the reference standard. The liver iron concentration (LIC) of each animal was determined at the end of the experiment in order to evaluate the influence of the studied iron sources on the liver iron stores. SFE-171-P presented BioFe, RBV and LIC values of (47 ± 7) %, 109% and (46.6 ± 3.4) mg/kg respectively, while the corresponding values for the reference standard were of (43 ± 7)%, 100% and (45.0 ± 4.7) mg/kg. These results show that the drying process used to produce the SFE-171-P does not affect its bioavailability, which is also adequate for the potential use of this product in food fortification or with pharmaceutical purposes.     

Bioavailability Studies of Stabilized Iron (II) Sulfate by Means of the Prophylactic-preventive Method

The bioavailability of stabilized ferrous sulfate was studied by means of the prophylactic-preventive test in rats. For comparative purposes, ferrous sulfate was used as reference standard. The test was performed in male weaned rats during 3 weeks, which were randomized into three groups of ten animals each. A control group received a basal diet of low iron content while the other groups received the same diet added with iron at a dose of 15 mg/kg as FeSO₄ 7H₂O and stabilized ferrous sulfate, respectively. Individual hemoglobin concentrations and weights were determined at the beginning and at the end of the study, and food intake was daily registered. Iron bioavailability (BioFe) of each source was calculated as the ratio between the amount of iron incorporated into hemoglobin during the treatment and the total iron intake per animal. A relative biological value was obtained as the ratio between the BioFe of stabilized ferrous sulfate and the reference standard given a value of 96%. Stabilized ferrous sulfate showed a high bioavailability, and when it is used to fortify dairy products as cheese and fluid milk in a dose of 15–20 mg of iron per kilogram, it behaved inertly in relation to the sensorial properties of the fortified food. These results suggest that this iron compound is a promising source to be use in food fortification.     

Iron supplementation moderates but does not cure the Belgrade anemia

Belgrade rats inherit microcytic, hypochromic anemia as an autosomalrecessive trait (gene symbol b). Erythrocytes and tissue are iron deficientin the face of elevated TIBC (total iron binding capacity) and percent ironsaturation; iron injections increased the number of erythrocytes but theirappearance remained abnormal. We have investigated iron supplements toimprove husbandry of b/b rats and to learn more about the underlying defectand its tissue distribution. Weekly IM (intramuscular) injections ofiron–dextran (Imferon at 30 mg kg) improved the anemia but did not alter thered cell morphology. Certain diets also improved the health of b/b rats whencompared to standard rat chows by the criteria of weight, survival toadulthood, hematology and reproduction. The critical nutritional factorturned out to be iron bioavailability, with ferrous iron added to the dietimproving the health of Belgrade rats without affecting the underlyingerythroid defect. Tissue iron measurements after dietary or parenteralsupplementation confirmed the iron deficient status of untreated b/b rats andestablished that dietary ferrous iron partially relieved this deficiency,with injections leading to greater amounts of tissue iron. Serum iron andTIBC were also found to be elevated in untreated b/b rats, with dietarysupplementation decreasing but not eliminating the elevation in TIBC. Thesestudies indicate that iron supplements can improve the health of b/b ratswithout altering the underlying defect and also suggest that the mutationcould alter iron uptake in the GI (gastrointestinal) tract.     

Effect of feeding supplemental copper on performance,fatty acid profile and on cholesterol contents and oxidative stability of meat of rabbits

One hundred and four rabbits, five weeks old at the beginning of the experiment, were divided into four groups according to a feed additive treatment. Rabbits of the 1st, 2nd, 3rd and 4th group were fed a basal granulated feed (control), basal feed supplemented with CuSO₄ · 5H₂O at 50mg Cu · kg^‐1, basal feed supplemented with 150mg Cu · kg^‐1, and the latter feed supplemented with 100mg · kg^‐1 vitamin E, respectively. The duration of the experiment was 42 days. Addition of Cu at 150mg · kg.^‐1 increased weight gain non‐significantly by 9.1%. This effect was the most pronounced in the first two weeks of fattening. The lowest mortality was observed in rabbits fed the highest amount of additives (7.7% vs. 19.2% in the control). Rabbits were slaughtered at the age of 11 weeks. Neither treatment influenced proportions of saturated, monounsaturated and polyunsaturated fatty acids in lipids extracted from the loin and hindleg muscles. In rabbits fed the highest amount of copper and vitamin E, the cholesterol concentration was significantly decreased by 13.6% and 17.9% in the loin and hindleg meat, respectively. Effects of Cu added at 50mg · kg^‐1 were marginal. Copper had no effect on the oxidative stability of meat, measured as thiobarbituric acid‐reactive substances in meat stored at 4°C for 0, 3 and 8 days. Vitamin E added in excess of nutritional requirement improved the oxidative stability of meat. In copper‐fed rabbits, Cu accumulated in the liver, but not in muscles. Feeding of the basal feed for 7 days to rabbits previously fed copper sulphate decreased the hepatic Cu concentration by 14.0 to 24.4%     

Influence of dietary protein type and iron source on the absorption of amino acids and minerals

The apparent digestibility coefficient (ADC) of amino acids and the balance of minerals (calcium, phosphorus, magnesium and iron) has been determined in rats fed four diets differing in the protein type (casein or soy protein) and iron source (ferrous sulphate or lactate) in order to study the possible interactions of these nutrients. The availability of amino acids, especially essential amino acids, was greater in the diet made with animal protein (casein). The iron source also affected the absorption of most amino acids in all the diets assayed with ferrous sulphate being greater. The balance of iron, magnesium and phosphorus was higher in the diets containing animal protein. The retention of calcium and magnesium was significantly greater when ferrous sulphate was used as iron source. These results demonstrate the important interaction between amino acids and minerals and between the minerals themselves, which must be carefully studied when selecting different types of protein or mineral sources in human or animal nutrition.     

Stabilized ferrous gluconate as iron source for food fortification: bioavailability and toxicity studies in rats

The iron bioavailability and acute oral toxicity in rats of a ferrous gluconate compound stabilized with glycine (SFG), designed for food fortification, was studied in this work by means of the prophylactic method and the Wilcoxon method, respectively. For the former studies, SFG was homogeneously added to a basal diet of low iron content, reaching a final iron concentration of 20.1 +/- 2.4 mg Fe/kg diet. A reference standard diet using ferrous sulfate as an iron-fortifying source (19.0 +/- 2.1 mg Fe/kg diet) and a control diet without iron additions (9.3 +/- 1.4 mg Fe/kg diet) were prepared in the laboratory in a similar way. These diets were administered to three different groups of weaning rats during 23 d as the only type of solid nourishment. The iron bioavailability of SFG was calculated as the relationship between the mass of iron incorporated into hemoglobin during the treatment and the total iron intake per animal. This parameter resulted in 36.6 +/- 6.2% for SFG, whereas a value of 35.4 +/- 8.0% was obtained for ferrous sulfate. The acute toxicological studies were performed in two groups of 70 female and 70 male Sprague-Dawley rats that were administered increasing doses of iron from SFG. The LD50 values of 1775 and 1831 mg SFG/kg body wt were obtained for female and male rats, respectively, evidencing that SFG can be considered as a safe compound from a toxicological point of view.     

Stabilized ferrous gluconate as iron source for food fortification

The iron bioavailability and acute oral toxicity in rats of a ferrous gluconate compound stabilized with glycine (SFG), designed for food fortification, was studied in this work by means of the prophylactic method and the Wilcoxon method, respectively. For the former studies, SFG was homogenously added to a basal diet of low iron content, reaching a final iron concentration of 20.1±2.4 mg Fe/kg diet. A reference standard diet using ferrous sulfate as an iron-fortifying source (19.0±2.1 mg Fe/kg diet) and a control diet without iron additions (9.3±1.4 mg Fe/kg diet) were prepared in the laboratory in a similar way. These diets were administered to three different groups of weaning rats during 23 d as the only type of solid nourishment. The iron bioavailability of SFG was calculated as the relationship between the mass of iron incorporated into hemoglobin during the treatment and the total iron intake per animal. This parameter resulted in 36.6±6.2% SFG, whereas a value of 35.4±8.0% was obtained for ferrous sulfate. The acute toxicological studies were performed in two groups of 70 female and 70 male Sprague-Dawley rats that were administered increasing doses of iron from SFG. The LD₅₀ values of 1775 and 1831 mg SFG/kg body wt were obtained for female and male rats, respectively, evidencing that SFG can be considered as a safe compound from a toxicological point of view.     

Trypanosoma brucei brucei: Effects of ferrous iron and heme on ecto-nucleoside triphosphate diphosphohydrolase activity

Trypanosoma brucei brucei is the causative agent of animal African trypanosomiasis, also called nagana. Procyclic vector form resides in the midgut of the tsetse fly, which feeds exclusively on blood. Hemoglobin digestion occurs in the midgut resulting in an intense release of free heme. In the present study we show that the magnesium-dependent ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) activity of procyclic T. brucei brucei is inhibited by ferrous iron and heme. The inhibition of E-NTPDase activity by ferrous iron, but not by heme, was prevented by pre-incubation of cells with catalase. However, antioxidants that permeate cells, such as PEG-catalase and N-acetyl-cysteine prevented the inhibition of E-NTPDase by heme. Ferrous iron was able to induce an increase in lipid peroxidation, while heme did not. Therefore, both ferrous iron and heme can inhibit E-NTPDase activity of T. brucei brucei by means of formation of reactive oxygen species, but apparently acting through distinct mechanisms.     

Non-heme Iron as Ferrous Sulfate Does Not Interact with Heme Iron Absorption in Humans

The absorption of heme iron has been described as distinctly different from that of non-heme iron. Moreover, whether heme and non-heme iron compete for absorption has not been well established. Our objective was to investigate the potential competition between heme and non-heme iron as ferrous sulfate for absorption, when both iron forms are ingested on an empty stomach. Twenty-six healthy nonpregnant women were selected to participate in two iron absorption studies using iron radioactive tracers. We obtained the dose?Cresponse curve for absorption of 0.5, 10, 20, and 50?mg heme iron doses, as concentrated red blood cells. Then, we evaluated the absorption of the same doses, but additionally we added non-heme iron, as ferrous sulfate, at constant heme/non-heme iron molar ratio (1:1). Finally, we compare the two curves by a two-way ANOVA. Iron sources were administered on an empty stomach. One factor analysis showed that heme iron absorption was diminished just by increasing total heme iron (P?<?0.0001). The addition of non-heme iron as ferrous sulfate did not have any effect on heme iron absorption (P?=?NS). We reported evidence that heme and non-heme iron as ferrous sulfate does not compete for absorption. The mechanism behind the absorption of these iron sources is not clear.     

Iron bioavailability from fortified petit suisse cheese determined by the prophylactic-preventive method

In this research, we measured the iron bioavailability of ferrous gluconate stabilized with glycine (SFG) when it is used to fortify petit suisse cheese using the prophylactic-preventive method in rats. Three groups of male, weaned rats received a basal diet (control diet; 5.2 ppm Fe), a reference standard diet (SO₄Fe; 9.2 ppm Fe), and a basal diet using iron-fortified petit suisse cheese as the iron source (cheese diet; 8.8 ppm Fe) for 22d. The iron bioavailability was calculated as the ratio between the mass of iron incorporated into hemoglobin and the total iron intake per animal during the treatment. These values (BioFe) were 68% and 72% for SFG and ferrous sulfate, respectively. The value of the Relative Biological Value (RBV) was 95% for SFG in petit suisse cheese. These results show that according to this method, the iron bioavailability from industrial fortified petit suisse cheese can be considered as a high bioavailability rate.     

High intakes of tin lower iron status in rats

The effects of relatively low (1, 10, and 50 mg/kg) and high (100 and 200 mg/kg) dietary concentrations of tin (added as stannous chloride) on iron status of rats were determined. After feeding the diets for 28 d, feed intake and body weights were not significantly affected. Iron concentrations in plasma, spleen, and tibia as well as percentage transferrin saturation were decreased in rats fed the diets supplemented with 100 or 200 mg tin/kg. In rats fed the diet containing 200 mg tin/kg, group mean hemoglobin, hematocrit, and red blood cell count were slightly lowered but total iron binding capacity was not affected. Iron status was not influenced by dietary tin concentrations lower than 100 mg/kg. If these results can be extrapolated to humans, then it may be concluded that tin concentrations in the human diet, which range from 2 to 76 mg/kg dry diet, do not influence iron status in humans.     

Effect of level of feed restriction during growth and/or fattening on fatty acid composition and lipogenic enzyme activity in heavy pigs

The influence of feeding system on growth performance, carcass characteristics, and fatty acids profile of subcutaneous backfat and lipogenic enzymes activity was studied. The following feeding systems were applied: HH: group of 12 pigs fed with a high feeding level during growth and fattening periods; HL: group of 12 pigs fed with a high feeding level during growth period and a low feeding level during fattening period; LH: group of 12 pigs fed with a low feeding level during growth period and a high feeding level during fattening period; LL: group of 12 pigs fed with a low feeding system during growth and fattening periods. The high and low feeding levels applied during growth and fattening periods were 70 and 35 g feed/kg live weight^0.75 (LW^0.75) day and 121 and 87 g feed/kg LW^0.75 day, respectively. The duration of the growth period was 163 days in all groups. The ages at slaughter of HH, HL, LH and LL pigs were approximately 383, 406, 439 and 503 days, respectively. The HH and LL pigs had the higher and lower average daily gain respectively, while the LH pigs presented the best feed conversion ratio during fattening period. At slaughter, the LL pigs had the higher value of loin width and the lower values of backfat thicknesses and Longissimus dorsi muscle intramuscular fat percentage (P<0.05). The subcutaneous backfat outer and inner layers from LH and LL pigs had a significantly (P<0.05) higher proportion of C18:1 n-9 and ΣMUFA (monounsaturated) acids than those from HH and HL pigs. The subcutaneous backfat inner layer from LL pigs had a lower C16:0 and ΣSFA (saturated) acids proportions than those from remaining groups of pigs. The activity of the fatty acid synthase (FAS) and glucose-6-phosphate dehydrogenase (G6PD) enzymes, measured in subcutaneous backfat, at the end of growing period was significantly (P<0.05) higher in pigs fed high feeding level during growing phase. An increase of feeding level during the fattening phase led to an increase in the activity of G6PD enzyme. In conclusion, a marked effect of feeding system was observed on pig productivity and fatty acid composition.     

Liver iron depletion and toxicity of the iron chelator Deferiprone (L, CP20) in the guinea pig

The use of the iron chelator deferiprone (L, CP20, 1,2-dimethyl-3-hydroxypyrid-4-one) for the treatment of diseases of iron overload and other disorders is problematic and requires further evaluation. In this study the efficacy, toxicity and mechanism of action of orally administered L were investigated in the guinea pig using the carbonyl iron model of iron overload. In an acute trial, depletion of liver non-heme iron in drug-treated guinea pigs (normal iron status) was maximal (approximately 50% of control) after a single oral dose of L1 of 200 mg kg, suggesting a limited chelatable pool in normal tissue. There was no apparent toxicity up to 600 mg kg. In each of two sub-acute trials, normal and iron-loaded animals were fed L (300 mg kg day) or placebo for six days. Final mortalities were 12/20 (L) and 0/20 (placebo). Symptoms included weakness, weight loss and eye discharge. Iron-loaded as well as normal guinea pigs were affected, indicating that at this drug level iron loading was not protective. In a chronic trial guinea pigs received L (50 mg kg day) or placebo for six days per week over eight months. Liver non-heme iron was reduced in animals iron-loaded prior to the trial. The increase in a wave latency (electroretinogram), the foci of hepatic, myocardial and musculo-skeletal necrosis, and the decrease in white blood cells in the drug-treated/normal diet group even at the low dose of 50 mg kg day suggests that L may be unsuitable for the treatment of diseases which do not involve Fe overload. However, the low level of pathology in animals treated with iron prior to the trial suggests that even a small degree of iron overload (two-fold after eight months) is protective at this drug level. We conclude that the relationship between drug dose and iron status is critical in avoiding toxicity and must be monitored rigorously as cellular iron is depleted.     

Bioavailability, absorption mechanism, and toxicity of microencapsulated iron (I) sulfate

The iron compounds used for food fortification have to meet certain requisites related to their bioavailability, absorption mechanism, and toxicity, since they will be consumed by a massive population group. With these purposes, we evaluated a new product used for the iron fortification of milk and lacteous derivatives, called SFE-171^TM, which is a ferrous sulfate, microencapsulated with phospholipids. The bioavailability studies were carried out using four groups of 30 female mice each. In two groups, we studied the absorption of ferrous ascorbate and ferrous sulfate, both in water as reference standards, which show absorptions of 13.1±4.9% and 13.2±4.3%, respectively. With the third group, we studied the absorption of ferrous sulfate in milk; its value, 7.9±3.2%, is significantly lower than that of the remaining groups, with ap<0.01. The studies with SFE-171^TM in milk, were performed on the fourth group, with a result of 11.6±4.5%, demonstrating that its absorption does not differ significantly from that of the reference standards. The absorption mechanism was determined by means of in vivo self-displacement studies of the ferrous ion and the SFE-171^TM, taking ferrous sulfate as the reference compound. For this study, 210 female mice were used, and no significant difference between the absorption mechanism of both products could be observed. Toxicity studies of the new product with regard to ferrous sulfate were carried out with two groups of 70 female mice each and two groups of 70 male mice each. The lethal dose 50% LD₅₀ for SFE-171^TM and for ferrous sulfate was 1200 and 680 mg/kg for female mice and 1230 and 670 mg/kg for male mice, respectively, demonstrating that the toxicity of the first product is substantially lower than that of the reference standard. We conclude that the iron product under study has a high bioavailability, an absorption mechanism equal to that of nonhemic iron, and lower toxicity than ferrous sulfate.     

Effect of probiotic CenBiot on the control of diarrhoea and feed efficiency in pigs

The production and testing of a probiotic formulated with a strain of Bacillus cereus (Probiotic CenBiot) is reported. The strain was grown in fed batch fermenters, desiccated, and mixed with cornflour. To test the effect of the probiotic in controlling piglet diarrhoea, two groups of sows and their respective litters received feed supplemented with probiotic CenBiot or Furazolidone; the control group received the same basic feed but not supplemented. Probiotic CenBiot was as effective as Furazolidone, reducing the prevalence of diarrhoea to half that in the control group. It also significantly improved feed conversion, daily weight gain and total weight gain in pigs in the nursery phase. Feed conversion ratios of weaned pigs were 1·904, 2·146 and 2·099 for the groups that received Probiotic CenBiot, a commercial probiotic, and basic feed, respectively. In terms of food consumption, the Probiotic CenBiot group consumed 12·6% less than the commercial probiotic group, and 10% less than the control group, to achieve the same weight.     

Effect of probiotic CenBiot on the control of diarrhoea and feed efficiency in pigs

The production and testing of a probiotic formulated with a strain of Bacillus cereus (Probiotic CenBiot) is reported. The strain was grown in fed batch fermenters, desiccated, and mixed with cornflour. To test the effect of the probiotic in controlling piglet diarrhoea, two groups of sows and their respective litters received feed supplemented with probiotic CenBiot or Furazolidone; the control group received the same basic feed but not supplemented. Probiotic CenBiot was as effective as Furazolidone, reducing the prevalence of diarrhoea to half that in the control group. It also significantly improved feed conversion, daily weight gain and total weight gain in pigs in the nursery phase. Feed conversion ratios of weaned pigs were 1.904, 2.146 and 2.099 for the groups that received Probiotic CenBiot, a commercial probiotic, and basic feed, respectively. In terms of food consumption, the Probiotic CenBiot group consumed 12.6% less than the commercial probiotic group, and 10% less than the control group, to achieve the same weight.     

Effects of time and dietary iron on tissue iron concentration as an estimate of relative bioavailability of supplemental iron sources for ruminants

Two experiments were conducted to investigate the effects of time and dietary Fe on tissue Fe concentrations following short-term, high level supplementation for use as a bioassay procedure for supplemental Fe sources for ruminants. In Experiment 1, 28 wethers were allotted randomly to four experimental diets which were fed for 15 or 30 days. The basal maize–soyabean meal–cottonseed hulls diet (193 mg kg⁻¹ Fe) was supplemented with 0, 400, 800 or1200 mg kg⁻¹ added Fe from reagent grade ferrous sulfate (FeSO₄·7H₂O). Iron concentrations in liver, kidney, and spleen increased (P<0.05) as dietary Fe increased; however, muscle, heart, and bone Fe concentrations were unaffected. A logarithmic transformation of liver or kidney Fe concentrations at 30 days regressed on added dietary Fe produced the best fits to a linear model. In Experiment 2, bioavailability of Fe from three feed grade ferrous carbonates known to differ (carbonates A, B, and C) was compared to that from reagent grade ferrous sulfate. The dietary treatments fed for 30 days included the above basal diet (90 mg kg⁻¹ Fe) supplemented with 0, 300, 600 or 900 mg kg⁻¹ added Fe from ferrous sulfate or 600 mg kg⁻¹ Fe from ferrous carbonates A, B, or C. Liver Fe concentrations from sheep fed ferrous sulfate were numerically greater than those of animals fed the carbonate sources or control diet. Kidney Fe concentrations from lambs fed ferrous sulfate at 600 mg kg⁻¹ Fe or carbonate-A were greater (P<0.05) than those fed carbonates B or C. Iron concentrations in spleen were lower (P<0.05) in lambs fed carbonate-B than for those fed 600 mg kg⁻¹ Fe as ferrous sulfate, but were similar to other carbonates. Overall average bioavailability estimates based on multiple regression slope ratios for the three tissues were ferrous sulfate 1.00, carbonate-A 0.55, carbonate-B 0.00, and carbonate-C 0.20. Estimates for carbonates A and C were similar to those based on hemoglobin concentrations reported previously for young swine supplemented at dietary concentrations near the requirement.