A Systematic Review and Meta-Analysis of the Effects of Various Sources and Amounts of Copper on Nursery Piglets.

This study evaluated the impact of different dietary levels and sources of copper on the growth performance of nursery piglets through a combination of systematic review and meta-analysis. The database for this study was created using articles selected from major electronic databases. Data analysis involved forest plots and analysis of variance using mixed-effects models. The database included 63 articles published between 1990 and 2021, comprising 21,113 piglets in 946 treatments. Positive effects of supranutritional levels of copper from both inorganic and organic sources on the growth performance of nursery piglets were detected using Forest plots and analysis of variance (p < 0.001). Using mixed models, it was observed that piglet performance is influenced by body weight (p < 0.001), age (p < 0.001), and copper intake (p < 0.001). Both organic and inorganic sources of copper at supranutritional levels (>81 mg Cu/kg of diet) improved the performance of nursery piglets, but levels higher than 201 mg Cu/kg of diet did not further improve growth performance compared to 80-200 mg Cu/kg of diet. The feed conversion was worse in piglets fed with inorganic Cu sources (p < 0.001). In conclusion, dietary Cu supplementation influenced the weight gain and feed conversion rate in weaned piglets, particularly during the first few weeks post-weaning. Levels of 81 and 200 mg Cu/kg improved growth performance, but no further benefits were obtained for higher levels.

Effects of high levels of zinc oxide and dietary zinc/copper ratios on the metabolism of iron in weaned pigs.

The present study compares the use different levels of dietary zinc oxide and zinc/copper ratios on the metabolism of iron (Fe) in weaned pigs. Two experiments were conducted using 120 and 160 weanling piglets (7.96 ±â€…1.17 kg and 7.81 ±â€…0.25 kg body weight, respectively) that were randomly assigned to the experimental treatments. Experiment I: diets supplemented with 100, 1,000, and 3,000 mg/kg of zinc (Zn) as ZnO (LZn, MZn, HZn) and 130 mg/kg of copper (Cu) as CuSO4; experiment II: diets supplemented with 100 or 3,000 mg/kg of Zn as ZnO (LZn and HZn) in combination with 6 or 130 mg/kg of Cu as CuSO4 (LCu and HCu). In both experiments, diets had similar levels of supplemental Fe (100 mg/kg of Fe as FeSO4). Piglets were slaughtered at d21 (weaning), d23 (experiment I), d28 (experiment II), d35, and d42 to assess whole blood, serum, and liver Fe concentrations, hemoglobin concentration, and the relative expression of key genes associated with Fe metabolism in jejunum and liver. Whole blood Fe and hemoglobin concentrations (experiment I) as well as serum Fe concentrations (experiments I and II) were not affected by dietary treatments (P ≥ 0.11). Liver Fe concentrations (experiment II) and total liver Fe content (experiments I and II) were lower (P ≤ 0.05) in HZn compared to LZn groups at d42. In both experiments, the mRNA expression of jejunal DMT1 was lowest and that of jejunal FTH1 was highest at d42 (P ≤ 0.04) for HZn piglets. In experiment II only, jejunal FTH1 and FPN1 expression were greater (P ≤ 0.04) in HCu compared to LCu groups at d42. The highest expression of hepatic FTH1 and FPN1 at d35 and d42 (P ≤ 0.02) was detected in HZn piglets in both experiments. For hepatic HAMP, expression values were greater (P = 0.04) at d42 in HZn groups. In conclusion, high dietary ZnO levels impair Fe metabolism but the effects are not intense enough to impact circulating Fe and hemoglobin concentrations.

Zinc oxide and copper sulfate are commonly used to prevent diarrhea in weaned piglets as an alternative to antibiotics. However, this strategy has been questioned due to environmental issues and public health concerns related to bacterial resistance. Recently, it was reported that high dietary zinc levels and/or high dietary zinc/copper ratios impair zinc and copper homeostasis and are detrimental to postweaning piglets' health. However, the eventual effects on iron metabolism are much less explored in pigs. Therefore, two experiments were conducted to evaluate the effects of increasing levels of dietary zinc (100, 1,000, and 3,000 mg/kg) and of different dietary zinc/copper ratios (3,000/130, 3,000/6, 100/130, and 100/6 mg/kg) on iron metabolism in weaned piglets. This study demonstrates that high dietary zinc levels, independently of dietary copper levels, impair iron metabolism by systemic and local (intestinal) mechanisms but also suggests that preweaning factors may interfere with postweaning iron homeostasis.

Effects of dietary zinc/copper ratios on the metabolism of zinc and copper in weaned pigs.

This study compared the effects of different dietary zinc/copper ratios on zinc (Zn) and copper (Cu) metabolism in weaned pigs. One hundred and sixty piglets (7.81 ±â€…0.25 kg; 21 d of age) were used in a completely randomized 2 × 2 factorial design composed with high (H) and low (L) levels of added dietary Zn (100 and 3,000 mg/kg) and dietary Cu (6 and 130 mg/kg). Piglets were slaughtered at 21, 28, 35, and 42 d of age for blood and tissues collection. Serum, jejunum mucosa, liver, and kidney concentrations of Zn and Cu were analyzed as well as tissues mRNA abundance of genes related to their metabolism. Serum and liver Zn concentrations increased at days 28, 35, and 42 in HZn groups compared to pre-treatment levels (day 21; P ≤ 0.01) but for LZn animals, values decreased at days 28, 35, and 42 in liver (P ≤ 0.01) but remained stable vs. day 21 levels in serum (P ≥ 0.37). Serum, jejunum mucosa, liver, and kidney Zn concentrations were greater in HZn groups from day 28 (P ≤ 0.01). In jejunum mucosa, the mRNA expression of ZIP4 was lower in HZn piglets at day 28 (P ≤ 0.01) and at day 42 whereas HCu supplementation increased ZIP4 expression in LZn but not in HZn diets (P = 0.05). For ZNT1, MT3, and MT1, values of relative mRNA expression were greater for HZn animals in jejunum mucosa, liver, and kidney (P ≤ 0.01) from day 28. In kidney (P < 0.01) at day 42, HZn supplementation increased MTs expression in both LCu or HCu groups. Serum and liver Cu concentrations decreased at days 35 and 42 in all treatments compared to day 21 (P ≤ 0.04), except LZnHCu in liver that was not different from day 21 (P ≥ 0.17). Serum Cu concentrations were lower in HZn and greater in HCu groups at days 35 and 42 (P ≤ 0.01) whereas hepatic Cu was reduced by HZn diets in both LCu and HCu groups at days 35 and 42 (P ≤ 0.01). Jejunum Cu concentrations were increased by HCu diets in HZn but not in LZn groups at days 28 and 42 (P ≤ 0.04). Renal Cu concentrations were greater in HZn groups at day 28 (P < 0.01) whereas at day 42 HZn diets increased Cu values in both LCu and HCu groups (P ≤ 0.01). The expression of ATP7A in kidney at day 42 was greater for HZn groups (P = 0.02). In conclusion, high dietary Zn levels were not efficiently regulated by homeostatic mechanisms and significantly impaired Cu homeostasis. Low dietary Zn/Cu ratios allow a more efficient regulation of the metabolism of these trace minerals in post-weaning piglets. The current official recommendations for Zn and Cu to post-weaning piglets apparently do not fulfill their requirements.

Zinc oxide and copper sulfate are commonly used as growth promoters and alternatives to antibiotics to prevent diarrhea in weaned piglets but their use in post-weaning pigs diets has been challenged due to environmental issues and concerns related to bacterial resistance to antibiotics and heavy metals. Recently, it was reported that high dietary zinc levels interfere with copper status and may be detrimental to post-weaning piglets' health. In fact, the optimal dietary zinc/copper ratios need to be determined. Therefore, this experiment was conducted to evaluate the effects of different dietary zinc/copper ratios (3,000/130, 3,000/6, 100/130, and 100/6 mg/kg) on zinc and copper metabolism in weaned piglets. This study demonstrated that high dietary zinc/copper ratios impaired zinc and copper homeostasis but also that 100 mg/kg of dietary zinc and 6 mg/kg of dietary copper are apparently not sufficient to fulfill the piglets' requirements during the first weeks post-weaning.

Effects of dietary zinc oxide levels on the metabolism of zinc and copper in weaned pigs.

This study compared different dietary zinc oxide (ZnO) levels on zinc (Zn) and copper (Cu) metabolism in weaned pigs. One hundred twenty weaned piglets (7.96 ± 1.17 kg; 21 d of age) were used in a completely randomized 3 × 4 factorial design composed with three levels of dietary ZnO at 100 (100Zn), 1,000 (1,000Zn), or 3,000 mg/kg (3,000Zn) and four ages at slaughter at 21 (day 21), 23 (day 23), 35 (day 35), and 42 d (day 42). Dietary Cu levels were constant at 130 mg/kg. Serum, jejunum, liver, and kidney levels of Zn and Cu as well as mRNA abundance of genes related to Zn and Cu metabolism were analyzed. Zinc levels were greatest in 3,000Zn piglets from day 35 in all tissues (P ≤ 0.01). In 3,000Zn piglets, mRNA expression of ZIP4 was reduced in jejunum whereas ZnT1 and MT3 were stimulated in jejunum and liver and MT1 in kidney (P ≤ 0.04) from day 35. Copper levels were greatest in jejunum (P = 0.06) and kidney (P ≤ 0.01; days 35 and 42 only) and lowest in liver and serum (P ≤ 0.01) of 3,000Zn piglets. In conclusion, the treatment containing 3,000 mg ZnO/kg triggered Zn homeostatic mechanisms in weaned pigs and impaired Cu metabolism through high enterocyte and kidney Cu sequestration.

Zinc oxide (ZnO) is commonly used in post-weaning pig diets as growth promoter alternative to antibiotics to prevent diarrhea. The use of supranutritional levels of ZnO in post-weaning pigs diets has been challenged due to environmental issues and concerns related to bacterial resistance to antibiotics and heavy metals. However, the limited knowledge of the consequences of high levels of dietary ZnO on the metabolism of trace minerals has hampered advances to replace this nutritional strategy without compromising piglets health. Therefore, this experiment was conducted to evaluate the effects of increasing levels of dietary ZnO (i.e., 100, 1,000, and 3,000 mg/kg) on Zn and Cu metabolism in weaned piglets. In this experiment, it was demonstrated that systemic Zn levels were not effectively regulated with supplementation levels at 3,000 mg of ZnO/kg of diet. In addition, this level of dietary ZnO increased the intestinal intracellular sequestration of Cu and impaired its renal reabsorption, negatively impacting hepatic, and systemic serum Cu concentrations. These results emphasize the potential risk of Cu deficiency under long-term supranutritional supplementation of dietary ZnO during the post-weaning period, with potentially detrimental impacts on piglets growth.

Effects of different sources and levels of dietary iron and selenium on the postprandial net portal appearance of these minerals in growing pigs.

The present study compares the net portal appearance of dietary iron (Fe) and selenium (Se) after meals containing different sources and levels of these minerals. Twelve pigs (55.1 ± 3.7 kg) were used in a cross-over design to assess the 11-h net portal-drained viscera (PDV) flux of serum Fe and Se after ingestion of boluses containing inorganic (I) or organic (O) dietary Fe and Se at industry average (A; 200 and 0.6 mg, respectively) or high (H; 400 and 1.2 mg, respectively) levels. Arterial serum Fe concentrations increased by an average of 158% within 6 h post-meal and gradually decreased thereafter (P < 0.001). Values were greater (P < 0.001) for I than for O until 6 h post-meal and greater (P ≤ 0.001) for A than for H from 4 to 8 h post-meal. For the whole post-prandial period (11 h), arterial serum Fe concentrations tended (P = 0.06) to be greater for I than for O and were lowest for HO (P ≤ 0.03). Net PDV flux of Fe tended to be greater for AI than for AO (P ≥ 0.07). Cumulative appearance of Fe in PDV serum (% of dietary intake) was greater for I than for O (2.43 vs. -0.76%; P = 0.02) and A tended to be greater than H (1.96 vs. -0.29 %; P = 0.09) until 3 h post-meal, but these effects further faded out (P ≥ 0.43). Arterial serum Se concentration decreased for all treatments (average of 7%) from premeal values (P < 0.001), and this was more pronounced for O than for I (P = 0.03). Irrespective of treatment, net PDV flux of Se was positive (different from 0, P ≤ 0.03) during the first 90 min post-meal, decreased to negative minimum values (different from 0, P = 0.03) at 5 h post-meal, and was not different from 0 thereafter (P ≥ 0.11). Cumulative appearance of Se in PDV serum (% of dietary intake) was greater for I than for O (20.0 vs. -3.8%; P = 0.04) only at 45 min post-meal. In conclusion, both dietary Fe and Se absorption are limited to the early post-meal period. Whereas for Fe, the level effect is in accordance with the known negative correlation between its dietary concentration and percentage of intestinal absorption, this was not the case for dietary Se. The postabsorptive availability of dietary I was greater than O for both minerals and, particularly for Fe, at low levels.

A survey of current levels of trace minerals and vitamins used in commercial diets by the Brazilian pork industry-a comparative study.

This study identified the levels of trace minerals and vitamins used in commercial diets by the Brazilian pork industry and compared these levels against the Brazilian reference tables and those from the North American pork industry. Animal feed companies (n = 15) and cooperatives/agro-industries (n = 15) from the Brazilian pork sector participated in this study. Levels of vitamin A, D, E, and K, thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folic acid, vitamin B12, choline, vitamin C, cobalt, copper, chromium, iodine, iron, manganese, selenium, and zinc were collected. Data were compiled by feeding phase to determine descriptive statistics. Ratios were calculated comparing the observed levels to those from the Brazilian reference tables and those from the North American pork industry. Average levels of trace minerals and vitamins used by the Brazilian pork industry were between 40% and 240% higher than the recommendations of the Brazilian reference tables. Compared to the North American pork industry, Brazilian levels for trace minerals were overall higher but for water-soluble vitamins they were significantly lower. Large between-companies variations were observed for most of the studied nutrients, especially for water-soluble vitamins. In conclusion, the Brazilian pork industry adds a significant margin of safety for trace minerals and vitamins supplementation. The large between-companies variations, especially for water-soluble vitamins, reflect the limited knowledge available on precise levels of supplementation for trace nutrients for pigs.

Impact of dietary zinc:copper ratio on the postprandial net portal appearance of these minerals in pigs1.

The present study compared the net intestinal absorption of zinc (Zn) and copper (Cu) after meals containing different dietary ratios among these trace elements. Ten 46-kg pigs were used in a cross-over design to assess the 10-h net portal-drained viscera (PDV) flux of serum Cu and Zn after ingestion of boluses containing ZnSO4 and CuSO4 in different Zn:Cu ratios (mg:mg): 120:20; 200:20; 120:8; and 200:8. Arterial Zn concentrations peaked within the first hour post-meal and responses were greater with 200 (0.9 to 1.8 mg/L) than with 120 mg (0.9 to 1.6 mg/L) of dietary Zn (dietary Zn × time, P = 0.05). Net PDV flux of Zn was greater (P = 0.02) with 200 than with 120 mg of dietary Zn and tended to be greater (P = 0.10) with 20 than with 8 mg of dietary Cu. The cumulative PDV appearance of Zn (% of dietary intake) was greater with 120 than 200 mg of dietary Zn from 8 h post-meal (P ≤ 0.04) and with 20 than 8 mg of dietary Cu from 7 h post-meal (P ≤ 0.05). At the end of the postprandial period (10 h), estimated PDV appearance of Zn was 16.0%, 18.4%, 12.0%, and 15.3% of Zn intake for 120:8, 120:20, 200:8, and 200:20 ratios, respectively. For Cu, irrespective of treatment, arterial values varied (P < 0.01) by less than 5% across postmeal times. Net PDV flux was not affected by treatments (P ≥ 0.12), but the value for ratio 120:20 was different from zero (P = 0.03). There was an interaction dietary Zn × dietary Cu on cumulative PDV appearance of Cu (% of dietary intake) at 30 min post-meal (P = 0.04) and thereafter at 3 h post-meal (P = 0.04). For the whole postprandial period (10 h), estimated PDV appearance of Cu was 61.9%, 42.1%, -17.1%, and 23.6% of Cu intake for 120:8, 120:20, 200:8, and 200:20 ratios, respectively. In conclusion, the present dietary amounts and ratios of Zn and Cu can affect the metabolic availability of both trace minerals for pigs. Ratios with 120 mg of dietary Zn maximized the postintestinal availability of both Zn and Cu.

Gene ontology analysis of expanded porcine blastocysts from gilts fed organic or inorganic selenium combined with pyridoxine.

BACKGROUND: Gene ontology analysis using the microarray database generated in a previous study by this laboratory was used to further evaluate how maternal dietary supplementation with pyridoxine combined with different sources of selenium (Se) affected global gene expression of expanded porcine blastocysts. Data were generated from 18 gilts randomly assigned to one of three experimental diets (n = 6 per treatment): i) basal diet without supplemental Se or pyridoxine (CONT); ii) CONT + 0.3 mg/kg of Na-selenite and 10 mg/kg of HCl-pyridoxine (MSeB610); and iii) CONT + 0.3 mg/kg of Se-enriched yeast and 10 mg/kg of HCl-pyridoxine (OSeB610). All gilts were inseminated at their fifth post-pubertal estrus and euthanized 5 days later for embryo harvesting. Differential gene expression between MSeB610 vs CONT, OSeB610 vs CONT and OSeB610 vs MSeB610 was performed using a porcine embryo-specific microarray. RESULTS: There were 559, 2458, and 1547 differentially expressed genes for MSeB610 vs CONT, OSeB610 vs CONT and OSeB610 vs MSeB610, respectively. MSeB610 vs CONT stimulated 13 biological processes with a strict effect on RNA binding and translation initiation. OSeB610 vs CONT and OSeB610 vs MSeB610 impacted 188 and 66 biological processes, respectively, with very similar effects on genome stability, ceramide biosynthesis, protein trafficking and epigenetic events. The stimulation of genes related with these processes was confirmed by quantitative real-time RT-PCR. CONCLUSIONS: Gene expression of embryos from OSeB610 supplemented gilts was more impacted than those from MSeB610 supplemented gilts. Whereas maternal OSeB610 supplementation influenced crucial aspects of embryo development, maternal MSeB610 supplementation was restricted to binding activity.

Pyridoxine (Vitamin B6) and the Glutathione Peroxidase System; a Link between One-Carbon Metabolism and Antioxidation.

Vitamin B6 (B6) has a central role in the metabolism of amino acids, which includes important interactions with endogenous redox reactions through its effects on the glutathione peroxidase (GPX) system. In fact, B6-dependent enzymes catalyse most reactions of the transsulfuration pathway, driving homocysteine to cysteine and further into GPX proteins. Considering that mammals metabolize sulfur- and seleno-amino acids similarly, B6 plays an important role in the fate of sulfur-homocysteine and its seleno counterpart between transsulfuration and one-carbon metabolism, especially under oxidative stress conditions. This is particularly important in reproduction because ovarian metabolism may generate an excess of reactive oxygen species (ROS) during the peri-estrus period, which may impair ovulatory functions and early embryo development. Later in gestation, placentation raises embryo oxygen tension and may induce a higher expression of ROS markers and eventually embryo losses. Interestingly, the metabolic accumulation of ROS up-regulates the flow of one-carbon units to transsulfuration and down-regulates remethylation. However, in embryos, the transsulfuration pathway is not functional, making the understanding of the interplay between these two pathways particularly crucial. In this review, the importance of the maternal metabolic status of B6 for the flow of one-carbon units towards both maternal and embryonic GPX systems is discussed. Additionally, B6 effects on GPX activity and gene expression in dams, as well as embryo development, are presented in a pig model under different oxidative stress conditions.

The importance of pyridoxine for the impact of the dietary selenium sources on redox balance, embryo development, and reproductive performance in gilts.

This study aimed to determine the effects of dietary pyridoxine and selenium (Se) on embryo development, reproductive performance and redox system in gilts. Eighty-four gilts were fed one of five diets: CONT) basal diet; MSeB60) CONT+0.3mg/kg of Na-selenite; MSeB610) diet 2+10mg/kg of HCl-pyridoxine; OSeB60) CONT+0.3mg/kg of Se-enriched yeast; and OSeB610) diet 4+10mg/kg of HCl-pyridoxine. Blood samples were collected for long-term (each estrus and slaughter) and peri-estrus (fourth estrus d -4 to d +3) profiles. At slaughter (gestation d 30), organs and embryos were collected. For long-term and peri-estrus profiles, Se level and source affected (P<0.01) blood Se concentration whereas B6 level increased (P<0.01) erythrocyte pyridoxal-5-phosphate concentration. A B6 level (P<0.05) effect was observed on long-term plasma Se-dependent glutathione peroxidase (Se-GPX) activity whereas peri-estrus Se-GPX was minimum on d -1 (P<0.01). Selenium level increased sows' organs and embryo Se concentration (P<0.01). Selenium source tended to enhance embryo Se content (P=0.06). Within-litter embryo Se content was increased by B6 level (P<0.01). Selenium level tended to affect Se-GPX and total GPX activities in organs mitochondria (P=0.09 and 0.07, respectively). Selenium source affected kidney ATP synthesis (P=0.05). In conclusion, B6 level affected the Se-GPX activity on a long-term basis, whereas the basal level of Se was adequate during the peri-estrus period. Embryo quality was not improved by dietary Se, and B6 impaired within-litter homogeneity.

Interaction between vitamin B6 and source of selenium on the response of the selenium-dependent glutathione peroxidase system to oxidative stress induced by oestrus in pubertal pig.

This study aimed to assess the interaction between vitamin B6 and selenium (Se) for the flow of Se towards the Se-dependent glutathione peroxidase (GPX) system in response to oxidative stress naturally induced by oestrus in a pubertal pig model. At first oestrus, forty-five gilts were randomly assigned to the experimental diets (n=9/group): basal diet (CONT); CONT+0.3mg/kg of Na-selenite (MSeB60); MSeB60+10mg/kg of HCl-B6 (MSeB610); CONT+0.3mg/kg of Se-enriched yeast (OSeB60); and OSeB60+10mg/kg of HCl-B6 (OSeB610). Blood samples were collected at each oestrus (long-term profiles), and daily from day -4 to +3 (slaughter) of the fourth oestrus (peri-oestrus profiles) after which liver, kidneys, and ovaries were collected. For long-term profiles, CONT had lower blood Se than Se-supplemented gilts (p<0.01) and OSe was higher than MSe (p<0.01). Lower erythrocyte pyridoxal-5-phosphate was found in B60 than B610 (p<0.01). No treatment effect was observed on GPX activity. For peri-oestrus profiles, treatment effects were similar to long-term profiles. Treatment effects on liver Se were similar to those for long-term blood Se profiles and OSe had higher renal Se concentrations than MSe gilts (p<0.01). Gene expressions of GPX1, GPX3, GPX4, and selenocysteine lyase in liver and kidney were greatest in OSeB610 gilts (p<0.05). These results suggest that dietary B6 modulate the metabolic pathway of OSe towards the GPX system during the peri-oestrus period in pubertal pigs.