1. Introduction
Iron is a crucial nutrient for animal growth, reproduction, and immunity. Hemoglobin (Hb), a component of red blood cells (RBCs), requires iron to facilitate the physiological process of oxygen transfer [
1]. Iron deficiency in pregnant animals can lead to decreased litter weight, increase in stillbirths, and neonatal infections [
2]. Furthermore, iron deficiency is often related to iron-deficiency anemia (IDA), which has negative health consequences and frequently results in early mortality in piglets. Due to their unique iron properties, newborn piglets are particularly vulnerable to IDA. Compared to other animals, newborn piglets have insufficient iron storage, increased iron consumption, and a lack of iron supply in breast milk [
3]. Iron supplements are typically given to animal diets to supplement iron elements, with inorganic iron supplements (ferrous sulfate) being the most commonly utilized. However, inorganic iron supplements are known to have low bioavailability, produce harmful side effects, and pollute the environment [
4]. Numerous studies on the efficacy of iron supplementation in pregnant sows have demonstrated that organic iron supplements, such as ferrous fumarate and amino-acid-chelated iron, have been confirmed to perform better than inorganic iron supplements in animal production applications [
1,
5,
6]. Iron storage in piglets increased significantly with the level of dietary glycine chelated and methionine iron during the early pregnancy and suckling periods [
7], and iron content in breast milk also increased significantly [
8]. However, it has also been reported that inconsistent results have been obtained, which may be related to the complexity of the existing organic iron processing technology and poor product stability [
9]. Therefore, it is necessary to find a new and more efficient iron supplement.
Nowadays, iron complexation of polysaccharides has recently become a popular topic in glycobiology research. Saccharide–iron complexes as novel iron supplements have aroused attention for the high iron absorption rate and no gastrointestinal irritation in oral doses. In addition, research on the biological activities of saccharide–iron complexes revealed that they also exhibited good abilities in treating anemia, eliminating free radicals, and regulating the immune response [
10,
11]. In this study, an organic iron complex generated from tapioca starch and ferric chloride was utilized. This product is a self-developed, plant-derived, natural compound that has a wide variety of sources and is cost-effective. The substantial potential of polysaccharide iron compounds has been reported [
10,
12,
13]. However, there has been no research on the effect of TpFe in iron supplementation using pig models. The National Research Council (NRC) recommends a minimum iron requirement of 80 mg/kg for late gestation and nursing sows [
14]. However, due to unidentified factors such as feed processing waste, it has been demonstrated that sows benefit from an exogenous iron supplement dose of around 100 mg/kg [
3,
15]. Thus, in this experiment, a half-dosage group of 50 mg/kg iron dextran was set up to determine if the half-dose could replace 100 mg/kg inorganic iron (ferrous sulfate monohydrate), as well as the benefits of 100 mg/kg iron dextran in the same dose group. In this study, we investigated the effects of maternal TpFe supplementation on sow reproductive performance, colostrum and cord blood active components, and growth performance of piglets, and for the first time determined the correlation between maternal dietary TpFe management and offspring growth performance.
4. Discussion
The use of inorganic iron in animal diets has various negative impacts in production, including gastrointestinal side effects, iron poisoning, and environmental pollution. Although several organic irons, such as ferrous fumarate and amino-acid-chelated iron, have superior effects and fewer side effects than inorganic iron, product stability is poor, and the complex synthesis process results in high production costs. TpFe, the iron preparation employed in this experiment, is a novel form of iron preparation with good stability, low cost, and independent innovation patents. In this study, we focused on the effects of maternal iron polysaccharide supplementation on the active components of colostrum and cord blood, as well as the correlation between these effects and offspring growth performance.
Inadequate feed consumption during lactation can reduce the number of nutrients available in milk production, which can restrict the growth and development of piglets [
18]. Increased feed intake by sows provides greater energy and nutrients for milk synthesis, leading to improved piglet development [
19,
20]. In the current study, the feed intake of sows during pregnancy did not change significantly, possibly because the sows did not have ad libitum access to feed during pregnancy, but rather a limit was placed on the amount they could consume. However, the feed intake of sows in the TpFe100 group was significantly increased during lactation. This is similar to previous studies in which adding organic iron to a typical lactation diet and feeding it to sows for the 26 days prior to farrowing increased their feed consumption [
21]. As we know, sow milk production increases with feed consumption during lactation, indicating that maternal TpFe supplementation provides a strong foundation for improving piglet development performance.
The effects of several organic iron chelates on sows are inconsistently described in the available literature. For example, the bioavailability of iron and the proportions of stillborn and mummified fetuses per litter were considerably enhanced by adding glycine chelate (62.5 mg/kg) to sow diets for 0, 2, 4, 6, or 8 weeks before farrowing [
9]. In contrast, sows administered an organic iron complex (120 mg/kg, Fe) throughout the entire pregnancy period had larger litters [
6]. However, when an organic iron chelate complex (80 mg/kg, Fe), consisting of 35% ferrous fumarate, 25% iron lactate, 37% glycine chelate iron, and 3% iron methionine chelate, was fed to sows from day 84 to parturition, the number of stillborn and mummified fetuses per litter was not reduced [
22]. In our study, TpFe (50 or 100 mg/kg, Fe) was used to supplement the sows’ feed from day 85 to parturition. The litter size, number of stillborn piglets, mummies, and duration of labor were not affected by TpFe administration, but a significant improvement in the number of live-born piglets, birth (alive) litter weight, and average weight of live-born piglets was observed under the 100 mg/kg Fe in TpFe treatment. This is similar to the effects of feeding sows a diet with 80 mg Fe/kg ferrous chelate, as reported by Wan et al. [
2]. These findings suggest that different reproductive effects in sows are influenced by the organic iron chelate source or structure, added concentration, and duration. However, the benefits of TpFe in improving sow reproductive performance are reflected in its ability to support fetal survival.
Colostrum synthesis begins considerably before parturition, and thus other mineral requirements in sow reproduction may impact its composition, particularly during the latter stages of fetal development during pregnancy [
23]. Previous studies have reported that changes in the chemical composition of colostrum, such as fat, protein, lactose, and non-milk fat solids, also reflect different physiological conditions of lactating sows [
24,
25]. In our findings, the total dry matter content, milk fat percentage, milk protein percentage, milk lactose percentage, and non-milk fat solid content of colostrum were not impacted, which is similar to a previous study [
26]. This discrepancy can be explained by the fact that the sow’s physical state and perpetual feeding strategy affect the nutritional content of colostrum [
27]. Furthermore, leukocytes and epithelial cells make up the majority of somatic cell types in milk and colostrum. According to Maurer et al. [
28], the somatic cell count (SCC) of milk is frequently used as a barometer for the well-being and quality of lactating animals, and the urea nitrogen concentration is a measure of urine nitrogen excretion that is connected to dietary crude protein (CP) intake and the ratio of degradable to undegradable protein [
29]. In this study, we found lower somatic cell counts and urea nitrogen concentrations in colostrum, which may indicate that maternal TpFe supplementation improved the quality of sow colostrum and had greater bioavailability than inorganic iron.
Moreover, colostrum and milk contain different amounts of iron, and the amount of iron absorbed by piglets varies from sow to sow. Piglets consume 0.5 to 1 L of milk per day. Piglets absorb 60–90% of lactoferrin in breast milk at concentrations of 0.2–4 mg/L [
30]. The iron content in colostrum can be increased by supplementing it with maternal iron chelates [
31] or lactoferrin [
3]. In contrast, the iron content of colostrum and breast milk remained constant at 0.15% with or without chelated iron [
32]. This is consistent with our current findings of no significant difference in colostrum iron content, possibly due to the short time interval between late gestation and parturient TpFe supplementation in sows.
Immunoglobulins from the colostrum are critical for the survival and development of piglets. Infants lack globulins and therefore rely on colostrum as their primary supply of antibodies. The primary source of innate immunity against bacterial infection is mainly immunoglobulins [
33]. Ma et al. [
34] reported that sows fed low-dose organic iron had no significant effect on colostrum IgM, IgA, and IgG. However, in our study, dietary supplementation of 100 mg/kg in the form of TpFe significantly increased the levels of IgM, IgA, and IgG in colostrum. This may indicate that TpFe is involved in the synthesis of colostrum immunoglobulins, which also improves the survival of newborn piglets.
Serum iron, or iron bound to transferrin in serum, has been used to qualitatively assess the bioavailability of iron supplements [
35]. Piglet serum iron significantly increased when Fe-Gly was added to sows’ diets, while FeSO
4·H
2O supplementation had some positive benefits, but they were not significant [
15]. High-iron diets during pregnancy have been linked to an increase in serum iron, according to Spruill et al. [
36], while iron from an amino acid complex has been linked to an increase in blood serum iron, according to Yu et al. [
37]. Moreover, a previous study reported that organic iron complexes increased sow serum iron concentration on day 1 of lactation compared to ferrous sulfate [
20]. In contrast, treatment with 100 mg/kg of TpFe significantly increased serum iron content in the current study. Dietary supplementation with 50 mg/kg of TpFe had no significant effect on serum iron content in sows, umbilical cord serum, or weanling piglets compared to the inorganic iron supplementation group. Surprisingly, the 100 mg/kg TpFe supplement group showed a consistent pattern of iron content changes in sow serum, cord blood serum, and weanling pig serum. This may be due to the fact that, like lactoferrin and heme iron [
38], TpFe has greater potential than inorganic iron to cross the placental barrier and enter the embryo. Unfortunately, we did not focus on the expression of genes and proteins involved in iron transport in sow placentas in this work, while additional in-depth mechanistic investigations may be required in the future.
There are few studies on the ability of polysaccharide iron to act as an antioxidant in mammals, and therefore, we can only compare it with other types of organic iron. Dietary supplementation of 200 mg/kg lactoferrin [
38] or 100 mg/kg glycine–iron complex [
3] to sows increased serum antioxidant parameters in pregnant sows and newborn piglets. Similarly to these results, the present study showed that dietary TpFe supplementation significantly increased the activities of T-AOC, GSH-px, and CAT in cord blood and serum of weaned piglets, suggesting that maternal iron supplementation can improve offspring antioxidant capacity. The process may be explained by how tapioca polysaccharide binds ferric ions and inhibits the production of free radicals. Fe3+ chelated with polysaccharide does not break the basic structure of polysaccharide [
39], and polysaccharide iron also has the basic structure of polysaccharide. Moreover, polysaccharide iron has more antioxidant activity than the original polysaccharide [
40,
41]. This is due to a change in the polysaccharide’s spatial structure after it was linked with iron, and the coordinating component interacts with the exposed active groups to promote free radical interaction, enhancing the ability to scavenge free radicals [
42]. However, this study did not evaluate changes in the placental antioxidant capacity of sows or the liver antioxidant capacity of piglets since the focus of this study was the maternal dietary TpFe on offspring performance through cord blood; further research is needed in this regard.
In our study, we focused on changes in immunoglobulin IgA, IgM, and IgG and found that maternal supplementation with 100 mg/kg of TpFe increased IgG levels in sow blood, cord blood, and piglets. This is consistent with recent findings that IgG levels in piglet plasma were substantially related to survival, and serum IgG concentrations were lower in deceased piglets than in living piglets [
43]. This also explained the differences in colostrum IgG found in the outcomes of the present research, indicating that maternal dietary supplementation with iron polysaccharide to enhance offspring performance involves the use of IgG as a crucial “bridge”.
Furthermore, RBCs play an essential role in tissue metabolism, and a sufficient number of RBCs are necessary to maintain tissue oxygenation and acid–base balance in the system [
44]. Hb is a protein present in red blood cells that mainly distributes oxygen to the body’s tissues, and iron is an essential element for Hb [
45]. Hb concentrations of 100 g/L or more show acceptable iron levels in an organism, while 80 g/L or higher is considered the anemia cutoff, and 60 g/L or higher is considered severe anemia [
46]. HCT is the proportion of entire blood occupied by red blood cells and is affected by Hb content in RBC. The mean corpuscular volume is a good indicator of anemia after birth. In piglets that did not receive iron, the cells become microcytic, and the MCV will decrease [
47]. Interestingly, compared to the inorganic-iron-fed sow group, the contents of RBC and Hb in cord blood and blood of piglets in the 100 mg/kg TpFe-supplemented sow group were significantly higher than the control group in our study. This was consistent with the findings of earlier research [
20]. Moreover, the contents of HCT and MCV of piglets were significantly increased. These results match Hb levels and suggest that sows had greater iron status or that nursing piglets had increased iron bioavailability.
Interleukin-2 (IL-2) is a Th1 cytokine mainly produced by activated T lymphocytes that plays a regulatory role by activating and enhancing various functions of immune cells. It plays an important immuno-physiological role in the immune response system [
15,
48]. Interleukin (IL)-6 is a pleiotropic cytokine that plays essential functions in immunological response, inflammation, and hematopoiesis control [
49]. The tumor necrosis factor alpha (TNF-α) is a cytokine with pleiotropic effects on numerous cell types. It has been identified as a significant regulator of inflammatory responses and is known to play a role in the etiology of various inflammatory and autoimmune diseases [
50]. In a recent double-blind randomized trial examining the impact of iron supplementation in anemic pregnant women, iron treatment significantly raised maternal IL-2 levels compared to the control group [
51]. In the present study, serum IL-2 levels in tapioca-iron-polysaccharide-fed sows significantly increased, although cord blood and piglet blood exhibited no change. Sow serum, cord blood, and piglet serum did not significantly differ in terms of IL-6 and TNF-. These results suggest that the effects of TpFe on serum inflammatory and immunological hormones were mainly maternal in nature and had little effect on passage.
Moreover, CEE and PRL can stimulate mammary gland growth and development, improve sow breastfeeding capacity, and improve reproductive success [
52,
53]. CCK is a typical brain and intestinal peptide that regulates the body to produce fullness signals and then inhibits eating activities in the digestive system and central and peripheral neurological systems [
54]. GH is required for postnatal somatic growth and the retention of lean tissue in the animal at maturity. In the current study, there were no significant increases in serum estrogen levels in sow serum or cord blood, while the PRL concentration was considerably higher in serum and cord blood of sows given polysaccharide iron. Similarly, the serum CCK of sows was significantly higher than that of inorganic iron groups, which might explain the higher feed intake of nursing sows. Furthermore, GH levels were significantly increased in cord blood and piglet blood corresponding to the group of sows fed polysaccharide iron, indicating that maternal supplementation with TpFe had an innate developmental advantage in piglets.
Organic iron supply boosts pig birth weight and fetal iron reserves, decreases postnatal pig mortality rates and stillbirths, and leads to larger weaning weights of pigs, according to some earlier research [
55]. Moreover, during the third trimester of pregnancy and lactation, sows fed an organic iron supplement (90 mg/kg) or an inorganic iron injection (60 and 80 mg/kg) were found to be more efficient, leading to a decrease in piglet mortality and higher weight at weaning [
56]. These findings support our findings that sows given 100 mg/kg of TpFe had improved litter weight and average body weight at weaning and reduced piglet mortality. However, one study had different results that dietary chelated iron supplementation of 150 mg/kg to sows had no effect on piglet weight at weaning [
57]. This may be related to the different types of iron chelates. In particular, the current study focused on a topic not covered in earlier research, the risk of diarrhea in suckling piglets, and found that maternal TpFe significantly reduced the probability of suckling piglets experiencing diarrhea. The cause of this may be the increased immunoglobulin levels in piglet serum and the observed better iron bioavailability of TpFe, which was consistent with the data discussed above.
The current data indicate a positive correlation between the live birth rate and the sows’ serum prolactin (PRL), which is consistent with earlier findings. An extremely strong and significant link between PRL and the number of live births (r = 0.670) suggested that improved litter size may be related to PRL [
58]. Indeed, there is evidence that maternal oxytocin concentrations increase with litter size [
59], possibly due to nest-building activity [
60]. Urea nitrogen concentration is usually used to reflect the digestion of dietary protein. Thus, it was easy to understand the results of our experiment indicating that the diarrhea rate of suckling piglets was positively correlated with the urea nitrogen content in colostrum, as a high-protein diet is usually associated with causing diarrhea in piglets [
61]. However, the level of colostrum IgM, IgG, and cord blood hemoglobin (Hb) was negatively correlated with the risk of diarrhea in suckling piglets. This is in agreement with the results of a previous study, which found a correlation between higher IgM and IgG group concentrations in sow colostrum and decreased incidence of diarrheal disease in newborn piglets [
62]. The development of Hb in cord blood would undoubtedly strengthen piglets’ inbuilt immunity. Immune globulins, primarily IgG and IgM, are thought to be the main components of colostrum protein, and they shield piglets from possible harm caused by pathogenic bacteria [
62].
Moreover, the mortality of suckling piglets was negatively correlated with the serum iron content of sows and IgM in colostrum, glutathione peroxidase (GSH-px), as well as IgG of cord serum. Although no study has directly focused on the connection between sows’ serum iron levels and piglet fatalities during lactation, it has been shown that sows fed organic iron had higher serum iron levels, which was followed by an increase in live litter size [
2]. It is possible that iron-rich maternal nutrition enhances piglets’ ability to adjust throughout lactation and boosts the iron stored in the liver. Similarly, the congenital adaptability of the piglets was also increased, and the chance of dying while nursing was decreased by a rise in IgM in colostrum, GSH-px, and IgG in cord blood. Furthermore, there was a trend of increased litter weight at weaning and piglet single-head weight with the enhancement of IgM and IgG concentrations in colostrum and milk [
63]. This was similar to the present study, where the average weight of weaning piglets at day 21 had a positive connection with colostrum IgM and IgG and cord serum red blood cell (RBC) count. This might be because the immunoglobulin system offers critical antimicrobial defense against a variety of diseases and can provide passive immunity to piglet development. Moreover, RBC and piglet weights revealed a similar relationship, as found in previous studies [
64]. This might be because red blood cells are necessary for tissue metabolism, and more of them are required to support the tissue oxygenation required for piglets’ fast growth.