Journal of Animal Science and Biotechnology
Research

Cinnamaldehyde supplementation in sows and their offspring: effects on colostrum and milk composition, performance, redox status and intestinal health

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Abstract

Background

Maternal nutrition significantly influences offspring development. This study investigated the effects of maternal or post-weaning cinnamaldehyde (CA) supplementation in sows and their offspring on reproductive performance and health. Sixty sows, selected based on body condition score and parity, were randomly allocated to control or CA (500 mg/kg) diets from d 107 of gestation to d 24 of lactation. At weaning, 128 piglets were assigned to four groups (n= 8) based on weight and source litter for a 21-d experiment. The four groups were CON-CON (both sow and piglet on CON), CON-CA (sow on CON, piglet on CA), CA-CON (sow on CA, piglet on CON), and CA-CA (both sow and piglet on CA).

Results

Maternal CA supplementation tended to improve body weight (+ 15%,P= 0.09) and average daily gain (+ 21%,P= 0.07) of suckling piglets, along with increased levels of milk IgG (P= 0.01) and IgM (P= 0.02), colostrum crude fat (P= 0.01), and plasma glutathione peroxidase (GSH-Px) activity (P= 0.02) at farrowing. Moreover, maternal CA supplementation significantly improved plasma antioxidant capacity, expressions of intestinal barrier and anti-inflammatory genes, and gut microbiota structure of piglets at the end of suckling. Additionally, maternal CA supplementation increased the apparent total tract digestibility (ATTD) of crude protein (P< 0.01), gross energy (GE;P= 0.03), and dry matter (P= 0.01), improved jejunal sucrase activity (P< 0.01), villus height (P= 0.03), the ratio of villi height to crypt depth (P= 0.02), and the expressions of intestinal barrier and anti-inflammatory genes in post-weaning piglets. Furthermore, post-weaning CA supplementation tended to decrease diarrhea scores of piglets during d 14–21 and increased the ATTD of GE (P= 0.02), activities of jejunal sucrase (P= 0.02), plasma catalase (P= 0.01), and total superoxide dismutase (P< 0.01) in piglets.

Conclusion

Maternal CA supplementation tended to increase the growth rate and weaning weight of suckling piglets, associated with improved antioxidant capacity and milk composition. Moreover, maternal CA supplementation or post-weaning CA supplementation improved nutrient digestibility, redox status, and intestinal function-related parameters of weaned piglets.

Background

In modern intensive pig farming, high-yielding sows in late pregnancy and lactation experience severe oxidative stress due to metabolic burden [1]. The accumulation of reactive oxygen species (ROS) from the placenta and mammary gland in sows exceeds the elimination capacity of both enzymatic and non-enzymatic antioxidants, which negatively affects sow performance, milk production, and the growth rate of suckling piglets [2, 3]. Additionally, the gut microbiota is closely associated with oxidative stress, milk quality, and piglet health [4, 5]. Recently, maternal nutrition strategies have been employed to improve the performance and health of both sows and their offspring [6,7,8].

Weaning piglets face challenges such as maternal separation, dietary changes, and environmental shifts, which possibly lead to weaning stress [9]. This stress disrupts digestive and barrier function, induces oxidative stress and pro-inflammatory responses, and results in higher diarrhea incidence, lower feed intake, and growth rates in piglets [10,11,12].

Cinnamaldehyde (CA), an active component extracted from cinnamon, exhibits antioxidant, anti-inflammatory, and antimicrobial effects [13]. It has been reported that CA can inhibit inflammation by regulating the nuclear factor kappa B (NF-кB) pathway and reducing pro-inflammatory cytokines [14, 15]. Moreover, CA can enhance antioxidant capability and inhibit ROS production and oxidative stress [16, 17]. In animal models, dietary CA supplementation has improved inflammatory responses by regulating the NF-кB signaling pathway in fish [18], enhanced intestinal barrier in chickens [19], and benefited nursery pigs [20]. However, previous studies have primarily focused on the growth and health responses of piglets to CA supplementation. In this study, therefore, we investigated the effects of maternal and/or post-weaning dietary CA supplementation on sow reproductive performance and weaned piglet growth performance. Specifically, CA was added to the sow diet from d 107 of gestation to d 24 of lactation to investigate the effects of CA on reproductive performance, milk composition, and physiological traits of sows. Furthermore, split-pot experiments were conducted to investigate the effects of CA supplementation at different stages on growth performance, immunity, and gut health of weaned piglets.

Materials and methods

Experimental design

A total of 60 sows (parity 2 to 3) were randomly allocated into two groups at d 107 of gestation based on body condition score and parity. The sows in two groups were fed the CON diet (basal diet) or CA diet with basal diet supplemented with CA at 500 mg/kg (purity ≥ 45%, supplied by Yangzhidao Feed Co., Ltd., Jiangsu, China). The sow diets were fed from d 107 of gestation to d 24 of lactation. The basal diet of sows was formulated to meet or exceed the recommendations of Nutrient Requirements of Swine [21], as shown in Table 1.

Table 1 Ingredients and nutrient levels of the basal diet fed to sows (as-fed basis)
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At the end of d 24 of lactation, a total of 128 piglets (initial body weight: 6.04 ± 0.10 kg) from 16 litters per treatment, according to body weight and source litter, were allotted into pens, and pens were assigned to dietary treatments in a split-plot design. Sows’ dietary treatment (CON diet or CA diet) served as the main plot and piglets’ dietary treatment (CON diet or CA diet) as a sub-plot. There were four groups with eight replicates and four piglets per replicate. The groups were as follows: 1) CON-CON (sows and piglets both fed CON diet); 2) CON-CA (sows fed CON diet and piglets fed CA diet); 3) CA-CON (sows fed CA diet and piglets fed CON diet); 4) CA-CA (sows and piglets both fed CA diet). The post-weaning experimental period lasted for 21 d, starting at weaning. The basal diet for piglets was formulated to meet or exceed the Nutrient Requirements of Swine [21], as presented in Table 2.

Table 2 Ingredients and nutrient levels of the basal diet fed to weaned piglets (as-fed basis)
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Feed management

On d 107 of gestation, sows were moved to farrowing crates (2.13 m × 0.66 m) in a farrowing room with environmental control systems (maintained at 24–26 °C). From d 107 to d 111 of gestation, sows were fed 3.2 kg of diet daily, split into two equal meals at 08:00 and 16:00. From d 112 of gestation until farrowing, the daily feed intake was reduced to 2.0 kg, also provided in two equal meals at the same times. On the day of parturition, sows were not fed; they received 2 kg the next day, with a gradual increase of 0.5 kg/d until ad libitum. Throughout the experimental period, CA was evenly mixed into the basal diet and manually administered to the sows’ troughs. All sows and their suckling piglets had free access to water. At the time of parturition, data on the number of total piglets born, stillborn piglets, mummified piglets, piglet weights, and farrowing duration were recorded. Within 48 h after birth, litter size was standardized to be 12 ± 1 piglets per sow through cross-fostering within the same group. All piglets were weighed at 48 h after birth and weekly during lactation to calculate average daily gain (ADG). During lactation, sow milk was the sole nutrient source for the suckling piglets. Meanwhile, feed intake of sows was recorded during lactation to calculate the average daily feed intake (ADFI).

On the morning of d 25 of lactation, a total of 128 suckling piglets were transferred to the nursery room. All weaned piglets had free access to water and were fed ad libitum in this period. Meanwhile, the body weight and feed intake of the weaned piglets were recorded weekly to calculate ADG, ADFI, and the ratio of ADFI to ADG (F:G). Diarrhea scores were monitored three times daily (at 08:00, 12:00, and 16:00). Fecal scores were determined based on the appearance of feces on the floor using a 4-point scale: 0 = solid and well-formed, 1 = soft and formed feces, 2 = fluid and yellowish feces, and 3 = watery and projectile feces. Diarrhea scores were calculated using the equation: diarrhea scores = the sum of diarrhea scores (numbers of piglets per pen × experimental days × assessed times per day).

Sample collection

Eight to ten sows were randomly selected from each group for sample collection. Blood samples (10 mL) were collected from the ear vein of ten sows in each group at 1 h after the onset of farrowing and on d 25 of lactation. The samples were then centrifuged at 3,000 × g for 15 min to separate plasma. Colostrum samples (20 mL) were manually collected from all functional teats of the sows within 1 h after the onset of farrowing. On d 8 of lactation, milk samples (20 mL) were collected by infusing the sows with 5 mL of oxytocin. Colostrum and milk samples were centrifuged at 12,000 × g for 1 h at 4 °C to obtain whey samples for further analysis. Fecal samples (2 g) were freshly collected from eight sows per group by rectal grab on d 25 of lactation. All plasma, colostrum, milk, colostrum whey, milk whey, and fecal samples were stored at −80 °C for further analysis. Eight piglets per treatment group at postnatal day 24 (PND 24) with body weight closest to the average were selected for plasma collection and then euthanized. The middle 4-cm segments of the ileum were dissected and rinsed in PBS, and the mucosa was scraped off with a glass slide before being stored in liquid nitrogen. Colonic contents were collected from the mid-colon segment of each piglet and stored at −80 °C for further analysis.

Chromic oxide, an indigestible marker, was added to diets at 0.3% for the digestion test. From d 12 to 14 after weaning, fresh fecal samples and diets (200 g each) were collected, then dried at 65 °C for 72 h, ground through a 0.42-mm screen, and stored at −20 °C for further analysis. Weaned piglets at 45 days of age were individually weighed, and those closest to the average weight (10.63 ± 0.12 kg) per pen were selected for plasma samples (n = 8 piglets/treatment) and then euthanized. About 5 cm segments of mid-jejunum were fixed with 4% paraformaldehyde solution for morphological analysis, while the remaining jejunal mucosa samples were scraped and collected using a sterile glass microscope slide. Ileal tissue samples were flushed with ice-cold saline and collected into liquid nitrogen. All samples were stored at −80 °C for further analysis.

Chemical analysis

The diets and fecal samples were analyzed for crude protein (CP), dry matter (DM), and ether extract (EE) following the methods of the Association of Official Agricultural Chemists [22]. The gross energy (GE) of diets and feces was determined using an automatic adiabatic oxygen bomb calorimeter (Parr Instrument Co., Moline, IL, USA). The Cr2O3 content was measured by a flame atomic absorption spectrophotometer (ContrAA 700, Analytikjena, Jena, Germany).

Milk composition

The concentrations of fat, protein, lactose, DM, and urea nitrogen in colostrum and milk were analyzed by an automatic milk composition analyzer (MilkoScan™ FT+, FOSS, Hilleroed, Denmark). The levels of immunoglobulin G (IgG) and immunoglobulin M (IgM) in colostrum whey and milk whey were determined using an automatic biochemical analyzer (Hitachi 3100, Tokyo, Japan) with corresponding commercial kits (Sichuan Maccura Biotechnology Inc., Chengdu, China).

Plasma parameters

The concentrations of glucose (catalog No. CH0101102), total cholesterol (TG, catalog No. CH0101151), urea (catalog No. CH0101051), non-esterified fatty acid (NEFA, catalog No. RB30951), total bile acid (TBA, catalog No. RB11301), C-reactive protein (CRP, catalog No. CH0105303), IgG (catalog No. IGM0011), IgM(catalog No. IGG0011), alanine aminotransferase (ALT, catalog No. CH0101201), aspartate amino transferase (AST, catalog No. CH0101202), gamma-glutamyl transpeptidase (GGT, catalog No. CH0101204), and complement C3 (C3, catalog No. C300011) in plasma were determined by using an automatic biochemical analyzer (Hitachi 3100, Tokyo, Japan) through corresponding commercial kits (Sichuan Maccura Biotechnology Inc., Chengdu, China).

The activities of total superoxide dismutase (T-SOD, catalog No. A001-1-2), catalase (CAT, catalog No. A007-1-1), glutathione peroxidase (GSH-Px, catalog No. A005-1-2), and the content of malondialdehyde (MDA, catalog No. A003-1-2) in plasma were measured using commercially available kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) following the manufacturer’s instruction.

The concentrations of IL-1β (catalog No. H002-1-2), interleukin-10 (IL-10, catalog No. A009- 1-2), and TNF-α (catalog No. H052-1-2) were determined by ELISA kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instruction.

Jejunal morphology

The jejunal samples from weaned piglets were taken out of the fixative solution, dehydrated, and embedded in paraffin. Each jejunal sample was sectioned at 5-μm thickness and stained with hematoxylin and eosin. Ten representative villi from each section were used to measure the villus height and crypt depth by magnification (Nikon, Tokyo, Japan). The ratio of villus height to crypt depth (VCR) was calculated.

Digestive enzyme analysis

The jejunal mucosa samples of weaned piglets were homogenized in ice-cold saline at the ratio of 1:9 (g/mL) and then centrifuged at 3,000 × g for 10 min at 4 °C to collect supernatant. The activities of maltase, sucrase, aminopeptidases A (APA), and aminopeptidases N (APN) were determined by commercial kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instructions.

RNA extraction and quantitative real-time PCR analysis

Total RNA was extracted from 0.1 g of ileal tissue of piglets at PND 24 and post-weaned day 21 (PWD 21) samples using TRIZOL reagent (TaKaRa Biotechnology, Dalian, China) following the manufacturer’s protocol. The concentration and purity of extracted RNA were determined by using a nucleic acid analyzer (Beckman DU-800; Beckman Coulter, Inc., Brea, CA, USA). Then, total RNA was reverse transcribed into cDNA using the PrimeScripte RT reagent kit (TaKaRa Biotechnology, Dalian, China) according to the manufacturer’s instructions. All real-time PCR was performed using SYBR Premix Ex Taq reagents (TaKaRa Biotechnology, Dalian, China) on a CFX96 Real-Time PCR System (Bio-Rad, Hercules, CA, USA). β-actin served as a housekeeping gene to normalize the mRNA expression levels of the target genes according to the 2−ΔΔCt method [23], and the primers were presented in Table 3.

Table 3 Primer sequences used for quantitative real-time polymerase chain reaction
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Sequencing of gut microbiota

Microbial genomic DNA was extracted from the feces of sows and colonic digesta of piglets at PND 24 using the CATB method. 1% (w/v) agarose gel electrophoresis was used to detect the integrity of extracted genomic DNA. The v4 hypervariable regions of 16S rRNA were amplified using primers 515F (5′-GTGYCAGCMGCCGCGGTAA-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′). The library was constructed by the NEB Next Ultra DNA library prep kit (Illumina, USA). Then, the library was sequenced on an Illumina HiSeq platform. Operational taxonomic units were generated based on the 97% sequence identity. The α-diversity was calculated from this output normalized data, and β-diversity was performed by principal coordinate analysis (PCoA).

Short-chain fatty acid analysis

The concentration of short-chain fatty acids (SCFAs) in sow fecal samples was determined using a CP-3800 gas chromatography instrument (Varian Medical Systems, Palo Alto, CA, USA). Specifically, the mixture of 0.5 g of colonic digesta samples with 1.5 mL of water was allowed to stand for 30 min and then centrifuged at 10,000 × g for 30 min at 4 °C to collect supernatant. 1 mL of supernatant was mixed with 0.2 mL of metaphosphoric acid and 23.3 μL of crotonic acid and centrifuged at 8,000 × g for 10 min. Then, 0.3 mL of supernatant was mixed with 0.9 mL of chromatographic methanol, centrifuged at 8,000 × g for 5 min, and filtered by a 0.22-μm filter membrane for analysis.

Statistical analysis

All data were checked for variance homogeneity and normality using the Shapiro–Wilk test and Levene’s test procedures of SAS software (Version 9.4, SAS Inst. Inc., Cary, NC, USA), respectively. Each sow served as an experimental unit, and suckling piglet was reported as a mean for the litter. The data were analyzed by an independent t-test of SAS. In the post-weaning study, the MIXED procedure in SAS for a split-plot arrangement with sow diet as the whole plot and piglet diet as the split plot was used to analyze data on growth performance, apparent total tract digestibility (ATTD), diarrhea scores, plasma biochemical parameters, gut morphology, barrier function, and microbiota data. Results were presented as means with pooled SEM. A P-value of less than 0.05 was considered significant, while a P-value between 0.05 and 0.10 was indicated as a tendency.

Results

Reproductive performance

Maternal CA supplementation had no significant effects on litter performance at parturition or ADFI of sows during lactation. However, maternal CA supplementation tended to increase ADG during lactation (P = 0.07) and the body weight (P = 0.09) of piglets at the end of suckling compared with the CON group (Table 4).

Table 4 Effects of maternal CA supplementation on sow performance
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Colostrum and milk composition

Maternal CA supplementation increased (P < 0.05) fat content in colostrum, IgG, and IgM in the milk compared with the CON group (Table 5).

Table 5 Effects of maternal CA supplementation on colostrum and milk
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Parameters related to metabolism and redox status of sows and offspring

Maternal CA supplementation increased the activity of plasma GSH-Px and tended to increase the plasma glucose concentration (P = 0.06) of sows at farrowing. In addition, maternal CA supplementation decreased the concentration of plasma TBA (P < 0.05) and tended to increase the activity of T-SOD (P = 0.05) of sows at the end of lactation, as compared with the CON group. Likewise, maternal CA supplementation increased (P < 0.05) the plasma activities of GSH-Px and CAT in piglets at PND 24, as compared with the CON group (Table 6).

Table 6 Effects of maternal CA supplementation on plasma parameters related to metabolism and redox status of sows and piglets at PND 24
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Inflammatory cytokines and immunoglobulins of sows and offspring

Maternal CA supplementation had no significant effects on the plasma concentrations of TNF-α, IL-1β, IL-10, CRP, IgG, and IgM of sows at farrowing or end of lactation, as compared with the CON group (Table 7). However, maternal CA supplementation tended to increase plasma IgM concentration (P = 0.05) in piglets at PND 24, as compared with the CON group (Table 7).

Table 7 Effects of maternal CA supplementation on immunological and inflammatory parameters in plasma of sows and piglets at PND 24
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Gene expressions of piglets at PND 24

Maternal CA supplementation increased mRNA expressions of OCLN (P < 0.05), ZO-1 (P = 0.09), SOD1 (P < 0.05), and IL-10 (P < 0.05) in the ileum, whereas GPX1 (P < 0.05), TLR-4 (P < 0.05), and TNF-α (P = 0.07) mRNA expressions in the ileum were decreased in piglets at PND 24 compared with the CON group (Fig. 1).

Fig. 1
figure 1

Effects of maternal CA supplementation on expressions of genes related to intestinal barrier, redox status and immunity of piglets at PND 24. A mRNA expressions of intestinal barrier-related genes in ileal tissue. B mRNA expressions of redox status-related genes in ileal tissue. C mRNA expressions of innate immunity-related genes in ileal tissue. D mRNA expressions of inflammatory genes in ileal tissue. CLDN1 Claudin 1, OCLN Occludin, TLR-4 Toll-like receptor 4, TLR-9 Toll-like receptor 9, Myd88 Myeloid differentiation factor 88, NF-κB Nuclear factor kappa B, ZO-1 Zonula occludens-1, SOD1 Superoxide dismutase 1, SOD2 Superoxide dismutase 2, GPX1 Glutathione peroxidase 1, CAT Catalase, TNF-α Tumor necrosis factor-α, IL-1β Interleukin 1-β, IL-6 Interleukin 6, IL-10 interleukin 10. CON Basal diet, CA Basal diet + 500 mg/kg cinnamaldehyde. Data expressed as mean ± a pooled SEM. *P < 0.05

Gut microbiota and fecal short-chain fatty acids of sows and offspring

There were no significant differences in Chao 1, Shannon, Simpson index, and PCoA of sows between CON and CA groups (Fig. 2A). At the phylum level, maternal CA supplementation increased the fecal Fibrobacteres abundance but decreased the Proteobacteria abundance of sows at the end of lactation (Fig. 2B). At the genus level, maternal CA supplementation decreased Christensenellaceae R-7 group abundance, but increased uncultured_Muribaculaceae abundance of sows at the end of lactation (Fig. 2B). LEfSe analysis showed that the CA group had higher relative abundances of g_Oscillospira, g_Blautia, and o_Fibrobacterales, while the CON group had higher relative abundances of g_Ruminococcus_1, g_Prevotellacease_UCG_001, p_Proteobacteria, c_Gammaproteobactria, g_Lachnospiraceae_UCG_009, and f_Enterobacteiaceae (Fig. 2C). In addition, there were no significant differences in fecal concentrations of acetate, propionate, butyrate, isobutyrate, isovalerate, valerate, and total SCFA between CON and CA groups (Fig. 2D).

Fig. 2
figure 2

Effects of maternal CA supplementation on gut microbiota and short chain fatty acid of sows and piglets at PND 24. A Alpha diversity index and principal coordinate analysis of sows. B Predominant microorganisms at phylum and genus level of sows. C LEfSe analysis of sows. D Concentrations of fecal SCFAs of sows. E Alpha diversity index and principal coordinate analysis of piglets at PND 24. F Predominant microorganisms at phylum and genus level, as well as LEfSe analysis of piglets at PND 24. CON Basal diet, CA Basal diet + 500 mg/kg cinnamaldehyde. Data expressed as mean ± a pooled SEM

Maternal CA supplementation increased the Shannon index (P < 0.05) and tended to increase Chao1 (P = 0.10) and Simpson (P = 0.06) index in piglets at PND 24 compared with the CON group (Fig. 2E). Meanwhile, there were significant differences in the colonic microbial composition of piglets at PND 24 from sows fed a CON diet or CA diet in beta diversity, as shown by PCoA (Fig. 2E). At the phylum level, maternal CA supplementation increased abundances of Firmicutes, Lentisphaerae, and Proteobacteria and decreased abundances of Epsilonbacteraeota and Fusobacteria in piglets at PND 24 (Fig. 2F). At the genus level, maternal CA supplementation decreased the abundances of Fusobacterium and Prevotella 2 in piglets at PND 24 (Fig. 2F). LEfSe analysis showed that piglets at PND 24 from sows fed the CON diet had higher relative abundances of g_Sutterella, f_Veillonellaceae, g_Campylobacter, and p_Epsilonbacteraeota, while piglets from sows fed the CA diet had higher relative abundances of g_horsej_a03, g_Pseudoflavonifractor, g_Intestinimonas, g_Oscillospira, g_Faecalibacterium, g_Subdoligranulum, g_Parasutterella, g_Phascolarctobacterium, g_Family_XIII_AD3011_group, g_daA_11_gut_group, g_Eubacterium_coprostanoligenes_group, g_Synergistes, f_Tannerellaceae, g_Parabacteroides, f_ Ruminococcaceae, and c_Clostridia (Fig. 2F).

Growth performance and diarrhea scores of weaned piglets

Diarrhea scores (P = 0.06) and F:G (P < 0.05) in weaned piglets from CA-fed sows were lower than those in piglets from CON-fed sows during d 8–14. Meanwhile, diarrhea scores in weaned piglets with post-weaning CA supplementation were lower than those in post-weaning CON piglets (P < 0.05) during d 15–21 (Table 8).

Table 8 Effects of dietary CA supplementation in sows or piglets on growth performance of weaned piglets
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ATTD of nutrients of weaned piglets

The ATTD of CP, GE, and DM in weaned piglets from CA-fed sows was higher than that in piglets from CON-fed sows (P < 0.05, Table 9). Meanwhile, the ATTD of GE in weaned piglets with post-weaning CA supplementation was higher than that in post-weaning CON piglets (P < 0.05).

Table 9 Effects of dietary CA supplementation in sows or piglets on apparent total tract digestibility of weaned piglets
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Activities of digestive enzymes and intestinal morphology of weaned piglets

The villus height, VCR, and sucrase activity of the jejunum in weaned piglets from CA-fed sows were higher than those in piglets from CON-fed sows (P < 0.05, Fig. 3A and B). Meanwhile, the sucrase activity of the jejunum in weaned piglets with post-weaning CA supplementation was higher than that in post-weaning CON piglets (P < 0.05, Fig. 3B).

Fig. 3
figure 3

Effects of dietary CA supplementation in sows or piglets on intestinal morphology and activities of digestive enzymes of weaned piglets. A Intestinal morphology of jejunal morphology. B Activities of digestive enzymes. CON-CON Sows and piglets both fed basal diet, CON-CA Sows fed basal diet and piglets fed basal diet + 500 mg/kg cinnamaldehyde, CA-CON Sows fed basal diet + 500 mg/kg cinnamaldehyde and piglets fed basal diet, CA-CA Sows and piglets both fed basal diet + 500 mg/kg cinnamaldehyde, VCR The ratio of villi height-to-crypt depth. Data expressed as mean ± a pooled SEM

Antioxidant capacity, inflammatory cytokines and immunoglobulins of weaned piglets

The plasma activities of T-SOD and CAT and the IgM plasma concentration in weaned piglets with post-weaning CA supplementation were higher than those in post-weaning CON piglets (P < 0.05, Fig. 4A and B). There was an interaction between the CA-supplemented diet for sows and the CA-supplemented diet for weaned piglets on increasing the plasma concentration of IgG (P < 0.05, Fig. 4B). Weaned piglets from the CA-CA group had higher IgG concentration in plasma compared with weaned piglets from the CON-CA or CA-CON group (P < 0.05, Fig. 4B).

Fig. 4
figure 4

Effects of dietary CA supplementation in sows or piglets on redox status, immunological and inflammatory response of weaned piglets. A Plasma redox status-related parameters of weaned piglets. B Plasma immunological parameters of weaned piglets. C Plasma inflammatory cytokine of weaned piglets. GSH-Px Glutathione peroxidase, MDA Malondialdehyde, CAT Catalase, T-SOD Total superoxide dismutase, TNF-α Tumor necrosis factor-α, IL-1β Interleukin 1-β, IgG Immunoglobulin G, IgM Immunoglobulin M, C3 Complement C3. CON-CON Sows and piglets both fed basal diet, CON-CA Sows fed basal diet and piglets fed basal diet + 500 mg/kg cinnamaldehyde, CA-CON Sows fed basal diet + 500 mg/kg cinnamaldehyde and piglets fed basal diet, CA-CA Sows and piglets both fed basal diet + 500 mg/kg cinnamaldehyde. Data expressed as mean ± a pooled SEM. a, bMean values with different superscripts letters are significantly different (P < 0.05)

Gene expressions

Weaned piglets from CA-fed sows had higher mRNA expressions of SOD2 and IL-10 (P < 0.05) in the ileum and lower mRNA expressions of GPX1 (P < 0.05), TLR-4 (P < 0.05), and MyD88 (P = 0.06) in the ileum than those in weaned piglets from CON-fed sows (Fig. 5B–D). Meanwhile, ileum mRNA expressions of SOD2 (P < 0.05) and IL-10 (P = 0.07) in weaned piglets with post-weaning CA supplementation were higher than those in post-weaning CON piglets (Fig. 5B and D). There was an interaction between the CA-supplemented diet for sows and the CA-supplemented diet for weaned piglets on up-regulating mRNA expression of OCLN (P = 0.08) and down-regulating (P < 0.05) mRNA expressions of TLR-9 and NF-κB in the ileum of weaned piglets (Fig. 5A and C). Weaned piglets from the CA-CON or CA-CA group had lower mRNA expressions of TLR-9 and NF-κB in the ileum compared with weaned piglets from the CON-CON or CON-CA group (P < 0.05, Fig. 5C).

Fig. 5
figure 5

Effects of dietary CA supplementation in sows or piglets on expressions of genes related to intestinal barrier, redox status and immunity of weaned piglets. A mRNA expressions of intestinal barrier-related genes in ileal tissue. B mRNA expressions of redox status-related genes in ileal tissue. C mRNA expressions of innate immunity-related genes in ileal tissue. D mRNA expressions of inflammatory genes in ileal tissue. CLDN1 Claudin 1, OCLN Occludin, TLR-4 Toll-like receptor 4, TLR-9 Toll-like receptor 9, Myd88 Myeloid differentiation factor 88, NF-κB Nuclear factor kappa B, ZO-1 Zonula occludens-1, SOD1 Superoxide dismutase 1, SOD2 Superoxide dismutase 2, GPX1 Glutathione peroxidase 1, CAT Catalase, TNF-α Tumor necrosis factor-α, IL-1β Interleukin 1-β, IL-6 Interleukin 6, IL-10 Interleukin 10. CON-CON Sows and piglets both fed basal diet, CON-CA Sows fed basal diet and piglets fed basal diet + 500 mg/kg cinnamaldehyde, CA-CON Sows fed basal diet + 500 mg/kg cinnamaldehyde and piglets fed basal diet, CA-CA Sows and piglets both fed basal diet + 500 mg/kg cinnamaldehyde. Data expressed as mean ± a pooled SEM. Mean values with different superscripts letters are significantly different (P < 0.05)

Discussion

In this study, the litter size at farrowing was not markedly affected by maternal CA supplementation, which is reasonable given the short period of CA supplementation, and the number of piglets per litter is generally determined by early ovulation rate and conception [24, 25]. However, maternal CA supplementation tended to increase the growth rate of suckling piglets, which may be related to improved milk composition, as the concentrations of fat in colostrum, IgG, and IgM in milk were increased by CA supplementation. Similarly, maternal supplementation with plant-derived bioactive components like Lonicera flos and Sucutellaria baicalensis has been found to improve milk composition [26]. As the sole nutrient source, colostrum and milk intake are vital for piglet growth and development [27, 28]. Milk fat provides energy for thermoregulation and body growth in newborn piglets [29], while milk immunoglobulins help develop passive immunity [30]. In this study, the optimized milk composition may reflect the beneficial effects of CA on sow health status and mammary gland development, as sow health is closely linked to milk composition [31]. Studies have reported that CA could reduce cytokine production (e.g., IL-6 and TNF-α) by regulating the NF-κB pathway [32]. Thus, the levels of IgG and IgM may be associated with CA-induced changes in the inflammatory environment of the mammary gland. Additionally, systemic oxidative stress in sows may damage mammary glands and reduce both milk quality and yield [33], while antioxidant supplementation has been shown to improve the redox status and milk composition of sows [34].

In this study, supportively, sows fed the CA diet had higher plasma activities of GSH-Px and T-SOD, indicating that the improved health status of sows fed with CA may partially account for the enhancement of milk composition. Acting as an agonist for nuclear factor erythroid 2-related factor 2 (Nrf2), CA facilitates the entry of Nrf2 into the cell nucleus, where it binds to antioxidant response elements, resulting in increased expression of antioxidant genes and enzymes [35, 36]. Additionally, oxidative stress may negatively impact the growth performance of piglets [37]. The excessive accumulation of ROS disrupts systemic oxidative homeostasis, inducing peroxidative damage to subcellular organelles and extracellular matrix components. The piglet intestine is primarily affected by ROS, leading to the destruction of intestinal structure, microbial imbalance, and nutrient absorption disorders, and ultimately results in lower feed intake and body weight gain, as well as systemic metabolic imbalances [38]. Antioxidant enzymes, such as GSH-Px, T-SOD, and CAT, could help reduce the levels of toxic ROS [39]. Our previous study also found a positive correlation between plasma and ileal antioxidant enzyme levels and the growth performance of piglets [40]. In this study, the higher activities of antioxidant enzymes and growth rates were observed in piglets at the end of suckling from CA-supplemented sows.

In mammals, the small intestine is crucial for nutrient digestion and absorption [41], and it also defends against pathogen invasion [42]. Claudin-1, ZO-1, and OCLN are key to the structure and function of epithelial tight junctions [43]. TLR recognizes endotoxins from Gram-negative bacteria, activating the MyD88/NF-кβ pathways and promoting cytokine expression, such as TNF-α, IL-1β, and IL-6 [44]. In this study, piglets from CA-supplemented sows had lower expressions of TLR-4 and TNF-α and higher expressions of OCLN and ZO-1 in the ileal mucosa at the end of suckling, indicating maternal CA supplementation benefits offspring’s intestinal function, though the underlying mechanism requires further investigation.

Diet significantly influences gut microbiota composition, function, and diversity [45]. In our study, maternal CA supplementation showed limited effects on sow gut microbiota at the end of lactation. However, LEfSe analysis revealed Escherichia-Shigella as the dominant bacterial species in CON sows. Notably, Escherichia-Shigella can be transmitted from the sows and colonized in the intestines of newborn piglets [46]. As a member of the Gram-negative Enterobacteriaceae family, Shigella causes severe watery diarrhea in hosts [47] by crossing the intestinal epithelium, triggering macrophage death, and disrupting the large intestinal epithelium [48]. The gut microbiota is crucial for maintaining intestinal homeostasis in newborn piglets [49]. Maternal sources like the vagina, milk, skin, and feces provide initial bacterial communities for piglet gut colonization [46]. In our study, maternal CA supplementation significantly increased Firmicutes abundance and the Firmicutes/Bacteroides ratio in piglet colonic digesta at the end of suckling. Firmicutes are more efficient than Bacteroidetes at energy extraction from food, potentially enhancing calorie absorption and weight gain [50]. Additionally, maternal CA supplementation significantly reduced Fusobacterium and Sutterella levels in piglet colonic digesta at PND 24. These pathogens are associated with severe intestinal damage [51, 52]. The reduction in pathogenic bacteria through maternal CA supplementation may be programmed to positively affect post-weaning growth and intestinal health. Consistently, maternal CA supplementation improved the post-weaning F:G, diarrhea scores, nutrient digestibility, sucrase activity, intestinal morphology, and mRNA expressions of anti-inflammatory-related and intestinal barrier genes in weaned piglets. Similar beneficial effects on intestinal health and development have been observed with maternal live yeast supplementation [53].

Post-weaning CA supplementation also tended to reduce diarrhea scores from d 14 to 21, which was consistent with previous results [54, 55]. Weaning stress can cause systemic oxidative stress and inflammation [56], leading to intestinal dysfunction and growth reduction in weaned piglets [57, 58]. However, CA enhances antioxidant defenses against ROS via Nrf2 activation [59] and reduces inflammation by inhibiting the NF-кB pathway and cytokine expression in rodent animals [60]. In this study, likewise, post-weaning CA supplementation improved plasma antioxidant enzyme activities and mRNA expressions of SOD and IL-10 in the ileal mucosa, while decreasing inflammation-related markers such as plasma IL-1β concentration and mRNA expressions of TLR-9 and NF-кB in the ileal mucosa, which are key factors in alleviating diarrhea and intestinal injury.

Conclusions

Maternal CA supplementation during d 107 of gestation to d 24 of lactation tended to increase the growth rate and weaning weight of suckling piglets, which may be related to the improvement of CA on antioxidant capacity and milk composition. Moreover, maternal CA supplementation or post-weaning CA supplementation tended to improve diarrhea scores, along with the improved nutrient digestibility, redox status and intestinal function-related parameters of weaned piglets.

Data Availability

The data used during the current study are available from the corresponding author upon reasonable request.

Abbreviations

  • ADFI:: Average daily feed intake
  • ADG:: Average daily gain
  • ALT:: Alanine aminotransferase
  • APA:: Aminopeptidases A
  • APN:: Aminopeptidases N
  • AST:: Aspartate amino transferase
  • ATTD:: Apparent total tract digestibility
  • C3:: Complement C3
  • CA:: Cinnamaldehyde
  • CAT:: Catalase
  • CLDN1:: Claudin 1
  • CP:: Crude protein
  • CRP:: C-reactive protein
  • DM:: Dry matter
  • EE:: Ether extract
  • FG:: The ratio of average daily feed intake to average daily gain
  • GE:: Gross energy
  • GGT:: Gamma-glutamyl transpeptidase
  • GPX1:: Glutathione peroxidase 1
  • GSH-Px:: Glutathione peroxidase
  • IgG:: Immunoglobulin G
  • IgM:: Immunoglobulin M
  • IL-1β:: Interleukin-1β
  • IL-6:: Interleukin-6
  • IL-10:: Interleukin-10
  • MDA:: Malondialdehyde
  • MyD88:: Myeloid differentiation factor 88
  • NEFA:: Non-esterified fatty acid
  • NF-κB:: Nuclear factor kappa B
  • OCLN:: Occludin
  • PCoA:: Principal coordinate analysis
  • PND:: Post-natal day
  • PWD:: Post-weaned day
  • SCFA:: Short-chain fatty acid
  • SOD1:: Superoxide dismutase 1
  • SOD2:: Superoxide dismutase 2
  • TBA:: Total bile acid
  • TG:: Total cholesterol
  • TLR-4:: Toll-like receptor 4
  • TLR-9:: Toll-like receptor 9
  • TNF-α:: Tumor necrosis factor-α
  • T-SOD:: Total superoxide dismutase
  • VCR:: The ratio of villus height to crypt depth
  • ZO-1:: Zonula occludens-1

References

  1. 1.Lee J, Shin H, Jo J, Lee G, Yun J. Large litter size increases oxidative stress and adversely affects nest-building behavior and litter characteristics in primiparous sows. Front Vet Sci. 2023;10.(2023)1219572. https://doi. org/10.3389/fvets.: 1219572.
    10.3389/fvets.2023.1219572
    Article|CAS|PubMed|Google Scholar
  2. 2.Sultana Z, Qiao Y, Maiti K, Smith R. Involvement of oxidative stress in placental dysfunction, the pathophysiology of fetal death and pregnancy disorders. Reproduction. 2023;166(2).(2023)org/10.1530/rep-22-0278.
    10.1530/rep-22-0278
    Article|CAS|PubMed|Google Scholar
  3. 3.Zhao Y, Kim SW. Oxidative stress status and reproductive performance of sows during gestation and lactation under different thermal environments. Asian-Australas J Anim Sci. 2020;33(5).(2020)19.0334.: 722.
    10.5713/ajas.19.0334
    Article|CAS|PubMed|Google Scholar
  4. 4.Wang H, Ji Y, Yin C, Deng M, Tang T, Deng B, et al. Differential analysis of gut microbiota correlated with oxidative stress in sows with high or low litter performance during lactation. Front Microbiol. 2018;9.(2018)1665. https://doi. org/10.3389/fmicb.: 1665.
    10.3389/fmicb.2018.01665
    Article|CAS|PubMed|Google Scholar
  5. 5.Wang K, Hu C, Tang W, Azad MAK, Zhu Q, He Q, et al. The enhancement of intestinal immunity in offspring piglets by maternal probiotic or synbiotic supplementation is associated with the alteration of gut microbiota. Front Nutr. 2021;8.(2021)686053. https://doi. org/10.3389/fnut.: 686053.
    10.3389/fnut.2021.686053
    Article|CAS|PubMed|Google Scholar
  6. 6.Llauradó-Calero E, Badiola I, Samarra I, Lizardo R, Torrallardona D, Esteve-Garcia E, et al. Eicosapentaenoic acid- and docosahexaenoic acid-rich fish oil in sow and piglet diets modifies blood oxylipins and immune indicators in both, sows and suckling piglets. Animal. 2022;16(10).(2022)100634. https://doi. org/10. 1016/j.animal.: 100634.
    10.1016/j.animal.2022.100634
    Article|CAS|PubMed|Google Scholar
  7. 7.Dowley A, O’Doherty JV, Mukhopadhya A, Conway E, Vigors S, Maher S, et al. Maternal supplementation with a casein hydrolysate and yeast beta-glucan from late gestation through lactation improves gastrointestinal health of piglets at weaning. Sci Rep. 2022;12.(2022)org/10.1038/s41598-022-20723-5.: 17407.
    10.1038/s41598-022-20723-5
    Article|CAS|PubMed|Google Scholar
  8. 8.Laviano HD, Gómez G, Escudero R, Nuñez Y, García-Casco JM, Muñoz M, et al. Maternal supplementation of vitamin E or its combination with hydroxytyrosol increases the gut health and short chain fatty acids of piglets at weaning. Antioxidants (Basel). 2023;12(9).(2023)org/10.3390/antiox12091761.: 1761.
    Article|CAS|PubMed|Google Scholar
  9. 9.Van Kerschaver C, Turpin D, Michiels J, Pluske J. Reducing weaning stress in piglets by pre-weaning socialization and gradual separation from the sow.(2023)a review.Animals (Basel).: 1644.
    Article|CAS|PubMed|Google Scholar
  10. 10.Bomba L, Minuti A, Moisá SJ, Trevisi E, Eufemi E, Lizier M, et al. Gut response induced by weaning in piglet features marked changes in immune and inflammatory response. Funct Integr Genomics. 2014;14(4).(2014)org/10.1007/s10142-014-0396-x.: 657.
    10.1007/s10142-014-0396-x
    Article|CAS|PubMed|Google Scholar
  11. 11.Boudry G, Péron V, Le Huërou-Luron I, Lallès JP, Sève B. Weaning induces both transient and long-lasting modifications of absorptive, secretory, and barrier properties of piglet intestine. J Nutr. 2004;134(9).(2004)9.2256.: 2256.
    10.1093/jn/134.9.2256
    Article|CAS|PubMed|Google Scholar
  12. 12.Ren W, Yu B, Yu J, Zheng P, Huang Z, Luo J, et al. Lower abundance of Bacteroides and metabolic dysfunction are highly associated with the post-weaning diarrhea in piglets. Sci China Life Sci. 2022;65(10).(2022)org/10.1007/s11427-021-2068-6.: 2062.
    10.1007/s11427-021-2068-6
    Article|CAS|PubMed|Google Scholar
  13. 13.Muhoza B, Qi B, Harindintwali JD, Koko MYF, Zhang S, Li Y. Encapsulation of cinnamaldehyde.(2023)an insight on delivery systems and food applications.Crit Rev Food Sci Nutr.: 2521.
    10.1080/10408398.2021.1977236
    Article|CAS|PubMed|Google Scholar
  14. 14.Li W, Zhi W, Zhao J, Li W, Zang L, Liu F, et al. Cinnamaldehyde attenuates atherosclerosis via targeting the IκB/NF-κB signaling pathway in high fat diet-induced ApoE-/-mice. Food Funct. 2019;10(7).(2019)org/10.1039/c9fo00396g.: 4001.
    Article|CAS|PubMed|Google Scholar
  15. 15.Almoiliqy M, Wen J, Xu B, Sun YC, Lian MQ, Li YL, et al. Cinnamaldehyde protects against rat intestinal ischemia/reperfusion injuries by synergistic inhibition of NF-κB and p53. Acta Pharmacol Sin. 2020;41(9).(2020)org/10.1038/s41401-020-0359-9.: 1208.
    10.1038/s41401-020-0359-9
    Article|CAS|PubMed|Google Scholar
  16. 16.Mao M, Zheng W, Deng B, Wang Y, Zhou D, Shen L, et al. Cinnamaldehyde alleviates doxorubicin-induced cardiotoxicity by decreasing oxidative stress and ferroptosis in cardiomyocytes. PLoS ONE. 2023;18(10).(2023)pone.0292124.
    10.1371/journal.pone.0292124
    Article|CAS|PubMed|Google Scholar
  17. 17.Sampath C, Chukkapalli SS, Raju AV, Alluri LSC, Srisai D, Gangula PR. Cinnamaldehyde protects againstP. gingivalisinduced intestinal epithelial barrier dysfunction in IEC-6 cells via the PI3K/Akt-mediated NO/Nrf2 signaling pathway. Int J Mol Sci. 2024;25(9).(2024)org/10.3390/ijms25094734.: 4734.
    Article|CAS|PubMed|Google Scholar
  18. 18.Faikoh EN, Hong YH, Hu SY. Liposome-encapsulated cinnamaldehyde enhances zebrafish (Danio rerio) immunity and survival when challenged withVibrio vulnificusandStreptococcus agalactiae. Fish Shellfish Immunol. 2014;38(1).(2014)15–24. https://doi. org/10. 1016/j.fsi.: 15.
    10.1016/j.fsi.2014.02.024
    Article|CAS|PubMed|Google Scholar
  19. 19.Huang Y, Lang A, Yang S, Shahid MS, Yuan J. The combined use of cinnamaldehyde and vitamin C is beneficial for better carcass character and intestinal health of broilers. Int J Mol Sci. 2024;25(15).(2024)org/10.3390/ijms25158396.: 8396.
    Article|CAS|PubMed|Google Scholar
  20. 20.Luise D, Correa F, Negrini C, Virdis S, Mazzoni M, Dalcanale S, et al. Blend of natural and natural identical essential oil compounds as a strategy to improve the gut health of weaning pigs. Animal. 2023;17(12).(2023)101031. https://doi. org/10. 1016/j.animal.: 101031.
    10.1016/j.animal.2023.101031
    Article|CAS|PubMed|Google Scholar
  21. 21.National Research Council (NRC). Nutrient Requirements of Swine. 11th ed. Washington, DC: The National Academic Press; 2012.
    Article|CAS|PubMed|Google Scholar
  22. 22.Association of Official Analytical Chemists (AOAC). Official Methods of Analysis. 18th ed. Rev. 2nd ed. In.(2007)Hortwitz W, Latimer Jr GW, editors.Gaithersburg: AOAC International;.
    Article|CAS|PubMed|Google Scholar
  23. 23.Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCTMethod. Methods. 2001;25(4).(2001)402–8. https://doi. org/10.1006/meth.: 402.
    10.1006/meth.2001.1262
    Article|CAS|PubMed|Google Scholar
  24. 24.King RH, Williams IH. The influence of ovulation rate on subsequent litter size in sows. Theriogenology. 1984;21(4).(1984)org/10.1016/0093-691x(84)90451-5.: 677.
    10.1016/0093-691x(84)90451-5
    Article|CAS|PubMed|Google Scholar
  25. 25.Foxcroft GR, Silva PV, Paradis F. Application of transcriptomic analyses to reproductive studies in contemporary commercial sows. Theriogenology. 2016;85(1).(2016)10.017.: 145.
    10.1016/j.theriogenology.2015.10.017
    Article|CAS|PubMed|Google Scholar
  26. 26.Wang L, Huo B, Huang L, Che L, Feng B, Lin Y, et al. Dietary supplementation with a mixture of herbal extracts during late gestation and lactation improves performance of sows and nursing piglets through regulation of maternal metabolism and transmission of antibodies. Front Vet Sci. 2022;9.(2022)1026088. https://doi. org/10.3389/fvets.: 1026088.
    10.3389/fvets.2022.1026088
    Article|CAS|PubMed|Google Scholar
  27. 27.Maciag SS, Bellaver FV, Bombassaro G, Haach V, Morés MAZ, Baron LF, et al. On the influence of the source of porcine colostrum in the development of early immune ontogeny in piglets. Sci Rep. 2022;12.(2022)org/10.1038/s41598-022-20082-1.: 15630.
    10.1038/s41598-022-20082-1
    Article|CAS|PubMed|Google Scholar
  28. 28.Quesnel H, Farmer C, Devillers N. Colostrum intake.(2012)Influence on piglet performance and factors of variation.Livest Sci.: 105.
    Article|CAS|PubMed|Google Scholar
  29. 29.Inoue R, Tsukahara T. Composition and physiological functions of the porcine colostrum. Anim Sci J. 2021;92(1).(2021)1111/asj.13618.
    10.1111/asj.13618
    Article|CAS|PubMed|Google Scholar
  30. 30.Gormley A, Jang KB, Garavito-Duarte Y, Deng Z, Kim SW. Impacts of maternal nutrition on sow performance and potential positive effects on piglet performance. Animals (Basel). 2024;14(13).(2024)org/10.3390/ani14131858.: 1858.
    Article|CAS|PubMed|Google Scholar
  31. 31.Chen J, Huang Z, Cao X, Zou T, You J, Guan W. Plant-derived polyphenols in sow nutrition.(2023)An update.Anim Nutr.: 96.
    10.1016/j.aninu.2022.08.015
    Article|CAS|PubMed|Google Scholar
  32. 32.Yang D, Liang XC, Shi Y, Sun Q, Liu D, Liu W, et al. Anti-oxidative and anti-inflammatory effects of cinnamaldehyde on protecting high glucose-induced damage in cultured dorsal root ganglion neurons of rats. Chin J Integr Med. 2016;22(1).(2016)org/10.1007/s11655-015-2103-8.: 19.
    10.1007/s11655-015-2103-8
    Article|CAS|PubMed|Google Scholar
  33. 33.Su G, Zhao J, Luo G, Xuan Y, Fang Z, Lin Y, et al. Effects of oil quality and antioxidant supplementation on sow performance, milk composition and oxidative status in serum and placenta. Lipids Health Dis. 2017;16(1).(2017)org/10.1186/s12944-017-0494-6.: 107.
    10.1186/s12944-017-0494-6
    Article|CAS|PubMed|Google Scholar
  34. 34.Laviano HD, Gómez G, Muñoz M, García-Casco JM, Nuñez Y, Escudero R, et al. Dietary vitamin E and/or hydroxytyrosol supplementation to sows during late pregnancy and lactation modifies the lipid composition of colostrum and milk. Antioxidants (Basel). 2023;12(5).(2023)org/10.3390/antiox12051039.: 1039.
    Article|CAS|PubMed|Google Scholar
  35. 35.Zhang Q, Liu J, Duan H, Li R, Peng W, Wu C. Activation of Nrf2/HO-1 signaling.(2021)An important molecular mechanism of herbal medicine in the treatment of atherosclerosis via the protection of vascular endothelial cells from oxidative stress.J Adv Res.: 43.
    10.1016/j.jare.2021.06.023
    Article|CAS|PubMed|Google Scholar
  36. 36.Huang TC, Chung YL, Wu ML, Chuang SM. Cinnamaldehyde enhances Nrf2 nuclear translocation to upregulate phase II detoxifying enzyme expression in HepG2 cells. J Agric Food Chem. 2011;59(9).(2011)org/10.1021/jf200579h.: 5164.
    10.1021/jf200579h
    Article|CAS|PubMed|Google Scholar
  37. 37.Tian J, Jiang Q, Bao X, Yang F, Li Y, Sun H, et al. Plant-derived squalene supplementation improves growth performance and alleviates acute oxidative stress-induced growth retardation and intestinal damage in piglets. Anim Nutr. 2023;15.(2023)386–98. https://doi. org/10. 1016/j.aninu.: 386.
    10.1016/j.aninu.2023.09.001
    Article|CAS|PubMed|Google Scholar
  38. 38.Hao Y, Xing M, Gu X. Research progress on oxidative stress and its nutritional regulation strategies in pigs. Animals (Basel). 2021;11(5).(2021)org/10.3390/ani11051384.: 1384.
    Article|CAS|PubMed|Google Scholar
  39. 39.Galasso M, Gambino S, Romanelli MG, Donadelli M, Scupoli MT. Browsing the oldest antioxidant enzyme.(2021)catalase and its multiple regulation in cancer.Free Radic Biol Med.: 264.
    10.1016/j.freeradbiomed.2021.06.010
    Article|CAS|PubMed|Google Scholar
  40. 40.Liu Y, Wu A, Mo R, Zhou Q, Song L, Li Z, et al. Dietary lysolecithin supplementation improves growth performance of weaned piglets via improving nutrients absorption, lipid metabolism, and redox status. J Anim Sci. 2023;101.(2023)org/10.1093/jas/skad293.
    Article|CAS|PubMed|Google Scholar
  41. 41.Beaumont M, Blanc F, Cherbuy C, Egidy G, Giuffra E, Lacroix-Lamandé S, et al. Intestinal organoids in farm animals. Vet Res. 2021;52(1).(2021)org/10.1186/s13567-021-00909-x.: 33.
    10.1186/s13567-021-00909-x
    Article|CAS|PubMed|Google Scholar
  42. 42.Yang S, Yang N, Huang X, Li Y, Liu G, Jansen CA, et al. Pigs’ intestinal barrier function is more refined with aging. Dev Comp Immunol. 2022;136.(2022)104512. https://doi. org/10. 1016/j.dci.: 104512.
    10.1016/j.dci.2022.104512
    Article|CAS|PubMed|Google Scholar
  43. 43.Otani T, Furuse M. Tight junction structure and function revisited. Trends Cell Biol. 2020;30(10).(2020)805–17. https://doi. org/10. 1016/j.tcb.: 805.
    10.1016/j.tcb.2020.08.004
    Article|CAS|PubMed|Google Scholar
  44. 44.Kim HJ, Kim H, Lee JH, Hwangbo C. Toll-like receptor 4 (TLR4).(2023)new insight immune and aging.Immun Ageing.: 67.
    10.1186/s12979-023-00383-3
    Article|CAS|PubMed|Google Scholar
  45. 45.Ross FC, Patangia D, Grimaud G, Lavelle A, Dempsey EM, Ross RP, et al. The interplay between diet and the gut microbiome.(2024)implications for health and disease.Nat Rev Microbiol.: 671.
    10.1038/s41579-024-01068-4
    Article|CAS|PubMed|Google Scholar
  46. 46.Liu H, Zeng X, Zhang G, Hou C, Li N, Yu H, et al. Maternal milk and fecal microbes guide the spatiotemporal development of mucosa-associated microbiota and barrier function in the porcine neonatal gut. BMC Biol. 2019;17.(2019)org/10.1186/s12915-019-0729-2.: 106.
    10.1186/s12915-019-0729-2
    Article|CAS|PubMed|Google Scholar
  47. 47.Kotloff KL, Riddle MS, Platts-Mills JA, Pavlinac P, Zaidi AKM. Shigellosis Lancet. 2018;391(10122).(2018)org/10.1016/s0140-6736(17)33296-8.: 801.
    10.1016/s0140-6736(17)33296-8
    Article|CAS|PubMed|Google Scholar
  48. 48.Baker S, The HC. Recent insights into Shigella. Curr Opin Infect Dis. 2018;31(5).(2018)1097/qco.0000000000000475.: 449.
    10.1097/qco.0000000000000475
    Article|CAS|PubMed|Google Scholar
  49. 49.Duarte ME, Kim SW. Intestinal microbiota and its interaction to intestinal health in nursery pigs. Anim Nutr. 2022;8(1).(2022)05.001.: 169.
    10.1016/j.aninu.2021.05.001
    Article|CAS|PubMed|Google Scholar
  50. 50.Magne F, Gotteland M, Gauthier L, Zazueta A, Pesoa S, Navarrete P, et al. The Firmicutes/Bacteroidetes ratio.(2020)a relevant marker of gut dysbiosis in obese patients? Nutrients.: 1474.
    Article|CAS|PubMed|Google Scholar
  51. 51.Yang Y, Weng W, Peng J, Hong L, Yang L, Toiyama Y, et al. Fusobacterium nucleatum increases proliferation of colorectal cancer cells and tumor development in mice by activating toll-like receptor 4 signaling to nuclear factor-κB, and up-regulating expression of microRNA-21. Gastroenterology. 2017;152(4).(2017)11.018.: 851-66.
    10.1053/j.gastro.2016.11.018
    Article|CAS|PubMed|Google Scholar
  52. 52.Kaakoush NO. Sutterella species, IgA-degrading bacteria in ulcerative colitis. Trends Microbiol. 2020;28(7).(2020)519–22. https://doi. org/10. 1016/j.tim.: 519.
    10.1016/j.tim.2020.02.018
    Article|CAS|PubMed|Google Scholar
  53. 53.Fu Y, Li E, Casey TM, Johnson TA, Adeola O, Ajuwon KM. Impact of maternal live yeast supplementation to sows on intestinal inflammatory cytokine expression and tight junction proteins in suckling and weanling piglets. J Anim Sci. 2024;102.(2024)org/10.1093/jas/skae008.
    Article|CAS|PubMed|Google Scholar
  54. 54.Zhao BC, Wang TH, Chen J, Qiu BH, Xu YR, Li JL. Essential oils improve nursery pigs’ performance and appetite via modulation of intestinal health and microbiota. Anim Nutr. 2024;16.(2024)10.007.: 174.
    10.1016/j.aninu.2023.10.007
    Article|CAS|PubMed|Google Scholar
  55. 55.Zhang Y, Li Q, Wang Z, Dong Y, Yi D, Wu T, et al. Dietary supplementation with a complex of cinnamaldehyde, carvacrol, and thymol negatively affects the intestinal function in LPS-challenged piglets. Front Vet Sci. 2023;10.(2023)1098579. https://doi. org/10.3389/fvets.: 1098579.
    10.3389/fvets.2023.1098579
    Article|CAS|PubMed|Google Scholar
  56. 56.Luo Z, Zhu W, Guo Q, Luo W, Zhang J, Xu W, et al. Weaning induced hepatic oxidative stress, apoptosis, and aminotransferases through MAPK signaling pathways in piglets. Oxid Med Cell Longev. 2016;2016.(2016)4768541. https://doi. org/10.1155/.: 4768541.
    10.1155/2016/4768541
    Article|CAS|PubMed|Google Scholar
  57. 57.Han H, Liu Z, Yin J, Gao J, He L, Wang C, et al. D-galactose induces chronic oxidative stress and alters gut microbiota in weaned piglets. Front Physiol. 2021;12.(2021)634283. https://doi. org/10.3389/fphys.: 634283.
    10.3389/fphys.2021.634283
    Article|CAS|PubMed|Google Scholar
  58. 58.Li X, Wang C, Zhu J, Lin Q, Yu M, Wen J, et al. Sodium butyrate ameliorates oxidative stress-induced intestinal epithelium barrier injury and mitochondrial damage through AMPK-mitophagy pathway. Oxid Med Cell Longev. 2022;2022.(2022)3745135. https://doi. org/10.1155/.: 3745135.
    10.1155/2022/3745135
    Article|CAS|PubMed|Google Scholar
  59. 59.Lin BH, Ma RX, Wu JT, Du SQ, Lv YY, Yu HN, et al. Cinnamaldehyde alleviates bone loss by targeting oxidative stress and mitochondrial damage via the Nrf2/HO-1 pathway in BMSCs and ovariectomized mice. J Agric Food Chem. 2023;71(45).(2023)jafc.3c03501.: 17362.
    10.1021/acs.jafc.3c03501
    Article|CAS|PubMed|Google Scholar
  60. 60.Qi L, Mao H, Lu X, Shi T, Wang J. Cinnamaldehyde promotes the intestinal barrier functions and reshapes gut microbiome in early weaned rats. Front Nutr. 2021;8.(2021)748503. https://doi. org/10.3389/fnut.: 748503.
    10.3389/fnut.2021.748503
    Article|CAS|PubMed|Google Scholar

Acknowledgements

We are grateful to Yuwei Zhang, Yifan Wang, Huaqi Song, Changqin Wang, Ruizhi Zhang, and Lingji Ye for their assistance during the animal trail.

Funding

This work was supported by the project of the earmarked fund for China Agriculture Research System (CARS-35), National Key Research and Development Program of China (2023YFD1300802).

Ethics Declaration

Ethics approval and consent to participate

All procedures in this experiment were approved by the Animal Care Committee of Sichuan Agricultural University (DKYB20121602) and performed in accordance with the National Research Council’s Guidelines for Care and Use of Laboratory Animals.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

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