Skip to main content
Download PDF

Dietary steviol glycosides mixture supplementation modulates the gene expression of gut chemoreceptors and enhances the antioxidant capacity in weaned piglets

Porcine Health Management volume 11, Article number: 6 (2025) Cite this article

Abstract

Background

Stevia glycosides (SGs) have been widely used as an ideal sugar alternative in the food industry. However, the potential application of SGs mixture in the diets of weaned piglets remains unexplored. This study aimed to investigate the effect of dietary SGs mixture supplementation on growth performance, gene expression of gut chemoreceptors, and antioxidant capacity in weaned piglets.

Methods

A total of 216 weaned piglets (Duroc × Landrace × Yorkshire, 7.36 ± 0.04 kg body weight) were randomly assigned to 6 groups (6 pens/group with 6 piglets/pen), and were fed with the basal diet supplemented with 0, 100, 150, 200, 250, or 300 mg/kg SGs mixture for 42 days. The serum, liver, longissimus thoracis, and jejunal samples were collected on day 43.

Results

The results showed that inclusion the SGs mixture in the diet did not have a significant impact on growth performance from days 1 to 28 (P > 0.05). But increasing the concentration of SGs mixture tended to linearly decrease the average daily gain from days 1 to 42 (P = 0.052). However, 150 mg/kg SGs mixture supplementation significantly increased the mRNA expression of taste receptor family 1 member 2 (T1R2) and glucose transporters 2 (GLUT2) in the jejunum (P < 0.05), while 150 and 200 mg/kg SGs mixture supplementation significantly increased T1R3 mRNA expression (P < 0.05). Moreover, 150 mg/kg SGs mixture supplementation significantly reduced serum malondialdehyde content (P < 0.05). Increasing the concentration of SGs mixture linearly and quadratically increased serum total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity, as well as hepatic T-SOD, GSH-Px activity, and muscle total antioxidant capacity contents (P < 0.05). Furthermore, piglets fed a diet supplemented with 100 mg/kg SGs mixture had higher serum T-SOD, CAT, and GSH-Px activities compared with the other treatments (P < 0.05).

Conclusions

Therefore, our results suggest that dietary 100 ~ 150 mg/kg SGs mixture supplementation modulates gene expression of sweet taste recognition receptors and glucose transporters, while also enhancing the antioxidant capacity of weaned piglets.

Background

Steviol glycosides (SGs), a group of diterpenoid glycosides derived from Stevia rebaudiana that are 200 ~ 300 times sweeter than sucrose, have become an ideal sugar alternative [1]. To date, 64 SGs have been identified in the leaves of Stevia rebaudiana. Among these, ten glycosides, including Stevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E, Rebaudioside F, Dulcoside A, Rubusoside, and Steviolbioside, are found in relatively high abundance [2]. The primary differences in the chemical structures of these SGs lie in the R1 and R2 groups, which are attached at the C13-hydroxyl and C19-carboxyl positions, respectively [2]. SGs are known for their heat stability, pH stability, non-caloric properties, and non-fermentative nature [3]. Over the past decades, SGs have been demonstrated to be non-toxic, devoid of side effects, non-carcinogenic, and safe for consumption, leading to their widespread use in the food and pharmaceutical industries [4, 5]. Beyond sweetness, SGs has multiple bioactivities such as anti-diabetes, anti-hypertension, anti-oxidation, anti-inflammation, anti-microbial, anti-cancer, and anti-diarrheal [3, 6,7,8,9], with potential performance benefits for livestock and poultry.

Weaning stress frequently leads to a decrease in feed intake in weaned piglets. The addition of sweeteners such as sucrose, glucose, lactose, and natural or artificial sweeteners to diets improves palatability and helps piglets cope with the reduced appetite that results from weaning stress [10,11,12,13]. Previous studies have shown that SGs have the ability to increase feed intake in broiler chickens and goats [14,15,16]. Wang et al. [17] found that increasing the dietary stevioside/rebaudioside A supplementation from 0 to 300 mg/kg led to a linear increase in average daily feed intake in weaned piglets. However, another study showed that dietary stevia addition had no beneficial effect on feed intake in newly weaned piglets [18]. The steviol glycosides products used in previous studies were nearly pure, exceeding 90%, and it is well-known that purifying products in the industry is a costly process. We hypothesize whether replacing these purified products with a SGs mixture, which is a more affordable option with a simpler production process, would positively impact food intake in weaned piglets. Moreover, in mammals, sweet molecules are typically detected by chemoreceptors, which then stimulate pathways associated with appetite and feed intake [19]. The role gut chemoreceptors play in SGs mixture sensing needs further research. On the other hand, weaning stress can induce the body to produce much free radicals, resulting in oxidative stress [20]. Besides, stevioside has been widely reported to have excellent antioxidant effect, which can alleviate the damage induced by oxidative stress [21]. Whether the SGs mixture could alleviate the oxidative stress in weaned piglets is still unclear. Therefore, this study was conducted to investigate the effects of dietary SGs mixture supplementation on performance, expression of gut chemoreceptors, and antioxidant capacity.

Methods

Steviol glycosides mixture compositions

The SGs mixture used in this study is a white powder provided by Dongtai Hirye Biotechnology Co. Ltd (Dongtai, China) with the product batch number M20220316. The compositions of SGs in the mixture were analyzed using HPLC following China National Standard GB 8270 − 2014, and detailed ingredients are provided in Table 1. The main components of the SGs mixture used in this study are Rebaudioside A (39.90%), Stevioside (30.40%), Rebaudioside C (12.40%), Rebaudioside F (2.00%), Rebaudioside D (1.00%), Rubusoside (0.70%), Rebaudioside B (0.60%), Dulcoside A (0.3%), and Steviolbioside (0.2%), and the total SGs is 87.50%. Sensory evaluation was conducted to assess the characteristics of the SGs mixture [22]. As depicted in Fig. 1, the SGs mixture exhibited a delayed onset of sweetness, with noticeable bitterness, a slight stringency, and an unpleasant metallic aftertaste when compared with sucrose in an iso-sweet water solution.

Table 1 Identified components and their contents in the steviol glycosides mixture used in this study (air-dry basis)
Full size table
Fig. 1
figure 1

Comparison of sensory attributes of steviol glycosides mixture (0.03%) and sucrose (6%) in iso-sweet water solutions

Full size image

Animal management and experimental design

A total of 216 weaned piglets (Duroc × Landrace × Yorkshire) aged 21 days with an average initial body weight (BW) of 7.36 ± 0.04 kg were randomly allocated to six treatments. Each treatment consisted of six replicates, with six pigs per replicate pen, including three barrows and three gilts. The experimental groups comprised a control group receiving a basal diet devoid of any sweeteners, and experimental groups received a basal diet supplemented with 100, 150, 200, 250, and 300 mg/kg SGs mixture (SGs100, SGs150, SGs200, SGs250, and SGs300), respectively. Before the commencement of this trial, the pigs underwent a 5-day acclimation period. The experiment lasted for 42 days. The basal diet (Table 2) was formulated according to the nutrient requirement recommendations for piglets by the NRC (2012). All diets were provided as pellet. Feed intake was recorded on a weekly basis in pen units to collectively measure consumption levels, while individual pig body weights were measured on day 29 and 43. These data facilitated the calculation of the average daily gain (ADG), average daily feed intake (ADFI), and feed to gain ratio (F/G) over the periods of 1 ~ 28 days, 29 ~ 42 days, and 1 ~ 42 days. All piglets were housed individually in pens within a temperature-controlled and enclosed nursery. The pens, measuring 1.2 × 2.1 m2, featured high-rise beds equipped with plastic slatted floors and an effective mechanical ventilation system. Each pen was outfitted with two stainless steel feeders and four nipple drinkers, ensuring that all pigs had ad libitum access to both feed and water.

Table 2 Composition and nutrient levels of the basal diet (air-dry basis)
Full size table

Sample collection

At the end of the experiment, all the pigs were weighed individually after 16-hours fasting. One pig from each pen (36 pigs total) was randomly selected for blood collection and sacrifice. A 10 mL blood sample was obtained from each pig by anterior vena cava puncture, and serum samples were separated by centrifugation at 1509.3 × g at 4 °C for 10 min. Then the pig was anesthetized by intravenous injection of pentobarbital sodium (30 mg/kg BW) and killed by bloodletting. The intestinal tissues were dissected on ice and categorized into the duodenum, jejunum, and ileum. The middle jejunum tissue was sampled, opened, and thoroughly washed with cold sterile phosphate-buffered saline. Mucosa samples were scraped from the inner side of the jejunum using a sterile slide, and then were collected. Approximately 0.5 g of the middle section of longissimus thoracis and the central portion of liver tissues from the same region were obtained to assess antioxidant capacity indicators. All samples were immediately transferred to liquid nitrogen for rapid freezing and subsequently stored at -80 °C for further analysis.

Appetite-associated hormones detection

Appetite-associated hormones such as ghrelin (GHRL), glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK), leptin (LEP), and insulin (INS) in serum were detected by commercial Elisa kits purchased from Jiangsu Meimian Industrial Co., Ltd (Jiangsu, China).

Real-time qPCR

The jejunal mucosal tissues were processed to extract total RNA using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). The concentration and purity of the RNA were evaluated using a NanoDrop ND-1000 Spectrophotometer (Nano-Drop Technologies, Rockland, DE), followed by monitoring RNA integrity on 1% agarose gels. Subsequently, the first-strand cDNA synthesis was conducted through reverse transcription of 1 µg total RNA utilizing the Prime Script RT reagent kit with gDNA Eraser (Takara, Tokyo, Japan). Real-time qPCR was conducted using the Bio-Rad CFX System in a final volume of 20 µL, comprising 2 µL of cDNA product (diluted at 1:9, v/v), 10 µL of iTaq Universal SYBR Green PCR Supermix (2×, Bio-Rad, Hercules, California, USA), 6.4 µL of RNase-free water, and 0.8 µL of each forward and reverse primers (10 µM/L) as detailed in Table 3. The PCR cycling conditions included an initial denaturation step (95 °C for 30 s) followed by 40 cycles of amplification and quantification (95 °C for 15 s, 60 °C for 30 s, and 72 °C for 30 s with a single fluorescence reading). All measurements were carried out in triplicate. The 2−ΔΔCT method was employed to analyze gene expression levels. β-actin was served as the housekeeping gene, and the data were normalized to the control group.

Table 3 Primers used for real-time PCR in this study
Full size table

Serum biochemical parameters measurement

Serum biochemical parameters including serum glucose (GLU), total protein (TP), albumin (ALB), urea (URE), triglyceride (TG), total cholesterol (CHO), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), alkaline phosphatase (ALP), and creatinine (CRE) were detected by commercial kits purchased from Biosino Biotechnology and Science inc. (Beijing, China) with an automatic biochemical analyzer (Selectra Pro XL, EliTechGroup, Puteaux, France).

Serum immunoglobulins, cytokine determination

Serum immunoglobulin A (IgA), IgG, and IgM, and cytokines interleukin-1 (IL-1), IL-1β, IL-6, IL-8, IL-10, IL-22, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and TNF-β were measured using commercial kits obtained from Jiangsu Meimian Industrial Co., Ltd (Jiangsu, China).

Antioxidant capacity evaluation

Malondialdehyde (MDA), total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were assessed using commercial kits purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Samples of liver and longissimus thoracis tissues were homogenized in physiological saline, and the supernatants were harvested after centrifugation for subsequent analysis. The final data were normalized to the total protein concentration in the tissues.

Statistical analysis

The data were analyzed using IBM SPSS Statistics V18.0 software (IBM Corp., Armonk, NY, USA). Results were presented as means with a pooled standard error (SEM). Prior to intergroup difference analysis, the normality of the data was assessed using the Shapiro-Wilk test. For variables with non-normal distributions, analysis was performed using one-way ANOVA followed by the Kruskal-Wallis test with false discovery rate (FDR) multiple corrections. When the data exhibited a normal distribution, one-way ANOVA analysis followed by the LSD test was employed. The linear and quadratic responses of parameters to different SGs mixture supplemental levels were evaluated using regression analysis with a curve estimation model. Each pen was considered as the experimental unit for the data of growth performance, while individual pigs served as the experimental unit for the other data. Statistical significance was considered at P < 0.05, with a significant trend noted at 0.05 ≤ P < 0.10.

Results

Growth performance

As illustrated in Table 4, increasing SGs mixture supplemental level from 0 to 300 mg/kg linearly (P < 0.05) and quadratically (P < 0.10) decreased ADG from days 29 to 42, as well as ADG from days 1 to 42 (P < 0.10). Furthermore, the ADFI from days 1 to 42 showed a trend of initially increasing, followed by a decrease as the level of dietary SGs mixture supplementation rose (P < 0.10), with the highest ADFI recorded in the group supplemented with 100 mg/kg of the SGs mixture. However, inclusion the SGs mixture in the diet did not have a significant impact on the ADG, ADFI, or F/G from days 1 to 28 in the piglets (P > 0.05).

Table 4 Effects of dietary steviol glycosides mixture supplementation on growth performance of weaned piglets
Full size table

Appetite-associated hormones

As displayed in Table 5, increasing SGs mixture supplementation resulted in a linear increase in the serum CCK content of weaned piglets (P < 0.05). However, there were no significant differences in serum GHRL, GLP-1, LEP, and INS levels among the different treatments (P > 0.05).

Table 5 Effects of dietary steviol glycosides mixture supplementation on serum appetite-related hormones in weaned piglets
Full size table

Gene expression of gut chemoreceptors

As shown in Figs. 2 and 150 mg/kg SGs mixture supplementation resulted in a significant up-regulation of the relative mRNA expression of T1R2 and GLUT2 in the jejunal mucosa compared with the control group (P < 0.05). In addition, the relative mRNA expression of T1R3 in the jejunal mucosa was significantly increased by 150 and 200 mg/kg SGs mixture supplementation (P < 0.05). Conversely, 250 mg/kg SGs mixture supplementation significantly decreased the mRNA expression of the jejunal mucosa SGLT1 gene compared with the control group (P < 0.05). However, the addition of SGs mixture did not affect the mRNA expression of the jejunal mucosa GLUT4 gene in weaned piglets (P > 0.05).

Fig. 2
figure 2

Bar graphs show the effect of dietary steviol glycosides mixture supplementation on the relative mRNA expression of chemoreceptors in the jejunum of weaned piglets. All data are expressed as the mean ± SEM (n = 6). Differences were determined by one-way ANOVA followed by LSD test. abc Means in the columns with different superscripts differ (P < 0.05)

Full size image

Biochemical parameters

As shown in Table 6, increasing SGs mixture supplementation linearly decreased serum GLU levels (P < 0.05). Furthermore, 100 and 150 mg/kg SGs mixture supplementation significantly reduced the concentration of serum TBIL compared with the control group (P < 0.05). However, the SGs mixture supplementation did not have a significant impact on the levels of serum TP, ALB, URE, TG, CHO, HDL-C, LDL-C, ALT, AST, ALP, and CRE (P > 0.05).

Table 6 Effects of dietary steviol glycosides mixture supplementation on serum biochemical parameters of weaned piglets
Full size table

Immunological function indicators

As shown in Table 7, increasing SGs mixture supplementation linearly decreased the proinflammatory cytokine IL-1β levels in serum (P < 0.05). However, there were no significant differences in the levels of serum IgA, IgG, IgM, IL-1, IL-6, IL-8, TNF-α, TNF-β, IL-10, and IL-22 among different treatments (P > 0.05).

Table 7 Effects of dietary steviol glycosides mixture supplementation on immunological function indicators of weaned piglets
Full size table

Antioxidant capacity

As shown in Tables 8 and 150 mg/kg SGs mixture supplementation resulted in a significant reduction in serum MDA content compared with the control group (P < 0.05). Increasing SGs mixture supplementation from 0 to 300 mg/kg linearly and quadratically increased serum T-SOD, CAT, and GSH-Px activities (P < 0.05). Piglets fed a diet supplemented with 100 mg/kg SGs mixture had higher serum T-SOD, CAT, and GSH-Px activities (P < 0.05). Moreover, increasing SGs mixture supplementation linearly increased hepatic T-AOC content (P < 0.05). Furthermore, increasing SGs mixture supplementation linearly and quadratically increased the hepatic T-SOD and GSH-Px activity, as well as muscle T-AOC content (P < 0.05). In contrast, increasing SGs mixture supplementation linearly and quadratically decreased the MDA content and T-SOD activity in the muscle (P < 0.05).

Table 8 Effects of dietary steviol glycosides mixture supplementation on the antioxidant capacity of weaned piglets
Full size table

Discussion

The utilization of SGs in livestock and poultry production has attracted significant attention due to their beneficial effects on enhancing production performance, feed efficiency, and the quality of animal products. For instance, research conducted by Jiang et al. [14] demonstrated that dietary 250 mg/kg stevioside supplementation significantly increased body weight, ADG, and ADFI in Ross 308 broiler chickens. Furthermore, another study revealed that including 80 mg/kg of stevia-based sweeteners (which contained 0.5% SGs) in the diets of Cobalt line broiler chickens for 42 days significantly enhanced final body weight and ADG [23]. However, it is worth noting that Wu et al. [24] reported no significant effect of stevioside on ADG, ADFI, F/G, and immune organ index in Arbor Acres broilers. In a study involving Shandong black goats, Han et al. [16] reported that adding 400 to 800 mg/kg of stevioside significantly increased both forage hay consumption and total feed intake. Furthermore, introducing 0.3% stevioside into the diet of Hanwoo cattle enhanced final weight, weight gain, and carcass crude protein content, while also reducing drip loss, shear force, and increasing meat color redness value in longissimus thoracis, thereby improving meat quality [25]. Additionally, including 0.3% stevia in the diet of growing pigs significantly improved ADG, feed utilization, body immunity, and carcass traits including backfat thickness [26]. A study indicated that increasing the concentration of stevioside or rebaudioside A from 0 to 300 mg/kg led to a linear increase in ADFI and ADG, while also resulting in a linear decrease in F/G from days 1 to 28 in weaned piglets [27]. Moreover, the addition of 167 mg/kg of stevia significantly boosted the ADG of weaned piglets in the second week post-weaning. However, varying proportions of stevia (0.0833%, 0.167%, or 0.334%) did not significantly affect feed intake or F/G in weaned piglets [18], which is consistent with similar findings from this experiment noting no significant effect of the SGs mixture on ADG, ADFI, and F/G in weaned piglets. Our research revealed a linear and quadratic relationship between the ADG and the SGs mixture supplemental levels, with ADFI exhibiting an initial increase followed by a decrease as the amount of SGs mixture in the diet increased. This implies that the effectiveness of SGs does not necessarily conform to a “more is better” principle.

The combination of certain sweeteners often leads to a synergistic sweetness effect [28]. A study conducted by Tian et al. [22] to assess the temporal perception of sweetness and bitterness for six commonly steviol glycosides indicated that Rubusoside and Stevioside display an immediate and pronounced bitter taste with a lingering aftertaste. In addition, SGs are commonly associated with a somewhat unpleasant bitter aftertaste, particularly at high concentration [29]. The main components of the SGs mixture used in this study are Rebaudioside A (39.90%) and Stevioside (30.40%). Therefore, the SGs mixture may show an unfavorable aftertaste experience for the piglets due to these reasons, despite the sweetness.

Sweet taste receptor cells were regulated by at least two signaling pathways, one mediated by a heterodimeric G-protein coupled receptor encoded by T1R2/T1R3 genes and another by glucose transporters and the ATP-gated potassium (KATP) channel [30]. The perception of sweetness is mainly facilitated by the sweet taste receptor T1R2/T1R3 present in taste cells of the lingual epithelium [31]. However, sweet taste receptors are also found in intestinal enteroendocrine cells [19, 32, 33]. Non-nutritive sweeteners have been shown to stimulate the mRNA expression of T1R2/T1R3 in the intestine of pigs [34]. Stevioside can enhance the function of the sweet taste transduction receptor called transient receptor potential melastatin 5 (TRPM5) [35], which is a calcium-activated cation channel present in type II taste receptor cells. In humans, stevioside was also reported to activate mRNA expression of T1R2/T1R3 [36]. Accordingly, our findings suggest that dietary SGs mixture supplementation can activate the mRNA expression of T1R2 and T1R3 in the jejunal mucosa of piglets.

A 2-dimensional organoid intestinal model experiment indicated that Rebaudioside A can significantly induce GLP-1 and CCK secretion [37]. GLP-1 is secreted from L cells in the intestine and exerts a strong incretin effect by enhancing insulin secretion in response to glucose levels and slowing down gastric emptying and motility [38]. Experiments with mouse and human intestinal enteroendocrine cell lines confirmed that Rebaudioside A stimulated GLP-1 release in a concentration-dependent manner via bitter taste signaling pathways. Contrary to this, we did not observe a rising in serum GLP-1 levels with increasing dietary SGs mixture supplementation. The discrepancy could be attributed to the short half-life of GLP-1 in the bloodstream. The levels of GLP-1 in plasma may not be a precise indicator of its local release in the intestine [39]. Consistently, we found a linear increase in serum CCK levels with increasing of SGs supplementation. CCK is released postprandially from the I cell of the small intestine into the bloodstream, and it has been reported to reduce food intake in both humans and rodents [40]. In the present study, the ADFI from days 1 to 42 decreased when the dietary supplementation of the SGs mixture reached 300 mg/kg. Our data suggest that the SGs mixture may function as an appetite suppressant when supplemented at a high concentration.

SGs are considered as a promising phytomedicine for managing diabetes. As a low-calorie, intensely sweet sugar substitute, it plays a pivotal role in improving the body’s blood glucose profile. Existing studies have indicated the effectiveness of stevia consumption in reducing postprandial blood glucose and insulin levels compared with aspartame and sucrose across lean and obese subjects [41]. When orally administered, SGs are resistant to degradation by gastric acid and digestive enzymes in the digestive tract. Only a small portion of SGs can be completely broken down into the aglycone steviol and glucose by the intestinal microflora in the lower intestinal tract [42,43,44]. Despite its sweetness, the limited absorption of SGs prevents a rapid spike in blood glucose levels post-ingestion. Deenadayalan et al. [45]. demonstrated that stevioside can effectively promote glucose uptake in diabetic gastrocnemius muscles by activating the insulin receptor (IR)/insulin receptor substrate-1(IRS-1)/Akt/GLUT4 pathway. Another study has suggested that SGs can mimic the effects of insulin by influencing GLUT translocation through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway [46]. Interestingly, we observed an upregulation of GLUT2 mRNA expression in the jejunum and a linear decrease in serum GLU content, but the underlying mechanism needs further research.

In broiler chickens, dietary SGs supplementation has been linked to elevated antibody levels against the Newcastle disease virus [23]. Insights into the mechanism of action of steviol glycosides have unveiled their modulatory effects on immune responses. These effects include the attenuation of pro-inflammatory factors like TNF-α, IL-1β, and IL-6 in mastitis-modeled mice by suppressing the toll-like receptor 2 (TLR2), nuclear factor kappa B (NF-κB), and MAPK pathways [47]. A study has indicated that SGs and steviol may have the potential to inhibit the release of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 induced by lipopolysaccharides (LPS) by modulating cytokine gene expression through the Iκ-Bα/NF-κB signaling pathway [48]. Consistent with these findings, our result demonstrates a significant linear decrease in serum IL-1β level with increasing SGs mixture supplementation, and indicates the potential of enhancing immune function and reducing inflammation of SGs mixture.

Weaning stress can damage the oxidation-antioxidant system and induce oxidative stress by decreasing the activity of SOD and increasing the concentration of MDA and NO [49]. Stevioside has been shown to mitigate diquat-induced cytotoxicity, inflammation, and apoptosis in IPEC-J2 cells, preserving cellular barrier integrity and combating oxidative stress by modulating the NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways [21]. In diabetic rats, SGs treatments have also demonstrated the ability to significantly enhance the activity of T-SOD and CAT in the liver [50]. Moreover, administration of 50 mg/kg stevioside has been reported to decrease oxidative stress markers such as 4-hydroxynonenal (4-HNE) and 3-nitrotyrosine (3-NT) and alleviate cisplatin-induced oxidative stress [51]. Furthermore, dietary stevioside supplementation normalized LPS-induced changes in protein expression of the antioxidant genes of nuclear factor-erythroid 2-related factor 2 (Nrf-2) and heme oxygenase-1 (HO-1), and ameliorated the redox damage by reducing MDA content and increasing total antioxidant capacity in boiler chickens [52]. In aged breeder hens, 0.25 g/kg stevioside supplementation significantly enhanced the antioxidant capacity of the ovary and shell glands by increasing the activity of CAT, SOD, or GSH-Px and reducing the MDA content [53]. Similarly, our findings also highlight that dietary SGs mixture supplementation can improve the antioxidant capacity of weaned piglets.

Conclusions

In conclusion, our findings indicate that dietary 100 ~ 150 mg/kg SGs mixture supplementation modulates gene expression of sweet taste recognition receptors and glucose transporters, while also enhancing the antioxidant capacity of weaned piglets.

Data availability

No datasets were generated or analysed during the current study.

Abbreviations

SGs:

Steviol glycosides

BW:

Body weight

ADG:

Average daily gain

ADFI:

Average daily feed intake

F/G:

Feed to gain ratio

GHRL:

Ghrelin

GLP-1:

Glucagon-like peptide-1

CCK:

Cholecystokinin

LEP:

Leptin

INS:

Insulin

SGLT-1:

Sodium glucose cotransporter-1

GLUT2:

Glucose transporters 2

GLUT4:

Glucose transporters 4

T1R2:

Taste receptor family 1 member 2

T1R3:

Taste receptor family 1 member 3

GLU:

Glucose

TP:

Total protein

ALB:

Albumin

URE:

Urea

TG:

Triglyceride

CHO:

Total cholesterol

HDL-C:

High-density lipoprotein cholesterol

LDL-C:

Low-density lipoprotein cholesterol

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

TBIL:

Total bilirubin

ALP:

Alkaline phosphatase

CHO:

Total cholesterol

CRE:

Creatinine

IgA:

Immunoglobulin A

IL-1:

Interleukin-1

IFN-γ:

Interferon-γ

TNF-α:

Tumor necrosis factor-α

MDA:

Malondialdehyde

T-AOC:

Total antioxidant capacity

T-SOD:

Total superoxide dismutase

CAT:

Catalase

GSH-Px:

Glutathione peroxidase

TLR2:

Toll-like receptor 2

NF-κB:

Nuclear factor kappa B

MAPK:

Mitogen-activated protein kinase

LPS:

Lipopolysaccharides

4-HNE:

4-hydroxynonenal

3-NT:

3-nitrotyrosine

Nrf-2:

Factor-erythroid 2-related factor 2

HO-1:

Heme oxygenase-1

References

  1. Mooradian AD, Smith M, Tokuda M. The role of artificial and natural sweeteners in reducing the consumption of table sugar: a narrative review. Clin Nutr ESPEN. 2017;18:1–8.

    Article  PubMed  Google Scholar 

  2. Wolwer-Rieck U. The leaves of Stevia rebaudiana (Bertoni), their constituents and the analyses thereof: a review. J Agric Food Chem. 2012;60(4):886–95.

    Article  PubMed  Google Scholar 

  3. Lemus-Mondaca R, Vega-Galvez A, Zura-Bravo L, Ah-Hen K. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: a comprehensive review on the biochemical, nutritional and functional aspects. Food Chem. 2012;132(3):1121–32.

    Article  CAS  PubMed  Google Scholar 

  4. Brusick DJ. A critical review of the genetic toxicity of steviol and steviol glycosides. Food Chem Toxicol. 2008;46(Suppl 7):S83–91.

    Article  CAS  PubMed  Google Scholar 

  5. Orellana-Paucar AM. Steviol glycosides from Stevia rebaudiana: an updated overview of their sweetening activity, Pharmacological Properties, and Safety aspects. Molecules. 2023;28(3):1258.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Chatsudthipong V, Muanprasat C. Stevioside and related compounds: therapeutic benefits beyond sweetness. Pharmacol Ther. 2009;121(1):41–54.

    Article  CAS  PubMed  Google Scholar 

  7. Gupta E, Purwar S, Sundaram S, Rai GK. Nutritional and therapeutic values of Stevia rebaudiana: a review. Acad Journals. 2013;7(46):3343–53.

    CAS  Google Scholar 

  8. Momtazi-Borojeni AA, Esmaeili SA, Abdollahi E, Sahebkar A. A review on the Pharmacology and Toxicology of Steviol glycosides extracted from Stevia rebaudiana. Curr Pharm Des. 2017;23(11):1616–22.

    Article  CAS  PubMed  Google Scholar 

  9. Chughtai MF, Pasha I, Zahoor T, Khaliq A, Ahsan S, Wu Z, et al. Nutritional and therapeutic perspectives of Stevia rebaudiana as emerging sweetener; a way forward for sweetener industry. CyTA - J Food. 2020;18:164–77.

    Article  Google Scholar 

  10. Clouard C, Val-Laillet D. Impact of sensory feed additives on feed intake, feed preferences, and growth of female piglets during the early postweaning period. J Anim Sci. 2014;92(5):2133–40.

    Article  CAS  PubMed  Google Scholar 

  11. Zhu L, Wang G, Dong B, Peng CC, Tian YY, Gong LM. Effects of sweetener neotame on diet preference, performance and hematological and biochemical parameters of weaned piglets. Anim Feed Sci Technol. 2016;214:86-94.

  12. Lee C, Yun W, Lee J, Kwak W, Oh H, An J et al. Effects of artificial sweeteners on feed palatability and performance in weaned pigs. Can J Anim Sci. 2019;99:307-14.

  13. Zhang W, He H, Gong L, Lai W, Dong B, Zhang L. Effects of sweetener sucralose on diet preference, growth performance and hematological and biochemical parameters of weaned piglets. Asian-Australasian J Anim Sci. 2020;33(5):802–11.

  14. Jiang J, Liu S, Jamal T, Ding T, Qi L, Lv Z, et al. Effects of dietary sweeteners supplementation on growth performance, serum biochemicals, and jejunal physiological functions of broiler chickens. Poult Sci. 2020;99(8):3948–58.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Jiang J, Qi L, Lv Z, Wei Q, Shi F. Dietary stevioside supplementation increases feed intake by altering the hypothalamic transcriptome profile and gut microbiota in broiler chickens. J Sci Food Agric. 2021;101(5):2156–67.

    Article  CAS  PubMed  Google Scholar 

  16. Han X, Chen C, Zhang X, Wei Y, Tang S, Wang J et al. Effects of Dietary Stevioside supplementation on feed intake, digestion, Ruminal Fermentation, and blood metabolites of goats. Anim (Basel). 2019;9(2).

  17. Wang LS, Shi Z, Shi BM, Shan AS. Effects of dietary stevioside/rebaudioside A on the growth performance and diarrhea incidence of weaned piglets. Anim Feed Sci Technol. 2014;187:104–9.

    Article  CAS  Google Scholar 

  18. Munro PJ, Lirette A, Anderson DM, Ju HY. Effects of a new sweetener, Stevia, on performance of newly weaned pigs. Can J Anim Sci. 2000;80(3):529–31.

    Article  Google Scholar 

  19. Steensels S, Depoortere I. Chemoreceptors in the gut. Annu Rev Physiol. 2018;80(1):117–41.

    Article  CAS  PubMed  Google Scholar 

  20. Novais AK, Deschêne K, Martel-Kennes Y, Roy C, Laforest JP, Lessard M, et al. Weaning differentially affects mitochondrial function, oxidative stress, inflammation and apoptosis in normal and low birth weight piglets. PLoS ONE. 2021;16(2):e0247188.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Xu Q, Liu M, Chao X, Zhang C, Yang H, Chen J, et al. Stevioside Improves Antioxidant Capacity and Intestinal Barrier Function while Attenuating Inflammation and Apoptosis by Regulating the NF-κB/MAPK Pathways in Diquat-Induced Oxidative Stress of IPEC-J2 Cells. Antioxidants. 2023;12(5):1070.

  22. Tian X, Zhong F, Xia Y. Dynamic characteristics of sweetness and bitterness and their correlation with chemical structures for six steviol glycosides. Food Res Int. 2022;151:110848.

    Article  CAS  PubMed  Google Scholar 

  23. Molina-Barrios RM, Aviles-Trejo CR, Puentes-Mercado ME, Cedillo-Cobian JR, Hernandez-Chavez JF. Effect of dietary stevia-based sweetener on body weight and humoral immune response of broiler chickens. Vet World. 2021;14(4):913–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Wu X, Yang P, Sifa D, Wen Z. Effect of dietary stevioside supplementation on growth performance, nutrient digestibility, serum parameters, and intestinal microflora in broilers. Food Funct. 2019;10(5):2340–6.

    Article  CAS  PubMed  Google Scholar 

  25. Shin Y, Rathnayake D, Mun H, Dilawar M, Pov S, Yang C-J. Sensory attributes, microbial activity, fatty acid composition and Meat Quality Traits of Hanwoo Cattle Fed a Diet supplemented with Stevioside and Organic Selenium. Foods. 2021;10:129.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Choi J-S, Jung D-S, Lee J-H, Choi YI, Lee J-JJKJFSAR. Growth Performance, Immune Response and Carcass Characteristics of Finishing Pigs by Feeding Stevia and Charcoal. 2012;32:228 – 33.

  27. WS Z, Shan LSBM. Effects of dietary stevioside/rebaudioside A on the growth performance and diarrhea incidence of weaned piglets. Anim Feed Sci Technol. 2014;187:104–9.

    Article  Google Scholar 

  28. Fujiwara S, Imada T, Nakagita T, Okada S, Nammoku T, Abe K, et al. Sweeteners interacting with the transmembrane domain of the human sweet-taste receptor induce sweet-taste synergisms in binary mixtures. Food Chem. 2012;130(3):561–8.

    Article  CAS  Google Scholar 

  29. Ye F, Yang R, Hua X, Shen Q, Zhao W, Zhang W. Modification of steviol glycosides using α-amylase. LWT - Food Sci Technol. 2014;57(1):400–5.

    Article  CAS  Google Scholar 

  30. Sukumaran SK, Palayyan SR. Sweet taste signaling: the Core pathways and Regulatory mechanisms. Int J Mol Sci. 2022;23(15):8225.

  31. Mack A, Singh A, Gilroy C, Ireland W. Porcine lingual taste buds: a quantitative study. Anat Rec. 1997;247(1):33–7.

    Article  CAS  PubMed  Google Scholar 

  32. Colombo M, Trevisi P, Gandolfi G, Bosi P. Assessment of the presence of chemosensing receptors based on bitter and fat taste in the gastrointestinal tract of young pig. J Anim Sci. 2012;90(Suppl 4):128–30.

    Article  PubMed  Google Scholar 

  33. Daly K, Al-Rammahi M, Arora DK, Moran AW, Proudman CJ, Ninomiya Y, et al. Expression of sweet receptor components in equine small intestine: relevance to intestinal glucose transport. Am J Physiol Regul Integr Comp Physiol. 2012;303(2):R199–208.

    Article  CAS  PubMed  Google Scholar 

  34. da Silva EC, de Jager N, Burgos-Paz W, Reverter A, Perez-Enciso M, Roura E. Characterization of the porcine nutrient and taste receptor gene repertoire in domestic and wild populations across the globe. BMC Genomics. 2014;15(1):1057.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Philippaert K, Pironet A, Mesuere M, Sones W, Vermeiren L, Kerselaers S, et al. Steviol glycosides enhance pancreatic beta-cell function and taste sensation by potentiation of TRPM5 channel activity. Nat Commun. 2017;8:14733.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Hellfritsch C, Brockhoff A, Stahler F, Meyerhof W, Hofmann T. Human psychometric and taste receptor responses to steviol glycosides. J Agric Food Chem. 2012;60(27):6782–93.

    Article  CAS  PubMed  Google Scholar 

  37. van der Wielen N, Ten Klooster JP, Muckenschnabl S, Pieters R, Hendriks HF, Witkamp RF, et al. The noncaloric sweetener Rebaudioside A stimulates Glucagon-Like peptide 1 release and increases enteroendocrine cell numbers in 2-Dimensional mouse Organoids Derived from different locations of the intestine. J Nutr. 2016;146(12):2429–35.

    Article  PubMed  Google Scholar 

  38. Nadkarni P, Chepurny OG, Holz GG. Regulation of glucose homeostasis by GLP-1. Prog Mol Biol Transl Sci. 2014;121:23–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Holst JJ. Glucagon and glucagon-like peptides 1 and 2. Results Probl Cell Differ. 2010;50:121–35.

    CAS  PubMed  Google Scholar 

  40. Cawthon CR, de La Serre CB. The critical role of CCK in the regulation of food intake and diet-induced obesity. Peptides. 2021;138:170492.

    Article  CAS  PubMed  Google Scholar 

  41. Anton SD, Martin CK, Han H, Coulon S, Cefalu WT, Geiselman P, et al. Effects of stevia, Aspartame, and sucrose on food intake, satiety, and postprandial glucose and insulin levels. Appetite. 2010;55(1):37–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Koyama E, Sakai N, Ohori Y, Kitazawa K, Izawa O, Kakegawa K, et al. Absorption and metabolism of glycosidic sweeteners of stevia mixture and their aglycone, steviol, in rats and humans. Food Chem Toxicol. 2003;41(6):875–83.

    Article  CAS  PubMed  Google Scholar 

  43. Geuns JM, Augustijns P, Mols R, Buyse JG, Driessen B. Metabolism of stevioside in pigs and intestinal absorption characteristics of stevioside, rebaudioside A and steviol. Food Chem Toxicol. 2003;41(11):1599–607.

    Article  CAS  PubMed  Google Scholar 

  44. Gardana C, Simonetti P, Canzi E, Zanchi R, Pietta P. Metabolism of stevioside and rebaudioside A from Stevia rebaudiana extracts by human microflora. J Agric Food Chem. 2003;51(22):6618–22.

    Article  CAS  PubMed  Google Scholar 

  45. Deenadayalan A, Subramanian V, Paramasivan V, Veeraraghavan VP, Rengasamy G, Coiambatore Sadagopan J et al. Stevioside attenuates insulin resistance in skeletal muscle by facilitating IR/IRS-1/Akt/GLUT 4 signaling pathways: an in vivo and in Silico Approach. Molecules. 2021;26(24).

  46. Rizzo B, Zambonin L, Angeloni C, Leoncini E, Dalla Sega FV, Prata C, et al. Steviol glycosides modulate glucose transport in different cell types. Oxid Med Cell Longev. 2013;2013:348169.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Wang T, Guo M, Song X, Zhang Z, Jiang H, Wang W, et al. Stevioside plays an anti-inflammatory role by regulating the NF-κB and MAPK pathways in S. aureus-infected mouse mammary glands. Inflammation. 2014;37(5):1837–46.

    Article  CAS  PubMed  Google Scholar 

  48. Boonkaewwan C, Burodom A. Anti-inflammatory and immunomodulatory activities of stevioside and steviol on colonic epithelial cells. J Sci Food Agric. 2013;93(15):3820–5.

    Article  CAS  PubMed  Google Scholar 

  49. Tang X, Xiong K, Fang R, Li M. Weaning stress and intestinal health of piglets: a review. Front Immunol. 2022;13:1042778.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Shivanna N, Naika M, Khanum F, Kaul VK. Antioxidant, anti-diabetic and renal protective properties of Stevia rebaudiana. J Diabetes Complications. 2013;27(2):103–13.

    Article  PubMed  Google Scholar 

  51. Potocnjak I, Broznic D, Kindl M, Kropek M, Vladimir-Knezevic S, Domitrovic R. Stevia and stevioside protect against cisplatin nephrotoxicity through inhibition of ERK1/2, STAT3, and NF-kappaB activation. Food Chem Toxicol. 2017;107(Pt A):215–25.

    Article  CAS  PubMed  Google Scholar 

  52. Jiang J, Qi L, Lv Z, Jin S, Wei X, Shi F. Dietary stevioside supplementation alleviates Lipopolysaccharide-Induced Intestinal Mucosal damage through anti-inflammatory and antioxidant effects in broiler chickens. Antioxid (Basel). 2019;8(12):575.

  53. Jingle J, Lina Q, Hongjian D, Chenhui H, Zengpeng L, Quanwei W et al. Dietary stevioside supplementation improves laying performance and eggshell quality through increasing estrogen synthesis, calcium level and antioxidant capacity of reproductive organs in aged breeder hens. Anim Feed Sci Technol. 2020;269:114682.

Download references

Acknowledgements

We would like to thank the staff at our laboratory for their ongoing assistance.

Funding

This work was supported by financial support from the Special Fund for Rural Revitalization Strategy of Guangdong (2023TS-3), Key Projects of Maoming Laboratory (2022ZD003), the earmarked fund for CARS-35, the Science and Technology Program of Guangdong Academy of Agricultural Sciences (202106TD), and the Open Project of Guangdong Provincial Key Laboratory of Animal Breeding and Nutrition (2022SZ02).

Author information

Author notes

  1. Yunxia Xiong and Zhentao He have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Animal Nutrition and Feed Science in South China Ministry of Agriculture, Heyuan Branch,Guangdong Laboratory for Lingnan Modern Agriculture, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong, 510640, China

    Yunxia Xiong, Zhentao He, Qiwen Wu, Hao Xiao, Shuting Cao, Xuefen Yang, Zongyong Jiang & Li Wang

  2. School of Animal Science and Technology, Foshan University, Foshan, 528225, China

    Zhentao He & Cui Zhu

  3. Dongtai Hirye Biotechnology Co., Ltd, Dongtai, 224200, China

    Yajing Li

Authors
  1. Yunxia Xiong
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Zhentao He
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Qiwen Wu
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Hao Xiao
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Shuting Cao
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Xuefen Yang
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Yajing Li
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Zongyong Jiang
    View author publications

    You can also search for this author inPubMed Google Scholar

  9. Cui Zhu
    View author publications

    You can also search for this author inPubMed Google Scholar

  10. Li Wang
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

LW, CZ, and ZYJ conceived and designed the trial. YJL analyzed the compositions of the SGs mixture. ZTH executed animal breeding and management. YXX, QWW, HX, STC, and XFY performed the sampling and parameter measurements. YXX and ZTH analyzed the data and wrote the manuscript. All authors approved the submitted version.

Corresponding authors

Correspondence to Cui Zhu or Li Wang.

Ethics declarations

Ethics statement

All animal experimental protocols used in the current study were according to the Chinese guidelines for animal welfare and approved by the Animal Care and Use Committee of the Guangdong Academy of Agricultural Sciences (GAASIAS-2022021). Written informed consent was obtained from the owners for the participation of their animals in this study.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xiong, Y., He, Z., Wu, Q. et al. Dietary steviol glycosides mixture supplementation modulates the gene expression of gut chemoreceptors and enhances the antioxidant capacity in weaned piglets. Porc Health Manag 11, 6 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s40813-024-00414-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s40813-024-00414-5

Keywords

Porcine Health Management

ISSN: 2055-5660

Contact us