Synbiotics for improving growth performance of weaned piglets and application of synbiotics

Abstract

The invention relates to a synbiotics for improving growth performance of weaned piglets and application thereof. The synbiotics comprise the following components: a) a probiotic component: bacterial powder of Bifidobacterium infantis B.infantis-zju (the preservation number is CCTCC NO: 20251039), the bacterial powder is prepared by culturing the bacterial strain in a TPY culture medium under anaerobic conditions, concentrating, freeze-drying and adding maltodextrin; and b) a prebiotic component: 3 '-sialylated milk oligosaccharide (3'-SL) with the purity of more than 95%, and the addition amount of the prebiotic component in the daily ration is 5g / kg. The synbiotics provided by the invention can significantly reduce the diarrhea rate of piglets after weaning, significantly improve the average daily gain of pigs in the nursing period, and reduce the feed-to-weight ratio. In addition, the synbiotics can also significantly reduce the level of proinflammatory factors in the serum of the weaned piglets and significantly improve the level of anti-inflammatory factors in the serum of the weaned piglets, which prompts that the synbiotics can reduce the level of intestinal inflammation and improve intestinal health, and has a great application prospect in healthy breeding of live pigs.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, and particularly relates to a synbiotic for improving growth performance of weaned piglets and use thereof. BACKGROUND

[0002] The pig industry is the backbone of China's animal husbandry. The weaning period of piglets is a critical stage in breeding. At this stage, the immune function and intestinal development of piglets are not yet perfect, and they are easily affected by factors such as feed conversion stress, environmental stress and intestinal flora imbalance, often resulting in growth retardation, high incidence of diarrhea and immune suppression, and causing huge economic losses. The weaning period is an important period for shaping the intestinal microorganisms, perfecting the digestive function and activating the immunity, and has a lasting impact on the later growth and development. Before and after weaning, piglets are prone to be infected by pathogenic bacteria due to the incomplete development of intestinal and immune functions, which can cause diarrhea and hinder growth, thereby reducing economic benefits.

[0003] How to take effective measures to prevent and control piglet diarrhea, reduce piglet mortality and improve piglet growth performance has become an important task for pig production. In view of the current breeding situation in China, how to realize the replacement of pig breeding feed antibiotics is of great significance to the healthy and sustainable development of animal husbandry. The research and development of probiotics and prebiotics and other feed additives are a candidate target that scientists at home and abroad are optimistic about, and have become a research hotspot in recent years. In recent years, more and more studies have shown that early intestinal microbial colonization and dynamic balance are one of the important factors affecting piglet intestinal health, and a stable microbial flora plays an important role in relieving post-weaning stress, achieving smooth transition after weaning and exerting piglet growth performance. Adding probiotics or prebiotics in the diet during the weaning period can promote the proliferation of beneficial microorganisms, inhibit the colonization of pathogenic bacteria, reduce the inflammatory response of the body, increase the weaning weight, and improve the adaptability of piglets after weaning. Importantly, increasing weaning weight significantly shortens the feeding time of the whole cycle and increases the growth performance in the later period. Therefore, in-depth understanding of the regulation mechanism of piglet intestinal development and barrier function is of great significance to piglet health, further strengthening the nutritional intervention during the weaning period, and fully exerting the growth performance and improving the economic benefits in the later period.

[0004] Synbiotics is a mixture of probiotics and prebiotics. It produces beneficial effects on host health by promoting the survival and colonization of live microbial supplements in the host gastrointestinal tract, and the combination of the two can overcome the limitations of single components. However, the design of synbiotics for weaned piglets needs to be based on two aspects: first, it needs to ensure the compatibility of prebiotics and probiotics to enhance their colonization ability and metabolic activity; second, it needs to design combinations based on the specific needs of the host to optimize the balance of the microbial flora and targeted efficacy. Existing studies have shown that synbiotics composed of lacto-oligosaccharides and bifidobacterium may produce synergistic effects and affect piglet health by regulating intestinal flora and the immune system.

[0005] Therefore, it is of great significance to develop a synbiotic for improving the growth performance of weaned piglets and to strengthen the nutritional intervention in the weaning stage, so as to fully exert the growth performance and improve the economic benefits in the later stage. SUMMARY

[0006] In order to overcome the shortcomings of the prior art, the purpose of the present application is to provide a synbiotic for improving the growth performance of weaned piglets and the use thereof.

[0007] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows: A synbiotic for improving the growth performance of weaned piglets, comprising the following components: a) probiotic component: infant Bifidobacterium longum subsp. B. infantis-zju (CCTCC NO: 20251039, preserved in China Center for Type Culture Collection, No. 299, Baying Road, Wuchang District, Wuhan City, Hubei Province on May 13, 2025) powder, which is prepared by adding malt dextrin to the powder of the strain after being cultured in TPY medium under anaerobic conditions, and then being concentrated, freeze-dried; b) prebiotic component: 3'-sialylated lacto-oligosaccharide (3'-SL), with a purity of more than 95%, and the addition amount thereof in the daily ration is 5 g / kg.

[0008] The strain is cultured in TPY medium under anaerobic conditions: Culture medium formula: hydrolyzed casein (10.0 g / L), yeast powder (2.0 g / L), plant peptone (5.0 g / L), glucose (5.0 g / L), dipotassium hydrogen phosphate (2.0 g / L), calcium chloride (0.15 g / L), magnesium chloride (0.5 g / L), ferric chloride (0.0001 g / L), zinc sulfate (0.25 g / L), L-cysteine (0.5 g / L), Tween 80 (1.0 g / L), pH value (6.5±0.1); Temperature condition: 37℃; gas condition: ternary mixed gas (10% H2, 10% CO2, 80% N2); The malt dextrin content in the powder is 5 g / 100 g.

[0009] The use of the synbiotic in the preparation of a product for reducing the diarrhea rate and improving the growth performance of weaned piglets, for reducing the diarrhea rate and improving the growth performance.

[0010] The use (a) diluting the probiotic powder with sterile PBS to a concentration of 1×10 9 CFU / mL; (b) orally gavaging 1 mL of the diluent to 21-day-old weaned piglets per day; (c) adding 5 g / kg of 3'-SL in the piglet diet, continuously using for 14 days.

[0011] 5. The use according to claim 3, which is applied to 21-day-old weaned piglets of 'Changbai' and 'Dabai', and is used for reducing the diarrhea rate of weaned piglets and improving the production performance of piglets; after weaning the piglets at the age of 21 days, 5 g / kg of 3'-sialylated milk oligosaccharide is added in the diet, and 1 mL of the diluent of probiotics is orally gavaged, so as to reduce the diarrhea rate of weaned piglets and the feed conversion rate in the rearing period, and improve the average daily gain of piglets in the rearing period.

[0012] The beneficial effects of the present application are: 1. The diarrhea rate of weaned piglets is significantly reduced, the average daily gain of piglets in the rearing period is significantly improved, and the feed conversion rate is reduced.

[0013] 2. The level of pro-inflammatory factors in the serum of weaned piglets is significantly reduced, the level of anti-inflammatory factors in the serum is significantly improved, and the immunity of the body is enhanced. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 It is a schematic diagram of the gavaging device of the bacterial solution.

[0015] Figure 2 It is the grouping situation of the test.

[0016] Figure 3A It is the influence of the synbiotic provided by the present application on the cytokines in the serum of weaned piglets (IL-1β).

[0017] Figure 3B It is the influence of the synbiotic provided by the present application on the cytokines in the serum of weaned piglets (IL-6).

[0018] Figure 3C It is the influence of the synbiotic provided by the present application on the cytokines in the serum of weaned piglets (IFN-γ).

[0019] Figure 3D It is the influence of the synbiotic provided by the present application on the cytokines in the serum of weaned piglets (TNF-α).

[0020] Figure 3E It is the influence of the synbiotic provided by the present application on the cytokines in the serum of weaned piglets (IL-10).

[0021] Figure 4A It is the influence of the synbiotic provided by the present application on the immunoglobulin in the serum of weaned piglets (IgA).

[0022] Figure 4BThe present application provides the influence of the synbiotic on serum immunoglobulin (IgM) of weaned piglets.

[0023] Figure 4C The present application provides the influence of the synbiotic on serum immunoglobulin (IgG) of weaned piglets. DETAILED DESCRIPTION

[0024] The present application is further described below in conjunction with the accompanying drawings and examples.

[0025] Example 1: Culture of probiotic bacterial strains The strains are cultured in TPY medium under anaerobic conditions: Medium formula: hydrolyzed casein (10.0 g / L), yeast powder (2.0 g / L), vegetable peptone (5.0 g / L), glucose (5.0 g / L), potassium phosphate dibasic (2.0 g / L), calcium chloride (0.15 g / L), magnesium chloride (0.5 g / L), ferric chloride (0.0001 g / L), zinc sulfate (0.25 g / L), L-cysteine (0.5 g / L), Tween 80 (1.0 g / L), pH value (6.5±0.1), temperature condition (37±0.5℃), gas condition: ternary mixed gas (10% H2, 10% CO2, 80% N2).

[0026] Process stability: after being cultured in TPY medium under strict anaerobic conditions, the strains are treated by concentration and freeze-drying process to ensure the stability of the activity of the strains, and a proper amount of malt dextrin is added as a protective agent during the freeze-drying process to improve the stability of the bacterial powder during storage, wherein the content of malt dextrin in the bacterial powder is 5 g / 100 g.

[0027] Example 2: Dilution and gavage of probiotics 1) Dilution operation Take 100 g of bacterial powder, dilute and mix with 100 mL of PBS to prepare a probiotic dilution bacterial solution, and transfer all to a veterinary gavage device (such as Figure 1 shown, the gavage device is a commercially available self-assembled device produced by Nantong Pin'ag Medical Instrument Co., Ltd., and the continuous syringe is a 1 mL A) for gavage.

[0028] 2) Gavage operation The probiotic dilution bacterial solution in the gavage device is gavaged to 21-day-old weaned piglets, and the test grouping design is shown in Figure 2 . The specific operation is as follows: one person carefully holds up the piglet, holds the head of the piglet slightly higher than the parallel ground, and the other person uses the gavage device to inject 1 mL of the dilution bacterial solution (10 9 CFU of live bacteria per piglet) or PBS into the root of the tongue of the piglet through the gavage head, and then releases the piglet after it completely swallows.

[0029] Example 3: Feeding trial to prevent diarrhea in weaned piglets Adding probiotics: Feed the probiotics into the diluted bacterial solution in the feeding device according to the feeding procedure described above.

[0030] Prebiotic addition: 3'-sialoyl lactoligosaccharide (3'-SL) was purchased from Shaanxi Panier Biotechnology Co., Ltd., with a purity greater than 95%, and was added to the diet at a fixed ratio of 5g / kg to ensure a stable intake for each piglet.

[0031] Data recording: Record data such as piglets' initial weight, final weight, feed intake, diarrhea status, and feed conversion ratio.

[0032] Feeding experiment: Thirty-two castrated male Landrace × Large White crossbred piglets of similar initial weight at 21 days of age in their second parity were selected and randomly divided into 4 groups (see details of grouping). Figure 2 Each group consisted of 8 animals, and the treatments were as follows: Control group, B. infantis group, 3'-SL group, and B. infantis+3'-SL group. The Control group served as the blank control group, the B. infantis group as the probiotic group, the 3'-SL group as the lactooligosaccharide group, and the B. infantis+3'-SL group as the synbiotic group. The Control group was orally administered 1 mL of PBS daily, and the B. infantis group was orally administered 1 × 10⁻⁶ PBS daily. 9 1 mL of CFU / mL Bifidobacterium infantis (B. infantis) was administered orally to the following groups during the experimental period: 3'-SL group received 5 g / kg of lactooligosaccharide 3'-SL in addition to the diet; B. infantis + 3'-SL group received 1 × 10 CFU / mL of B. infantis + 3'-SL in addition to the 5 g / kg lactooligosaccharide 3'-SL in the diet. 9 1 mL of CFU / mL Bifidobacterium infantis (B. infantis) was administered. Feed intake, diarrhea, and other data were recorded throughout the process. At the end of the nursery stage, the body weight of each group was measured to assess the impact of the synbiotic on the growth performance of piglets.

[0033] Diarrhea rate in piglets during the Biostime application trial = Total number of piglets experiencing diarrhea during the trial period / (Total number of piglets in the trial × Number of days in the formal trial) × 100% Table 1. Effects of Example 3 of the present invention on growth performance and diarrhea rate of weaned piglets.

[0034] Note: Different shoulder note letters in the same line indicate significant differences. P < 0.05).

[0035] As shown in Table 1, there was no significant difference in initial body weight (6.52-6.59 kg) among the experimental groups of piglets. P Based on the baseline of > 0.05, growth performance showed significant changes after different interventions. Compared with the control group (8.16±0.55 kg), the final body weight of the 3'-SL group (9.05±0.40 kg) and the B. infantis + 3'-SL synbiotic group (9.34±0.68 kg) was significantly increased. P < 0.05, with the Synbiotic group showing the best performance. Regarding nutrient utilization efficiency, the average daily feed intake (352.25±21.38 g / d) and daily weight gain (191.81±31.25 g / d) of the Synbiotic group were significantly higher than those of the control group (289.51±17.45 g / d, 120.78±22.23 g / d), while the feed conversion ratio (1.83±0.03) was 23.4% lower than that of the control group (2.39±0.07). Health indicators showed that the incidence of diarrhea in the Synbiotic group (5.82±0.27%) was significantly lower than that in the control group (7.73±0.38%), and further improved compared to the group supplemented with *B. infantis* alone (6.68±0.36%) and the 3'-SL group (6.95±0.12%). The data indicate that the synergistic effect of *B. infantis* and 3'-SL can significantly improve the growth performance of piglets. P < 0.05), indicating that synbiotics have a synergistic effect in promoting weight gain, improving feed conversion rate, and reducing the risk of diarrhea.

[0036] Figures 3A-3E This invention relates to Example 3 of the effect of synbiotics (B. infantis combined with 3'-SL) on cytokines in the serum of weaned piglets. The present invention systematically evaluated the regulatory effect of synbiotics on inflammatory responses by analyzing the concentrations of cytokines IL-1β, IL-6, IL-10, IFN-γ, and TNF-α in the serum of weaned piglets. As shown in the figure, the synbiotic group (B. infantis + 3'-SL) significantly reduced the levels of pro-inflammatory cytokines IL-1β, IL-6, IFN-γ, and TNF-α, with the most significant inhibitory effect on pro-inflammatory cytokines IL-6 and TNF-α, significantly lowering these levels compared to B. infantis or 3'-SL treatment alone. P< 0.05), and it most effectively increased the content of the anti-inflammatory factor IL-10, exhibiting the strongest anti-inflammatory effect, suggesting that B. infantis and 3'-SL have a synergistic effect in regulating inflammatory pathways. These results indicate that synbiotics can restore inflammatory balance, alleviate weaning stress-induced systemic inflammatory responses, and increase the immune function of piglets by targeting and inhibiting the release of pro-inflammatory factors (IL-1β, IL-6, IFN-γ, and TNF-α) and promoting the increase of the anti-inflammatory factor IL-10.

[0037] Figure 4A , Figure 4B , Figure 4C This invention relates to Example 3 of the effect of synbiotics on serum immunoglobulins in weaned piglets. This invention systematically evaluated the regulatory effect of synbiotics on humoral immune function by analyzing serum immunoglobulin levels in weaned piglets. As shown in the figure, compared with the control group, the B. infantis+3'-SL synbiotic group significantly increased serum IgA concentration (…). P <0.05), while the effects of adding B. infantis or 3'-SL alone on IgA were weak. In IgM expression, B. infantis, 3'-SL, and the combined group B. infantis+3'-SL were all significantly higher than the control group ( ). P < 0.05), but there were no statistically significant differences among the treatment groups. P > 0.05). Furthermore, there was no statistically significant difference in IgG expression among the treatment groups compared to the control group ( P > 0.05), indicating that synbiotics may not have a direct regulatory effect on the IgG secretion pathway. These results suggest that combined intervention with B. infantis and 3'-SL can selectively promote the increase of serum IgA and IgM through synergistic effects. This finding echoes the results of previous studies on the targeted inhibition of pro-inflammatory factors by synbiotics, together revealing its potential mechanism of alleviating weaning stress by reshaping immune balance.

[0038] In summary, the synbiotic provided by this invention can significantly reduce the diarrhea rate in piglets after weaning, significantly increase the average daily weight gain during the nursery period, and reduce the feed conversion ratio. Furthermore, the synbiotic can also significantly reduce the levels of pro-inflammatory factors in the serum of weaned piglets, significantly increase the levels of anti-inflammatory factors in the serum of weaned piglets, and enhance the body's immune function.

[0039] The above embodiments can be further combined or replaced, and the embodiments only describe the preferred embodiments of the present application, and do not limit the concept and scope of the present application. Without departing from the design idea of the present application, various changes and improvements of the technical solutions of the present application made by those skilled in the art shall fall within the protection scope of the present application. The protection scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. A synbiotic for improving growth performance of weaned piglets, characterized in that, Comprise the following components: a) probiotic component: B. infantis-zju (CCTCC NO: 20251039) powder, which is prepared by concentrating, freeze-drying and adding malt dextrin after the strain is cultured in TPY medium under anaerobic conditions; b) prebiotic component: 3'-sialylated lactose oligosaccharide (3'-SL), purity greater than 95%, and the addition amount in the daily ration is 5g / kg.

2. The synbiotic of claim 1 for improving the growth performance of weaned piglets, characterized in that: The strain is cultured in TPY medium under anaerobic conditions: Medium formula: hydrolyzed casein (10.0 g / L), yeast powder (2.0 g / L), plant peptone (5.0 g / L), glucose (5.0 g / L), dipotassium hydrogen phosphate (2.0 g / L), calcium chloride (0.15 g / L), magnesium chloride (0.5 g / L), ferric chloride (0.0001 g / L), zinc sulfate (0.25 g / L), L-cysteine (0.5 g / L), Tween 80 (1.0 g / L), pH value (6.5±0.1); Temperature condition: 37℃; gas condition: ternary mixed gas (10% H2, 10% CO2, 80% N2); The malt dextrin content in the powder is 5g / 100g.

3. Use of the synbiotic of claim 1 in the manufacture of a product for reducing the incidence of diarrhea and improving growth performance in weaned piglets, characterized in that, For reducing the diarrhea rate and improving the growth performance.

4. The use of claim 3, characterized in that: (a) the probiotic bacteria powder is diluted in sterile PBS to a concentration of 1 x 10 9 CFU / mL; (b) orally gavage 1mL of the diluted solution to 21-day-old weaned piglets per day; (c) add 5g / kg of 3'-SL in the piglet diet, continuously use for 14 days.

5. Use according to claim 3, characterized in that, Applied to "Changbai ×Dabai" 21-day-old binary weaned piglets, for reducing the diarrhea rate of weaned piglets and improving the production performance of piglets; after weaning at 21 days of age, add 5g / kg of 3'-sialylated lactose oligosaccharide in the diet, and orally gavage 1mL of the probiotic diluted solution, to reduce the diarrhea rate of piglets after weaning and the feed conversion ratio during the nursery period, and to improve the average daily gain of pigs during the nursery period.

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