CN107811128A - A feed additive that can regulate intestinal flora of piglets - Google Patents

Abstract

The invention relates to a feed additive capable of regulating the intestinal flora of piglets, wherein the weight ratio of the active ingredient galactomannan oligosaccharide to ligustalactone A is (10:1)-(1:10). Compared with common feed additives, the feed additive of the invention has simple preparation method, reasonable compatibility, remarkable curative effect and can significantly improve intestinal flora of piglets. Since there are no western medicine ingredients in the additive, the feed additive has no toxic side effects and no drug resistance.

Description

A feed additive that can regulate intestinal flora of piglets

technical field

The invention relates to the field of feed, in particular to a feed additive for piglets.

Background technique

Since the discovery of the growth-promoting effect of antibiotics on animals in the middle of the 20th century, antibiotics have been widely used as feed additives in the aquaculture industry; in view of the negative effects of antibiotics and people’s growing concern about food health, many countries have enacted laws to ban antibiotics. and the laws and regulations on the use of chemically synthesized drugs. In recent years, research on the development of safe, non-toxic, and side-effect-free alternatives to antibiotics has begun.

Studies have confirmed that fructooligosaccharides can improve the production performance and disease resistance of pigs, chickens, cattle and other animals; mannan oligosaccharides can improve the production performance of mice, pigs, chickens, cattle, aquatic animals and other animals and promote beneficial bacteria in the intestines (such as bifidobacterium, lactobacillus, etc.) proliferation, and inhibit the reproduction of harmful bacteria (such as Escherichia coli), and improve the body's immunity.

Galactomannan-oligosaccharide (GMOS), also known as galactomanno-oligosaccharide, is an oligosaccharide composed of galactomannose and glucose connected by glycosidic bonds. It is a functional oligosaccharide variety independently developed in my country. According to reports, galactomannan oligosaccharides have the following functions: 1. By optimizing the intestinal micro-ecosystem, forming an intestinal flora with Bifidobacterium and Lactobacillus as the dominant bacteria, and playing a beneficial health care function for the host. 2. It has the functions of identifying, adhering and eliminating pathogenic microorganisms. 3. It can be an immunomodulator and has the effect of activating the response of the immune system. This feature has also aroused great interest. It is generally believed that mannan oligosaccharides can enhance the non-specific immunity of animals. In recent years, many researchers have verified its application effect on heterogeneous gibel carp juveniles, broiler chickens, meat rabbits, early weaned piglets, conventional weaned piglets, growing and finishing pigs and lactating sows.

At present, the main raw materials for the production and preparation of GMOS are galactomannan gums, such as squash gums. Squat gum is derived from the inner latex of Squatia seeds, and its chemical composition is galactomannan with a relative molecular mass of about 200,000 Da. Contains a small amount of protein, insoluble in various organic solvents, soluble in water. Squat gum is composed of mannose units to form the main chain, galactose units to form branch chains, and the ratio of galactose to mannose units is 1:2. The mannose units are connected by α-1,4 glycosidic bonds, and the galactose units are connected on the mannose backbone by β-1,6 glycosidic bonds. The production and preparation of galactomannan oligosaccharides are mainly obtained by enzymatically degrading squash gum.

120 heterogeneous gibel crucian carp were fed for 10 weeks. The results showed that adding 0.2% GMOS to the basal diet could significantly increase the weight gain rate and specific growth rate of heterogeneous gibel carp juveniles. It reduces the bait coefficient and can significantly improve the non-specific immune function of fish. In terms of weight gain rate, the 0.1%, 0.2% and 0.3% groups increased by 2.33, 30.0 and 18.1 percentage points respectively compared with the blank control group. In terms of specific growth rate, the 0.2% group and 0.3% group increased by 19.8 and 9.5 percentage points respectively compared with the blank control group; while the feed coefficient of 0.1%, 0.2% group and 0.3% group respectively decreased by 11% compared with the blank control group .1, 13.9 and 9.5 percentage points.

The test results show that GMOS can replace broiler dietary antibiotics (50mg/kg aureomycin + 40mg/kg salinomycin) to a certain extent, and can significantly increase the serum Newcastle disease antibody titer level, thereby enhancing broiler humoral and cellular immunity.

30 50-day-old New Zealand meat rabbits were fed for 20 days. The results showed that compared with the control group supplemented with 0.005% olaquindox, the survival rate of rabbits in the experimental group supplemented with 0.25% GMOS increased by 7.5%, and the feed-to-weight ratio and diarrhea rate decreased. 6.1% and 38.15%. In addition, the evisceration rate of the test group was increased by 2.22% compared with the control group after slaughter.

Studies have shown that GMOS has a preventive effect on diarrhea caused by Escherichia coli, and the effect is better than chitosan. GMOS can improve the growth potential of 14-day-old early-weaned piglets, as shown in: it can increase the concentration of insulin-like growth factor-1 (IGF-1) in serum, the concentration of growth hormone and the gene expression level of IGF-1 in liver and eye muscles, thereby through Mediated to play a growth-promoting role: GMOS can improve the immune function of 14-day-old early-weaned piglets. It is manifested in: the concentration of IgG, IgA, IgM, IL-2, IL-6 and IL-1β immune factors in piglet serum is increased, and the concentration of piglet peripheral blood mononuclear cytokine IL-1β, jejunal mucosa IL-1β and intestinal The expression level of IL-1β gene in the model lymph nodes can enhance the body's cellular immunity and humoral immunity. Studies have shown that feeding different concentrations of GMOS can reduce the number of ileal microbial populations in early-weaned piglets, inhibit harmful bacteria such as Staphylococcus aureus and Clostridium perfringens, and promote the reproduction of some beneficial bacteria such as Lactobacillus and Bifidobacterium; The colon segment can also reduce the number of E. coli and significantly increase the number of Lactobacillus and Bifidobacteria in the colon.

Experiments with 72 28-day-old weaned piglets showed that adding 0.12% GMOS to the diet could significantly increase the average daily feed intake and average daily gain, and reduce the incidence of diarrhea. Tests have shown that GMOS can completely replace chlortetracycline in 9.4-15kg piglet diets and achieve better growth performance, but the effect on 15-27kg piglets is inferior to 75mg/kg aureomycin.

45 Du×Yuan pigs weighing 25.3±2.3kg were fed for 94 days. The results showed that a certain proportion of yeast culture and GMOS was added to the diet of 25-90kg growing and finishing pigs. It can increase daily weight gain, reduce feed-to-weight ratio, and reduce diarrhea rate. Obtained good economic and social benefits. 0.10% GMOS was used to feed growing pigs for 96 days, and the results showed that compared with adding 50mg/kg aureomycin, it could increase the daily gain and reduce the incidence of gastrointestinal diseases such as diarrhea, but it had no improvement effect on respiratory diseases such as asthma; In terms of serum physiological and biochemical indicators, GMOS has a tendency to reduce serum cholesterol and triglyceride, can reduce serum aspartate aminotransferase and alanine aminotransferase activities to varying degrees, play a role in protecting the liver, and can also significantly increase serum GH, In terms of slaughter performance, GMOS has a tendency to reduce skin thickness, backfat thickness, eye muscle area and increase lean meat percentage. There is almost no effect on the growth and slaughter rate; in terms of pork quality, GMOS can significantly reduce the indole content in the eye muscle, improve the pork flavor, and also have a tendency to increase the water retention.

The test showed that when the nutritional level of the sow diet was the same and no antibiotics were added, the addition of GMOS could significantly increase the milk production, reduce the feed-to-milk ratio, and regulate the endocrine function of the sows compared with the non-supplemented group; When antibiotics are added, GMOS can also increase sow milk production, reduce feed-to-milk ratio, and can also significantly increase the weaned individual weight of piglets, and the improvement effect is significantly better than that of the control group and kitasamycin, which is equivalent to virginiamycin; Compared with the two antibiotic groups, the milk protein content of the galactomannooligosaccharide group was the highest, with an average increase of 14.9%.

Chinese medicine preparations are rich in various active ingredients such as polysaccharides, which play a significant role in improving the body's immunity and disease resistance. It has been proved that hundreds of Chinese herbal medicines have various immune activities, such as promoting the growth and development of immune organs, strengthening the body Cellular immunity and humoral immunity can promote the production of interferon-γ, interleukin-2 and other cytokines, and it is green, safe and not easy to produce drug resistance. Therefore, the development and utilization of Chinese herbal medicine has become popular.

Ligusticolides belong to lactone compounds, mainly including Lisantrolactone A, Lisantrolactone H, Lisantrolactone I, Lisantrolactone D, Lisantrin I, Lisantrine F, Lisantrin J , chuanxionglide M, chuanxionglide Q, etc., are relatively high in the traditional Chinese medicine Chuanxiong, Angelica and other Umbelliferae plants, are the main active components and metabolic components, and have a variety of pharmacological activities. Chuanxiong lactone compounds have various pharmacological effects such as anti-oxidative damage, anti-inflammatory and analgesic, anti-coagulation and anti-platelet aggregation, vasodilation, and cytotoxicity.

After incubating cardiomyocytes for 55 minutes with ligustilide A, hypoxia for 2 hours, and reoxygenation for 1 hour, lactate dehydrogenase (LDH) and malondialdehyde (MDA) in the supernatant were significantly reduced. The activity of superoxide dismutase (SOD) was increased, and compared with the blank control group, it could also reduce the activity of mitogen-activated protein kinase (MAPK) in cardiomyocytes, indicating that ligustilide A had no effect on neonatal rat cardiomyocytes cultured in vitro. Oxygen/reoxygenation injury has a protective effect, and the MAPK signaling pathway may be involved in this protective effect.

It has also been reported in the literature that pretreatment with Ligustilide A has a protective effect on hypoxia/reoxygenation (H/R) of cardiac microvascular endothelial cells, and can significantly increase the number of viable cells. NO and nitric oxide synthase in the cell culture medium (iNOS) activity increased, endothelin activity decreased, iNOS mRNA expression increased and endothelin mRNA expression decreased, and the mechanism of action may be related to up-regulation of iNOS mRNA expression and down-regulation of endothelin mRNA expression in endothelial cells. In addition, the Langedorff isolated heart perfusion method was used to perfuse the heart with KH perfusate and different concentrations of ligustonin A under constant temperature and pressure conditions. The results showed that pretreatment with ligustilide A could increase the coronary flow rate of isolated rat hearts during reperfusion stoppage, reduce the concentration of IL-1β in the coronary effluent and the ratio of TXB ₂ /6-Keto-PGF _1α , the incidence of ventricular tachycardia and ventricular fibrillation were all reduced, and the duration was also shortened; at the same time, the contents of LDH and MDA in the myocardial homogenate were reduced, and the SOD activity was increased. The injury of vascular endothelial cells caused by ischemia-reperfusion has a protective effect.

Ligusticolide A has a certain relaxing effect on rat isolated aortic vessels. It has inhibitory effects on PGF _2α , phenylephrine, serotonin, and KC1-induced vasoconstriction, and adenine nucleotide cyclase inhibitors, soluble guanylate cyclase inhibitors, non-selective Potassium ion channel blocker tetraethylamine and removal of endothelium have no significant effect on the relaxation effect of chuanxionglide A, and its specific mechanism needs to be further studied.

Ligustilide A is cytotoxic to colon cancer cell line HT-29, and can inhibit its cell proliferation, with an IC ₅₀ of (10.40±0.98) μg/mL. The inhibitory effect on the survival of HT-29 by citrolactone A was higher than that of CCD-18Co in normal colon cells, with an IC ₅₀ of (20.95±4.22) μg/mL, and citrolactone A was associated with butylphthalide, z - Ligustilide has an integrative effect on HT-29 cells and a dose-dependent inhibition on colon cancer cells.

At present, there is no literature report on the combination of galactomannan oligosaccharides and ligustonin A as a feed additive for piglets to regulate intestinal flora.

Contents of the invention

The technical problem to be solved by the present invention is to provide a feed additive for piglets that can effectively regulate intestinal flora and has low toxic and side effects.

The technical scheme adopted by the present invention to solve the above problems is to provide a feed additive for regulating the intestinal flora of piglets, wherein the weight ratio of the active ingredient galactomannooligosaccharide to ligusticolactone A is (10:1)～( 1:10).

Preferably, the weight ratio of galactomannan-oligosaccharide to ligustonin A in the feed additive is (9:1)˜(1:1).

More preferably, the weight ratio of galactomannan-oligosaccharide to ligustonin A in the feed additive is (5:1)˜(3:1).

Most preferably, the weight ratio of galactomannan-oligosaccharide to ligustonin A in the feed additive is 4:1.

Preferably, the feed additive further includes other active ingredients.

More preferably, the other active ingredients are selected from one or more of amino acids, vitamins and the like.

Most preferably, the amino acid is glycine, alanine, leucine, valine, cysteine, methionine, threonine, serine, phenylalanine, tyrosine, tryptophan , methionine, glutamic acid, arginine and/or histidine.

Most preferably, the vitamins are vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin H, vitamin K and/or vitamin P.

The present invention also provides an application of the above-mentioned feed additive in preparing feed for regulating intestinal flora of piglets.

Preferably, said 100 parts by weight of feed contains 1-5% by weight of feed additives.

The present invention has positive and beneficial effects:

Surprisingly, the present invention unexpectedly found that the combined use of galactomannan oligosaccharides and ligusticolactone A has a synergistic effect on regulating the intestinal flora of piglets, and significantly reduces the dosage of active ingredients. Other active ingredients work better.

Compared with ordinary feed additives, the piglet feed additive of the present invention has simple preparation method, reasonable compatibility and remarkable curative effect, and the experiment proves that it has a good effect on regulating intestinal flora of piglets; since there is no western medicine in the additive, the feed additive has no toxic and side effects, no drug resistance.

Detailed ways

The present invention will be further described below in conjunction with examples, but the embodiments of the present invention are not limited thereto. The experimental methods used in the following examples are conventional methods unless otherwise specified.

Embodiment 1, the preparation of feed additive

Take 9 parts by weight of galactomannooligosaccharide and 1 part by weight of ligustalactone A, and mix them to prepare a feed additive S1 with a weight ratio of 9:1 between galactomannan-oligosaccharide and listonin A.

Take 5 parts by weight of galactomannooligosaccharide and 5 parts by weight of ligustalactone A, and mix them to prepare a feed additive S2 with a weight ratio of 1:1 between galactomannan-oligosaccharide and lisantrolactone A.

Take 8 parts by weight of galactomannooligosaccharide and 2 parts by weight of ligustalactone A, and mix them to prepare a feed additive S3 with a weight ratio of 4:1 between galactomannooligosaccharide and lisantrolactone A.

Embodiment 2, the application of the feed additive of the present invention in regulating the intestinal flora of weaned piglets

1. Experimental animals and grouping treatment

Choose 120 weaned piglets of Duroc × (Landrace × Large White) three-way hybridization 28 days old, all boars castrated, according to body weight, sex, consanguineous principle, be divided into 6 groups immediately, promptly the composition group 1-3 of the present invention (implementation) Feed additives S1-S3) prepared in Example 1, galactomannan oligosaccharide group, ligustonin A group and control group, 20 piglets in each group. In the diet of S1-S3 composition group, galactomannan oligosaccharide group, and Ligustilide A group, add the above-mentioned feed additive according to 1% of the total weight of the feed (the total weight of the active ingredients added by each group is the same), the The diet was pig feed of the same weight without adding the above-mentioned feed additives. The piglets in each group were fed 3 times a day. During the test period, they had free access to food and provided sufficient drinking water. Other daily management measures were carried out according to the routine management of the farm. , Regular immunization and deworming, the test period is 30 days.

2. Determination of intestinal flora

At the end of the experiment, the piglets of each test group were slaughtered, and the colon segment was immediately ligated by laparotomy. After each ligation port was disinfected with alcohol cotton balls, they were put into sterilized plastic bags and transported to the microecology laboratory for detection of Escherichia coli, lactic acid, etc. The number of bacilli and bifidobacteria.

Medium: Eosin-methylene blue medium (EMB) for Escherichia coli, MRS medium for Lactobacillus, and BLb medium for Bifidobacterium.

Counting: Bacteria counting was carried out by plate spreading method. Use sterilized physiological saline to perform multiple dilutions to 10 ^-1 , 10 ^-2 , 10 ^-3 --- until 10 ^-8 , then use a microsampler to take 0.02m1 of the dilution solution starting from the high-fold dilution, and drop them into each Select the culture medium and make three dilutions for each sample (10 ^-4 , 10 ^-5 , 10 ^-6 for Escherichia coli ; 10 ^-5 , 10 ^-6 , 10 ^-7 for Lactobacillus; 10 ^-5 for Bifidobacterium , 10 ^-6 , 10 ^-7 ), each dilution was repeated three times, and the average was taken. After the inoculation, use a curved glass stick to mix well, culture E. coli aerobically for 24 hours, and culture Lactobacillus and Bifidobacterium anaerobically for 48 hours. Count the numbers of Escherichia coli, Lactobacillus and Bifidobacteria respectively. Results are expressed as the common logarithm of cfu (logcfu).

3. Test results

The effects of feed additives in each test group on the intestinal flora of weaned piglets are shown in Table 1.

Table 1 The impact of feed additive of the present invention on the intestinal flora of weaned piglets (logcfu/g intestinal content)

As can be seen from Example 2, compared with the control group, the feed additive of the present invention can significantly reduce the number of Escherichia coli in the intestinal flora of piglets, increase the number of lactobacilli and bifidobacteria, and is an excellent intestinal bacteria for piglets. flora regulator. In particular, the combined use of galactomannan oligosaccharides and ligustalactone A produced a synergistic effect and reduced the amount of drugs used, among which the feed additive with a weight ratio of galactomannan oligosaccharides and listonin A of 4:1 had the most prominent effect , resulting in unexpected excellent results.

Claims (8)

1. A feed additive capable of regulating the intestinal flora of piglets, characterized in that the weight ratio of the active ingredient galactomannan-oligosaccharide to chuanxionglide A is (10:1) to (1:10). 2 . The feed additive according to claim 1 , characterized in that the weight ratio of galactomannan oligosaccharides to liguscaractone A is (9:1)˜(1:1). 3. The feed additive according to claim 2, characterized in that the weight ratio of galactomannan oligosaccharide to liguscaractone A is (5:1)-(3:1). 4. The feed additive according to claim 1, characterized in that the feed additive further comprises other active ingredients. 5. The feed additive according to claim 4, characterized in that, the other active ingredients are selected from one or both of amino acids and vitamins. 6. feed additive according to claim 5, is characterized in that, described amino acid is selected from glycine, alanine, leucine, valine, cysteine, methionine, threonine, serine , phenylalanine, tyrosine, tryptophan, methionine, glutamic acid, arginine and/or histidine, the vitamins are selected from vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, Vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin H, vitamin K and/or vitamin P. 7. Use of the feed additive according to any one of claims 1-6 in the preparation of feed for regulating intestinal flora of piglets. 8. The use according to claim 7, characterized in that 100 parts by weight of the feed contains 1-5% by weight of feed additives.