KR101164927B1 - Feed additive including quercetin from onion as a antibiotic alternative, feed composition and its feeding method - Google Patents

Abstract

The present invention relates to a feed additive prepared by extracting the antioxidant quercetin from onion and mixed with excipients in a proportion to replace the antibiotic for feed additives, and the same. The broilers fed the feed additives according to the present invention had a higher weight gain compared to the non-antibiotics and vitamin E supplements of similar properties, and showed a similar degree in basic physiological activity with the antibiotic supplements. In order to replace antibiotics, there should be no toxicity and no immunity to other physiological activities. The feed additives showed such effects as shown in the experimental results. In addition, it has been shown that the content of antioxidants in the blood increases the overall health of the animal.

Quercetin, onion, antibiotic replacement, broiler, productivity

Description

Feed additive including quercetin from onion as a antibiotic alternative, feed composition and its feeding method

The present invention relates to a feed additive prepared by extracting the antioxidant quercetin from onion and mixed with excipients in a proportion to replace the antibiotic for feed additives, and the same.

The use of growth-promoting antibiotics in livestock feed is aimed at maximizing livestock productivity, preventing high-density breeding and disease outbreaks due to poor breeding conditions. However, with the emergence of antibiotic residues and the emergence of resistant bacteria in livestock products, regulations on the use of growth-promoting antibiotics have been tightened, and interest in non-antibiotic livestock products has increased in response to increasing competitiveness for imported livestock products and consumer demand for safe livestock products. Is rising. However, the elimination of antibiotics and raising livestock productivity and disease increase are a problem, so the countermeasures are urgent.

Antibiotics added to feed are completely digested and discharged after 2 to 3 months, and when shipped and edible before such a drug holiday period, there is a problem of lowering the immunity of the human body ingesting the meat products due to antibiotics remaining in livestock. In addition, the transfer of plasmid, which is a genetic material containing resistance, to bacteria that are resistant to antibiotics can be transferred to other bacteria, which causes the occurrence of Salmonella, etc. Done. As a result, the increase in these antibiotic-resistant bacteria always carries the risk of metastasis to the human body.

For these reasons, with the recent rise of antibiotics in meat products, these antibiotic-resistant bacteria have been raised, and research on antibiotic substitutes is being actively conducted. As antibiotic substitutes, substances that activate the body's immune function There is an active research on.

More specifically, probiotics, powders, enzymes, mineral supplements, and oligosaccharides are examples. Recently, antimicrobial peptides (AMPs) extracted from animals (Zasloff, 1987; Skerlavaj et al., 1999; Henzler et al., 2003;) and plants (Taniguchi and Kubo, 1993; Broekaert et al., 1995;) have been discovered. It is getting attention. Antimicrobial peptides are low molecular weight proteins that act on the innate immune defense system and are found in all living organisms (Boman and Hultmark, 1987; Boman et al., 1991; Hoffmann, 1995). Antimicrobial peptides also have a broad spectrum of antimicrobial effects, including bacteria (Gazit et al., 1994; Saberwal and Nagaraj, 1994; Steiner et al., 1998), viruses, and fungi (Quang et al., 1992; Lee et al., 1995; De Lucca and Walsh, 1999; Lee). Et al., 1999;) have the potential as novel antibiotic replacements by inhibiting growth and improving immunity.

In addition, various substances using useful substances in plants have been used as antibiotic substitutes, but the effects are not satisfactory. In general, antimicrobial properties are widely used as antibiotic replacements, and antimicrobial properties are used for the purpose of suppressing intestinal harmful microorganisms, and antioxidant components are used for the purpose of strengthening immunity. However, in the case of antibiotic substitutes currently being developed, even if they are effective, if the manufacturing cost is high, or if the cost is low, there is a problem that it is difficult to commercialize due to the minimal antibiotic replacement effect.

Under such technical background, the present inventors have developed an antibiotic replacement feed additive having excellent economical efficiency, easy production method, and no limitation on feeding of animals to substantially replace the existing antibiotic.

After all, it is an object of the present invention to provide a feed composition for extracting quercetin from onion and formulating it in a form that is easy to feed livestock, thereby improving the immunity and productivity of the livestock.

In order to solve the above problems, the present invention provides a method for preparing a feed additive by extracting quercetin from onion and formulated in powder form.

According to one aspect of the invention, the step of obtaining a liquid extract and solid content of the onion by pressing the onion; Dissolving the obtained onion solids in a 1: 1 weight ratio with an organic solvent, stirring and filtering to obtain an extract; Mixing the obtained extract and the liquid extract of onion, and adding and stirring ethanol so that the weight ratio of ethanol is 1: 2; Removing ethanol from the mixture to obtain a solid; And it may be provided a method for producing a feed additive comprising the step of completing the powder formulation by adding silica and powder and soda ash to the obtained solid content.

According to one embodiment, the organic solvent for melting the obtained onion solids may be ethanol, the stirring may be carried out at 200 ~ 400rpm at 35 ~ 45 ℃ 20 to 40 minutes.

According to one embodiment, the powder formulation may be prepared by adding onion extract solid content of 0.40 to 1.0% by weight, silica 35 to 45% by weight, powder 35 to 45% by weight and soda ash 10 to 20% ratio.

According to one embodiment, it is possible to provide an antibiotic replacement feed composition having excellent immuno-enhancing and productivity by adding the feed additive to 5 to 50% by weight of the total feed composition.

According to another aspect of the present invention, there can be provided a method of administering a feed composition to enhance the immunity and productivity of the livestock, characterized in that for administering the antibiotic replacement feed composition excellent in the immune boosting and productivity.

The broilers fed the feed additives according to the present invention had a higher weight gain compared to the non-antibiotics and vitamin E supplements of similar properties, and showed a similar degree in basic physiological activity with the antibiotic supplements. In order to replace antibiotics, there should be no toxicity and no immunity to other physiological activities. The feed additives show these effects as shown in the experimental results. Appeared to be sufficient. In addition, it has been shown that the content of antioxidants in the blood increases the overall health of the animal.

Hereinafter, the present invention will be described in more detail.

Feed additives used in the present invention means processed and formulated materials other than the material of the general feed for addition to the feed composition. In the present invention, it refers to a state in which the mulberry extract is processed and powdered by mixing silica, powder, soda ash and the like.

Feed composition refers to the final mixture that feed additives are added to a common feed to feed for livestock. In the present invention, it means a state in which the feed additive is mixed in a predetermined ratio to the general feed.

The present invention relates to a feed additive that can replace synthetic antibiotics using onion-derived quercetin. To extract quercetin, onions were extracted with ethanol, and ethanol was prepared as a solid. It was prepared as a feed additive using powder, silica and soda ash. It is a step of obtaining a liquid extract and solid content of the onion by pressing the onion; Dissolving the obtained onion solids in a 1: 1 weight ratio with an organic solvent, stirring and filtering to obtain an extract; Mixing the obtained extract and the liquid extract of onion, and adding and stirring ethanol so that the weight ratio of ethanol is 1: 2; Removing ethanol from the mixture to obtain a solid; And adding silica, powder and soda ash to the obtained solid to complete the powder formulation.

Here, the organic solvent for dissolving the obtained onion solid content may be ethanol, quercetin can be dissolved well, it is easy to purchase, any of the livestock can be edible.

 In addition, for the extraction of quercetin, it is preferable to stir at 200 to 400 rpm for 20 to 40 minutes at 35 to 45 ° C. At temperatures higher than 45 ° C, the solvent may evaporate. The extraction may not occur sufficiently, which is undesirable. The stirring time and the stirring speed are sufficient to allow quercetin to be extracted in a stable state. If the stirring is too long for more than 40 minutes, there is a risk of growth and contamination of various bacteria, which is not suitable, and the stirring is performed at a high speed of 400 rpm or more. When the vibration of the solvent is too severe, the yield may be rather deteriorated.

According to an embodiment of the present invention, the powder formulation is prepared by adding a feed additive of 0.4 to 1.0% by weight of silica, 35 to 45% by weight of silica, 35 to 45% by weight of powder and 10 to 20% by weight of soda ash. When the content of onion extract solids exceeds 1.0% by weight, it exceeds the content necessary for enhancing oil-based productivity and immunity, which is undesirable in terms of economics, and when added in an amount less than 0.40% by weight, it shows the best effect in terms of productivity and immunity. It is also undesirable. Silica, powder, and soda ash are most preferably formulated at the above ratios. When the content of silica and powder exceeds 45% by weight, foreign substances increase in feed intake, which is undesirable. It is not desirable to have a side effect of reducing feed intake due to off-flavor.

According to one embodiment of the present invention, a feed composition for improving antibiotics and an excellent antibiotic boosting agent having a feed additive added to 0.5 to 1.0% by weight of the total feed composition may be provided. As described above, the feed additive is 1.0 weight. If it exceeds the percentage, the resveratrol is contained more than necessary for productivity and immunity enhancement, which is not economically desirable.

The feed composition comprising the feed additive of the present invention can be used as an excellent antibiotic substitute in terms of immunological enhancement and productivity, and by administering a feed composition for replacing antibiotics with high immunity and productivity to the livestock, the immunity of the livestock is improved and productivity is improved. You can.

Hereinafter, the present invention will be described in more detail with reference to Examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.

The feed prepared as described above was examined for the broiler for five weeks, and the effects of antibiotics and other nutrients on biomaterial content were investigated.

Example  1. Preparation of Quercetin-containing Feed Additives Using Onions

1 kg of dried onion was crushed and 1,000 ml of ethanol was added thereto to block the light for 72 hours, soaked and extracted at room temperature. The extract was filtered under reduced pressure, and the residue was extracted twice more in the same manner. The filtrates were then combined, concentrated under reduced pressure at 40 ° C., and lyophilized at −50 ° C. to obtain an ethanol extract solid (69.89 g).

The ethanol extract solids were extracted three times with a mixture of hexane (n-hexane) and water (1: 1) for separation and extraction according to polarity, and the water layer from which the hexane layer was removed was again extracted with chloroform and water (1: 1). Three times). In addition, the water layer from which the chloroform layer was removed was extracted three times with a mixture of ethylacetate (ethylacetate) and water (1: 1), and each layer was concentrated under reduced pressure and lyophilized in the same manner as above, and the water layer from which ethyl acetate was removed was The extract was extracted three times with a mixture of butanol (n-butanol) and water (1: 1), and each layer was concentrated under reduced pressure and lyophilized in the same manner as described above. Hexane fraction (0.83 g), chloroform fraction (0.20 g), ethyl acetate fraction (0.74 g), butanol fraction (5.04 g) and water fraction (56.64 g) were obtained.

 Quercetin content in the fractions was analyzed using high performance liquid chromatography (HPLC). The instrument used was Agilent 1100 series, column was henomenex Luna C18 (II) 5um 4.6 X 250mm, and mobile phase was dissolved in water with 1% formic acid. 1% formic acid dissolved in acetonitrile was mixed and detected at 375 nm.

Quercetin in each fraction contained 6.02 mg in the hexane fraction, 1.2 mg in the chloroform fraction and 77.5 mg in the ethyl acetate fraction, but was not detected in the butanol fraction and the water fraction.

Therefore, the content of quercetin contained in the extract of 1 kg of dried onions with ethanol was 84.72 mg, and the onion extract was dried to obtain a solid content for feed formulation, wherein the ratio of quercetin content of solid content was 81 weight. Appeared in%. A feed additive having a concentration range suitable for feeding broilers was prepared. This, 0.4% by weight of the ethanol extract solid, 40% by weight silica, 45% by weight powder, soda ash 146% by weight, the quercetin content of the feed additive thus prepared is 400ppm.

Example  2. Broiler Feed Test of Quercetin-Containing Feed Compositions

In order to determine the antibiotic replacement effect of quercetin prepared in Example 1, non-antibiotic, antibiotic and vitamin E spheres were used as a comparison, and the example was placed in the quercetin supplement. Antibiotics were combined with abilamycin and salinomycin, with vitamin E at 200 IU and quercetin at 200 ppm. All additives were mixed to equal content relative to kg of feed weight.

The reason for the treatment of the vitamin E phrase is that because quercetin is a fat-soluble antioxidant, it is fed a similar fat-soluble antioxidant vitamin.

Table 1 shows the productivity of broilers, broiler chickens (Ross) 1 day-old cockerel was used, 160 times, 4 repetitions per treatment, 10 numbers per repetition is shown and the results of the productivity.

Comparative example Example
(Quercetin) Antibiotic Antibiotic Vitamin E  Weight gain (g) 1,984 2,035 2,004 2,031 Feed Intake (g) 3,282 3,279 3,120 3,264 Feed rate 1.65 1.61 1.56 1.60

The weight gain of quercetin-treated diet was similar to that of antibiotic-treated diet, and was higher than that of non-antibiotic and vitamin E diet (see FIG. 3). Feeding rate of quercetin-treated diet was similar to that of antibiotic-treated diet and improved compared to non- antibiotic. The weight gain of quercetin-treated diet showed very good results when referring to the comparison with the various groups used as the comparative example, and it can be seen that it is very suitable for use as a feed additive.

In addition, Figure 4 shows the results of measuring the antioxidant enzyme-like activity (SOD like activity) in order to identify the antioxidant properties in the blood. The higher the levels of these antioxidant enzymes, the stronger the antioxidant properties. Antioxidant enzyme levels in quercetin-treated diets were significantly higher than those in antibiotic-free and antibiotic-treated diets, which may be attributed to the antioxidant function of quercetin.

In addition, Figure 5 shows the MDA (Malondialdehyde) content of the lipid peroxide in the blood, the lower the value means that the state of the animal is good. Quercetin-fed diets were found to have lower concentrations than antibiotic- and antibiotic-fed diets.

Quercetin showed similar results to vitamin E in antioxidant properties, and similar results to antibiotics in terms of productivity, which contributed to improving the health of livestock through antioxidant reactions.

Table 2 shows the results of biochemical composition in blood.

division
Comparative example Example
(Quercetin) Antibiotic Antibiotic Vitamin E  Creatinine (mg / dL) 0.27 0.28 0.24 0.25 Urea Nitrogen (BUN)
(mg / dL) 2.80a 2.40ab 2.20b 2.60a Total protein (g / dL) 3.38 3.36 3.12 3.22 albumin
(g / dL) 1.66 1.64 1.59 1.52  Globulin (g / dL) 1.72 1.72 1.53 1.70  Albumin / globulin 0.97 0.96 1.04 0.89 AST
(U / dL) 178.6a 170.6ab 164.8b 167.6b ALT
(U / dL) 6.20 5.80 5.40 5.45

The biochemical composition of blood components described in Table 2 has the following meanings.

1. Creatine: As a product of muscle metabolism, this increase is related to renal failure or dehydration, and is used as an effective indicator of renal failure determination along with blood urea nitrogen concentration.

2. Urea Nitrogen (BUN): It indicates kidney function and increases in case of disorder.

3. Total protein: There are more than 500 kinds of proteins in the blood, and they are responsible for the movement of various substances, iron metabolism, immunity, and blood coagulation. The total protein is an indicator of such a living state, and if it is lower or higher than the normal value, the synthesis or degradation of the protein may be suspected or the protein absorption disorder may be suspected.

4. Albumin (Albumin): Albumin is a protein that accounts for two-thirds of the total protein in the blood, plays an important role in maintaining blood pressure, and is responsible for transporting various nutrients and hormones.

5. Globulin: Determination of immune system abnormalities by measuring the amount of proteins responsible for immune function in the blood.

6. Albumin / globulin ratio: As a criterion for determining the abnormality of blood proteins, if decreased, it may indicate chronic inflammation, digestive disease, kidney disease, and the like.

7. AST: It is an enzyme in the liver that increases when liver cells are damaged.

8. ALT: ALT is an enzyme in the liver that increases when hepatocytes are damaged by hepatitis.

In other words, creatine, BUN (Blood urea nitrogen) levels as a substance showing renal function, and albumin / globulin levels, AST and ALT values indicating liver function were investigated.

As shown in Table 2, globulin, an immunoprotein, showed similar results to the comparative example, and AST and ALT levels, enzymes indicating liver function, were improved in quercetin-treated groups compared to antibiotic- and antibiotic-treated groups. Showed.

The broilers to which the feed composition according to the present invention was administered had a higher weight gain compared to the non-antibiotic feed and vitamin E fed group having similar properties, and showed a similar degree in basic physiological activity as the antibiotic fed group.

In order to replace antibiotics, there should be no toxicity and no immunity to other physiological activities. The feed additives show these effects as shown in the experimental results. Appeared to be sufficient. In addition, it has been shown that the content of antioxidants in the blood increases the overall health of the animal.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

1 is a schematic diagram showing a process for extracting and formulating quercetin from onion.

2 shows formulated quercetin.

Figure 3 is a graph of the weight gain comparison of the feed composition feed sphere and the comparative example according to the present invention.

Figure 4 is a graph comparing the antioxidant properties (SOD like activity) in the blood of the feed composition feed spheres and the comparative example according to the present invention.

Figure 5 is a graph comparing the antioxidant properties (Malondialdehyde) in the blood of the feed composition feed sphere and the comparative example according to the present invention.

Claims (7)

Method for preparing a feed additive comprising the following steps: Squeezing the onion to obtain a liquid extract and solid content of the onion; Dissolving the obtained onion solids in a 1: 1 weight ratio with an organic solvent, stirring and filtering to obtain an extract; Mixing the obtained extract and the liquid extract of onion, and adding and stirring ethanol so that the weight ratio of [the mixture of the obtained extract and liquid extract of onion: ethanol] becomes [1: 2]; Removing ethanol from the mixture to obtain a solid; And Adding silica, powder and soda ash to the obtained solid to complete the powder formulation. The method of claim 1, wherein the organic solvent for dissolving the obtained onion solid is ethanol. The method of claim 1, wherein the agitation is a method of preparing a feed additive, characterized in that the 200 to 400rpm is carried out for 20 to 40 minutes at 35 ~ 45 ℃. According to claim 1, wherein the powder formulation is obtained after removing the ethanol in the final step after the ethanol treatment, 0.4 ~ 1.0% by weight of onion solid, 35 to 45% by weight of silica, 35 to 45% by weight of powder and soda ash 10 to 20 Method for producing a feed additive, characterized in that the production by adding in a proportion by weight. A feed composition for replacing antibiotics with high immunity and productivity, wherein the feed additive prepared according to any one of claims 1 to 4 is added to 5 to 50% by weight of the whole feed composition. A method of feeding a feed composition to enhance the immunity and productivity of livestock, characterized by administering a feed composition for replacing antibiotics with excellent immunity and productivity of claim 5. An antibiotic replacement feed additive prepared according to any one of claims 1 to 4.

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