KR20190074100A - A feed additive containing complex strains and earthworm cast and manufacturing method therof - Google Patents

Abstract

The present invention relates to a feed additive for poultry comprising a complex strain isolated from earthworm casts and earthworm casts, and a method for producing the same. The feed additive for poultry according to the present invention can replace common probiotics added to conventional poultry feed; can promote the growth of poultry and improve the productivity of poultry by promoting beneficial microbial growth and inhibiting harmful microbial growth in the intestine; can contribute to increasing farm household income by improving the quality of poultry meat and eggs, at the same time; and ultimately has the effect of providing a high quality and safe eco-friendly livestock food.

Description

TECHNICAL FIELD [0001] The present invention relates to a feed additive containing feedstock,

The present invention relates to a feed additive for poultry containing a complex strain isolated from earthworm fungi and an earthworm-containing fungus, and a method for producing the same.

Chicken and egg are low calorie nutrient sources including proteins, fatty acids and minerals. Especially, eggs are low in blood pressure, contain antioxidants, and are rich in phospholipids and lecithin, which lower cholesterol. They help prevent cardiovascular diseases and decrease blood lipids And brain food that helps prevent Alzheimer's dementia.

Probiotics are live microorganisms that are beneficial to host animals when supplied in sufficient quantities. Probiotics can help the growth of beneficial bacteria in the intestinal tract and can completely eliminate pathogenic bacteria, and secrete some enzymes that help the digestive tract wall function, regulation of intestinal mucosal immune system, production of antimicrobial agent and digestion of feed, And beneficial effects on host animals through changes and absorption of food for intestinal microorganisms. Probiotics not only increase innate immunity and increase the protective wall of the gastrointestinal mucosa, but also stimulate the growth of livestock by stimulating the secretion of antimicrobial β-defencin. The bacterium in the intestine has the ability to prevent metabolism, immunity and disease, and produces SCFA and vitamin K to exert its metabolic control.

Feeding of lactic acid bacteria as a probiotics in broilers and laying hens increased the productivity of broiler chickens, egg weight and egg production rate, and the feeding of probiotics with mixed lactic acid bacteria improved broiler productivity, quality of chicken, egg production, feed efficiency, egg weight and egg quality, Rate and feed efficiency were improved and egg yolk cholesterol was reduced by about 18.8%. However, Watkins and Kratzer (1984) reported that the addition and feeding of probiotics did not affect the productivity of livestock, and the Lactobacillus strains in the laying hens did not show differences in egg production, egg production, (Gooding et al., 1987). The differences in the results of these studies may be due to inaccurate culture of the live cells, differences due to the species and the method of addition, and differences in the number of viable cells.

Accordingly, the present inventors have completed the present invention by developing poultry productivity and techniques for improving chicken and egg quality by feeding broiler and fermented feed additives prepared by using three complex strains including Bacillus subtillis to broilers and laying hens.

Korean Patent No. 10-0407073

The object of the present invention is to provide a feed additive for poultry containing at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm- .

Another object of the present invention is to provide a poultry feed comprising the poultry feed additive.

It is another object of the present invention to provide a method for producing the feed additive for poultry.

In order to achieve the above object,

The present invention provides a feed additive for poultry including at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm feces do.

The present invention also provides a poultry feed comprising the poultry feed additive.

Further, the present invention relates to a method for producing a microorganism which comprises 1) preparing at least one strain selected from the group consisting of a Bacillaceae strain, a Streptomyces strain and a Sphingobacteriaceae strain isolated from an earthworm- (Step 1); And

2) heat-treating the earthworm fungi and inoculating the strain of step 1 (step 2);

And a feed additive for poultry.

The poultry feed additive of the present invention is prepared by inoculating a strain isolated from earthworm-fecal soil into an earthworm-infested soil and is a feed additive capable of replacing common probiotics added to ordinary poultry feed. It can promote the growth of poultry, Promoting microbial growth and inhibiting harmful microbial growth, thereby improving the productivity of poultry, improving the quality of poultry meat and eggs, contributing to the increase in farm income, and ultimately providing an environmentally safe and livelihood food product with excellent quality and safety .

Fig. 1 is a graph showing the results of the comparison between the feed (FCD5) containing the general feed (control C) of the present invention, 0.2% of the general probiotics, CP, 0.3% of the mixed- Of the egg yolk of the laying hens.

Hereinafter, the present invention will be described in detail.

Complex strain  And earthworms Feces  Feed additives and feed for poultry

The present invention provides a feed additive for poultry including at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm feces do.

In the present invention, the feed additive for poultry including the strain and the earthworm-containing feces can be a composite prophylactic agent prepared by inoculating the combined strain isolated from the earthworm-infested soil into the earthworm-infested earthworm medium. In the present invention, The feed additive for poultry was named as a fungus feed additive.

In the feed additive for poultry according to the present invention, the strain may be a strain isolated from an earthworm-infested soil. The Bacillus subtilis strain may be Bacillus subtillis (KCTC2213), the Streptomyces sp . Strain may be Streptomyces galilaeus (KCTC1919), and the Ssp. Sphingobacteriaceae spp., BR5-29).

The feed additive for poultry according to the present invention comprises at least one strain selected from the group consisting of the above Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, ⁴ to 1.0 x 10 < ⁸ > cfu / g, preferably 1.0 x 10 < ⁵ > to 1.0 x 10 < ⁷ > cfu / g. According to one embodiment of the present invention, a poultry feed additive of the present invention is Bacillus subtilis (Bacillus subtillis, KCTC2213), mouse (the Streptomyces Streptomyces galril La galilaeus , KCTC1919) and Sphingobacteriaceae spp. (BR5-29) at 1.0 x 10 ⁶ cfu / g, respectively.

In the feed additive for poultry according to the present invention, the poultry may be at least one kind selected from the group consisting of broiler chickens, laying hens, native chickens, ducks, turkeys, quails and goats, preferably at least one selected from broilers and laying hens .

The feed additive for livestock according to the present invention may promote the growth of poultry and improve the quality of poultry meat and the productivity of poultry. In one embodiment of the present invention, when the feed additive for livestock according to the present invention is fed, the growth of broiler and laying hens is promoted, the beneficial microorganism growth promotion and harmful microorganism growth inhibition are shown in the intestines, At the same time, it was confirmed that chicken meat quality and egg quality of broiler chicks could be improved (see Examples 1 to 4).

The present invention also relates to a feed additive for poultry comprising at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm- And a poultry feed.

The feed additive for poultry according to the present invention may include 0.01 to 10 parts by weight of the feed additive for poultry, relative to 100 parts by weight of the feed for poultry. Preferably 0.1 to 1.0 part by weight, more preferably 0.1 to 0.7 part by weight, particularly preferably 0.3 to 0.5 part by weight.

Complex strain  And earthworms Feces  For preparing feed additives for poultry containing

The present invention relates to a method for producing (1) a step of preparing at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain isolated from earthworm feces Step 1); And 2) inoculating the strain of step 1 after heat-treating the earthworm fecal soil (step 2); The present invention relates to a method for producing a feed additive for poultry including one or more strains selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm- ≪ / RTI >

In the manufacturing method according to the present invention, the heat treatment of the earthworm-modified soil may be performed at 1000 ° C to 2000 ° C for 30 minutes to 3 hours. Preferably 1200 占 폚 to 1800 占 폚 for 30 minutes to 2 hours, and more preferably 1300 占 폚 to 1700 占 폚 for 30 minutes to 1 hour and 30 minutes.

In the preparation method according to the present invention, the Bacillus strain may be a Bacillus subtilis (Bacillus subtillis, KCTC2213), Streptomyces sp may be a mouse (Streptomyces galilaeus, KCTC1919) in Streptomyces galril La , The spp. Bacterium sp. Strain may be Sphingobacteriaceae spp., BR5-29.

In the manufacturing method according to the present invention, the step 2 is a step of adding a feed additive for poultry, which is finally produced, to a feed additive comprising the Bacillaceae strain, the Streptomyces strain, and the Sphingobacteriaceae strain to contain one or more strains selected from the group 1.0 × 10 ⁴ to 1.0 × 10 ⁸ cfu / g, respectively may be produced by controlling the amount of strain in the vaccinated earthworms bunbyeonto. Preferably, it may be prepared so as to contain 1.0 10 ⁵ to 1.0 10 ⁷ cfu / g of each strain. According to one embodiment of the present invention, a poultry feed additive according to the production method of the present invention is a mouse Bacillus subtilis (Bacillus subtillis, KCTC2213), Streptomyces galril La (Streptomyces galilaeus , KCTC1919) and Sphingobacteriaceae spp. (BR5-29) were prepared to contain 1.0 × 10 ⁶ cfu / g, respectively.

In the production method according to the present invention, the poultry may be at least one kind selected from the group consisting of broiler chickens, laying hens, native chickens, ducks, turkeys, quails and goats, and preferably at least one species selected from broilers and laying hens .

In the production method according to the present invention, the strain of step 1 may be isolated by the following steps:

3) heat-treating the earthworm fern (step 3);

4) separating the Bacillaceae strain from the heat-treated earthworm feces of step 3 (step 4);

5) re-inoculating the separated Bacillus sp. Strain of step 4 in the heat-treated earthworm feces of step 3 (step 5); And

6) isolating the Streptomyces strain and the Sphingobacteriaceae strain from the earthworm-infested soil of step 5 (step 6).

In the manufacturing method according to the present invention, the heat treatment of the earthworm-modified soil may be performed at 1000 ° C to 2000 ° C for 30 minutes to 3 hours. Preferably 1200 占 폚 to 1800 占 폚 for 30 minutes to 2 hours, and more preferably 1300 占 폚 to 1700 占 폚 for 30 minutes to 1 hour and 30 minutes.

Hereinafter, the content of the present invention will be described in more detail with reference to Examples and Experimental Examples. It is to be understood, however, that these examples and experiments are illustrative only for the understanding of the present invention and are not to be construed as limiting the scope of the present invention, and the following embodiments may be constructed such that all or some of them are selectively combined to make various modifications .

< Example  1> The complex strain  Preparation of Fermented Feed Additives Using Fertilized Soil

Bacillus subtilis isolates isolated from earthworms (KCTC2213), Streptomyces galilaeus (KCTC1919), Sphingobacteriaceae spp . (BR5-29) were used.

Specifically, the earthworm fecal soil was subjected to heat treatment at a high temperature of 1500 ° C for 1 hour to kill the harmful bacteria. Thereafter, Bacillus subtillis was isolated from the feces , and then the strain was inoculated again to the heat-treated feces and Streptomyces galilaeus (KCTC1919) and Sphingobacteriaceae spp . (BR5-29) strain.

(KCTC2213), Streptomyces ( Bacillus subtilis), and the like were obtained by inoculating the above-mentioned three strains in the same amount to the earthworm fungi killed with noxious bacteria by performing heat treatment at a high temperature of 1,500 &lt; galilaeus (KCTC1919) and Sphingobacteriaceae spp . (Hereinafter referred to as &quot; powdery soil fermented feed additive &quot;) comprising a complex strain (BR5-29).

The above-mentioned fermented feed additives of Bacillus subtilis (KCTC2213), Streptomyces galilaeus (KCTC1919) and Sphingobacteriaceae spp . (BR5-29) of 1.0 x 10 ⁶ cfu / g, respectively.

< Experimental Example  1> Preparation of compound feed using fermented feed additives

For the feeding experiment, the powdery fermented feed additives or the common probiotics of Example 1 were added to the general feeds of the following Table 1 to prepare compounded feeds. The level of addition was determined according to the compounded feed addition standard, The amount of yellow corn added to the feed was controlled by the amount of probiotics added. The general diet was formulated to meet the nutrient requirements recommended by the Korean Livestock Specification Standard Poultry (2012). The formula and chemical composition of the general diet are shown in Table 1 below.

Ingredients % Chemical composition % Yellow corn 58.86 Moisture 10.83 Soybean oil meal 16.80 Crude protein 17.88 Distillers dried grains with
solubles (HP-DDGS) 4.10 Crude fat 4.12 Feather meal 3.20 Crude fiber 2.94 Rapeseed oil meal 2.00 Crude ash 13.17 Wheat bran 3.00 Calcium 4.01 Tallow 1.00 Available phosphorous 0.31 Limestone 9.70 Lysine 0.75 Salt 0.25 Methionine 0.38 Shell powder 0.50 Methionine + Cystine 0.68 Calcium phosphotate monobasic 0.40 Metabolizable energy (ME), kcal / kg 2,750 Vit-min mix ^{1 )} 0.10 Methionine 0.09 Total 100.00

¹⁾ Vitamin mix (Vit-min mix) provides the following nutrients per kg: vit.A 10,000 KIU; vit.D3 3,500 KIU; vit.E 15.0 KIU; Vit K3 2,000 mg; vit.B1 1,500 mg; vit.B2 5,000 mg; vit.B6 3,000 mg; vit.B12 20 mg; niacin 25,000 mg; Pantothenic acid 6,000 mg; folic acid 500 mg; biotin 50 mg; Cu 9,000 mg; I 1,500 mg; Mn 80,000 mg; Zn 80,000 mg; Se 250 mg; Fe 50,000 mg; Co 100 mg.

The experimental treatments were as follows: control (C, control: general feed feed without addition of fermented feed additives or general probiotics), 0.3% of fermented feed additives (FCD3, fermented cast diet 0.3% 0.3% by weight), 0.4% (FCD4, fermented cast diet 0.4%) of powdered fermented feed additive, 0.4% by weight of powdered fermented feed additive to the total weight of the mixed feed, 0.5% (FCD5, fermented cast diet 0.5%) and general probiotics 0.2% (CP, commercial probiotics 0.2%) compared to the total weight of the compounded feed, 0.2% by weight).

< Experimental Example  2> Experiment design and specification management

On the day of hatching of the Ross system (Ross 308), 240 male broilers were completely randomized into 4 treatments (C, CP, FCD3 and FCD5) and kept for 35 days. A total of 60 treatments were used and 3 replicates (20 per repetition) were applied to analyze the broiler performance and cecal microbial changes.

On the day of hatching of the Ross system (Ross 308), 225 male broilers were completely randomly assigned to three treatments (C, CP and FCD4) and kept for 35 days. A total of 75 treatments were used for each treatments and 3 treatments (25 treatments per iteration) were used to evaluate the quality of chicken.

One hundred fifty-week-old brown Hyline brown were purchased from Eunbang Farm (Anseong, Gyeonggi-do) and used for the experiment for 6 weeks (51-56 weeks) after a week of adaptation period in iron three-stage egg cage. Four different treatments (C, CP, FCD3, and FCD5) were used, and 60 eggs per each treatment were randomly assigned and one egg per cage was used to analyze productivity and eggs quality of laying hens.

All broilers and laying hens were fed unlimited amounts of water and regular feeds, and 18 hours (08: 30-24: 30) were lit. All experiments were in compliance with the regulations of the Animal Experiment Ethics Committee of Kangwon National University.

< Experimental Example  3> Productivity of broiler and quality evaluation of chicken

3-1. Broiler  Specification Grade Analysis

The broiler (broiler) fed broiler (broiler) diets containing the general feed (control (C)), the compound feed containing the fermented feed additive (FCD3 and FCD5) or the common feed preparation (CP) for 35 days after the hatching of Experimental Example 2 Weight gain, feed intake and feed efficiency) were measured and shown in Table 2 below.

Gruops Item C CP FCD3 FCD5 Body weight, g / head 1,718 ± 38,29 1,834 ± 43,88 2,032 ± 37.55 2,087 ± 38,01 Feed intake, g / head 2,867 ± 12.80 3,015 ± 23.01 3,255 ± 33.17 3,215 ± 26.77 Feed efficiency ratio 0.60 + 0.03 0.61 + 0.02 0.62 + 0.03 0.65 + 0.02

As shown in the above table, the body weight and feed intake of the control group (C) were the lowest in the order of FCD5, FCD3 and CP (p <0.05) The highest in the treatments. The feed efficiency ratio was the highest in FCD5 compared to the control (p <0.05).

3-2. Broiler  Analysis of cecal microorganism change

After the completion of the analysis of the test results of Experimental Example 3-1, the cecum was collected from the sacrificed broiler by aseptic method and stored anaerobically. Then, the total number of bacteria of Lactobacillus, Escherichia, Salmonella, Coliform bacteria and Total aerobic bacteria were examined. Specifically, 1 g of aseptically collected cecum contents was mixed with 9 mL of sterile saline (PBS 0.1M, pH 7.0), diluted 10-fold (1: 9, wt / vol) . All procedures were performed in an anaerobic condition in the anaerobic chamber (5% hydrogen, 5% CO 2, balanced nitrogen), the culture is selected by the frequency divider ssikeul each 100 uL in 10 ^-2 to 10 ^-7 dilution sterile plate medium that is Salmonella (SS agar, Difco); E. coli (McConkey purple agar, Difco); Coliform bcteria (Violet red agar, Difco); Total aerobic bacteria medium (Nutrient agar, Difco). The aerobic microorganisms were incubated for 24 hours in an incubator at 37 ℃. The anaerobic microorganism cultures were incubated for 48 hours in anaerobic condition with 37 ℃ CO ₂ incubator. The number of microbial communities measured is shown in Table 3 below as the number of microcapsules per g of the contents of the cecum, with base-10 logarithm colony-forming units, log ₁₀ cfu / g of fresh cecal content.

Item (log ₁₀ cfu / g content) Groups C CP FCD3 FCD5 Lactobacillus 7.07 ± 0.16 ^{c1 )} 7.65 ± 0.13 ^b 7.97 ± 0.22 ^a 8.01 + - 0.31 ^a Escherichia 4.52 ± 0.28 ^a 3.97 ± 0.03 ^b 3.71 ± 0.02 ^c 3.68 ± 0.08 ^c Salmonella 4.80 + 0.17 ^a 3.97 ± 0.06 ^b 3.55 + - 0.23 ^c 3.57 0.06 ^c Coliform bacteria 6.52 + 0.22 ^a 6.11 ± 0.27 ^b 5.07 ± 0.26 ^c 5.07 ± 0.27 ^c Total aerobic bacteria 6.07 ± 0.43 ^a 5.12 ± 0.38 ^b 4.22 ± 0.27 ^c 4.12 + - 0.22 ^c

As shown in Table 3, Lactobacillus showed higher FCD treatment (FCD3 and FCD5) than control (C), and FCD5 was the highest . On the other hand, Escherichia, Salmonella, Coliform bacteria and Total aerobic bacteria were the highest in the control (C) and significantly lower in the FCD5, FCD3, and CP order (p <0.05). Therefore, it can be considered that the growth of broiler chickens was promoted by the feeding of FCD3 or FCD4 in Experimental Example 3-1 because of the change of microorganism.

3-3. Fatty acid composition analysis of chicken meat

In order to evaluate the quality of chicken according to the type of feed, a general feed (control), a mixed feed (FCD4) containing 0.4% of a fermented feed additive of the fecal soil, or a common feed preparation (CP) for 35 days after hatching in Experimental Example 2 After the ingested broiler (broiler) was sieved, the leg flesh was pulverized to analyze the fatty acid composition. The lipid of the chicken was modified by the method of Folch et al . ( J. Biol . Chem . , 226, 497 (1957)) and the method of Morrison and Smith ( J. Lipid Res, 5, 600 (%) Of total fatty acids was calculated and the results are shown in Table 4 below.

Name
(% of total fatty acids) Groups C FCD4 CP 8: 0 Octanoic acid 0.21 0.07 ^a 0.12 0.02 ^b 0.04 0.02 ^b 10: 0 Decanoic acid 0.02 0.02 ^b 0.06 0.02 ^a 0.02 ± 0.01 ^b 12: 0 Lauric acid 0.04 0.07 ± 0.01 0.05 ± 0.03 14: 0 Myristic acid 0.83 0.01 ^b 0.93 + 0.03 ^a 0.83 0.08 ^b 16: 0 Palmitic acid 25.67 ± 0.11 ^a 24.10 ± 0.50 ^b 25.66 ± 1.02 ^a 16: 1 n-9 Palmitoleic acid 10.24 ± 0.16 ^b 7.13 ± 0.18 ^c 10.79 ± 0.24 ^a 18: 0 Stearic acid 6.90 ± 0.16 7.20 ± 0.16 6.85 ± 0.25 18: 1n-9 Oleic acid 43.14 + 0.04 ^a 39.23 ± 0.19 ^b 42.51 ± 0.82 ^a 18: 2n-6 Linoleic acid 11.22 + 0.08 ^b 18.31 + - 0.32 ^a 11.62 ± 0.89 ^b 18: 3n-3 Linolenic acid 0.67 ± 0.01 ^b 1.67 ± 0.09 ^a 0.68 ± 0.12 ^b 20: 0 Arachidic acid - - - 22: 0 Behenic acid 0.25 + 0.04 0.22 ± 0.06 0.20 + 0.02 22: 1 Erucic acid 0.81 + 0.03 0.95 + 0.24 0.76 + 0.07 24: 0 Lignoceric acid - - - Total 100 100 100 Saturated fatty acid 33.92 + 0.18 32.70 + - 0.50 33.65 ± 0.90 Unsaturated fatty acid 66.08 ± 0.18 67.30 ± 0.50 66.35 ± .090 n-6 / n-3 ratio 16.84 + - 0.10 ^a 10.94 + - 0.71 ^b 17.00 ± 1.67 ^a

Omega - 3 fatty acids were significantly increased in FCD4 compared to control (p <0.05), but there was no difference between control (C) and CP treatments. The omega-6 / omega-3 ratio (n-6 / n-3 ratio) was not significantly different between control and CP treatments, but was significantly decreased in FCD4 treatment (p <0.05).

Omega Fatty Acid is a fatty acid which shows the position of the first double bond when the carbon atom number is given from the methyl group (-CH3) located at one end of the fatty acid. It is classified into n-6 and n-3. Fatty acids. In general, the proper balance of fatty acids accumulated in blood and tissues contributes to maintenance and promotion of health. However, unbalance, that is, a high n-6 / n-3 ratio causes various metabolic diseases, 3 Higher intake of foods may increase LDL-C in the blood, causing cardiovascular disease deaths.

Thus, from the above results, it was confirmed that the feed of the fodder-based fermented feed additive of the present invention can increase the omega-3 fatty acid content of chicken, and reduce the n-6 / n-3 ratio, thereby improving the quality of broiler chickens.

< Experimental Example  4> Estimation of productivity and eggs quality in laying hens

4-1. Egg Productivity and Specimen Performance Analysis in Laying Hens

The fertilized eggs (control) were fed to the brown laying hens (Hyline brown) of Experimental Example 2, fed with the feed containing the fermented feed additives (FCD3 and FCD5) or the common feed additives (CP) Egg production,%) and feed intake (g / head / day) were measured and are shown in Table 5 below.

Items Groups C CP FCD3 FCD5 Egg production,% 81.67 ± 5.01 ^b 89.33 ± 4.23 ^ab 91.67 ± 3.64 ^a 92.00 ± 2.78 ^a Feed intake, g / head / day 102.9 ± 6.81 ^b 144.6 + 4.05 ^a 144.7 + 5.01 ^a 147.8 ± 6.55 ^a

As a result, the egg production rate of FCD treated group was 91.67-92.00%, which was significantly increased to 12.65% compared to the control (C) 81.67% (p <0.05). There was no statistically significant difference between the FCD3 and FCD4 treatments and there was no significant difference between the control and CP. The egg production rate reached a constant level of plateau that did not increase any more in CP. Feed intake was also significantly higher in FCD3 and FCD5 than in the control (p <0.05).

These results indicate that supplementation with 0.3% ~ 0.5% of FCD feed to the laying hens can increase the feed intake and the egg production rate significantly. This is due to the maintenance of bacterial forests in the cecum due to the action of the combined probiotics containing B. subtilis in the powdery soil fermented feed additive, which promotes the growth of beneficial microorganisms, inhibits the growth of harmful microorganisms, and improves the egg production rate can see.

4-2. Egg quality analysis

Each feed was fed to the brown laying hen (Hyline brown) of Experimental Example 2, and eggs produced were collected to evaluate egg quality. Specifically, eggs produced from day 21 after feeding were collected and the average daily egg weight was calculated during the experiment. The quality of eggs (egg unit, eggshell thickness, blue intensity, yolk yellow index And changes in the height of the deep egg white and improvement rate) were measured and expressed as an average value.

The Haugh unit (HU) represents the freshness of the eggs. The height of albumin height and the weight of eggs (Egg weight) were measured by using a HouUn unit (FHK Co., Japan) (H + 7.57-1.7W ^0.37 )]. Where H is the height of the thick egg white (mm), and W is the weight of the egg (g). The yellowish color was measured using Roche egg color fan (Germany) and the yellowish color was expressed as an average index. Eggshell thickness and breaking strength were measured using an egg quality measuring machine (FHK, Co., Japan). Egg thickness was calculated by measuring the egg tip, middle portion, and tip portion of the egg shell four times, respectively, in the eggs used for HU and egg yolk measurements. The measured values are shown in Table 6 below.

Items Groups C CP FCD3 FCD5 Egg weight, g 67.52 ± 3.77 67.57 + - 4.03 67.38 + - 2.72 67.48 ± 3.01 Haugh unit (HU) 92.52 ± 1.10 ^b 93.08 ± 0.93 ^b 94.09 ± 0.78 ^a 94.17 ± 1.123 ^a Eggshell thickness, mm 0.37 + 0.07 ^b 0.40 0.06 ^b 0.45 ± 0.08 ^a 0.46 ± 0.05 ^a Breaking strength, kg / cm ² 3.11 ± 0.03 ^b 3.39 ± 0.07 ^b 3.81 ± 0.05 ^a 3.94 ± 0.06 ^a Egg yolk index 7.88 ± 0.33 ^b 8.01 ± 0.23 ^b 9.12 ± 0.18 ^a 9.20 ± 0.20 ^a Albumin high, mm 8.24 + 0.03 ^c 9.05 ± 0.05 ^b 9.73 ± 0.07 ^a 9.83 + 0.02 ^a

As a result, egg HU, egg shell thickness, blue intensity, egg yolk index and albumen height were significantly higher than those of control (C) by 1.78, 24.32, 26.69, 23.66 and 14.35% (P <0.05), respectively. Egg quality, especially egg shell thickness, was significantly increased (0.45-0.46 mm) compared to control (0.37 mm) (p <0.05). Egg yolk of FCD was significantly darker than control (C) and CP (Fig. 1). On the other hand, there was no difference in egg quality except for the increase of albumin height (p <0.05) between control and CP treatments.

The increase in egg quality by FCD feeding, especially egg shell thickness, compared to the control, enhances the utilization rate of nutrients by small intestinal villus activation due to the action of the combined probiotic containing B. subtilis in the fecal fermented feed additive of the present invention And the preservation of bacterium in the intestine increased the absorption rate of calcium.

4-3. Analysis of small intestine villi and intestinal gland length in laying hens

The development of duodenal villi can reflect the health status of the small intestine in birds. New epithelial cells are produced in the duodenal mucosal glands of the small intestine and move to the top along the villi. The histological structure of the avian digestive tract is similar to that of the mammalian digestive tract. On the surface of the intestinal mucosa, the small intestine is a finger-like protruding structure with the intestinal glands located below. Damage of the intestinal glands leads to loss of small villi and functional loss leading to metabolic disorders such as diarrhea. In addition, malnutrition and nutrient uptake due to dehydration and electrolyte imbalance can be lowered.

In order to confirm whether the result of egg quality improvement according to the FCD feeding of Experimental Example 4-2 was due to small villus activation, we observed the duodenum villus and the crypt depth of the laying hens fed diets Are shown in Table 7 below.

Items Groups C CP FCD3 FCD5 Duodenum villus, μm 1178 ± 35.63 ^d 1394 + - 43.17 ^c 1488 ± 62.12 ^b 1694 + - 45.7 ^a Crypt depth, μm 267 ± 25.1 ^d 292 ± 26.8 ^c 364 ± 22.7 ^b 404 ± 32.1 ^a

As shown in Table 7, the small villus length and the length of the intestine were significantly increased in order of FCD5, FCD3 and CP compared to the control (p <0.05). That is, it was confirmed that the feeding of the fermented feed additive of the present invention to the small intestine villi activates the egg, thereby improving the absorption rate of nutrients and improving the absorption of calcium due to the maintenance of intestinal bacterial forest.

4-4. Analysis of egg yolk fatty acid composition

To analyze the fatty acid composition of egg yolks, eggs collected from the 30th day were collected and randomly selected at intervals of 10 days for each treatment group. Egg lipids were obtained by adding 5 mL of egg yolk separated from boiled eggs to 200 mL of a mixed organic solvent (chloroform: methanol = 2: 1) and 6 mL of 0.88% KCl and stirring at 2,500 rpm using an ultra turrax T-25 homogenizer (US / IKA) After vigorously stirring for 3 minutes and centrifuging, the lipid layer was firstly separated and then the process was repeated three times to concentrate the extracted lipid by nitrogen gas. The process of methylation of lipids is as follows. 10 mg of the concentrated lipid fraction was placed in a reaction vessel for saponification and 1 mL of freshly prepared 0.5 N methanolic NaOH (2 g NaOH / 100 mL methanol) was added and the mixture was heated for 15 minutes and then cooled. 2 mL of reagent BF3-methanol for methylation was added and heated for another 15 minutes. After cooling sufficiently to room temperature, 1 mL of heptane and 2 mL of NaCl saturated solution were added and mixed for 1 minute and left at room temperature for 30 minutes. 1-2 μL of the supernatant was taken and injected into GLC for fatty acid analysis (ACEM 6000 model, Youngin Scientific, Korea) to analyze the fatty acid profile. As the standard solution used for the fatty acid analysis, 37 components FAME Mix (Sigma-Aldrich Co., St. Louis, Mo.) were used. The column was a SPTM-2560 Capillary GC Column (L x I.D. 100mx0.25mm, df 0.20um Omegawax 320 capillary column, USA). The starting temperature was programmed to 150 DEG C for 8 minutes and then increased by 2 DEG C per minute to 150-190 DEG C and the final temperature was fixed at 190 DEG C. [ Helium was controlled at a flow rate of 40 mL per minute with carrier gas and the split ratio was 100: 1. The injection temperature was 250 ° C and the detector temperature was 265 ° C. The peak area of the reference material and the peak area of the sample are compared to calculate the fatty acid content and are shown in Table 8 below.

Name
(% of total fatty acids) Groups C CP FCD3 FCD5 8: 0 Octanoic acid 0.03 ^b 0.43 + 0.11 ^a 0.41 + 0.02 ^a 0.41 + 0.04 ^a 10: 0 Decanoic acid - 0.45 ± 0.02 ^a 0.18 0.04 ^b 0.03 0.02 ^c 12: 0 Lauric acid 0.01 ^c 0.09 + 0.01 ^a 0.03 ± 0.01 ^b 0.03 ± 0.01 ^b 14: 0 Myristic acid 0.52 + 0.03 ^b 0.62 ± 0.03 ^a 0.49 + 0.01 ^b 0.54 + 0.03 ^b 16: 0 Palmitic acid 25.77 + - 0.10 ^a 25.21 ± 0.29 ^b 24.95 ± 0.07 ^b 23.92 + 0.17 ^c 16: 1 n-9 Palmitoleic acid 4.33 + 0.03 ^a 4.35 ± 0.08 ^a 4.03 0.06 ^b 4.44 ± 0.13 ^a 18: 0 Stearic acid 8.87 ± 0.07 ^b 9.42 + 0.04 ^a 8.76 ± 0.09 ^b 6.51 + 0.03 ^c 18: 1n-9 Oleic acid 45.51 ± 0.20 ^a 44.47 ± 0.16 ^b 44.82 + - 0.12 ^b 44.43 ± 0.39 ^b 18: 2n-6 Linoleic acid 14.38 ± 0.35 ^b 14.26 ± 0.05 ^b 14.46 ± 0.08 ^b 16.84 ± 0.51 ^a 18: 3n-3 Linolenic acid 0.08 ± 0.01 ^a 0.05 ± 0.02 ^b 0.03 0.02 ^b 0.08 0.02 ^a 20: 0 Arachidic acid 0.34 0.03 ^c 0.43 0.02 ^c 1.52 + - 0.10 ^b 2.46 ± 0.09 ^a 22: 0 Behenic acid 0.02 ± 0.01 0.02 ± 0.01 0.03 ± 0.01 0.03 ± 0.01 22: 1 Erucic acid 0.02 ± 0.01 ^b 0.06 0.02 ^a 0.03 ± 0.01 ^b 0.02 ^b 24: 0 Lignoceric acid 0.12 + 0.01 ^b 0.14 + 0.01 ^b 0.25 0.05 ^a 0.25 0.06 ^a Total 100 100 100 100 Saturated fatty acid 35.42 ± 0.19 ^b 36.43 ± 0.26 ^a 35.13 ± 0.18 ^b 31.81 0.09 ^c Unsaturated fatty acid 64.58 ± 0.19 ^b 63.57 + - 0.26 ^c 64.87 ± 0.18 ^b 68.19 + 0.09 ^a n-6 / n-3 ratio 42.46 ± 2.84 ^a 33.21 ± 1.65 ^b 9.57 ± 0.72 ^c 6.85 + - 0.46 ^c

Egg omega-3 fatty acid (n-3) was significantly higher than control (C) and CP treatment (p <0.05). In contrast, the n-6 / n-3 ratio was significantly lowered (P <0.05) as compared with control (C) 42.06 and CP treatment 33.23, with FCD treatment being 9.88-9.95. On the other hand, the omega-3 fatty acids in the eggs between control (C) and CP were higher in CP but not in n-6 / n-3 ratio (p <0.05).

The ideal ratio of n-6 / n-3, which is generally beneficial for health, is known to be less than 4: 1. Therefore, the above results show that when a feed supplemented with the powdered fermented feed additive of the present invention is fed to a laying hens, high-quality high-quality egg production, in which omega-3 fatty acid content and n-6 / n-3 ratio are lowered to an ideal 4: This is possible.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (11)

A feed additive for poultry comprising at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain, and Sphingobacteriaceae strain, and earthworm feces. The method according to claim 1,
Wherein the strain is a strain isolated from earthworm-infested soil. The method according to claim 1,
The strain of the genus Bacillus is Bacillus subtillis (KCTC2213), the strain of Streptomyces galilaeus (KCTC1919) is Streptomyces galileeus , the strain of Streptomyces galilaeus is KCTC1919 , ( Sphingobacteriaceae spp . , BR5-29). The method according to claim 1,
Wherein the feed additive for poultry comprises at least one strain selected from the group consisting of the Bacillaceae strain, the Streptomyces strain and the Sphingobacteriaceae strain at a concentration of 1.0 × 10 ⁴ to 1.0 × 10 ⁸ cfu / g. The method according to claim 1,
Wherein the poultry is at least one selected from the group consisting of broiler chickens, laying hens, native chickens, ducks, turkeys, quails and goji. The feed additive for poultry according to claim 1, wherein the feed additive for livestock promotes the growth of poultry and improves the quality of poultry meat and the productivity of poultry. A feed for poultry comprising the feed additive for poultry of claim 1. [7] The feed for poultry according to claim 7, wherein the feed additive for poultry is included in an amount of 0.01 to 10 parts by weight based on 100 parts by weight of the whole feed for poultry. 1) preparing at least one strain selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain isolated from earthworm feces, ; And
2) heat-treating the earthworm fungi and inoculating the strain of step 1 (step 2);
The present invention relates to a method for producing a feed additive for poultry including one or more strains selected from the group consisting of Bacillaceae strain, Streptomyces strain and Sphingobacteriaceae strain, and earthworm- Way. 10. The method of claim 9,
A method for producing a feed additive for poultry, characterized in that the strain of step 1 is isolated by the following steps:
3) heat-treating the earthworm fern (step 3);
4) separating the Bacillaceae strain from the heat-treated earthworm feces of step 3 (step 4);
5) re-inoculating the separated Bacillus sp. Strain of step 4 in the heat-treated earthworm feces of step 3 (step 5); And
6) isolating the Streptomyces strain and the Sphingobacteriaceae strain from the earthworm-infested soil of step 5 (step 6). 11. The method according to claim 9 or 10,
Wherein the heat treatment of the earthworm-modified feces is performed at 1000 ° C to 2000 ° C for 30 minutes to 3 hours.

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