KR20180011824A - Tablet of livestock probiotics for oral administration improving delivery efficiency to digestive tract and preparation method thereof - Google Patents

Abstract

The present invention relates to a tablet of livestock probiotics for oral administration. Even if thermal resistant probiotics for livestock, P. acidlactici PAGS, with improved antibacterial activity are formulated with only a pH-sensitive hydroxypropyl methylcellulose phthalate (HPMCP) single compound, provided is a tablet of livestock probiotics with improved delivery efficiency to digestive tracts and storage abilities of probiotics.

Description

Technical Field [0001] The present invention relates to a tablet for livestock prophylaxis for oral administration and a method for preparing the same,

More particularly, the present invention relates to a method for producing a livestock prophylactic tablet for oral administration, which comprises the steps of: preparing a livestock prophylactic agent for livestock by a tablet using only a single compound of pH-sensitive hydroxypropylmethylcellulose phthalate (HPMCP) The present invention relates to a heat-resistant livestock prophylactic < RTI ID = 0.0 > tablets, < / RTI >

Feed additives can be defined as non-nutrient supplement materials that are added to feed in small amounts for the purpose of improving productivity or improving meat quality. Antibiotics, probiotics, enzymes, organic acids, flavors, sweeteners, antioxidants, various natural and functional substances Can be classified as a feed additive. According to FDA, a concentrated feed additive is defined as a high concentration feed additive that can not be directly fed to an animal, which must be diluted with compound feeds and general feed additives. That is, It is known that about 0.5-5.0kg should be added to 1 ton of compound feed (Yang Yoon-mo, 2014, Advanced Technology for Biological Resource Information Service, Final Report of Feed Additive Patent Analysis, Korea Patent Information Center).

In recent decades, our livestock industry has been using antibiotics in order to improve the productivity of livestock. However, the number of microorganisms that are resistant to use frequently increases, the efficacy of the antibiotics is lowered and they are left in livestock products, In Korea, the use of antibiotics for feed additions since July 2011 was prohibited in Korea.

As an alternative to the prohibition of the use of antibiotics, probiotics such as lactic acid bacteria have already been used, and they are well known as antibiotic substitute agents because they have no antibiotic resistance and are highly effective in inhibiting pathogens and stabilizing microorganisms in the gastrointestinal tract.

However, when the probiotics are administered orally to livestock, they are not able to withstand the low pH of the stomach and have a problem in that a considerable number of viable cells are reduced during the propagation / storage of the probiotic. Many studies have been conducted to solve these problems, but there is a lack of effective strategies.

Therefore, the inventors of the present invention have developed a prodrug formulation capable of enduring at the lower pH and releasing at a neutral pH of the small intestine, while improving the shelf life of the probiotic.

In the prior art of the present invention, a method for producing a micro-coated probiotic micro-organism using vegetable mucin and water-soluble dietary fiber separated from the microorganism is disclosed in Korean Patent No. 10-1487431, The present invention relates to a probiotics formulation having a physiological function and viability of a living organism when the composition is orally administrated by microfabricating dietary fiber as a coating binder. In addition, a composition for an enteric hard capsule and an enteric hard capsule prepared using the composition are disclosed in Korean Patent Laid-Open No. 10-2012-0100305. However, it is also known that hydroxypropylmethylcellulose phthalate (HPMCP) and hydroxypropylmethylcellulose (HPMC), a water-soluble divalent salt, and an alkali agent to improve mechanical strength and salt retention, thereby improving storage stability. In addition, a method for manufacturing an enteric hard capsule and an enteric hard capsule are disclosed in Korean Patent No. 10-1182827. However, it is also known that hydroxypropylmethylcellulose phthalate (HPMCP) and hydroxypropylmethylcellulose acetate succinate (HPMCAS) capsule forming aid HPMC (hydroxypropyl methyl cellulose) and a neutralizing agent MC (methyl cellulose).

Therefore, in the above-mentioned Patent Documents, a livestock prophylactic agent tablet formulation in which a prodrug for livestock is formulated into a tablet using only a single compound of a pH sensitive polymer, hydroxypropylmethylcellulose phthalate (HPMCP) Is not known or implied.

Accordingly, it is an object of the present invention to provide a method for producing a heat-resistant livestock product which can formulate a heat-resistant livestock prophylactic agent with only pH-sensitive hydroxypropylmethylcellulose phthalate (HPMCP) To provide a prophylactic tablet formulation.

The above object of the present invention is achieved by preparing a livestock tablets according to the present invention by mixing a livestock prod- uct for livestock with hydroxypropyl methylcellulose phthalate (HPMCP) in a ratio of 1: 1 (w / w) and increasing the hardness to 3 to 10 KP This study was accomplished by evaluating the storage of heat - resistant high - hardness probiotics and the digestive tract delivery efficiency.

The present invention provides a livestock prophylactic tablet formulation in which the heat-resistant livestock prod- uct for livestock is formulated using only pH-sensitive single-component hydroxypropylmethylcellulose phthalate (HPMCP) It has excellent effect.

1 is a graph showing survival rates of probiotics carried on a livestock proliferative tablet prepared according to the present invention.
FIG. 2 is a graph showing the stability of a livestock food-grade tablet prepared according to the present invention in a gastrointestinal environmental condition (SGF).
3 is a graph showing survival rates of probiotics in a gastrointestinal environmental condition (SGF) of a livestock prophylactic tablet prepared according to the present invention.
4 is a graph showing survival rate of probiotics in the gastric environment condition (SGF + pepsin) of the livestock proliferative tablets prepared according to the present invention.
FIG. 5 is a graph showing stability in small intestinal environmental condition (SIF) of a livestock proliferative tablet prepared according to the present invention.
FIG. 6 is a graph showing the degree of release of probiotics carried on a livestock-produced tablets prepared according to the present invention in an in vitro gastrointestinal environment (SGF) and a small intestine environment (SIF).
FIG. 7 is a graph showing the survival rate of probiotics carried on a livestock-produced tablets prepared according to the present invention in an in vitro gastric environment (SGF) and a small intestine environment (SIF).
FIG. 8 is a graph showing the viability of a live-bacterium tablet prepared according to the present invention for 6 months at 4 ° C.
FIG. 9 is a graph showing viability of a livebye tablets for livestock produced according to the present invention at room temperature for 6 months.
FIG. 10 is a graph showing the efficiency of digestion intestinal tract delivery after 3 hours of administration to a broiler system for livestock prophylactic tablets prepared according to the present invention.
11 is a graph showing the digestion efficiency of the digestive tract after 6 hours of administration to the broiler system for the livestock tablets prepared according to the present invention.

The probiotic prophylactic agent tablets according to the present invention were prepared by combining the heat-resistant probiotics invented by the present inventors with a single compound hydroxypropylmethylcellulose phthalate (HPMCP) in a 1: 1 equivalent ratio (w / w) Which is characterized in that it is improved.

In the present invention, the type of the probiotics may be any conventional livestock prophylactic agent, but it is preferable that the developers of the present invention, among lactic acid bacteria, yeast, and bacillus, Using Pedicoccus acidlactici PA GS (KCTC 18229P).

Hereinafter, the present invention will be described in more detail with reference to embodiments and drawings. However, the following examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

Example 1. Production of Probiotic Tablet for Livestock of the Present Invention

In order to prepare a livestock active tablet for livestock according to the present invention, GS1 lactic acid bacteria (Genome shuffled 1), which is a lactic acid bacteria commonly used as a probiotics, and P. acidlactici, which is irradiated with ultraviolet rays, ) Was purchased from Shin-Etsu Chemicals Ltd. (Tokyo, Japan).

The method of manufacturing a livestock prophylactic tablet for livestock according to the present invention comprises mixing the lyophilized GS1 lactic acid bacteria and hydroxypropylmethylcellulose phthalate (HPMCP) powder in the same weight ratio (w / w) ratio, (Lactic Acid Bacillus: 12.5 mg: HPMCP: 12.5 mg) and the total hardness was 3, 6, and 10 KP (Kilo Pound). The hardness of the tablet can increase the hardness of the tablet by further increasing the tabletting pressure.

Experimental Example 1 The survival rate of probiotics carried on the live-frozen tablets for the present invention was confirmed

Survival rates of the probiotics carried on the three kinds of probiotics tablets of 3, 6 and 10 KP prepared in Example 1 were confirmed. As a control, probiotic powder was used.

Each tablet was dissolved in 1 mL of phosphate buffered saline (PBS), diluted stepwise by 10 times, plated on an MRS agar plate, incubated at 37 ° C for 24-36 hours, and then viable cells were counted . The viable count was expressed as colony forming unit (cfu).

As shown in Fig. 1, the survival rate of the probiotics supported on the livestock prophylactic tablets of the present invention showed 90% or more of all three types of probiotics of hardness 3, 6 and 10 KP, and the tendency was slightly decreased as the hardness of the tablet increased This is because the heat generated as the tablet pressure increases during tablet production lowers the heat resistance of the probiotic agent and reduces the survival rate.

Experimental Example 2 Stomach condition stability test

Stability of the 10KP probiotic tablet of the present invention was confirmed in pH 2 gastric environment, which is a strong acidic condition.

10KP probiotic tablets were immersed in simulated gastric fluid (SGF) and examined for changes in morphology according to time (0 min, 15 min, 30 min, 60 min, and 120 min).

As shown in FIG. 2, it was confirmed that the 10 mg KP tablets of the present invention did not disintegrate for 120 minutes even when treated with the activated gastric juice (SGF), and maintained its complete shape and maintained stability under gastrointestinal conditions.

Experimental Example 3. Probiotic survival rate in gastrointestinal environment in vitro

In an in vitro gastrointestinal environment, the survival rate of the probiotics carried on the hardness 3,6, 10KP probiotic tablet of the present invention was confirmed. As a control, probiotic powder was used.

Each tablet was shake-cultured in 5 ml of activated gastric juice (pH 2.0) or activated gastric juice (SGF) supplemented with pepsin at 37 ° C and 100 rpm for 2 hours, 15 minutes, 30 minutes, 60 minutes, and 120 minutes). Each sample was diluted stepwise by 10 folds, plated on MRS agar plate, cultured at 37 ° C for 24-36 hours, and viable cell counts were then measured. The number of viable cells was expressed in terms of the bacterial unit (cfu).

As a result, as shown in FIG. 3, when the hardness of 3,6,2 10 KP probiotic tablet of the present invention was cultured for 2 hours under the condition of activated gastric juice (SGF), the survival rate of the probiotics was significantly higher than that of the control group. Survival rate of probiotics was more than 70% even in gastric environment.

Also, as shown in FIG. 4, even in the gastrointestinal environmental condition in which activated gastric juice and pepsin were simultaneously treated, the hardness of the 3,6,10 KP probiotic tablet of the present invention was much higher than that of the control group.

Experimental Example 4. Small intestinal stability test

The stability of the present hardness 3, 6, 10KP probiotic tablet of the present invention was confirmed in a pH 7.2 small environment. Each tablet was placed in 5 ml of phosphate buffered saline (PBS) to determine the time at which the tablet was completely disintegrated.

As shown in FIG. 5, when the stimulated intestinal fluid (SIF) was applied to the hardness 3,6, 10 KP probiotic tablet of the present invention, the hardness 3 KP tablet disintegrated within 30 minutes, while the hardness 10 KP tablet was deformed for more than 1 hour Respectively. As a result, it was confirmed that the tablet compression time in the small intestine environment increased as the tablet pressure increased.

Experimental Example 5 The probiotic release test carried on the live-frozen tablets for the present invention

In the in vitro gastrointestinal environment (SGF) and small intestine environment (SIF) in which the digestive physiological conditions of livestock were similar, the degree of release of the probiotics carried on the hardness 3,6, 10KP probiotic tablet of the present invention was measured.

Each tablet was placed in the gastrointestinal environment (SGF) for 1 hour, then in the intestinal environment (SIF) for 4 hours thereafter, every 30 minutes for the first 3 hours, then every hour thereafter, Were sampled. Each sample was diluted stepwise by 10 folds, plated on MRS agar plate, cultured at 37 ° C for 24-36 hours, and viable cell counts were then measured. The viable cell counts were expressed in terms of bacterial growth units (cfu).

As shown in FIG. 6, the hardness 3,6,10 KP probiotic tablet of the present invention was preserved in the tablet in the gastrointestinal environment (SGF) for 1 hour without release of the probiotic agent and in the small intestine environment (SIF) 20% to 50% of the probiotics and 100% of the probiotics were released after 5 hours. In addition, the degree of release of the probiotics carried on the probiotic microbial tablet of the present invention showed a tendency to be slower as the hardness of the tablet was higher.

Experimental Example 6 . In vitro in the gastrointestinal and small intestinal environment , Survival rate of probiotics supported

* Survival rate of probiotics carried on the hardness 3, 6, 10KP probiotic tablet of the present invention was confirmed in the in vitro gastric environment (SGF) and small intestine environment (SIF) as in Experimental Example 5 above.

Each tablet was shaken for 2 hours in the gastrointestinal environment (SGF) and in the small intestine environment (SIF) for 4 hours at 37 ° C and 100 rpm, and sampling was performed every hour. Each sample was diluted stepwise by 10 folds, plated on MRS agar plate, cultured at 37 ° C for 24-36 hours, and viable cell counts were then measured. The number of viable cells was expressed in terms of the bacterial unit (cfu).

As shown in FIG. 7, the survival rate of the probiotics carried on the hardness 3,6, 10KP probiotic tablet of the present invention decreased to 65-85% over time in the gastrointestinal environment (SGF), but stable in the small intestine environment (SIF) Respectively. In particular, the survival rate of the probiotics carried on the hard 10KP probiotics tablet of the present invention showed 80% or more in the gastrointestinal environment and the small intestine, showing a very high probiotic protective effect.

Experimental Example 7. Assay of Probiotic Storage of Probiotic Tablet for Livestock of the Present Invention

In order to evaluate the efficacy stability of the hardness 3,6, 10KP probiotic tablet of the present invention during the storage period, the survival rate of the probiotics carried on the tablet was measured after storing the tablet of the present invention for 4 months at room temperature and room temperature for 6 months.

As shown in FIGS. 8 to 9, the shelf life of the three hardness tablets of 3, 6 and 10 KP according to the present invention was 6.5 log CFU / tablt after 6 months in both the 4 ° C refrigeration environment (FIG. 8) Respectively.

Experimental Example 8. In vivo assay of the livestock prophylactic tablet of the present invention

In order to investigate the in vivo delivery efficiency of the probiotics carried on the probiotic tablet according to the present invention, the survival rate of probiotics in the digestive tract was measured after oral administration of the livestock probiotics tablet of the present invention to a broiler house of 11 days old at 6 days per group. The group of animals was divided into a control group (control), a solution containing GS1 liquid lactic acid bacteria, and a treatment group (Tablets) containing the 10 KP probiotic tablet of the present invention. The dose of the G1 lactic acid bacterium was 2 x 10 ⁸ CFU for 5 times. The G1 lactic acid bacteria administration time is 0, 4, 24, 28, 48 hours after the start of the test. To confirm the survival rate of lactic acid bacteria in digestive tract, proximal, intestine and cecum of broiler chickens were harvested 3 hours and 6 hours after the last administration of lactic acid bacteria, and the contents were cultured in Pediococcus selective medium.

As shown in Figs. 10 to 11, survival rates of lactic acid bacteria in digestive gut in the case of livestock tablets treated according to the present invention for 3 hours (Fig. 10) and 6 hours (Fig. 11) after oral administration were significantly higher than those of control and liquid lactic acid bacteria Especially in small intestine (DI) and cecum (CE). Thus, it was confirmed that when the high-hardness tablet of the heat-resistant probiotics for livestock according to the present invention is orally administered to the livestock, the probiotics carried on the tablet can be remarkably efficiently delivered into the gastrointestinal tract.

As described above, the high-hardness tablet prepared from the same ratio of the heat-resistant livestock bacterium to the single compound HPMCP is an extremely useful invention in the animal feed and livestock industry because it has excellent digestive tract transmission efficiency and storability of probiotics at the same time.

Claims (2)

(Pediococcus acid lactic acid PAGS: KCTC18229P) is selected from among the probiotics based on 100 parts by weight of the whole of the probiotic tablet composition and 50 parts by weight of hydroxypropylmethylcellulose phthalate (HPMCP) is mixed with 50 parts by weight of the antibacterial agent to form a tablet having a hardness of 3 to 10 kp Gt; < tb >< / TABLE >
A heat resistant livestock tablets for livestock produced by the method of claim 1.

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