KR20060062740A - Novel lactic bacterium, and a method for preparing the fermented cactus - Google Patents

Abstract

The present invention relates to a novel lactic acid bacteria strain isolated from the fruit of a cactus, in particular palm cactus, a method for producing the lactic acid bacteria fermentation of the cactus using the same, and a method of breeding flounder using the produced lactic acid bacteria fermentation.

Cactus, fermented product, Lactobacillus plantarum, olive flounder, feed additive

Description

Novel Lactic Bacterium, And A Method for preparing the fermented Cactus}

1 is a result obtained by sequencing of 16S rRNA, showing which strain belongs to a novel lactic acid bacteria CNU001 strain isolated from cactus fruit.

Figure 2 is a view showing the change in sugar by fermentation period when the cactus fermentation using the novel lactic acid bacteria CNU001 strain according to the present invention.

Figure 3 is a view showing the acidity change by the fermentation period when the cactus fermentation using the novel lactic acid bacteria CNU001 strain according to the present invention.

Figure 4 is a view showing the change in the number of lactic acid bacteria by fermentation period obtained when inoculated into the cactus fruit juice of the novel lactic acid bacteria CNU001 strain according to the present invention.

5 is a view showing the results of the analysis of the total sugar content of each fermentation period obtained when fermenting a novel lactic acid bacteria CNU001 strain according to the present invention.

6 is a view showing the change of the organic acid content in the cactus fruit juice fermented by the novel lactic acid bacteria CNU001 strain according to the present invention.

Figure 7 is a view showing how much the strain of the present invention is settled in the intestine of the flounder when the novel lactic acid bacteria CNU001 strain according to the present invention to the flounder.

8 is a view showing the antimicrobial effect of the novel lactic acid bacteria CNU001 strain according to the present invention (A: effect on Streptococcus, B: effect on Edward, C: effect on Vibrio).

9 is a diagram showing a 16S rRNA nucleotide sequence of Lactobacillus plantarum CNU001.

The present invention relates to a novel lactic acid bacteria strain isolated from the fruit of a cactus, in particular palm cactus, a method for producing the lactic acid bacteria fermentation of the cactus using the same, and a method of breeding flounder using the produced lactic acid bacteria fermentation.

The palm cactus (opuntia ficus-indica) is called baeknyeoncho. It is a native plant of Jeju Island, grows well in relatively poor soil, grows widely in difficult places, and contributes to small-scale income in farms.

The effectiveness of palm cactus is known to have functions such as antibacterial and antiviral, and has recently been reported on immune function increase and antidiabetic effect. This effect is mostly taken from palm cactus fruit or processed by itself, and the active ingredient is covered by mucus, so there is a weak point in terms of effective intake of the active ingredient, and at the same time, humans are disgusted with smell and high viscosity. It's coming up. As a method for solving this problem, functional extracts have been developed by adding extracts from palm cactus fruits and stems and adding fructose and the like.

However, due to the high viscosity when making the extract, when added as an additive to functional materials for other uses, such as aquaculture feed, it is difficult to uniformly mix, there is a disadvantage that the recovery rate is low and the active ingredient contained in the fiber bundle is lost. .

In order to solve this problem, Korean Patent Laid-Open Publication No. 2003-94879 discloses a step of selecting and washing a palm cactus fruit; Grinding the cactus fruit with a mixer to obtain a homogeneous liquid; Mixing a fermentation accelerator such as molasses or starch syrup and useful microbial agents such as lactic acid bacteria in the homogeneous liquid; Fermenting at least 2 weeks at room temperature to obtain a fermentation broth; And it describes a method for producing a cactus fermentation comprising the step of preparing the fermentation broth as a lyophilized powder, and a method of farming flounder using the same.

However, useful microorganisms used in the above technique are not excellent in fermentation performance, and thus there has been a need in the art for useful microorganisms having better fermentation performance.

Therefore, the present inventors also have excellent performance in fermenting cactus, especially palm cactus, and research to find a novel microorganism which can further improve the growth of the flounder when the flounder is raised by using the fermented product of the prepared cactus. As a result, a fermented product of cactus, in particular, palm cactus, was prepared using the Lactobacillus plantarum CNU001 isolated from the cactus fruit by the present inventors, and the fermented product was used as a feed additive to raise the flounder. In this case, it was found that the very good flounder growth can be achieved, and completed the present invention.

Accordingly, it is an object of the present invention to provide a novel lactic acid bacteria strain isolated from the fruit of a cactus, in particular palm cactus.

In addition, another object of the present invention is to provide a method for producing a lactic acid bacteria fermentation product of cactus using the isolated novel lactic acid bacteria strain.

In addition, another object of the present invention, and to provide a method of raising the flounder by the lactic acid bacteria fermented product prepared using the isolated novel lactic acid bacteria strain.

In order to achieve the above object, as a novel microorganism with excellent performance in fermenting a cactus, especially a palm cactus, Lactobacillus plantarum CNU001 (KCTC18105P) isolated from cactus fruit is provided.

The microorganism is used to ferment cactus, especially palm cactus, to produce a cactus fermentation product, and in particular when breeding flounder using it, it can further promote the growth of the flounder, which can be used as a feed additive for the flounder growth. have.

As a novel microorganism discovered by the present inventors, Lactobacillus plantarum CNU001 was strained on October 14, 2004 to the Korea Institute of Bioscience and Biotechnology, located at 52, Eeun-dong, Yuseong-gu, Daejeon, Korea. Received a strain accession number of -18105P.

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

As a novel microorganism developed by the inventors, Lactobacillus plantarum CNU001 can be cultured in MRS liquid medium or agar medium for Lactobacillus commonly known in the art. As to conditions and methods for culturing the microorganism according to the present invention, culture conditions and methods commonly known in the art may be used for culturing Lactobacillus plantarum. For example, according to the method described in Example 1, Lactobacillus plantarum CNU001 which is the microorganism of the present invention can be cultured.

In addition, the microorganism Lactobacillus plantarum CNU001 according to the present invention is a bacillus, stained gram positive, and has a catalase negative reaction. In addition, the microorganism of the present invention exhibits sugar fermentation properties having 99.9% homogeneity with Lactobacillus plantarum (ATCC 14917). More specifically, the microorganism CNU001 according to the present invention is L-arabinose, ribose, galactose, glucose, fructose, mannose, mannitol, sorbitol, α-methyl-D-mannoside, N-acetyl-glucosamine, amigdaline, Sugar sources such as arbutin, esculin, salicylic acid, cellulose, maltose, lactose, melibiose, sucrose, trehalose, merezitose, raffinose and D-turanose can be used. .

In addition, the strains according to the invention have 99% similarity in Lactobacillus plantarum DSM 20205 and 16S rRNA sequences.

However, although the microorganism CNU001 according to the present invention exhibits significant species similarity compared to those of the Lactobacillus plantarum family, cacti, in particular, compared to the known Lactobacillus plantarum family of species or several other lactic acid bacteria. It shows a very good performance in fermentation properties of palm cactus, and also shows a significant improvement in the growth effect of the flounder using the produced cactus fermentation. The fermentation performance of these microorganisms and the effect of improving the flounder growth has not been reported so far in the art.

A fermentation product of a cactus, in particular a palm cactus, using the microorganism Lactobacillus plantarum CNU001 according to the present invention is a method similar to a method of preparing a fermentation product of a cactus, in particular, a palm cactus, using a lactic acid bacterium, for example, a patent publication. It may be prepared using a method similar to that described in Publication 2003-94879. For example, selecting and washing the palm cactus fruit; Grinding the cactus fruit with a mixer to obtain a homogeneous liquid; Mixing the homogenate with a fermentation accelerator such as molasses or starch syrup and Lactobacillus plantarum CNU001 which is a strain according to the present invention; Fermentation at room temperature for 1 minute to 2 weeks, or more to obtain a fermentation broth; And the method comprising the step of preparing the fermentation broth lyophilized powder can be used for the production of cactus fermentation. In the above preparation, various strains for obtaining cactus lactic acid bacteria fermented products, such as the amount of Lactobacillus plantarum CNU001 as the strain for fermentation, the amount of fermentation accelerators such as molasses or starch syrup used, fermentation conditions, and methods for producing lyophilized powder Conditions may be referred to from Japanese Patent Laid-Open Publication No. 2003-94879 or obtained through routine experimentation.

On the other hand, according to the present invention, as a problem caused by simply extracting and using cactus, in particular palm cactus, inconvenience of use or storage due to high viscosity; Heterogeneous mixing that occurs when used as an additive in functional materials for other uses, such as aquaculture feeds; The problems of low recovery rate of active ingredient in cactus and loss of active ingredient in cactus can be solved by using Lactobacillus plantarum CNU001 as the lactic acid bacterium according to the present invention.

In addition, according to the present invention, the Lactobacillus plantarum CNU001 strain according to the present invention is very high in viability, and excellent in intestinal fixation ability, when preparing a cactus ferment using the same, strain CNU001 according to the present invention Is present in the fermented product in a live state, and in the flounder ingested such a cactus fermented product, the strain CNU001 according to the present invention can be delivered and settled in the living state to the intestine. And their beneficial effects can improve the intestinal health of the flounder, which can significantly improve the flounder's rearing effect.

On the other hand, the cactus ferment produced using the strain according to the present invention can be used as a feed additive for the culture of the flounder.

When the cactus fermentation product according to the present invention is used as a feed additive, it exhibits a high antibacterial effect against various microorganisms such as streptococci, Edwards bacteria and Vibrio bacteria which should be avoided in the flounder culture. By using the cactus fermentation according to, it is possible to obtain a high breeding effect by improving the breeding environment of the flounder.

In addition, when the cactus fermentation is used as an additive for feed, the cactus fermentation may exhibit antiviral action such as suppression of proliferation of EHV virus and invasive gastrointestinal virus.

Due to the advantages described above, the cactus fermentation product according to the present invention prepared using Lactobacillus plantarum CNU001 as a novel strain can be used as a feed additive of flounder.

On the other hand, a technique for producing a feed using the cactus fermentation according to the present invention as an additive for feed, can be prepared in the same manner as the technique known in the art for producing a feed by adding an additive known in the art to the feed have.

When the flounder is cultivated using the feed containing the cactus fermentation product according to the present invention as the feed additive of the flounder, the flounder may exhibit a significant growth effect in both the fry and the stage.

The amount of cactus fermentation added to the feed may vary depending on the production method of the feed, the animal to be bred, the health condition of the animal, etc., and the appropriate amount may be easily recognized by those skilled in the art through repeated experiments.

The cactus fermented product according to the present invention can be used as an additive for feed of a flounder, but also as an additive for feed of various animals such as pets such as dogs and cats, aquatic animals such as various fishes, various reptiles, various amphibians and various insects, in addition to the flounder. Can be used, which is also included within the scope of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

In the following examples, what is described as a cactus is intended to mean the palm cactus, unless otherwise noted.

EXAMPLE

Example 1 Isolation and Identification of New Lactic Acid Bacteria

1) Isolation of bacteria

Naturally aged cactus fruit was aseptically homogenized and juiced, then diluted in sterile saline solution (0.9% NaCl), plated in MRS medium containing 0.5% CaCO _3, and incubated at 30 ° C. for 3 days. . After 3 days of incubation, the strain with the most transparent ring around the cells was isolated. The isolated strain was passaged in the same medium to obtain a single strain. This strain was identified as belonging to the genus Lactobacillus plantarum through subsequent experiments, and the strain was named Lactobacillus plantarum CNU001 (hereinafter, often referred to as 'CNU 001'). The strain was deposited on October 14, 2004 at the Korea Biotechnology Research Institute Biological Resource Center, 52, Eeun-dong, Yuseong-gu, Daejeon, and received a strain accession number of KCTC-18105P.

2) Cultivation of CNU001 Strains

Lactobacillus MRS liquid medium and Lactobacillus MRS agar medium (Difco Lacboratories, MD USA) were used to culture the isolated strain CNU001. In the case of liquid medium, 55 g of medium is added to 1 L of distilled water and then boiled for 2 minutes to completely dissolve all compounds, followed by sterilization for 15 minutes at 121 ° C. In the case of solid medium, 70 g of medium is added to 1 L of distilled water and then boiled for 2 minutes. Completely dissolved and then sterilized for 15 minutes at 121 ℃ was used. The preparation of the culture was inoculated with a single colony of the strain isolated in a liquid medium divided into 100mL each 500mL Erlenmeyer flask and shake cultured at 30 ℃, 150rpm for 2-3 days.

3) Identification of Strain CNU001

-Morphological characteristics of bacteria

Some of the liquid cultures cultured at 30 ° C. for 2 to 3 days using Lactobacillus MRS liquid medium were found to be bacilli, and gram staining resulted in gram positive bacteria. In addition, as a result of examining catalase production, a catalase negative reaction was observed.

-Physiological characteristics of bacteria and utilization of carbon source

In order to investigate the physiological properties and carbon source availability of the strain, the sugar fermentation properties were investigated using the API 50CHL (Biomerieux, France) system according to the manufacturer's instructions. The isolated strains were incubated at 30 ° C. for 3 days and then inoculated onto the substrate of the kit for Lactobacillus, and as a result the sugar fermentation characteristics of the isolated strains in the present invention are shown in Table 1 below. As shown in the table below, the CNU001 strain was found to have 99.9% homology with Lactobacillus plantarum . In Table 1, the species of ATCC 14917 is a species of the Lactobacillus plantarum family.

-Sequence analysis of the bacterium 16S rRNA

The isolated strains were inoculated in Lactobacillus MRS liquid medium (Difco, USA) and incubated at 30 ° C. for 18 hours, followed by centrifugation at 10,000 × g for 10 minutes to harvest the cells, followed by NucleoSpin Tissue Kit (ClonTech, Lab. Inc.). Genomic DNA was isolated using the manufacturer's method. PCR was performed using 27f (5'-AGAGTTTGATCCTGGCTCAG-3 ') and 1522r (5'-AAAGAGGTGATCCARCCGCA-3') primers to amplify 16S rDNA from the isolated whole DNA. PCR conditions were denatured at 95 ° C. for 5 minutes, and then cycled 33 times in cycles of 95 ° C.-1 min, 55 ° C.-1 min, and 72 ° C.-1 min, and finally reacted at 72 ° C. for 10 minutes. The amplified PCR product was confirmed by electrophoresis on 1% agarose gel. The identified PCR product was recovered and purified using a QIAEX ^R II Gel Extraction kit, ligated to pGEM-T Easy Vector (Promega, Madison. Wis.), Transformed to E. coli JM109, and then 40ug / mL. Was cultured at 37 ° C. for 18 hours using X-gal and LTG agar medium containing IPTG and 50 ug / mL ampicillin, and then white colonies were selected, followed by analysis of gene sequences using SP6 and T7 primers. It was. The analyzed gene sequence is as shown in FIG.

Gene sequencing was performed by Big-Dye Cycle Sequencing kit (PE Applied Biosystems, Fster City, CA) and ABI Prims 310 automatic sequencer (PE Applied Biosystems, Foster City, CA). Analysis of the identified sequences was performed using the DNAstar program and GeneBank's blast search database. In order to elucidate the phylogenetic relationship of the bases, the analysis was performed using Lasergene's Megalign program, which was performed by the Cluster method. The results identified through 16S rDNA sequencing are shown in FIG. 1. As shown in FIG. 1, the CNU001 strain was found to have 99% similarity with Lactobacillus plantarum DSM 20205.

Example 2. Characteristics of Isolated Lactobacillus CNU001

1) Cactus fermentation assay of isolated strain

After removing the thorns of cactus fruit, washed with water, put 1L of distilled water in 10Kg, steamed at 90 ℃ and then juiced to remove the fruit juice for fermentation was prepared.

Fermented fruit juice of the prepared cactus was fermented using Lactobacillus plantarum CNU001, which is a strain of the present invention. That is, 1 mL of the strain solution was inoculated into 100 mL of sterilized MRS liquid medium and incubated at 30 ° C. for 20 hours. The culture solution was inoculated with 0.2% of cactus fruit juice, dispensed into 2L culture bottles, sealed, and incubated at 25 ° C. for 7 days, and the pH, sugar content, titratable acidity, and lactic acid bacteria count were examined for each fermentation period.

-Sugar content by fermentation period

The obtained fermentation broth was measured for each fermentation period using a refractometer (Hand refractometer, No. 507-1, NOW, Japan). The results are shown in FIG.

As shown in FIG. 2, the change in sugar content during the culture period was only slightly changed from 4.0 Brix to 3.2 to 3.4 Brix, but no significant change was confirmed.

-Measurement of pH and total acidity of fermentation broth by fermentation period

The pH was measured by a pH meter taking a certain amount of the sample, the total acidity was calculated by the following equation by adding 5 mL of sterile water to 5 mL of fermentation broth and titrating to pH 8.3 with 0.1N NaOH. The results are shown in FIG.

As shown in Figure 3, when the fermentation is in progress, the pH change sharply decreased from the first day of culture, the pH change at the end of fermentation was found to be pH3.50. The change in total acidity produced acid rapidly as fermentation progressed, and the acidity was 0.85% at the end of fermentation.

-Lactic acid bacteria count

0.5 mL of the fermentation broth prepared above was suspended in 4.5 mL of 0.9% NaCl solution, diluted 10-fold, and then pour plating using Lactobacillus MRS agar medium containing 0.5% CaCO _3. Incubated at 3 ° C. for 3 days, the colonies were counted to form a clear and relatively clear zone, and the same experiment was repeated three times to calculate an average value. The results are shown in FIG.

As shown in Figure 4, the CNU001 strain was inoculated in the cactus fruit juice solution and fermented at 25 ℃ as a result of measuring the change in the number of lactic acid bacteria as a log value, each inoculation amount 6.1 ~ 6.9 CFU / mL growth characteristics according to the fermentation period As a result, only one day of culture increased significantly to reach 8.4 CFU / mL and maintained similar level from the second day until the end of fermentation.

-Analysis of sugars and organic acids in fermentation broth

After the prepared fermentation broth was centrifuged, the supernatant was collected and filtered with a 0.2 μm Millipore membrane filter. The filtrate was analyzed and quantified by the HPLC using the sugars of the fermentation broth. At this time, HPLC was used as a water carbohydrate column, the detector was ELSD2000, the eluent was acetonitrile: H ₂ O (75:25), the flow rate was 1.0 mL / min, and the injection volume was 10 μl. The organic acid analysis of the fermentation broth was performed using a SupelcogelTM C-610H column, the detector was UV (210 nm), the eluate was 0.1% phosphoric acid, the flow rate was 0.5 mL / min, and the injection volume was 5 μl. The results are shown in FIGS. 5 and 6.

As shown in FIG. 5, as a result of analyzing the total sugar content according to the fermentation period, the glucose content rapidly decreased from 4.6 mg / mL to 1.0 mg / mL until the third day of fermentation and gradually decreased from the third day. Fructose tended to decrease gradually as fermentation progressed. And the sucrose content was found to be almost unchanged.

In addition, as shown in Figure 6, the organic acid content of the cactus fruit juice juice fermented by Lactobacillus plantarum CNU001, mainly citric acid, malic acid, lactic acid, acetic acid and the like was separated. While the citric acid was fermented at about 1.8 mg / mL before fermentation, there was almost no change, but lactic acid was not detected before fermentation but increased as the fermentation progressed. In particular, after the end of the fermentation, the content of lactic acid was detected at 3.87 mg / mL.

Example 3 Preparation of Cactus Fermentation Using CNU001

The cactus fruit juice was calibrated to pH 6.6 before fermentation and inoculated with lactic acid bacteria for 3-4 days. The maltose was added to the fermentation broth to 10% concentration and homogenized. The skim milk was added to the fermentation broth as a cryoprotectant at a concentration of 10%, homogenized by mixing, and then frozen at -70 ° C for 3 days, and then dried in a freeze dryer. The dried sample was ground and powdered.

Example 4. Component Analysis of Cactus Ferment Prepared Using CNU001

In order to confirm the fermentation efficacy of the strain of the present invention, the content of various components before and after fermentation of the cactus using the strain CNU001 according to the present invention was analyzed.

The results of analysis of the contents of total components, amino acid components and lipid components of cactus before and after fermentation using Lactobacillus plantarum CNU001 strain as fermentation strains are shown in Tables 1, 2 and 3, respectively.

As shown in the following results, it can be seen that through the fermentation using the strain of the present invention, the content of mucus components such as fibers in cactus, which has been a problem in the past, has been greatly reduced.

Analytical Ingredient Before fermentation After fermentation moisture(%)  93.18 96.86 Crude Protein (%) 0.07 0.07 Crude Lipids (%) 0.15 0.08 Crude fiber (%) 0.11 0.04 (%) 0.53 0.49 Ca (ppm) 236.03 188.02 P (ppm) 52.45 35.53 Vitamin C (ppm) 4.45 Not detected Carotenoids (ppm) 0.04 0.06 Flavonoids (ppm) 70.24 37.20 Vitamin E (a-tocopherol) (ppm) 2.31 0.74

Analytical Ingredients Before fermentation (ppm) After fermentation (ppm) Aspartic Acid Threonine Serine Glutamate Proline Glycine Alanine Valine Isoleucine Leucine Tyrosine Phenylalanine Histidine Lysine Arginine Cysteine Methionine Tryptophan 39.64 10.11 28.01 74.14 24.44 25.02 15.18 15.06 14.66 13.19 110.81 2.66 87.45 10.90 9.53 121.23 22.11 13.40 26.00 6.63 17.21 67.41 94.35 13.38 14.91 18.25 23.57 3.90 37.10 20.95 78.28 21.11 5.03 139.68 31.20 12.28

Analytical Ingredient Before fermentation (g / 100g) After fermentation (g / 100g)  Myristic acid C14: 0 Pentadecanoic acid C15: 0 Palmitic acid C16: 0 Palmitic acid C16: 1 Tridecanoic acid C13: 0 Margarine C17: 0 Margaoleic acid C17: 0 Stearic acid C18: 0 Oleic acid C18: 1 Linole Reinic acid C18: 2n6 Linolenic acid C18: 4n3 Arachidic acid C20: 0 Eicosenoic acid C20: 1 Arachidonic acid C20: 4n6 EPA C20: 5n3 Behenic acid C22: 0 Docosateranoenoic acid C22: 4n6 DPA C22: 5n3 DHA C22 : 6n3 capric acid nonadecanoic acid C19: 0 lauric acid C12: 0 eicosaprienenoic acid C20: 3n6 erucic acid C22: 1  0.20 0.10 14.03 0.87 0.03 0.12 0.08 4.36 13.43 59.75 1.50 0.78 0.40 0.21 0.52 0.56 0.30 0.07 0.07 0.35 0.37  0.21 13.21 0.93 0.13 0.10 4.71 12.56 60.79 0.68 0.84 0.67 0.52 0.57 0.14 0.28 0.29 0.17

In addition, the contents of the total component, amino acid component and lipid component of the cactus, analyzed after performing the lyophilization process after adult fermentation, are shown in Tables 1-2, 2-2, and 3-2, respectively.

TABLE 1-2

Analysis item unit Analysis moisture % 14.02 Crude protein % 6.02 George Room % 1.32 Joe Island Yu % 7.61 Inquiry minutes % 10.85 calcium % 1.85 sign % 0.02 Vitamin c ppm 22.34 Vitamin E (α-tocopherol) ppm 101.11 Carotenoids ppm 17.89 Flavonoids ppm 712.80

Table 2-2

Analysis item Analysis result (g / 100g) Aspartic acid 0.40 Threonine 0.17 Serine 0.18 Glutamic acid 0.63 Proline 0.42 Glycine 0.30 Alanine 0.28 Valine 0.21 Isoleucine 0.12 Leucine 0.22 Tyrosine 0.18 Phenylalanine 0.14 Histidine 0.21 Lee Sin 0.17 Arginine 0.21 Cysteine 0.15 Methionine 0.06 Tryptophan 0.04

Table 3-2

Analysis item Analysis result (g / 100g fatty acid) Lauric acid C _{12: 0} 0.69 Myristic acid C _{14: 0} 1.90 Myristic oleic acid C _{14: 1} 0.86 Palmitic acid C _{16: 0} 17.89 Palmitic Acid C _{16: 1} 2.35 Stearic Acid C _{18: 0} 3.63 Oleic Acid C _{18: 1} 13.43 _Linoleic acid C _{18: 2 n6} 49.47 Linolenic Acid C _{18: 3n3} 7.02 Arachidic acid C _{20: 0} 0.65 Eicosadienoic acid C _{20: 2} 0.46 Behenshan C _{22: 0} 0.71 DPA C _{22: 5 n3} 0.93 Nonpublic 0.01

Example 5 Intestinal Bacterial Distribution of Cactus Fruit Fermentation Solution Prepared Using CNU001 Strains through the Olive Flounder

In the present embodiment, using the Lactobacillus plantarum CNU001 strain of the present invention, the cactus fermentation was prepared according to Example 3, and the prepared fermented product was prepared in EP feed at a concentration of 1% by weight. Japan), and then ingested into the flounder for about 3 months, when the strain was administered to the flounder, it was carried out to confirm the extent of the strain of the present invention in the intestine of the flounder.

The results are shown in FIG. That is, as shown in Figure 7, Lactobacillus plantarum The fermented cactus fermented with CNU001 strain was continuously administered to the flounder, and the change in the intestinal bacteria on the flounder was measured by viable cell count. In the group, the intestinal bacterial count was reduced by 23.6% compared to the control group, and the lactic acid bacteria was present at 3.3 Log10 CFU per 1g of intestine at the administration.

Example 6. Antibacterial Effect of Cactus Fermentation According to the Present Invention

In the present embodiment, when the fermentation of cactus fermentation prepared according to Example 3 to flounder, the experiment was conducted to check the antimicrobial effect of the flounder (Paralichthys olivaceus), and to check the antibacterial effect in the flounder culture Streptococci, Edwards bacteria and Vibrio bacteria that require sterilization were used.

1) Streptococcus

To investigate the effect of cactus fermentation on the growth of streptococci, the growth rate of bacteria in liquid medium was compared. First, the freeze-dried cactus ferment prepared according to Example 3 was suspended in BHI (Brain Heat Infusion; Difco) at a final concentration of 2% (w / v), mixed at room temperature for 30 minutes, and then adjusted to pH 7.0. . The mixture was placed in an ice box and sonicated for 15 seconds and allowed to stand for 15 seconds for 8 times. After sonication, the insoluble material was removed with a filter paper, and then used as an experimental group by passing through a syringe filtration sterilizer (0.45 um pore size). BHI without cactus fermentation was used as a control.

Streptococci were inoculated at 1.2 and 10 ⁴ CFU / ml levels for 18 hours in BHI in the experimental and control groups, and then at 0, 2, 4, 8, 12 and 24 hours after inoculation while standing at 22 ° C. The number of bacteria was measured. The bacterial count was measured by diluting 100 µl of the culture solution in sterile saline solution and then smearing in TSA (Tryptic Soy agar; Difco) added with 2% NaCl at each measurement time in the experimental and control groups. It was measured. The results are shown in Table 4 and FIG. 8A. As shown in Table 4 and Figure 8A, there was a significant bacterial growth inhibitory effect compared to the control. That is, the control group without the cactus was grown to 9.2 log10 CFU / ml after 24 hours in the first 4.1 log10 CFU / ml, but in the experimental group to which the cactus ferment was added, 4.7 log10 CFU / ml after 24 hours of culture. The number of bacteria was shown and no growth of bacteria was observed.

Changes in Cell Numbers Over Time 0 hours 2 hours 4 hours 8 hours 12 hours 24 hours Cactus Ferment 11800 11800 10500 17800 15000 60000 Control 11800 11800 39500 1480000 125200000 1775000000

In addition, pathogenic experiments of streptococci were performed as follows. The olive flounder used in the experiment was a 6cm sized fish that was fed with a feed added with 1.0% of cactus fermentation for 3 months in a FRP round water tank (ㆈ 74 cm × 80cm) installed in the Jeju Fisheries Research and Development Institute. During the breeding period, the water temperature was 17.4 ~ 21.6 ℃, salinity concentration 33.0 ~ 35.4%, DO 5.84 ~ 7.12 mg / l, and pH ranged from 7.57 ~ 8.04.

As a specific experimental method, the flounder fed with cactus fermentation (EP feed containing cactus fermentate at a concentration of 1% by weight) and the flounder fed EP feed were divided into five experimental groups and five control groups, respectively. Placed. Each group consisted of four flounder and activated Streptococci were cultured in BHI liquid medium, and 9.8, 9.8 × 10 ² , 9.8 × 10 ⁴ , 9.8 × 10 ⁶ , 9.8 × 10 ⁸ Streptococci per ml 0.1 ml each was inoculated. During the 14 days after inoculation, mortality and appearance symptoms were checked daily. Lung and expression fish were dissected to observe lesions and cultured in liver and kidney. The results are shown in Table 5 below.

group Inoculation concentration (cell / ml) Number of bodies after administration Number / test number of dead flounder Mortality rate (%)  One 2 . . 7 8 9 10 11 12 13 14 Experimental group One 9.8 0/4 0 2 9.8 × 10 ² 0/4 0 3 9.8 × 10 ⁴ 0/4 0 4 9.8 × 10 ⁶ One 1/4 25 5 9.8 × 10 ⁸ 2 2/4 50 Control One 9.8 One 1/4 25 2 9.8 × 10 ² One One 2/4 50 3 9.8 × 10 ⁴ One One 2/4 50 4 9.8 × 10 ⁶ 0/4 0 5 9.8 × 10 ⁸ 2 One 3/4 78

As shown in Table 5 below, in the non-fed flounder fed with the cactus fermentation, 78% of the control group inoculated with 1 × 10 ⁹ viable cells (streptococci) died in 9 days; The control group inoculated with 1 × 10 ³ viable cells died 50% in 12 days; The control group intraperitoneally injected with 1 × 10 ¹ viable cells died 25 days after 10 days. In addition, in the control group inoculated with 1 × 10 ⁷ viable cells, there was no mortality, but ascites and ocular protrusion showed clinical symptoms, and causative bacteria such as inoculation bacteria were detected in liver and kidney.

However, flounder fed feed supplemented with cactus ferment died 50% in 8 days in the experimental group inoculated with 1 × 10 ⁹ viable cells; In the experimental group inoculated with 1 × 10 ⁷ viable cells, 25% died in 13 days.

Therefore, the cactus fermentation extract according to the present invention was confirmed to exhibit an antimicrobial effect against cocci.

2) Edward bacteria

In order to investigate the effect of cactus fermentation on the growth of Ed Ward bacteria, the growth rate of bacteria in liquid medium was compared and analyzed. First, the freeze-dried cactus ferment prepared according to the method of Example 3 was suspended in BHI at a final concentration of 2% (w / v), mixed at room temperature for 30 minutes, and then adjusted to pH 7.0. The mixture was placed in an ice box and sonicated for 15 seconds and allowed to stand for 15 seconds for 8 times. After sonication, the insoluble material was removed with a filter paper, and then used as an experimental group by passing through a syringe filtration sterilizer (0.45 um pore size). BHI without cactus was used as a control.

Streptococci were inoculated at 1.2 and 10 ⁴ CFU / ml levels for 18 hours in BHI in the experimental and control groups, and then at 0, 2, 4, 8, 12 and 24 hours after inoculation while standing at 22 ° C. The number of bacteria was measured. Bacterial counts were inoculated in BHI agar containing 2% NaCl after diluting 100 µl of the culture solution in sterile saline at each measurement time in the experimental and control groups. Each experiment was repeated twice. The results are shown in Table 6 and FIG. 8B.

As shown in Table 6 and FIG. 8B, the control group showed a bacterial count of 8.4 log10 CFU / ml after 12 hours of cultivation and 11.0 log10 CFU / ml after 24 hours, and 12 hours of culturing for the experimental group to which fermented cactus fruit fermentation powder was added. After 4.5 log10 CFU / ml, after 24 hours 9.5 log10 CFU / ml showed a bacterial count inhibited the growth of Ed Ward bacteria up to 12 hours was able to see the effect of inhibiting bacterial growth.

Changes in Cell Numbers Over Time 0 hours 2 hours 4 hours 8 hours 12 hours 24 hours Cactus Ferment 0 0 0 430 1700 121000000 Control 0 0 910 3500 26000 121000000

In addition, the pathogenic experiment of Edward was carried out as follows. The olive flounder used in the experiment was 6cm sized and used as a feed supplemented with 1.0% of the cactus fermented product for 20 days in a FRP circular tank (φ74 cm × 80cm) installed in the Jeju Fisheries Research and Development Institute. During the breeding period, the water temperature was 17.4 ~ 21.6 ℃, salinity concentration 33.0 ~ 35.4%, DO 5.84 ~ 7.12 mg / l, and pH ranged from 7.57 ~ 8.04.

As a specific experimental method, the flounder fed with cactus fermentation (EP feed containing cactus fermentate at a concentration of 1% by weight) and the flounder fed only EP feed were divided into five experimental groups and five control groups, respectively. Placed. Each group consisted of four flounders. The activated Edvar bacteria were incubated in BHI liquid medium, and the grown cells were suspended in sterile saline solution to 9.8, 9.8 × 10 ² , 9.8 × 10 ⁴ , 9.8 × 10 ⁶ , 9.8 x 10 ⁸ Ed Ward bacteria were inoculated intraperitoneally by 0.1 ml. During the 20 days after inoculation, mortality and appearance symptoms were checked daily. Lung and expressed fish were dissected to observe lesions and cultured in liver and kidney. The results are shown in Table 6-2 below.

Table 6-2

Control Inoculation concentration (cell / ml) Number of carcasses after administration Number / test number of dead flounder Mortality rate (%)  1-6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 One 1 × 10 ¹            1 1 2/4 50 2 1 × 10 ³            One 1/4 25 3 1 × 10 ⁵                                   One 1/4 25 4 1 × 10 ⁷                                   One 1/4 25 5 1 × 10 ⁹            2 2 4/4 100

Experimental group Inoculation concentration (cell / ml) Number of carcasses after administration Number / test number of dead flounder Mortality rate (%)  1-6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 One 1 × 10 ¹ 0/4 0 2 1 × 10 ³ 0/4 0 3 1 × 10 ⁵ 0/4 0 4 1 × 10 ⁷ 0/4 0 5 1 × 10 ⁹            2 in 1 3/4 75

As shown in Fig. 6-2, the flounder without the feed containing the cactus fermentation was 100% dead in 9 days in the control group inoculated with 1 × 10 ⁹ viable cells and 1 × 10 ³ ~ 1 × 10 ⁷ In control group inoculated with living cells, 25% died in 16 days, and 50% in 12 days in control group intraperitoneally injected with 1 × 10 ¹ cells. The flounder fed the feed containing the cactus fermentation showed 100% mortality between 8 and 14 days in both the powder and liquid phase inoculated with 1 × 10 ⁹ viable cells. There was no significant difference in the mortality rate between the experimental group and the control group inoculated with 1 × 10 ⁷ viable cells.

On the other hand, the flounder infected with Ed Ward bacteria showed blackening in appearance and showed major symptoms such as hernia, eye protrusion and cloudiness, and mucus on body surface. Anatomically, there was a lot of thin mucous ascites and hepatic congestion and hypertrophy of kidney and spleen, similar to natural infected fish. The kidney tissues of the flounder infected with Edward bacteria were found to have delamination, necrosis and urinary tubules. In some livers, necrotic cells and their populations were identified in the hepatic pancreas and adjacent connective tissue areas.

3) Vibrio bacteria

To investigate the effect of cactus fermentation on the growth of Vibrio bacteria, the growth rate of bacteria in liquid medium was compared. First, the freeze-dried cactus fermentation product prepared according to the method of Example 3 was suspended in BHI at a final concentration of 2% (w / v), mixed at room temperature for 30 minutes, and then adjusted to pH 7.0. The mixture was placed in an ice box and sonicated for 15 seconds and allowed to stand for 15 seconds for 8 times. After sonication, the insoluble material was removed with a filter paper, and then used as an experimental group by passing through a syringe filtration sterilizer (0.45 um pore size). BHI without cactus was used as a control.

The experimental and control groups were inoculated with Vibrio bacteria previously incubated for 18 hours in BHI at 1.2 × 10 ⁴ CFU / ml, followed by standing at 22 ° C., at 0, 2, 4, 8, 12 and 24 hours after inoculation. The number of bacteria was measured. Bacterial counts were inoculated in BHI agar containing 2% NaCl after diluting 100 µl of the culture solution in sterile saline at each measurement time in the experimental and control groups. Each experiment was repeated twice. The results are shown in Figure 8C below.

As shown in FIG. 8C, the control group showed a bacterial count of 9.2 log10 CFU / ml after 12 hours of incubation and 11.1 log10 CFU / ml after 24 hours, and 3.6 log10 after 12 hours of incubation in the experimental group to which the palm cactus fruit fermentation powder was added. After 24 hours of CFU / ml, 6.4 log10 CFU / ml showed the number of bacteria, which inhibited the growth of vibrio bacteria up to 24 hours.

In addition, pathogenic experiments of Vibrio bacteria were performed as follows. The flounder used in the experiment was a 6cm sized fish that was fed with a feed of 1.0% of cactus fermented for 1 month in a FRP round water tank (φ74 cm × 80cm) installed in the Jeju Fisheries Research and Development Institute. During the breeding period, the water temperature was 17.4 ~ 21.6 ℃, salinity concentration 33.0 ~ 35.4%, DO 5.84 ~ 7.12 mg / l, and pH ranged from 7.57 ~ 8.04.

As a specific experimental method, each group was composed of four flounder, and the activated Vibrio bacteria were cultured in BHI liquid medium, and the grown cells were suspended in sterile physiological saline to 1 × 10, 1 × 10 ³ , 1 × 10 ⁵ , 1 × 10 ⁷ , 1 × 10 ⁹ Vibrio bacteria were inoculated into the abdominal cavity by 0.1 ml each. During the 31 days after inoculation, mortality and appearance symptoms were checked daily. Lung and expressed fish were dissected to observe lesions and cultured in liver and kidney. The results are shown in Table 7 below.

Control Inoculation amount (cell / ml) Number of bodies after administration Number / test number of dead flounder Mortality rate (%)  1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 1 × 10 ¹ 0/4 0 1 × 10 ³ 0/4 0 1 × 10 ⁵                One 1/4 25 1 × 10 ⁷    2 in 1 3/4 75 1 × 10 ⁹  4 4/4 100

Experimental group

Inoculation amount (cell / ml) Number of bodies after administration Number / test number of dead flounder Mortality rate (%)  1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 1 × 10 ¹ 0/4 0 1 × 10 ³ 0/4 0 1 × 10 ⁵ 0/4 0 1 × 10 ⁷      1 1 2/4 50 1 × 10 ⁹  4 4/4 100

As a result, as shown in Table 7, in the flounder that did not eat the feed containing the lactic acid bacteria of the present invention, 100% died in one day when 1 × 10 ⁹ viable cells were inoculated; 75% died in 3 days when inoculated with 1 × 10 ⁷ viable cells; Inoculation of 1 × 10 ⁵ viable cells killed 25% in 8 days.

In addition, as shown in Table 7, in the flounder fed feed containing the cactus fermentation, 100% was killed in 1 day when 1 × 10 ⁹ viable cells were inoculated; Inoculation of 1 × 10 ⁷ viable cells resulted in 50% death in 4 days. Ascites and ocular protrusion showed clinical symptoms, and causative organisms such as inoculation bacteria were detected in liver and kidney.

On the other hand, olive flounder infected with Vibrio bacteria showed blackening in appearance and showed major symptoms such as hernia, eye protrusion and cloudiness, and mucus on body surface. Anatomically, there was a lot of thin mucous ascites and hepatic congestion and hypertrophy of kidney and spleen, similar to natural infected fish. Renal tissues of flounder infected with Vibrio bacteria were observed to have delamination and necrosis of urinary tubular epithelial cells and a marked decrease in urinary tubules. In some livers, necrotic cells and their populations were identified in the hepatic pancreas and adjacent connective tissue areas.

Example 7 Antiviral Effect of Cactus Fermentation According to the Present Invention

1) Inhibitory Effect of Cactus Fermentation on Equine Herpes Virus (EHV)

Equine herpesvirus (EHV) is a DNA virus belonging to the herpesvirus family.If it is infected, it is a non-pneumonia that causes transient fever and nasal juice during the initial infection of the follicle. The result is known to cause pseudoacids. This experiment was carried out to confirm whether the cactus fermentation according to the present invention has a proliferation inhibitory effect on EHV.

As a specific experimental method, MDBK-cells were grown in monolayers in 96-well microtiterplates. After removing the cell culture solution, each well was infected with 50 PFU / 50 μl of EHV virus for 1 hour, and 100 μl of the cactus ferment prepared in Example 3, which was diluted twice, was dispensed into each well to control the growth of the virus. Observed through the ratio of the number of plaques to. Each experiment was performed four times and the average value was calculated.

As a result, it was confirmed that in the plate treated with the cactus fermented product prepared in Example 3, no virus was propagated, no plaque was produced at all, and an inhibitory effect of 100% was obtained.

In addition, to confirm whether the cactus fermentation prevents the EHV virus, the following experiment was performed. That is, MDBK-cells were grown in monolayers on 96-well microtiter plates. After removing the cell culture solution, 50 µl of the cactus ferment prepared in Example 3 diluted twice-fold was dispensed into each well and incubated for 1 hour. After removing the cactus ferment diluted in each well, each well was infected with 50 PFU / 50 μl of EHV virus for 1 hour. Virus propagation was observed through the ratio of the number of plaques in the test group to the number of plaques in control wells without cactus extract. Each experiment was performed four times and the average value was calculated. Each experiment was performed four times and the average value was calculated.

As a result, it was confirmed that in the plate treated with the cactus fermented product prepared in Example 3, no virus was propagated, no plaque was produced, and a 100% prophylactic effect was obtained.

2) Growth Inhibition Effect of Cactus Ferment against Bobbin Herpes Virus (BHV)

Bobbin herpes virus (BHV) is an infection of the DNA virus belonging to the herpes virus family. It is an infectious disease of bovine respiratory and genital malformations. It is known to be characterized by. This experiment was carried out to check whether the cactus fermentation according to the present invention has a proliferation inhibitory effect on BHV.

As a specific experimental method, MDBK-cells were grown in monolayers in 96-well microtiter plates. After removing the cell culture solution, each well was infected with 50 PFU / 50 μl of BHV virus for 1 hour, and 100 μl of the cactus ferment prepared in Example 3, which was diluted twice, was dispensed into each well, and the control well was checked for virus growth. Observed through the ratio of the number of plaques to. Each experiment was performed four times and the average value was calculated.

As a result, it was confirmed that in the plate treated with the cactus fermented product prepared in Example 3, no virus was propagated, no plaque was produced at all, and an inhibitory effect of 100% was obtained.

In addition, in order to determine whether there is a preventive effect on the BHV virus of the cactus fermentation, the following experiment was performed. That is, MDBK-cells were grown in monolayers on 96-well microtiter plates. After removing the cell culture solution, 50 µl of the cactus ferment prepared in Example 3 diluted twice-fold was dispensed into each well and incubated for 1 hour. After removing the cactus ferment diluted in each well, each well was infected with 50 PFU / 50 μl of EHV virus for 1 hour. Virus propagation was observed through the ratio of the number of plaques in the test group to the number of plaques in control wells without cactus extract. Each experiment was performed four times and the average value was calculated. Each experiment was performed four times and the average value was calculated.

As a result, in the plate treated with the cactus fermentation product prepared in Example 3, the virus did not grow, no plaque was produced, and it was confirmed that 100% of the prophylactic effect appeared.

Example 8 Breeding of Flounder Using Cactus Fermentation According to the Present Invention

In this embodiment, when used as a feed additive of the cactus fermentation according to the present invention, it was carried out to confirm that excellent growth effect can be seen in the flounder cultivated.

At the time of the flounder fry, the PP fish tank (1 ton) was used to adsorb 1% by weight of the cactus fermentation broth into the EP feed (Japanese-Chinese Marubeni feed, Japan) and feed fish twice a day for 3 months. It was supplied sufficiently until it was not seen. The length, weight, and survival rate of the flounder before and after the experiment were examined, and the results are shown in Table 8 below.

group Early final Survival rate (%) Flounder TL (cm) BW (g) Flounder TL (cm) BW (g) Control 200 7.54 ± 0.07 3.70 ± 0.02 112 16.32 ± 0.66 46.30 ± 9.57 56.00 ± 49.50 One % 200 7.56 ± 0.23 3.81 ± 0.02 193 17.95 ± 0.38 59.50 ± 2.14 96.50 ± 0.71 * The values are expressed as mean ± standard deviation. * In the final size of the fry, the control group showed a significant difference of P <0.05 compared with the experimental group. * TL represents the length of the flounder and BW represents the weight of the flounder. * 1% indicates that 1% cactus fermentation was added to the EP feed.

On the other hand, for the flounder tuna, MP feed (freeze-dried cactus fermented powder using a PP small water tank (1 ton) manufactured by the inventors is made by mixing 90% of horse mackerel and 10% fish meal) Adsorption was carried out at various concentrations of 0.02 to 0.16% by weight, and the test fish was sufficiently supplied twice a day for 8 months until no feeding activity was observed. The length, weight, and survival rate of the flounder before and after the experiment were examined, and the results are shown in Table 9 below.

group Early final Survival rate (%) Flounder TL (cm) BW (g) Flounder TL (cm) BW (g) 0.00% 60 32.40 ± 0.08 394.52 ± 8.03 55 40.42 ± 0.85 ^a 825.62 ± 65.60 ^a 91.67 ± 7.64 ^ab 0.02% 60 32.45 ± 0.35 400.94 ± 10.73 57 40.76 ± 0.86 ^ab 843.47 ± 56.68 ^a 95.00 ± 5.00 ^ab 0.04% 60 32.20 ± 0.22 393.76 ± 3.33 58 40.77 ± 0.66 ^ab 856.40 ± 52.11 ^a 96.67 ± 2.89 ^ab 0.08% 60 32.07 ± 0.13 391.64 ± 36.46 53 42.03 ± 0.67 ^b 968.60 ± 40.48 ^a 88.33 ± 2.89 ^a 0.16% 60 32.32 ± 0.21 399.70 ± 11.10 59 40.92 ± 0.87 ^ab 857.57 ± 70.35 ^a 98.33 ± 2.89 ^b

Example 9 Breeding Effect of the Flounder Compared to the Feed Additive According to the Present Invention and the Feed Additive Comprising the Cactus Fermented Extract Prepared Using Other Microorganisms

The effects of fermentation broth fermented with five kinds of fermentation strains (EM), photosynthetic bacteria, and lactic acid bacteria as fermentation strains for the fermentation of cactus were investigated. Then, in a similar manner to Example 8, the growth effect of the flounder bred using each feed additive was compared, and the results are shown in Table 10 below.

As shown in Table 10, the rearing effect was confirmed to be the largest effect in Lactobacillus plantarum CNU001.

group                                              Early final Survival rate (%) Flounder TL (cm) BW (g) Flounder TL (cm) BW (g) Control 300 7.07 ± 0.34 3.33 ± 0.85 214 15.83 ± 0.14 ^a 39.12 ± 0.30 71.33 ± 5.13 EM 300 7.13 ± 0.17 3.50 ± 0.62 229 16.90 ± 0.35 ^b 46.43 ± 2.34 76.33 ± 15.37 A 300 7.08 ± 0.62 3.24 ± 1.22 204 16.47 ± 0.82 ^ab 44.16 ± 6.30 68.00 ± 5.29 B 300 7.10 ± 0.35 3.41 ± 0.91 232 16.08 ± 0.73 ^ab 42.46 ± 5.67 77.33 ± 3.06 C 300 7.07 ± 0.24 3.26 ± 0.45 224 15.81 ± 0.57 ^a 38.24 ± 5.96 74.67 ± 10.02 D 300 6.95 ± 0.05 3.18 ± 0.24 224 16.15 ± 0.29 ^ab 41.66 ± 2.99 74.67 ± 1.53 E 300 6.95 ± 0.11 3.17 ± 0.22 210 16.39 ± 0.31 ^ab 44.28 ± 3.60 70.00 ± 10.44 F 300 7.15 ± 0.04 3.53 ± 0.10 214 16.54 ± 0.67 ^ab 47.13 ± 4.97 71.33 ± 5.69 A: Lactobacillus brevis IMSNU 1011075 B: Rhodoschudomonas palustris DSM123 C: Lactobacillus plantarum IMSNU 10124 D: Leukonoschenk menceteroides IMSNU 10146 E: Pediococcus dextrinicus KCTC 3506 F: Lactobacillus plantar Room CNU001 * The values are expressed as mean ± standard deviation.

As described above, when using the Lactobacillus plantarum CNU001 isolated from the cactus fruit as a strain of the present invention to produce a fermentation of cactus, in particular palm cactus, and breeding flounder using the prepared fermentation, It is possible to achieve a very excellent flounder growth, it can be seen that the cactus fermentation extract using the strain of the present invention can be used as a feed additive of various animals.

Claims (7)

A method for producing a cactus lactic acid bacteria fermentation product comprising the step of fermenting a cactus with Lactobacillus plantarum CNU001 (KCTC18105P) as lactic acid bacteria. The method of claim 1, wherein the steps are as follows: Selecting and washing the cactus fruit; Grinding the cactus fruit with a blender to obtain a homogeneous liquid; Mixing the homogenate with a fermentation accelerator such as molasses or starch syrup and Lactobacillus plantarum CNU001 which is a strain according to the present invention; And Fermentation at room temperature for 1 minute to 2 weeks, or more to obtain a fermentation broth; And preparing the fermentation broth into lyophilized powder. Method for producing a cactus lactic acid bacteria fermentation, characterized in that it comprises a. The method of claim 2, wherein the cactus is a palm cactus. Lactic acid bacteria fermented product of a cactus manufactured by the method of any one of Claims 1-3. 5. The lactic acid bacteria fermentation product of cactus according to claim 4, wherein the lactic acid bacteria fermentation product of the cactus is used as a halibut feed additive. Flounder feed method comprising the intake of the flounder containing the cactus lactic acid bacteria fermented product prepared by the method according to any one of claims 1 to 3 as a flounder feed additive. . Lactobacillus strain, Lactobacillus plantarum CNU001 (KCTC18105P)

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