KR100951273B1 - Feed additive containing an extract of bamboo and microorganism cultivate,for prevention of Porcinediarrhea caused by enterotoxigenic enterobacteria - Google Patents

Abstract

The present invention relates to a bamboo leaf microbial fermentation broth having intestinal harmful bacterium inhibitory activity and having a diarrhea inhibitory effect on livestock, and a feed additive comprising the same. The feed additive of the present invention is a bamboo leaf fermentation broth which is dried and fermented in a culture solution of Lactobacillus plantarum , which is a kind of lactic acid bacteria after drying. By adding 0.1% to 5% to the feed of the livestock and then fermenting it by secondary fermentation, it has the effect of inhibiting the growth of intestinal harmful bacteria and reducing diarrhea and may be useful as a feed additive for improving the health of the livestock.

Bamboo leaf extract, Lactobacillus plantarum, livestock diarrhea, stalk

Description

Feed additive containing an extract of bamboo and microorganism cultivate, for prevention of Porcinediarrhea caused by enterotoxigenic enterobacteria}

The present invention relates to a microbial active liquid feed additive containing a scavenger extract having an intestinal harmful bacterium inhibitory activity and a diarrhea inhibitory effect.

Bamboo is an evergreen perennial plant belonging to a monocotyledonous plant (Paceae), a family of Bambusoideae, and there are four kinds of four kinds of bamboo in Korea. There are king genus, haejangjuk, genus genus and genus genus. Among them, the shortest and thinnest bamboo, but the strongest is genus genus.

Sasa quelpaertensis Nakai is a native species of Korea, native to Jeju Island, less than 80cm in height, about 3 ~ 4mm in diameter, green without hairs, nodes thick, and slightly purple. The leaves of stalks contain nutrients such as carotene, vitamin A, B1, B2, B12, and aspartic acid, iron, calcium, protein, and the polysaccharide bamphorin, which act to strengthen cell membranes and improve immunity. The main ingredients, Friedelin, Glutinol, and Isoorientin, have been reported to have high effects on antibacterial and antioxidant properties. It is said to have an effect on fever, conflict, and resolution. Heo Jun's consent is said to have mysterious effects on stroke, mental stability, and fatigue recovery. Recently, due to such pharmacological function and nutritional effect, research is underway to develop the food as a functional food by vacuum-packing the leaves of the stalks and freezing them or making them into powder and adding them to food.

Lactic acid bacteria is a microbiological classification included in Eubacteriales and Lactobacillus plantarum belonging to the Lactobacilleae family, Lactobacillus plantarum decomposes sugars such as glucose and lactate. acid) is a bacterium that produces lactic acid bacteria. Lactic acid, an organic acid produced by lactic acid fermentation, inhibits the growth of pathogens and harmful bacteria, and densely setstles in the gut gut wall of the animal, and helps to use it as an energy source by decomposing nutrients and fibrin ingested by animals. In addition, it is known to play an important role in maintaining the health of the intestine by producing lactic acid and antibiotics to suppress the occurrence and growth of intestinal harmful bacteria. Young animals that do not have complete intestinal flora are required for lactic acid bacteria to form the intestinal flora, mainly due to beneficial strains. Lactobacillus is also required to suppress the occurrence of. Intestinal harmful bacteria include Salmonella sp ., Shigella sp ., E. coli , and Enterococcus sp . Infection with Salmonella can lead to sepsis, chronic diarrhea, septic colitis and chronic. Lactobacillus is one of the predominant floras of mammalian intestinal microorganisms, and has been reported for beneficial effects such as resistance to infection, anticancer effect, control of blood cholesterol content and intestinal effect (Hentges D. Hosons A. et al ., Agr. Biol . Chem ., 1990, 54, 7,1639-1643). In addition, it is known to play an important role in promoting animal growth and improving feed utilization, increasing resistance to disease, inhibiting growth of harmful bacteria, reducing mortality, inhibiting the production of toxic toxins, and producing various vitamins.

Conventionally, the technique of using bamboo leaves has been limited mainly to the food, pharmaceutical, and cosmetic raw material fields until now (registered patent: 10-0722674) (published patent: 2000-17469, 2003-0084393, 2003-0021640). In addition, in the case of using Lactobacillus as a livestock bioactive agent, Korean Patent No. 211529 discloses a novel acid resistant Lactobacillus microorganism having a harmful microbial inhibitory activity and a livestock bioactive agent containing the same. A case of a feed additive comprising bamboo liquid bamboo vinegar and microbial culture as an active ingredient is disclosed in Korean Patent Registration No. 10-0573519. However, this is a liquid organic acid liquid that collects and cools the smoke generated in the process of carbonizing bamboo in a charcoal kiln. The main components are compounds such as acetic acid, organic acid, and phenol, which are water-soluble liquids generated during carbonization. It is unclear whether nutrients such as carotene, vitamin A, B1, B2, B12, and aspartic acid of bamboo leaves, and iron, calcium, protein, and polysaccharide bamphorin. Therefore, in addition to strengthening cell membranes and enhancing immunity, it is not possible to identify major ingredients such as Friedelin, Glutinol, and Isoorientin, which show high effects on antibacterial and antioxidant effects. . In addition, bamboo vinegar is mixed with microbial culture and fed to livestock to produce high-quality meat. However, there are no cases where bamboo leaves were fermented in lactic acid bacteria culture medium to extract useful components of bamboo leaves without heat or chemical denaturation and applied as livestock diarrhea inhibitors. In addition, the method using only the strains known in the prior art is not very effective in inhibiting harmful bacteria in the intestine, and diarrhea due to the occurrence and proliferation of harmful bacteria in the intestine is frequently occurring.

Therefore, the present inventors studied the feed additives that increased the inhibitory effect of intestinal harmful bacteria, and confirmed that the fermentation broth fermented by adding lactic acid leaf to the lactic acid bacteria culture medium inhibited the intestinal harmful bacteria and proliferated and reduced the diarrhea of livestock. The invention was completed.

The present invention relates to a novel probiotic feed additive that can be used as a diarrhea inhibitor in pigs using a feed composition containing the same as a fermentation broth of Lactobacillus plantarum fermented with a sasa quelpaertensis Nakai leaf culture. Feed additive of the present invention can be applied to prevent diarrhea and health promotion of livestock such as pigs, cows, chickens, ducks, and more particularly, the present invention is a probiotic feed additive for preventing and inhibiting diarrhea in pigs containing fermented broccoli Lactobacillus plantarum It is about.

It is an object of the present invention to provide a fermentation broth Lactobacillus plantarum fermentation broth (trade name YC2004. Hereinafter referred to as YC2004) excellent in inhibiting intestinal harmful bacteria and inhibiting diarrhea in livestock, and a feed additive having the same as an active ingredient.

The YC2004 of the present invention was found to be excellent in inhibitory activity of harmful intestinal bacteria, but did not have any negative effect on beneficial bacteria, and the diarrhea inhibitory effect in pig pigs and growing pigs. Therefore, YC2004 of the present invention can be easily used as a feed additive having an inhibitory effect of diarrhea caused by the generation and propagation of harmful bacteria in the intestine of pigs.

In order to achieve the above object, the present invention relates to a novel probiotic feed additive that can be used as a diarrhea inhibitor of pigs using a fermentation broth, fermentation broth fermented to Lactobacillus plantarum culture medium, feed composition containing the same. Feed additive of the present invention can be applied to prevent diarrhea and health promotion of livestock such as pigs, cows, chickens, ducks, and more particularly, the present invention is a probiotic feed additive for preventing and inhibiting diarrhea in pigs containing fermented broccoli Lactobacillus plantarum It is about.

Bamboo leaves, especially bamboo leaves of the genus Littung selected in the present invention is known to be rich in various nutrients, that is, carotene, vitamins, aspartic acid.

In addition, iron, calcium and protein, polysaccharides contain bhamphorin has the effect of strengthening the cell membrane and improve the immunity of animals. In addition, it contains a large amount of antibacterial and antioxidants, and it is said in ancient literature that it is effective for tumor, antipyretic, conflict, and resolution. In the present invention, in order to be able to extract the useful components of bamboo leaves without damage, a fermentation method through microorganisms was used to avoid denaturation by avoiding heat or chemical extraction methods. Fermentation refers to the process of decomposing organic substances using enzymes owned by microorganisms. Fermentation and decay reactions are carried out by similar processes, but when decomposition results in substances that are useful for our lives, they are called fermentation. When harmful substances are made, they are called decay. In addition, various organic substances decomposed through the fermentation process are decomposed into high molecular weight substances in the polymer material, so that the absorption is quick and the effect is exerted quickly. Temperature and fermentation environmental conditions and selection of fermentation bacteria are important for promoting fermentation. If the fermentation process is too fast, only a part of the active ingredient can be selectively extracted, so that the useful ingredients can be evenly extracted by lowering the temperature slightly to about 25 ° C. and increasing the fermentation time to 7-10 days. In addition, drying and cutting to a certain size to extract useful components. In order to enlarge the contact area of fermented bacteria and to extract efficiently. Fermented bacteria were selected from Lactobacillus plantarum among lactic acid bacteria having intestinal effect and organic acid producing ability, and produced lactic acid by decomposing sugar such as glucose. At this time, lactic acid, an organic acid produced, inhibits the growth of pathogenic harmful bacteria, settles in the gut wall of the intestine, and decomposes nutrients and cellulose, and serves as an energy source. The useful components of bamboo leaves are extracted in the fermentation process of these lactic acid bacteria and present in the culture medium, and the interaction between bamboo leaf components and lactic acid bacteria inhibits the occurrence and reproduction of intestinal harmful bacteria. In addition, the results of the experiment are preferably confirmed that there is no inhibitory effect on the enterobeneficial bacteria, suggesting the applicability for the health promotion of livestock.

Based on the antagonistic ability of YC2004, which is characteristic of YC2004, 1 ~ 2% of feed weight is added to the feed by diluting 100 ~ 500 times diluted solution of YC2004. Induced fermentation of organics. It is the secondary fermentation process of feed according to YC2004. The fermented feed containing YC2004 produced through this process was fed to weaning pigs and growing pigs of pigs with the greatest risk of diarrhea, and the diarrhea inhibitory effect was observed. In addition, fermented feed containing YC2004 was fed to pigs, growing pigs, and sows for diarrhea-producing pigs. As a result of the test, it was confirmed that the diarrhea inhibitory effect of YC2004 livestock, that is, pigs, was clearly exerted. Experimental contents regarding the diarrhea inhibitory effect in piglets, growing pigs, and sows in the manufacturing process of YC2004 of the present invention will be described in detail below.

<Example 1> Intestinal harmful bacterium inhibition of six medicinal plants

In order to select the bamboo leaf plants used in the present invention, six kinds of plant juices such as Medicinal plants, hawthorn, camphor tree, goblin needles, pear herbaceous leaves, and pine needles were extracted to examine the inhibitory effects of intestinal harmful bacteria. First, 200 ml of distilled water was added to 200 g of each non-dried plant, and then ground using a blender for 3 to 5 minutes, finely ground, and left for about 1 hour. This process was repeated 2-3 times to obtain a supernatant of the plant grinding solution obtained to prepare a plant juice. Is as follows: Typical enteropathogenic bacteria Escherichia coli (E.coli), Salmonella (Salmonella), Shigella B (Shigella B), Enterococcus (Enterococcus sp.), And the like Staphylococcus brother Sears (Staphylococcus auseus) in the optimal medium Each bacterium was cultured at optimal temperature in a nutrient medium. After diluting the bacterial culture suspension to a concentration of 2.0 × 10 ⁵ CFU / ml, 100 μl of the agar plate (Nutrient agar is composed of 3.0 g of beef extract (beef extract), 5.0 g of peptone, 15.0 g of agar and 1.0 of distilled water). Consisting of L). 70 [mu] l of each plant juice was smeared onto the smeared agar plate and soaked in a paper disk and attached by a paper disk method. Then, the antimicrobial activity of each plant was confirmed by the size of the inhibitory ring formed during 24 hours of incubation at the optimum temperature of each strain, and the results are shown in (Table 1). Among the 6 medicinal plants, the antimicrobial activity of bamboo leaf was the best, and Songjeol and ear needles showed antimicrobial activity against some bacteria.

<Example 2> Collection and pretreatment of bamboo leaves

Through the experiment of <Example 1> confirmed the antimicrobial activity of the intestinal harmful bacteria of bamboo leaves and went through the collection and pre-treatment process for using the bamboo leaves in the present invention will be described in detail below.

Bamboo species is a genus of southern and Jeju islands, also known as Sanjuk (山竹). It is known to be small in Korea and strong in weakness. In the treatment process, bamboo leaves are collected from the bamboo stalks, the foreign material on the epidermis is removed, washed with water, and dried in a well-ventilated place with good shade. After drying, bamboo leaves are cut to about 4 ~ 5cm in diameter. It is a treatment method for inducing fermentation promotion and easy extraction of active ingredients. Drying is usually about 7 to 10 days in a place where the ventilation of the shade room temperature is desired. Dried leaves are prepared by subdividing about 1.0kg into a tightly spaced mesh (less than 0.5mm in diameter).

<Example 3> Confirmation of inhibitory ability of enteric harmful bacteria of six microorganisms

In <Example 2> of the present invention, six microorganisms, such as Bacillus and Lactobacillus, which were beneficial bacteria, were cultivated under respective culture conditions in order to select a microbial species for extracting the active ingredient of bamboo leaves prepared through the pretreatment process without damage. Each strain was activated by subcultured at least twice in an optimal medium and assayed for growth inhibitory activity against five major intestinal harmful bacteria used in <Example 1>. Intestinal beneficial bacteria include Bacillus subtilis , Pseudomonas sp ., Lactobacillus acidophilis , Lactobacillus plantarum , and Saccharomyces cerevisiae. Saccharomyces cerevisiae ) and Acinetobacter baumannii strains were used. In order to confirm the antimicrobial activity, the concentration of 5 harmful bacteria was diluted to a density of 2.0 × 10 ⁵ CFU / ml in the same manner as in <Example 1>, and 100 μl of the agar plate (Nutrient agar) was used. 3.0 g of beef extract, 5.0 g of peptone, 15.0 g of agar and 1.0 L of distilled water). Each agar was smeared with 70 microliters of intestinal fungal stock on agar disk, and the paper disk method was performed. After culturing each strain in an optimal culture condition for 24 hours, the antimicrobial activity was confirmed for each strain by the size of the inhibitory ring formed, and the results are shown in Table 2. Lactobacillus and Bacillus showed excellent activity against intestinal harmful bacteria, but as a result, Lactobacillus plantarum had the highest antibacterial activity.

<Example 4> Culture of lactic acid bacteria

Lactobacillus Lactobacillus plantarum ( Lactobacillus plantarum : KCTC 3106) species were purchased from Korea Research Institute of Bioscience and Biotechnology was used for the production of lactic acid bacteria culture. Lactobacillus plantarum ( Lactobacillus plantarum ) is a lactobacillus ( Lactobacillus ) lactobacillus, rod-shaped bacilli and gram-positive bacteria. Its size is 0.7-1.0 X 3.0-8.0㎛ and growth titration temperature is 29-33 ℃ and it is aerobic bacteria. Lactobacillus plantarum is composed of arabinose, glucose, fructose, fructose, galactose, maltose, sucrose, dextran, and raffinose. Ferhalose (trehalose) is fermented to produce lactic acid. It is mainly separated from milk, cheese, butter, kefir, fermented sausage, fermented potato, grains, acid dough of bread, sesame like pickle or kimchi, tomato, etc. One of the lactic acid bacteria. In particular, the lactic acid bacteria that play an important role in the fermentation of kimchi in Korea, salt resistance is about 5.5%. There are about 30 kinds of various lactic acid bacteria reported in Kimchi of Korea. It is grown mainly in the late fermentation of Kimchi and has strong acid and bile resistance. Lactobacillus plantarum (KCTC 3106) was incubated for 16 hours at 33 ℃ in the liquid medium (MRS, see Table 3) to allow the cell number to a density of 2.0X10 ⁸ CFU / ㎖ or more. The composition of the medium for Lactic acid bacteria (MRS) is as follows.

<Example 5> Preparation method of Lactobacillus plantarum fermentation broth (trade name YC2004) of Sasa quelpaertensis Nakai leaf

The method for preparing YC2004, which is a fermentation broth of Sasa quelpaertensis Nakai leaf Lactobacillus plantarum, is added to the Lactobacillus plantarum broth prepared in <Example 4> after the washing and drying of the sap leaf in <Example 2>. It is made by fermentation process. The dried body of walnut leaves is cut into 4 ~ 5cm in diameter and subdivided 1.0kg into nets with holes less than 0.5mm in diameter. The optimum microbial density of Lactobacillus plantarum cultures is more than 2.0 × 10 ⁸ CFU / mL. If the germicidal density is low, the fermentation period can be increased to increase the fermentation efficiency. In 300L of Lactobacillus plantarum culture solution, 1.0 kg of dried dried soybean paste, that is, 0.3% of the dried dried soybean paste, which is 0.3% of the weight of the culture, is added to the net in a state subdivided into the fermentation. The fermentation process is anaerobic and fermented under a matt environmental condition at a temperature of about 25 ° C. for 7-10 days. In the fermentation process, a odor of mixed bitter odor of soybean leaves and sour and vile smell of lactic acid bacteria is produced. Finally, YC2004 is made of a mixture of color and off-white color of the greenish green color of bamboo leaf leaves. It is made of weakly acidic liquid of pH 3.6. In addition, the shape of the leaf of the fermentation broth is changed to boiled dark brown mainly in which the fiber in the leaf is present. When the fermentation is fully processed, YC2004 is produced, which has a microbial density of less than pH3.6 of 2.0 × 10 ⁸ CFU / ml. There is no.

<Example 6> Confirmation test for growth inhibition of enteric bacteria of Lactobacillus plantarum fermentation broth (YC2004)

Agar Hall method (Yousten, A, A. and EW Davidson, Appl. Environ. Microbial., 1982, 44, 1449-1455; Ju, Young-Kee Chung, Korean Journal of Biotechnology and Biotechnology, 2000, 15, 6: 584-588). Specifically, the dried bamboo leaves were dried and then inoculated with Lactobacillus plantarum on L. medium (Table 3), which is a lactic acid bacteria culture medium, and dried bamboo leaves in the lactic acid bacteria culture medium cultured at 35 ° C. for 16 hours under anaerobic conditions. The leaves were fermented for 7-10 days. 60 µl of the thus fermented YC2004 was obtained. Agar plate (Nutrient agar consists of 3.0g of beef extract, 5.0g of peptone, 15.0g of agar and 1.0L of distilled water) Enterococcus , vancomycin -ResistantEnterococci resistant to pathogens And methicillin -resistant Staphylococcus aureus ( Methicillin-Resistant Staphylococcus aureus ), respectively, after smearing 60 μl of the YC2004 of the present invention obtained above by the agahole method and incubating for 24 hours to determine the diameter of the transparent ring. By measuring the growth inhibition against the intestinal bacteria. As a result, the pathogen was not resistant to YC2004 of the present invention, and a transparent ring was formed around the YC2004 liquid of the present invention. Therefore, it was confirmed that YC2004 of the present invention has growth inhibitory activity against pathogens. In addition, in the same manner as above, using the YC2004 of the present invention, intestinal harmful bacteria in addition to the above bacteria, Klebsiella sp ., Escherichia coli , E. coli , Serratia sp . Staphylococcus aureus ( evaluate the growth inhibitory effect on Staphylococcus auseus like), parent Nella Mo going (Morganella morgani), Salmonella (Salmonella), Shigella B (Shigella B), Shigella d, Enterococcus (Enterococcus sp.). As a result (Table 4), the YC2004 of the present invention was confirmed to have an excellent inhibitory effect against enteric harmful bacterium, and particularly the growth inhibitory effect on Enterococcus. Therefore, it is expected to be effective in suppressing livestock diarrheal diseases caused by intestinal harmful bacteria. Intestinal beneficial bacteria Bacillus subtilis , Pseudomonas sp ., Lactobacillus acidophilis , Lactobacillus plantarum , Saccharomyces cerevisiae cerevisiae ) and Acinetobacter baumannii were also evaluated for antimicrobial activity in the same manner as described above. (Table 5) It was confirmed that YC2004 of the present invention had no effect on intestinal beneficial bacteria growth. Therefore, it was confirmed that YC2004 of the present invention has growth inhibitory activity against pathogens, but does not adversely affect the growth of enteric beneficial bacteria. The antagonism of pathogens of YC2004 is considered to be sufficiently used as an inhibitor of diarrhea caused by the infection and proliferation of intestinal harmful bacteria in animals. Livestock diarrhea can be divided into external environment (stress, feed, temperature, etc.), and bacteria such as Escherichia coli and Salmonella, and viruses such as TGE and PED. If severe diarrhea continues for any reason, the body's absorption of nutrients will be impaired and the body's tissues will be deficient in water and electrolytes, leading to dehydration and electrolyte imbalance, leading to animal death and dehydration rather than diarrhea itself. Is caused. For the treatment of diarrheal disease, it is very important to replenish the water and electrolytes as well as to remove the pathogen. In particular, diarrhea caused by the infection and outbreak of harmful enterobacterial bacteria, which are directly caused by the hygiene environment, feed condition and feeding process of livestock raising facilities, is the most likely cause of livestock diarrhea. In fact, in order to confirm whether YC2004 of the present invention exerts the inhibitory effect on livestock diarrhea, the amount of feed was added to the feed for pigs to determine the incidence of diarrhea by feeding after a certain period of fermentation. Diarrhea in pigs usually occurs in piglets with low resistance, especially in premature piglets. So when it happens, almost all of them die and they interfere with growth, resulting in economic losses. Therefore, the prevention and suppression of diarrheal disease in livestock is an important task to determine the success and failure of the pig farming business.

<Example 7> Comparison of YC2004 content (0.1% to 5%) intestinal harmful bacterium inhibitory activity

In the present invention, the results of <Example 6> confirmed the growth inhibitory effect on the harmful bacteria in the enterobacteriaceae of the bamboo leaf fermentation broth and the positive effect on the beneficial bacteria. In order to determine the effects of diarrhea in livestock, the inhibitory effect on the major intestinal harmful bacteria whose concentration of YC2004 is 0.1% to 5% in order to confirm the maximum growth inhibitory effect on harmful enteric bacteria at what percentage of YC2004 Was evaluated. Five major intestinal harmful bacteria were cultured in nutrient liquid medium (Nutrient Broth consists of beef extract 3.0g, peptone 5.0g, distilled water 1.0L) for 24 hours at optimum temperature, and the strain density was 2.0X10 ⁸ After adjusting the culture density closer to the CFU / ml, the strain density was measured 48 hours after the addition of a different concentration of YC2004. Of Escherichia coli (E.coli) major enteropathogenic bacteria, Salmonella (Salmonella), Shigella (Shigella sp.), Confirmed the Staphylococcus brother Sears (Staphylococcus auseus) inhibited the growth of the specific content, such as effect YC2004. Confirmation of the growth of enterohazardous bacteria was confirmed by measuring the optical density (OD) at the UV wavelength of 660 nm using a spectrophotometer to determine the growth density of bacteria grown on nutrient liquid medium (NB). At the concentration of 1 ~ 2% of YC2004, the growth of most harmful bacteria was suppressed and the antimicrobial activity was less than 0.5%. More than 2% showed antimicrobial activity similar to that of 1%. Therefore, it can be seen that the concentration of about 1% is the maximum antibacterial activity against harmful bacteria.

<Example 8> Inhibition effect of diarrhea occurrence of pig piglets of Lactobacillus plantarum fermentation broth (YC2004)

This study revealed 150 pigs of LYxD ternary hybrid hybrids that are being raised on Y farm in Jeju. YC2004 was diluted 100-500 times with water in 1% and 2% of the feed weight of the general compound feed (Table 7) of the present invention and then fed after fermentation at around 25 ° C. for 7 days. The application test for pigs was primarily given to 30 heads of weaning pigs with low resistance to diarrhea, and the rate of diarrhea was observed continuously after feeding. The test period for weaning pigs was 45 days from August 7, 2008 to September 21, 2008. After adding 1% and 2% of YC2004 to the feed, the treated and control groups were fed Norufu. The incidence of diarrhea was compared between the treatment of knightly (gentamic acid sulfate) and vitamin supplements (2.0 times more vitamin A, D3, B2, B12, and E). As a result, diarrhea rate was 26.7% for weaning pigs, but less than 3% for YC2004 and 16.7% for gentamicin sulfate. The incidence rate was 23.3%. These results confirm that YC2004 has a definite inhibitory effect on diarrhea caused by enterotoxic bacteria of weaning piglets.

<Example 9> Confirmation of diarrhea inhibitory effect of pig growth pigs of Lactobacillus plantarum fermentation broth (YC2004)

This test revealed 100 pigs of LYxD ternary hybrid hybrid pigs being raised in Y farm in Jeju city. After adding YC2004 of the present invention to the feed, it was confirmed whether diarrhea incidence of pig-growing pigs fed fermented feed was suppressed. Feed was fed after diluting YC2004 with 100% -500 times dilution of water in 1% and 2% of feed weight in general formula feed for feeding (Table 7), followed by secondary fermentation at around 25 ° C for 7 days. The application test for pigs was primarily conducted on weaned pigs with low resistance to diarrhea and secondly on 100 pigs raised.

The test period for growing pigs was carried out for 50 days from August 10, 2008 to September 29, 2008. After adding 1% of the feed weight of YC2004 to the feed, it was fermented. The incidence of diarrhea was compared between the Shin (gentamic acid) supplements and vitamin supplements (2.0 times added vitamin A and D3, B2, B12, E).

The results showed that diarrhea rate was 20.0% when fed only the feed for growing pigs, and 0% when YC2004 was used and there was one case of leprosy. 15.0% of the gentamicin sulfate-based drugs were fed, and 20.0% of the four vitamins were added. These results confirm that YC2004 has a definite inhibitory effect on diarrhea caused by enterotoxic bacteria of raised pigs.

<Example 10> The recovery effect of Lactobacillus plantarum fermentation broth (YC2004) on pig diarrhea

This study revealed 100 diarrheal pigs among LYxD ternary hybrid pigs raised in Y farm in Jeju city. YC2004 of the present invention was added to feed to feed diarrhea incidence and confirmed the change of diarrhea symptoms.

The feed was diluted with 100% -500-fold dilutions of 1% and 2% of YC2004 in the general formula feed for feeding (Table 7), followed by secondary fermentation at around 25 ° C for 7 days and then fed. For pigs, pigs were fed a diet containing 1% YC2004 and pigs and sows fed a diet containing 2% YC2004. The pay period was 10 days after receiving the initial symptoms of diarrhea. The results of the test began to improve after 2 days of salary, and after about 7 days, more than 80% of diarrhea pigs recovered. In the case of piglets and sows, diarrhea did not stop continuously in the untreated areas, and after 10 days or more, the feed containing 2% of YC2004 was uniformly fed for 2-3 days.

1 is a process diagram illustrating a method for preparing fermented broth of Lactobacillus plantarum (YC2004) according to the present invention and a process for preparing fermented feed containing the same.

Figure 2 is a photograph showing the growth inhibitory activity of the enteric harmful bacteria ( E. coli, Salmonella sp. ) According to the concentration (%) of Lactobacillus plantarum fermentation broth.

3 is a graph showing the density (O.D) change of harmful bacteria by concentration (0.1% to 5%) of Lactobacillus plantarum fermentation broth (YC2004).

Claims (6)

Feed additives for suppressing diarrhea in pigs fermented for 7 to 10 days after adding 0.3% of the dried leaves of Sasa quelpaertensis Nakai to the Lactobacillus plantarum culture medium with a bacterial density of 2.0X10 ⁸ CFU / mL or more. How to manufacture delete delete delete delete delete

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