KR102216577B1 - Extruded pellet expanded fermentation feed for fish that has the effect of promoting immune activity, and promoting growth and a method for producing the same - Google Patents

Abstract

The present invention relates to an extruded dry expanded fermentation mixed feed for fish having digestion promotion, immunity activation, growth promotion effect by adsorbing and fermenting pellet feed for fish using Lactobacillus, and a method for producing the same. The method for producing an extruded dry expanded fermentation mixed feed for fish comprises: a first process of producing a mixed solution by mixing an effective microorganism culture medium containing the Lactobacillus parafarraginis KCTC12357BP strain and oligosaccharides; a second process of producing an adsorbate having 37 to 52 wt% of moisture content by inputting pellet feed for fish in the mixed solution and stirring the same to make the mixed solution adsorbed; and a third process of producing the extruded dry expanded fermentation mixed feed for fish by conducting post fermentation of the adsorbate at a temperature of 25 to 30°C for 24 to 48 hours while the adsorbate is sealed. According to the present invention, provided are an extruded dry expanded fermentation mixed feed for fish having digestion promotion, immunity activation, growth promotion effect by increasing protein content, a digestion rate, and immunity with a method for adsorbing and fermenting an existing pellet feed for fish with an effective microorganism containing Lactobacillus strains, and a method for producing the same.

Description

(Extruded pellet expanded fermentation feed for fish that has the effect of promoting immune activity, and its manufacturing method, which has the effect of adsorbing and fermenting the pellet feed for fish to promote immune activity and growth. promoting growth and a method for producing the same}

The present invention relates to a fermented fermented mixed feed for fish that has the effect of adsorbing and fermenting a pellet feed for fish using lactic acid bacteria to promote digestion, immune activity, and growth promotion, and to a method for manufacturing the same, and more specifically, lacto It is a method of adsorbing and fermenting useful microorganisms including strains of the genus Bacillus. It relates to a fermented fermented feed for fish with digestion promotion, immune activity, and growth promoting effects, and a method of manufacturing the same.

Fish in aquaculture farms Raw feed (MP), Moisture Pellet (MR), Extruded Pellet (EP: Extruded Pellet), Soft Extruded Pellet (SEP), compressed Gun injury is divided into food.

In the aquaculture field, raw feed is a combination of raw feed and MP feed, and mixed feed is compressed dry feed (EP), semi-humid feed (SEP), and compressed dry feed (floating on water, so-called'EP injury feed'). Include. Combined feed is characterized by a small amount of loss when administering food and can protect fry, and has high stability of feed supply and high water quality protection effect.

Among them, compressed dry pallet feed has the advantage of not being released while floating on the water for about 12 hours, so field fishermen tend to use compressed dry pellet feed a lot among compound feeds.

However, these compressed dried pellet feeds have problems in low feed efficiency and digestibility due to the destruction of nutrients as high temperatures occur during feed production, so it is a reality that the current flatfish farms are avoiding them.

Raw feed is a feed that is pelletized by crushing raw meat, but it is high in feed efficiency and digestibility, but when about 30% of feed is dissolved in water when feeding, it deteriorates the water quality by 2 to 5 times, and food safety is not inspected when used. There is a problem with high disease incidence rate. In particular, these diseases are mainly caused by infection by pathogenic bacteria, viruses, and parasites. Recently, the use of raw feed has been obligated to prohibit its use from 2022.

For this reason, a compressed gun pellet feed that can replace raw feed containing antibiotics is being developed.However, such conventional compressed gun pellet feed threatens human health due to the accumulation of antibiotics in fish due to the indiscriminate abuse of antibiotics and excessive feed administration. There are other problems, such as the spread of antibiotic-resistant strains and export disruption due to the detection of antibiotics in live export fish.

Accordingly, several studies have been conducted on compressed dried pallet feed containing natural herbs instead of antibiotics, but this is limited in practical use due to the increase in the unit cost of compressed dried pallet feed due to the use of expensive natural herbs. Existing compressed dry pellet feeds have low protein content and low digestibility, so there are still limitations in applying them as substitutes for raw feed.

1. Korean Patent Registration No. 10-1423293'High-efficiency halibut EP feed composition for replacing raw feed with controlled protein ingredients' 2. Korean Patent Registration No. 10-1302299'EP Feed for Eel'

It is an object of the present invention to provide a fermented fermented blended feed for a compressed gun for fish having an effect of promoting digestion, promoting immune activity and promoting growth by enhancing protein content, digestibility and immunity by supplementing the disadvantages of conventional compressed gun pellet feed There is.

In order to achieve the above object, the method for preparing a fermented fermented feed for a compressed gun wound for fish having an effect of promoting digestion, promoting immune activity and promoting growth of the present invention is a useful microorganism including the Lactobacillus parafarraginis KCTC12357BP strain. A first step of preparing a mixed solution by mixing the culture solution and oligosaccharide, a second step of preparing an adsorbate having a moisture content of 37 to 52% by weight by adding and stirring the fish pellet feed to the mixed solution so that the mixed solution is adsorbed, and And a third process of preparing a fermented fermented feed for fish floating in a compressed gun by post-ripening fermentation at 25 to 30° C. for 24 to 48 hours in a sealed state.

In particular, the Lactobacillus parafarraginis ( Lactobacillus parafarraginis ) is characterized by having a salt resistance and a nucleotide sequence represented by SEQ ID NO: 1.

In addition, the mixed solution of the first step is characterized by consisting of 80 to 90% by weight of the useful microorganism culture solution and 10 to 20% by weight of the oligosaccharide.

Another aspect of the present invention is that the fermented blended fermented feed for fish, which has the effect of promoting digestion, promoting immune activity, and promoting growth, is manufactured by the above manufacturing method.

The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems that are not mentioned will be clearly understood by those of ordinary skill in the technical field to which the present invention belongs from the following description. I will be able to.

In the present invention, in order to improve the low protein content and low digestibility, which are disadvantages of the existing compressed dry pellet feed, the introduction of specific useful microorganisms with high antibacterial activity against pathogenic bacteria and the introduction of a post-ripe fermentation process makes it soft like wet feed, thus reducing digestion time. It is shortened and fermented for a long time, so that the nutrient absorption rate is increased, so that it is possible to provide a fermented fermented feed with a compressed gun wound for fish that has the effect of promoting digestion, promoting immune activity and promoting growth.

In addition, disease prevention is possible without the introduction of additional antibiotics, so that it can be more economically and stably commercialized.

1 is a diagram showing a manufacturing process diagram of a fermented fermented feed for a compressed gun wound for fish having an effect of promoting digestion, promoting immune activity, and promoting growth according to Example 1 of the present invention.
2 is a diagram showing a graph comparing hydrogen peroxide-producing ability by homogeneous strains.
3 is a graph showing a comparative graph of antibacterial activity of L. gavieae , a pathogenic bacterium in each supernatant treated with catalase and untreated with homogeneous strains.
4 is a graph showing the degree of bacterial growth according to the salt concentration targeting mixed strains.

Hereinafter, preferred embodiments and experimental examples of the present invention will be described in detail with reference to the accompanying drawings, and detailed descriptions of known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted.

The present invention relates to a method of manufacturing a fermented blended feed for fish with a compression gun wound having an effect of promoting digestion, promoting immune activity and promoting growth, and the existing conventional pellet feed for fish (EP wound feed) additionally contains antibiotics or Practical use is limited because it contains herbal medicines. In particular, conventional compressed dry pallet feed for fish has a problem in applying it as a substitute for raw feed due to its low protein content and low digestibility. Accordingly, the inventors of the present invention compensate for the disadvantages of the conventional compressed gun pellet feed for fish to exhibit similar efficacy to raw feed and at the same time prevent disease without the introduction of additional antibiotics, so that it is economically and stably practical. As a result of several studies to resolve the distrust of the quality of the blended feed of the field fishermen, first, the conventional compressed gun pellet feed for fish is used as it is, but the above conventional conventional feed is added to a mixture consisting of useful microorganism culture solution and oligosaccharide. It was found that the present invention can achieve the above-described effects of the present invention when the compressed gun pellet feed for fish is added and stirred to adsorb the mixed solution to the feed, and the feed is prepared through only a post-fermentation step.

In more detail, as shown in Figure 1, the mixed solution production step (S10), the adsorbate production step (S20) and the production step of the fermented blended feed for the compression gun for fish having the effect of promoting digestion, promoting immune activity and promoting growth (S30) ), which will be described in detail as follows.

<Production process of fermented fermented feed for compressed gun wound for fish having the effect of promoting digestion, promoting immune activity and promoting growth of the present invention>

1. First process: Preparation of mixed solution (S10)

In this process, Lactobacillus parafarraginis ( Lactobacillus parafarraginis ) KCTC12357BP strain is characterized by mixing the useful microorganism culture medium and oligosaccharides to prepare a mixture.

In other words, the useful microorganisms settle in the intestine and prevent disease by preventing pathogenic bacteria from attaching to the epithelium of the digestive tract, and antibiotics of microbial metabolites generated by useful microorganisms kill or prevent the growth of fish pathogenic bacteria. Lactobacillus parafarraginis is applied as a strain of the genus Lactobacillus that has high antibacterial activity that allows it to inhibit, and in more detail, Lactobacillus parafarraginis has a nucleotide sequence represented by SEQ ID NO: 1 and has flame resistance ( It is best to apply Lactobacillus parafarraginis ) KCTC12357BP.

Here, the culture medium may mean a material including all substances contained in the culture medium in which the strain is cultivated, for example, it may mean a material including metabolites or secretions that are the result of culturing the strain, or a lysate thereof, and the strain itself It may be contained in culture. In addition, the culture medium may be cultured according to a method commonly used in the art.

As the medium, a conventional medium may be used, and after inoculating the Lactobacillus paraparaginis strain in a liquid medium, it is cultured at 35 to 37° C. for 24 to 72 hours to achieve a concentration of 1×10 ⁸ CFU/ml or more. It is characterized by preparing a microbial culture solution, and more preferably, after inoculating the microbial strain in the liquid medium, aeration and agitation of 0.05 to 0.4 vvm (vol/vol/min) for 24 to 72 hours at 35 to 37°C It is cultured at a speed of 100 to 200 rpm (rotary/min) and an internal pressure of 0.1 to 1 kgf/㎠, and the pH of the cultured Lactobacillus paraparaginis strain culture solution is preferably 3.0 to 5.0.

In addition, the culture medium may be configured to further include the following strains in addition to the Lactobacillus paraparaginis strain.

In other words, Lactobacillus para casei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus Francisco tareom (Lactobacillus plantarum), Lactobacillus person noseom switch (Lactobacillus rhamnosus) the culture medium is in addition to Lactobacillus para para jiniseu strain , Lactobacillus reuteri , Lactobacillus acidophilus , Lactobacillus heterohiochii, Lactobacillus brevis , Lactobacillus bulgaricus , Lactobacillus bulgaricus By applying any one or more of salivarius ( Lactobacillus salivarius ), Lactobacillus fermentum ( Lactobacillus fermentum ), these strains are applied in a mixed state in the same amount as the Lactobacillus paraparaginis strain, and these strains are Korea Life Insurance Anyone can easily get pre-sale from the Engineering Research Institute and the Korea Microbial Conservation Center.

In addition, the Lactobacillus parafarraginis ( Lactobacillus parafarraginis ), Lactobacillus paracasei ( Lactobacillus paracasei ) strains are major pathogenic bacteria that frequently occur in marine fish compared to other homologous strains ( Eddosiella tarda , Streptococcus sp ., Vibrio sp . , Pseudomonas anguilliseptica, etc.), shows high antibacterial activity, has salt resistance, and has a high survival rate in the intestine of fish.

The oligosaccharide is added to improve the intake of fish, and as the type, any one or more of fructo-oligosaccharide, soybean oligosaccharide, malto-oligosaccharide, galacto-oligosaccharide, allulose oligosaccharide, glucose, syrup, and fructose is applied. It is preferable to do so, more preferably, it is most suitable to apply allulose oligosaccharides.

In other words, the allulose oligosaccharide is widely used as a substitute for substances called nutrient sweeteners such as glucose, fructose, and syrup, and has a lower calorie taste than other sweeteners while having a taste close to natural sugar. Allulose oligosaccharide is known as a substance that is applied as an almost zero-calorie sweetener because it has only about 5% of sugar (4 kcal per gram) while producing a clean sweet taste close to sugar.

Accordingly, in the present invention, it is most suitable to use allulose oligosaccharides to increase the fish intake rate by increasing the taste without changing the body calories maintained by the active ingredients of the blended feed.

In this process, the useful microorganism culture solution and the oligosaccharide are mixed to form a mixed solution, characterized in that 80 to 90% by weight of the useful microorganism culture solution and 10 to 20% by weight of the oligosaccharide based on the total weight of the mixed solution. It is desirable. That is, if the content of the useful microbial culture medium is less than 80% by weight, the content is too small and disease prevention is not properly performed, and there is a risk of being still exposed to fish pathogenic bacteria, and if it exceeds 90% by weight There is a concern that the content of oligosaccharides is relatively low, and thus the ease of consumption of fish is degraded.

In addition, if the content of the oligosaccharide is less than 10% by weight, there is a fear that the intake of fish will also decrease, and if the content of the oligosaccharide exceeds 20% by weight, the content of the oligosaccharide is relatively large, so that the following adsorbates are usually prepared. There is a concern that adsorption may not be performed properly on the pellet feed for fish.

2. Second process: adsorbate manufacturing (S20)

In this process, pellet feed for fish is added to the mixture prepared in the first process and stirred for 2 to 5 minutes to prepare an adsorbate.

In other words, the conventional pellet feed for fish is a feed that is already known and used in the aquaculture industry, and its density is reduced by controlling moisture, temperature, and pressure so that starch in the feed is gelatinized, and surface treatment of materials required for feed floatation. As it expands, it is a floating feed that allows it to float on the water for about 12 hours. It has the disadvantages that the absorption rate of crude protein in the fish body is somewhat low and the digestibility is low. Nevertheless, in the current aquaculture industry, water pollution is low and feed efficiency verification is accurate, so it is dependent on such pellet feed for fish.

Therefore, as in this process, the conventional fish pellet feed is used as it is, but the mixed liquid is adsorbed thereto to compensate for the disadvantages of the existing fish pellet feed. Add 1 to 3 ℓ of the mixed solution per 20 kg of compressed dry pellet feed into a stirrer and agitate at a speed of 200 to 500 rpm for 2 to 5 minutes so that the mixed solution is adsorbed to the conventional fish pellet feed to prepare an adsorbate, 37 to 52 It is characterized by preparing an adsorbate to have a moisture content of% by weight. In other words, if the mixed solution exceeds 3ℓ, the moisture content of the adsorbed material exceeds 52% by weight, and the shape of the fermented blended feed for fish floating may be disturbed, resulting in a water quality deterioration problem. If it contains less than 1 ℓ, the mixture is not properly adsorbed to the conventional fish pallet feed, so disease prevention is not properly performed, and there is a concern that the present invention is still exposed to fish pathogenic bacteria. There is a fear that the desired effect may not be obtained.

In addition, even when the agitation time or the stirring speed is out of the above conditions, there is a concern that the desired effect of the present invention may not be obtained due to the hassle of re-molding due to excessive adsorption conditions, or the adsorption of the mixed solution is not made completely. As possible, it is most preferable to keep the stirring time or stirring speed as possible.

In addition, since the moisture content of the adsorbate exceeds 52% by weight, it becomes excessively large, making it difficult to eat fish, making it impossible for the user as a feed, or having to go through a separate drying process, resulting in loss of useful microorganisms. When the moisture content of the adsorbed material is less than 37% by weight, the fermentation is not properly performed during the following post-ripening fermentation, and there is a fear that the desired effect of the present invention may not be obtained. do.

3. 3rd process: Manufacture of fermented blended feed for final fish floating with compression gun (S30)

In this process, the adsorbate is subjected to post-ripening fermentation at 25 to 30°C for 24 to 48 hours in a sealed state to produce a final fermented fermented feed for fish with a compressed gun of the present invention.

In other words, when the adsorbate is used as it is, it prevents diseases of fish and improves the intake of fish without additional antibiotics due to the useful microorganisms and oligosaccharides contained therein. However, there is still a problem that the digestibility is low.

Accordingly, the adsorbed material must be used after passing through the post-ripening effect tablet. At this time, the post-ripening fermentation should be performed only for 24 to 48 hours at 25 to 30°C, and the post-ripening effect tablet is completely sealed to avoid exposure to air. Should be.

In other words, if over-fermentation is performed outside the above temperature and time, the useful microorganisms are destroyed, or the intake rate of fish decreases due to excessive fermentation, and if the fermentation is performed less, the digestibility is still lowered, which is the disadvantage of conventional fish pellet feed. It is difficult to see that it has been supplemented.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples, but these Examples and Experimental Examples are for illustrative purposes only and are not intended to limit the protection scope of the present invention.

<Example 1> Preparation of fermented fermented feed 1 with a compression gun for fish of the present invention

After preparing a useful microorganism culture solution consisting of Lactobacillus parafarraginis KCTC12357BP culture solution (cultivated in MRS broth) having the nucleotide sequence represented by SEQ ID NO: 1, add 100 ml of allulose oligosaccharide to 900 ml of this useful microorganism culture solution. 1 L was prepared.

Into 1 L of the prepared mixed solution, 20 kg of the flatfish pellet feed from W Company was put in a stirrer like a bucket and agitated at a stirring speed of 500 rmp for 5 minutes, so that the mixed solution was sufficiently adsorbed to the pellet feed for flounder so that it had a moisture content of 37%. Water was prepared.

The thus prepared adsorbate was subjected to post-ripening fermentation at 25° C. for 48 hours to prepare a fermented fermented feed 1 floating with a compressed gun for fish of the present invention.

<Example 2> Preparation of fermented fermented feed 2 with a compression gun for fish of the present invention

Prepared in the same manner as in Example 1, but instead of the single strain, Lactobacillus parafarraginis ( Lactobacillus parafarraginis ) culture solution (cultured in MRS broth) and Lactobacillus paracasei ( Lactobacillus paracasei ) culture solution (cultured in MRS broth) 1 After fermentation at 30° C. for 24 hours using a useful microorganism culture solution consisting of mixed strains mixed at a :1 weight ratio, a fermented fermented feed 2 of the present invention was prepared.

<Comparative Example 1> Preparation of fermented blended feed 3 with a compression gun for fish

It was prepared in the same manner as in Example 1, but instead of the useful microorganisms, conventional EM microorganisms (including strains of the genus Saccharomyces) were applied to prepare a fermented fermented feed 3 with a compressed gun for fish.

<Comparative Example 2> Preparation of fermented blended feed 4 with a compression gun for fish

It was prepared in the same manner as in Example 1, but when preparing the mixture, 250 ml of allulose oligosaccharide was added to 750 ml of the useful microorganism culture solution and mixed to prepare a fermented fermented feed 4 with a compressed gun for fish.

<Comparative Example 3> Preparation of fermented blended feed 5 with a compression gun for fish

It was prepared in the same manner as in Example 1, but when preparing the mixed solution, 50 ml of allulose oligosaccharide was added to 950 ml of this useful microorganism culture solution and mixed to prepare a fermented fermented feed 5 with a compressed gun for fish.

<Comparative Example 4> Preparation of fermented blended feed 6 with a compression gun for fish

It was prepared in the same manner as in Example 1, but by putting 20 kg of the W company's pellet feed for flatfish in 5ℓ of the mixed solution, a compressed gun-floating fermented blended feed 6 for fish having a moisture content of 60% by weight was prepared.

<Comparative Example 5> Preparation of fermented fermented feed 7 with compression gun for fish

It was prepared in the same manner as in Example 1, but by putting 20 kg of the W's pellet feed for flatfish in 0.7 L of the mixed solution, a compressed gun-floating fermented blended feed 7 for fish having a moisture content of 30% by weight was prepared.

<Experimental Example 1> Confirmation of antimicrobial activity by homologous strains of Lactobacillus against fish pathogenic bacteria

1. Experiment method

Four strains cultured in MRS broth were inoculated with 5 μl of MRS agar, dried at room temperature for 30 minutes, and cultured at 37°C. Then, to investigate the antimicrobial effect of fish pathogenic microorganisms on these strains, L. garvieae , S. parauberis , E. tarda and A. salmonicida , the major fish pathogenic microorganisms frequently occurring in marine fish, were TSB at 37°C. After cultivation in 5 ml of fresh soft agar, 100 µl of each plate was inoculated, and after culturing at 37°C for 24 hours, the antibacterial effect was identified by measuring the clear zone around the colonies of the strains. .

2. Experiment result

Pathogens Inhibition zone diameter (mm) mean±SD L. paracacei L. farraginis L. vini L. parafarraginis L. garvieae 24.2±0.3 22.8±0.8 20.3±0.3 24.8±0.3 S. parauberis 44.2±0.3 40.0±1.0 40.5±0.7 44.7±0.3 E. tarda 38.3±1.4 34.8±0.3 30.8±0.6 37.5±0.7 A. salmonicida 41.8±0.3 40.0±1.8 37.0±1.4 43.0±1.4

As shown in Table 1 above, the antimicrobial effect exhibited against fish pathogenic microorganisms was slightly different even for the same Lactobacillus genus, as shown in Table 1, but antibacterial activity was shown in all lactic acid bacteria genus strains, among which the most antibacterial activity It was confirmed that these two excellent strains were Lactobacillus paracasei and Lactobacillus parafarraginis .

<Experimental Example 2> Confirmation of the antimicrobial mechanism (hydrogen peroxide production ability) for each strain of the same species in Lactobacillus

1. Experiment method

The pathogen used for the identification of the antibacterial effect of Experimental Example 1 was L. garvieae, and the culture solution was suspended in TSB so as to be 0.2 at OD 600 nm, and then catalase (1 mg/ml) treated group and untreated group were respectively Each 100 µl was added to confirm the growth of L. garvieae every 1 hour for 6 hours.

2. Experiment result

As shown in Figs. 2 and 3, the supernatant of L. vini and L. paracacei in the catalase-treated and untreated groups showed no significant difference in the growth of L. garvieae , but in the case of L. paraffaraginis and L. farraginis , the two were It was confirmed that there was a distinct difference in the growth of the pathogenic bacteria L. gavieae in the group. In other words, it can be seen that there is a difference in the ability to produce hydrogen peroxide in the four strains of the Lactobacillus genus homogenous strain, which is closely related to the antibacterial effect.

<Experimental Example 3> Confirmation of salt resistance to mixed strains of Lactobacillus genus

1. Experiment method

In order to check the growth degree of bacteria by salt (NaCl) concentration, 3, 3.5, 4, 5, 10 ‰ of NaCl was added to 100 ㎖ of LB broth (Yeast extract 0.5%, Tryptone 1%), and amplified one day before the experiment. After culturing for a total of 3 days by adding 500 µl of L. parafarraginis culture, the growth of bacteria was analyzed by counting the number of bacteria using optical density (600 nm) measurement.

2. Experiment result

As shown in FIG. 4, the experimental results confirmed that the L. parafarraginis culture medium has high resistance to salt concentration, and it can be seen that it is suitable for use as a material for fish feed.

<Experimental Example 4> Confirmation of fish intestinal viability against mixed strains of the genus Lactobacillus-Confirmation of bile acid resistance

1. Experiment method

In order to confirm the viability of the fish intestine in the L. parafarraginis culture solution in Experimental Example 1, resistance to bile acids secreted while digested by cattle was confirmed.

At this time, the bile acid was extracted from the cow's gallbladder with a syringe and the pH of the bile acid was measured. After centrifuging 1 ml of the mixed strain, the supernatant was discarded and suspended in 1 ml of bile acid. After preculture at 30° C. for 2 hours, 100 μl of MRS The viability of the mixed strains was investigated by comparing the number of colonies after 24 hours of smearing on and PDA plate medium.

2. Experiment result

10 ^- ¹ 10 ^- ² 10 ^- ³ 10 ^- ⁴ 10 ^-5 10 ^-6 10 ^-7 PDA > > 43 3 - - - MRS > > 386 44 7 - - >: Uncountable due to many colonies
-: No number of colonies

As a result of the experiment, as shown in Table 2, it was found that the mixed strain is viable in bovine bile acids and has excellent bile resistance. In other words, when the L. parafarraginis culture solution is orally administered to fish, it shows resistance to bile acids, so that the microorganisms are expected to settle in the intestine, and it is predicted that the viability will also be high in the fish intestine.

<Experimental Example 5> Analysis of feed ingredients by microbial type

1. Experiment method

Contents of crude protein, crude fat, crude fiber, crude powder, calcium, phosphorus by requesting a component analysis agency for the fermented fermented feed 2 with the compressed gun for fish of Example 2 and fermented fermented feed 3 for the fish with the compressed gun of Comparative Example 1 Was examined.

2. Experiment result

Sample: 200g Inspection items unit Example 2 Comparative Example 1 Crude protein % 45.60 40.89 Crude fat % 9.16 8.97 Crude fiber % 0.18 0.92 Minutes % 8.58 7.99 calcium % 1.76 1.79 sign % 1.33 1.29

As a result of the above experiment, as shown in Table 3, Comparative Example 1 is a fermented feed for fish to which previously known EM microorganisms (Saccharomyces sp. strain) were applied, and the content of crude protein was found to be 40.89% even after passing through the post-ripening effect tablet. On the other hand, Example 2 is obtained by post-ripening fermentation of a mixed feed containing a mixed solution containing a useful microbial culture medium containing Lactobacillus parafarraginis , which has the highest antimicrobial activity applied in the present invention, and the crude protein content is 45.60%. It was confirmed that the overall component was increased compared to Comparative Example 1. In particular, since the content of crude protein is increased a lot, it can be seen that the problem of the existing pellet feed for fish having a low crude protein content can be solved through the specific manufacturing process of the present invention.

<Experimental Example 6> Confirmation of the increase rate and feed efficiency of the fermented blended feeds from the compressed gun for fish by post-ripening fermentation conditions

1. Experiment method

Each of 30 flounders of 100-120g produced in the nursery were fed for 2 weeks, respectively, to check the gain rate and feed efficiency of the fermented fermented feeds with compressed guns for fish by post-ripe fermentation conditions.

Feeding was supplied twice a day (9 am, 5 pm) close to full. In order to supply oxygen to the culture tanks smoothly, an air stone was installed in each culture tank and the water temperature was maintained at 19~24℃.

At this time, each feed was prepared by applying the same manufacturing process as in Example 1, and applying only different fermentation temperature and time after ripening.

2. Experiment result

Conditions for fermentation after ripening Increase rate (%) Feed efficiency (%) At 20℃ for 50 hours 50% 55% At 25℃ for 48 hours 90% 85% At 30℃ for 24 hours 92% 90% At 33℃ for 20 hours 45% 50%

As shown in Table 4, it was confirmed that even in the same fermented fermented feed with a compressed gun for fish, there was a large difference in the gain rate and feed efficiency of fish according to the post-ripe fermentation conditions. In other words, in the case of the compressed gun-floating fermentation blended feed for fish prepared in Example 1 or Example 2 of the present invention, the conventional fish pellet feed was post-ripe fermented for 24 to 48 hours at a specific condition of 25 to 30°C. As a result, it shows a significantly higher weight gain and feed efficiency than the existing pellet feed for fish (gain rate: 79%, feed efficiency: 68.39%) that has not been fermented after ripening. It can be seen that it improves the digestibility of fish.

On the other hand, in the case of post-fermentation under conditions other than these, the fermentation conditions are not met, so the useful microorganisms contained in the fermented blended feed for fish are lost, or the feed itself is decayed, and the existing fish that have not been fermented after fermentation. It can be seen that the weight gain and feed efficiency performance is lower than that of the compressed gun pellet feed. Therefore, it is most important to do it for 24 to 48 hours at 25 to 30°C when fermenting after ripening.

<Experimental Example 7> Confirmation of mortality rate, weight gain, and feed efficiency according to the application of the compressed gun-floating fermented blended feeds for fish of Examples 1 and 2 and the fermented blended feed for fish-compressed guns of Comparative Examples 1 to 5

1. Experiment method

The mortality rate, increase rate, and feed efficiency of the fermented fermented feeds for fish of Examples 1 and 2 and Comparative Examples 1 to 5 were checked by feeding 30 flatfish of 100 to 120 g each produced in the nursery for 2 weeks. . Feeding was supplied twice a day (9 am, 5 pm) close to full. In order to supply oxygen to the culture tanks smoothly, an air stone was installed in each culture tank and the water temperature was maintained at 19~24℃.

At this time, as a control group, the existing fish pallet feed (flour W company feed) was applied.

2. Experiment result

Death rate (%) Increase rate (%) Feed efficiency (%) Control 30% 79% 68.39% Example 1 8% 90% 85% Example 2 5% 92% 90% Comparative Example 1 15% 60% 55% Comparative Example 2 50% 82% 85% Comparative Example 3 8% 75% 65% Comparative Example 4 20% 50% 45% Comparative Example 5 25% 45% 40%

As shown in Table 5 above, the mortality rate, body weight gain rate, and feed efficiency were all increased higher than that of the control group in both the compressed gun-floating fermented blended feed 1 for fish of Example 1 and the fermented fermented blended feed for fish flotation of Example 2. Bar, it can be seen that it can be sufficiently used as a feed for fish growth and is sufficiently applied as a substitute for raw feed because it improves the low crude protein and low digestibility, which are the disadvantages of conventional fish pallet feed, and further increases immunity.

On the other hand, in the case of the fermented fermented feed 3 with the compressed gun for fish of Comparative Example 1, an existing EM microorganism different from that of Example 1 was applied, and when the post-ripening fermentation did not meet the conditions, the feed efficiency was lowered compared to the control group. It was also confirmed to fall. In the case of the fermentation blended feed 4 with a compressed gun for fish of Comparative Example 2, the content of allulose oligosaccharides is relatively high, so that the useful microbial culture medium is not properly adsorbed to the conventional fish pellet feed, and the useful microbial culture medium is relatively high. Due to its low content, it was confirmed that the mortality rate was high due to exposure to fish pathogenic bacteria. In the case of the fermented mixed feed 5 for fish in Comparative Example 3, the content of oligosaccharide was relatively low, so the intake of fish was inferior, and it was confirmed that the gain rate and feed efficiency were similar to those of the existing fish pellet feed (control). I did.

In the case of the fermented fermented feed 6 with the compression gun for fish of Comparative Example 4, the moisture content of the adsorbate itself consisting of useful microorganisms and oligosaccharides was too large, and when feeding to fish, the form of the feed was disturbed and it was difficult for flounder to eat easily. In the case of the fermentation blended feed 7 with a compressed gun for fish of Comparative Example 5, it was confirmed that the moisture content of the adsorbate itself composed of useful microorganisms and oligosaccharides was not properly fermented, so that the gain rate and feed efficiency were also reduced.

As described above, in the present invention, the low protein content and digestibility, which are the disadvantages of conventional fish pellet feeds, are relatively improved through the introduction of specific useful microorganisms having high antibacterial activity against pathogenic bacteria and post-ripening fermentation, while increasing immune activity. It was possible to provide a fermented blended feed for fish with a compressed gun wound, as well as disease prevention without the introduction of additional antibiotics, and it was found that it was possible to commercialize it more economically and stably.

The present invention has been looked at around preferred embodiments, and those of ordinary skill in the art to which the present invention pertains will implement the detailed description and other forms of embodiments within the essential technical scope of the present invention. I will be able to. Here, the essential technical scope of the present invention is shown in the claims, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Korea Research Institute of Bioscience and Biotechnology KCTC12357BP 20130123

<110> Lbio technology co.,ltd <120> Extruded pellet expanded fermentation feed for fish that has the effect of promoting digestion, immune activity, and promoting growth and a method for producing the same <130> Lbio-03 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1137 <212> DNA <213> Lactobacillus parafarraginis <400> 1 acatgcaagt cgaacgaggc aaccataatg aagttaggtg cttgcattta actgatttaa 60 cattgagacg agtggcgaac tggtgagtaa cacgtgggta acctgccctg aagtggggga 120 taacacttgg aaacaggtgc taataccgca taacaacgaa aaccacatgg ttttcgtttg 180 aaagatggct tcggctgtca cttttggatg gacccgcggc gtattagctt gttggtgagg 240 taacggctca ccaaggccat gatacgtagc cgacctgaga gggtaatcgg ccacattggg 300 actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc acaatggacg 360 aaagtctgat ggagcaacgc cgcgtgagtg atgaagggtt tcggctcgta aaactctgtt 420 gttggagaag aacaggtgat agagtaactg ttatcatctt gacggtatcc aaccagaaag 480 ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggat 540 ttattgggcg taaagcgagc gcaggcggtt ttttaggtct gatgtgaaag ccttcggctt 600 aaccggagaa gggcatcgga aaccgggaga cttgagtgca gaagaggaca gtggaactcc 660 atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag gcggctgtct 720 ggtctgtaac tgacgctgag gctcgaaagc atgggtagcg aacaggatta gataccctgg 780 tagtccatgc cgtaaacgat gagtgctaag tgttggaggg tttccgccct tcagtgctgc 840 agctaacgca ttaagcactc cgcctgggga gtacgaccgc aaggttgaaa ctcaaaggaa 900 ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcta cgcgaagaac 960 cttaccaggt cttgacatct tctgctaacc taagagatta ggcgttccct tcggggacgg 1020 aatgacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 1080 caacgagcgc aacccttatt gtcagttgcc agcatttagt tgggcactct ggcgaga 1137

Claims (5)

A mixture was prepared by mixing 80 to 90% by weight of useful microorganism culture solution containing the strain of Lactobacillus parafarraginis KCTC12357BP and 10 to 20% by weight of oligosaccharides having salt resistance and having a nucleotide sequence represented by SEQ ID NO: 1. The first step to do;
The fish pellet feed is added to the mixed solution and stirred so that the mixed solution is adsorbed, but 1 to 3 L of the mixed solution per 20 kg of the fish pellet feed is put into a stirrer and stirred for 2 to 5 minutes under conditions of 200 to 500 rpm so that the mixed solution is adsorbed A second step of preparing an adsorbate having a moisture content of 37 to 52% by weight and,
Including; a third step of producing a fermented fermented feed with a compressed gun for fish by post-fermenting the adsorbed material at 25 to 30°C for 24 to 48 hours in a sealed state.
A method for producing a fermented fermented feed with a compressed gun for fish that has the effect of promoting fish immune activity and promoting growth.
delete delete The method of claim 1,
The useful microbial culture medium of the first step includes Lactobacillus paracasei , Lactobacillus casei , Lactobacillus plantarum , Lactobacillus rhamnosus , Lactobacillus rhamnosus , Lactobacillus reuteri , Lactobacillus acidophilus , Lactobacillus heterohiochii , Lactobacillus brevis , Lactobacillus bulgaricus salivacillus, Lactobacillus salivacillus Lactobacillus salivarius ), Lactobacillus fermentum ( Lactobacillus fermentum ) characterized by further comprising any one or more of,
A method for producing a fermented fermented feed with a compressed gun for fish that has the effect of promoting fish immune activity and promoting growth.
It is manufactured by the manufacturing method of any one of claims 1 or 4,
It is characterized by having an effect of promoting immune activity and promoting growth of fish,
Compressed gun floating fermented blended feed for fish.

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