KR101187731B1 - Lactobacillus parafarraginis strain having deodorization activity and water purification ability in eel farm and uses thereof - Google Patents

Abstract

The present invention is Lactobacillus parafarraginis strain having a deodorizing activity and the water purification function of the eel farm, the microorganism preparation for water purification of the deodorant and eel farm containing the strain or its culture as an active ingredient, the culture of the strain Method for preparing a microorganism preparation for water purification of deodorizing and eel farm comprising a step, the method for removing the odor by treating the strain or its culture solution in a wastewater treatment plant, barn, compost factory or toilet and the strain or its culture solution eel Provides a way to purify the water by treating the farm.

Description

Lactobacillus parafarraginis strain having deodorization activity and water purification ability in eel farm and uses according to deodorizing activity and water purification of eel farms

The present invention relates to a Lactobacillus parafarraginis strain having a deodorizing activity and water purification function of an eel farm, and more particularly, to a Lactobacillus paraparagigi having a deodorizing activity and a water purification function of an eel farm. Nice strain, the microorganism preparation for water purification of deodorizing and eel farm containing the strain or its culture as an active ingredient, a method for producing a microbial preparation for water purification of deodorizing and eel farm comprising the step of culturing the strain, the strain or The present invention relates to a method for removing odor by treating the culture solution thereof in a wastewater treatment plant, a barn, a compost factory, or a bathroom, and to purify the water quality by treating the strain or its culture solution in an eel farm.

Recently, as the standard of living improves, the noise, vibration or dust that causes unpleasantness in daily life as well as odor are regulated as pollution. Odors are typically acidic odors such as hydrogen sulfide, methylmercaptan, methyl sulfide and methyl disulfide; Basic odors such as ammonia, trimethylamine and skatole; Or neutral odors such as acetaldehyde and styrene. Conventional deodorization methods for removing such odors include a chemical deodorization method (masking method) in which a smell is concealed by using aromatic substances such as perfumes or fragrances; Chemical deodorization which decomposes using oxidation, reduction, neutralization, addition condensation and ion exchange reaction; Physical adsorption method for collecting by using porous materials such as activated carbon, zeolite and silica gel; Or it is roughly classified into a biological deodorization method that inhibits using microorganisms, enzymes and preservatives. However, chemical deodorization is likely to generate new odor or toxicity depending on natural or synthetic ingredients, and the physical adsorption method is effective, but the odor is regenerated according to environmental conditions or long-term use, and the chemical deodorization method can be developed in various ways. There is a limit to the total deodorizing power.

Specifically, odorous substances include ammonia, methylmercaptan, H ₂ S, sulficmethyl, disulfic methyl, trimethylamine, CH ₃ COOH, and styrene. In particular, representative substances are ammonia and amines, which are a major cause of odor. Ammonia is primarily irritating to eye irritation and has a particularly high incidence of respiratory disease. Nitrate and ammonia nitrogen and organic substances contained in a large amount act as a major odor causing factors causing many side effects in the odor removal process, such as waste water treatment plant, barn, manure treatment or composting process. In addition, chronic exposure has a detrimental effect on human health as well as livestock because it acts as a stressor. Therefore, there is a continuing interest in developing a method for removing simple and efficient odors.

On the other hand, microbial probiotics are an important part of the complex food world and play a very beneficial role in the health of freshwater and sea creatures such as fish, shrimp and farmed fish species. The microbial probiotic of the present invention is a feed additive and a water purifying agent, and may include living microorganisms. In the present invention, the microorganisms of the probiotic and the metabolites produced by the microorganisms promote the health of the farmed fish species and remove water pollution and odor. We've seen whether the function can be performed at the same time.

Korean Patent No. 10-1029346 discloses a microbial fermentation broth having a odor gas reduction and antibacterial effect and a method of using the same, and Korean Patent No. 10-0654427 discloses a Lactobacillus planta having an odor generating and deodorizing activity. Room CU03KACC 91103 is disclosed, but Lactobacillus paraparaginais strains having deodorizing activity and water purification functions of eel farms as in the present invention and their use are not known.

The present invention has been made in accordance with the above requirements, the inventors of the present invention, when the liquid sample containing the Lactobacillus parafarraginis strain to a high concentration of ammonia when the ammonia concentration is fast through the detection gas detector When the strain was mixed with a certain amount of feed and orally administered to the eel and treated at the same time as the eel farm, the amount of nitrite and ammonia gas generated in the tank during the conventional feed administration was remarkably relieved, and a small amount was generated. The gas removal effect is also excellent, so that the water temperature and pH are kept constant even at low return rates (less than 10%) in eel farms that required 100% return at least once a day. The invention was completed.

In order to solve the above problems, the present invention provides a Lactobacillus paraparaginais strain having a deodorizing activity and water purification function of the eel farm.

The present invention also provides a microorganism preparation for water purification of deodorant and eel farm containing the strain or its culture as an active ingredient.

In addition, the present invention provides a method for producing a microorganism preparation for water purification of deodorant and eel farm comprising the step of culturing the strain.

In addition, the present invention provides a method for removing the odor by treating the strain or its culture solution in a waste water treatment plant, barn, compost factory or toilet.

The present invention also provides a method for purifying water quality by treating the strain or its culture in an eel farm.

Since the Lactobacillus paraparaginanis strain of the present invention has a very good deodorizing effect of ammonia, which is a major component of malodor production, the composition containing the Lactobacillus paraparaginanis strain or its culture solution as an active ingredient in wastewater treatment plant, barn, compost It can be very useful not only for industrial use but also for home use because it can remove odor easily in a fast time by treating the place where there is a lot of odors such as a factory or a bathroom.

In addition, when the strain of the present invention is mixed orally administered to the eel and treated in the eel farm at the same time, the water temperature and pH are kept constant for a long time even with a minimum return rate of the tank, which can greatly contribute to the economics of the eel farm industry. have.

Figure 1 shows the change in concentration of ammonia over time after treatment with Lactobacillus parafarraginis strain.
Figure 2 shows the pH change of the tank according to the microbial feed amount and microbial input in the pom-poms. The microbial feed amount is a feed containing the Lactobacillus paraparaginanis of the present invention, the microorganism feed is a lactobacillus paraparaginais input of the present invention.
Figure 3 shows the return of the tank and the fry mortality with the increase of the microbial feed input in the fry culture (ⓐ, fry mortality; ⓑ, microbial feed dosage).
Figure 4 shows the changes in pH, water temperature and return of the tank according to the microbial feed amount and microbial input in the eel culture (up to about 50 days eel culture).
Figure 5 shows the change in pH, water temperature and return of the tank according to the microbial feed amount and microbial input in the eel culture (from about 50 days to about 100 days eel culture).
Figure 6 shows the change in pH, water temperature and return of the tank according to the microbial feed amount and microbial input in the eel culture (from about 100 days to about 140 days eel culture).
Figure 7 shows the change in the amount of water and pH of the tank when the normal feed in the eel culture.

In order to achieve the object of the present invention, the present invention provides a Lactobacillus paraparaginais strain having deodorizing activity and water purification function of the eel farm.

Lactobacillus parafarraginis strain of the present invention was isolated and identified in plants, and the strain was deposited on February 16, 2012 to the National Institute of Agricultural Science, Agricultural Genetics Center (Accession Number: KACC91710P).

"Deodorizing activity" means the ability to remove the odor, the odor causing substances are odors such as ammonia, methylmercaptan, H ₂ S, sulfic methyl (sulfic methyl), disulfic methyl, trimethylamine, CH ₃ COOH, styrene and the like, preferably ammonia, but is not limited thereto.

The water purification function of the eel farm of the present invention means the removal of nitrous acid (NO ₂ ) or ammonia (NH ₃ ) gas caused by the secretion of eel or decay of feed fed into the tank, but is not limited thereto.

The present invention also provides a microorganism preparation for water purification of deodorant and eel farm containing the strain or its culture as an active ingredient.

The deodorizing microbial preparation of the present invention may be a powder or liquid formulation, and the microbial powder formulation is used by suspending in sterile physiological saline, spraying the liquid formulation directly on an object, or generating the odor by putting the liquid formulation in a container. It may also be used as a method of placing in the wealth, but is not limited thereto.

The microbial preparation for water purification of the eel farm of the present invention may be a powder preparation or a liquid preparation, and may be used by suspending the microbial powder preparation in sterile physiological saline, or by spraying the liquid preparation directly on an eel farm. This is not restrictive.

In addition, the present invention provides a method for producing a microorganism preparation for water purification of deodorant and eel farm comprising the step of culturing the strain. The method of culturing the strain of the present invention may be cultured according to methods commonly used in the art, and preferably, but not limited to, using plate count agar (PCA) and tryptic soy agar (TSA) medium.

In addition, the present invention provides a method for removing the odor by treating the strain or its culture solution in a waste water treatment plant, barn, compost factory or toilet.

The barn may be a space for raising mammals such as pigs, cattle, goats, fish such as carp, goldfish, and poultry such as pheasants, chickens, ducks, turkeys, etc., but is not limited thereto.

The present invention also provides a method for purifying water quality by treating the strain or its culture in an eel farm.

In order to purify the water quality by treating the eel farm of the present invention, the strain of the present invention or its culture solution can be directly treated to the eel farm, and at the same time, when mixed with a feed to a certain concentration and orally administered to the eel, the water purification effect May be used, but is not limited thereto.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example 1.Lactobacillus paraparaginis of the present invention ( Lactobacillus parafarraginis Isolation of Strains

In order to separate the microorganism strain having deodorizing activity from the plant, the liquid sample provided by the plant was diluted by a continuous dilution method, plated in a medium for microbial separation, and incubated at 25 ° C. for 3 to 7 days. As the medium for microbial separation, PlateAgar (PCA), Tryptic Soy Agar (TSA), MRS (De Man, Rogosa, Sharpe), and YM (yeast separation) agar medium were used. Bacteria appeared almost similar in the three kinds of media, and in the MRS medium, which is a lactic acid bacterium separation medium, only yeast and bacteria can be distinguished, and only the counts were used. 5 to 10 strains were selected in consideration of the back. Yeast was also selected seven strains based on the size, shape and color of the colony.

More detailed description of the strain selection method, inoculated by 12 dilution by continuous dilution method cultured in microbial separation medium to Tryptic Soy Broth each shaking incubation for 9-48 hours at a temperature of 25 ~ 45 ℃ deodorized The most active and few colonies were selected, and the agar medium and green tea extract were nano-ized using an ultrasonic crusher, and the second culture was selected for 9 to 40 hours at a temperature of 25 to 40 ° C.

Example 2. Lactobacillus paraparaginas of the present invention ( Lactobacillus parafarraginis Identification of Strains

Identification was carried out using 16S rDNA sequencing to isolate the strain selected in Example 1 by molecular biological methods. As a result of 16S rDNA sequence similarity analysis, it was determined that the bacteria belong to the genus Lactobacillus . Lactobacillus is present in 140 species and 9 subspecies (as of December 2010). Among them, Lactobacillus parafarraginis standard strain and 16S rDNA sequence showed a very high similarity (99.731%). ). In addition to L. rapi , Lactobacillus hilgadi ( L. hilgardii ) and L. parabuchneri ( L. parabuchneri ) The high degree of similarity with the standard strains, such as 97% or more. The strain was analyzed by 16S rDNA nucleotide sequence of 1,114bp, the phylogenetic analysis using the nucleotide sequence showed that the strain of the present invention forms the same branch as the Lactobacillus paraparaginais standard strain. Homology analysis of the DNA sequence of SEQ ID NO: 1 obtained as a result of the analysis of the bacteria was performed using the NCBI BLASTN program (http: //www.ncbi.nlm.nihgov/blast/), which was the Lactobacillus paraparagi Nice ( Lactobacillus parafarraginis ) was named.

Example 3 Analysis of Ammonia Deodorization Activity Following Treatment of Lactobacillus Paraparaginas

In order to confirm whether the strain of the present invention has a deodorizing activity against ammonia, the concentration of ammonia over time after the strain treatment was examined. The test was carried out in the state in which the strain of the present invention was not treated, and a blank was used as a control, and the liquid sample containing the strain of the present invention was treated as a sample (Table 1). The sample was diluted 5 times with the distilled water, and 20 ml of the strain was placed in a 5 L reactor and sealed. The initial ammonia concentration as a test gas was injected into the blank and the sample at 50 μmol / mol, the temperature was maintained at 18 to 28 ° C, the humidity was maintained at 40 to 60%, and the concentration of the test gas was initially (0 minutes) and 30 minutes. At 60 minutes, 90 minutes and 120 minutes. The concentration of the test gas was measured by a detector tube gas meter (KS I 2218: 2009) (Table 1). The removal rate of test gas at each time slot was calculated by the following equation; Test gas removal rate (%) = [{(blank concentration)-(sample concentration)} / (blank concentration)] x 100. As a result, the ammonia concentration in the blank was almost unchanged, but in the sample, the ammonia concentration measured after 30 minutes decreased to 1 μmol / mol, showing a 98% deodorization rate, and after 90 minutes, the ammonia was removed to an undeterminable amount. It could be seen (Figure 1). These results can deodorize ammonia in a faster time than the previous invention, it can be said that the strain of the present invention is excellent in ammonia deodorizing activity efficiency.

Test Items
Test result
Experiment method Blank concentration
(μmol / mol) Sample concentration
(μmol / mol) Deodorization rate (%) Deodorization test
ammonia
(NH ₃ )
0 min 50 50 0.0 Requester
Presentation method
30 minutes 49 One 98.0 60 minutes 49 One 98.0 90 minutes 49 N.D. N.D. 120 minutes 49 N.D. N.D. N.D. (not detected): Not detected (detection limit 0.2μmol / mol)

Example 4 pH Change of Aquarium with Microbial Feeding and Microbial Feeding in Eels

The tank administration of the Lactobacillus paraparaginanis microorganism of the present invention was initially administered more than the experimental standard for the stabilization of the water quality of the tank and can be confirmed after the stable microbial administration (Fig. 2). In addition, when looking at the feed amount containing lactobacillus paraparaginanis of the present invention, three different types (inoculated with lactobacillus paraparaginanis microorganisms in the degreasing river, powdered probiotic type, microorganisms are injected into the general dough feed) The change in feed amount for the conversion of the feed of one liquid state type, the type pelletized by spraying liquid microorganisms) was confirmed. Even in the microbial feed dose and microbial dose it was confirmed that the pH value is maintained within a certain range (Fig. 2).

Example 5 Water Recovery and Poor Mortality of Fish Tanks with Increasing Microbial Feeding Rates in Eels

With the increase and decrease of feed amount according to the growth of eel fry, the amount of mixed lactobacillus paraparaginanis microbial feed of the present invention was also increased and decreased, and an average of 60% of the tank water was recovered due to water quality-sensitive fry (FIG. 3). Based on this situation, the mortality rate of fry decreased as feed volume increased, and it was confirmed that no mortality occurred from the mid-term. Comparing the moving average line ⓐ and ⓑ of the microbial feed dose and mortality, it was possible to compare and confirm the water quality stabilization situation is significantly changed (Fig. 3).

Example 6 Changes in pH, Water Temperature, and Return of Fish Tanks with Microbial Feeding and Microbial Feeding in Eel Culture

Figure 4 shows the change in pH, water temperature and return of the water tank according to the feed amount and microorganism feed containing the strain of the present invention from the start of the eel culture from about 50 days. The first screening process, which started with 110 kg of 35,640 animals, was able to confirm the stable water temperature, microbial administration in a constant tank, and feed volume increased with eel growth. The return of the tank was adjusted from the maximum 17% to the minimum 4%. In general, the water quality of the tank in the eel farming is a direct influence on the mortality, etc. It is common to perform a large amount of return, in the present invention was able to maintain a stable water quality using microorganisms to maintain a minimum return rate (Fig. 4). .

In order to investigate the effect of the microorganism of the present invention on improving the water quality through the decomposition of eel waste (nitrous acid and ammonia gas) and residual floating feed, a total of 12 ponds including 9 total 30 pyeong ponds and 3 90 pyeong ponds It was diluted at the time of feeding to 220,000 animals and mixed orally with general feed (product name, eel sacred feed; manufacturer, dominant feed) and microorganisms in ponds to observe the multipurpose effect including water removal in proportion to the food dosage. As a result of injecting from 57 tons to 282 tons of water from 500cc to 2L, as shown in Table 2 and Table 3, nitrous acid and ammonia gas were removed drastically, stable pH was maintained, and the daily return amount was also significantly reduced. It could be confirmed (Table 2 and Table 3).

Figure 5 shows the change in pH, water temperature and return of the tank according to the microbial feed amount and microorganism input from about 50 days to about 100 days of eel culture, the water temperature was maintained stably based on 29 ℃, About 5-8% of the conditions were maintained similar to the primary screening mode. Several spikes in return and a decrease in feed volume were caused by reasons such as tank cleaning. Feed volume increased continuously as the eel grew, and microbial administration in the tank was also administered at regular intervals. It was found that the pH value was maintained within a stable range due to the increase in feed and the introduction of microorganisms (FIG. 5).

Figure 6 shows the change in pH, water temperature and return of the tank according to the microbial feed amount and the microorganism input from about 100 days to about 140 days eel culture, the variation curve of the graph is the same as that of FIG. It turned out that it was. However, the amount of feeding was in line with the growth of the eel, the increase in the amount of microorganisms mixed, and the increase in the depth of the eel were slightly variable. The decrease in feed volume and the increase of water return on the graph were changed due to the cleaning of the tank. It was found that pH and water temperature were kept stable.

Figure 7 shows the change in the amount of water and pH of the tank when the general feed in the eel culture, it can be seen that the return of the tank proceeds to 100% for water quality management from April 2, 6th day of the tank injection of the eel have. It is estimated that 100% of the tanks were returned due to the eel manure and corruption of the feed. In addition, according to the increase and decrease of the feed it was confirmed that the pH is also increased or decreased in the same pattern. The pH value in the tank was found to be directly related to the feed volume.

Agricultural Biotechnology Research Institute KACC91710P 20120216

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Claims (5)

Lactobacillus parafarraginis strain (KACC91710P) having deodorizing activity and water purification function of eel farm. The microorganism preparation for water purification of deodorant and eel farm containing the strain of claim 1 or a culture thereof as an active ingredient. A method for preparing a microorganism preparation for water purification of deodorant and eel farm comprising the step of culturing the strain of claim 1. Method of removing odor by treating the strain of claim 1 or its culture solution in a waste water treatment plant, barn, compost factory or toilet. A method for purifying water quality by treating the strain of claim 1 or a culture thereof in an eel farm.

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