JPS5884094A - Purifying agent for fresh water - Google Patents

Abstract

PURPOSE:To obtain a water purifying agent which maintains an excellent water purifying effect for a long period of time by culturing saprophytes having less pathogenicity which are >=1 kind among inorg. or org. nitrogen compd. decomposing bacteria, cellulose decomposing bacteria and oil and fat decomposing bacteria artificially and putting the same into a dormant state. CONSTITUTION:1 or >=2 kinds selected among inorg. or org. nitrogen compd. decomposing bacteria, cellulose decomposing bacteria and oil and fat decomposing bacteria are cultured in a fermenting tank of large capacity made of stainless steel 1. The medium compsn. in this case is, for example, 3% soybean meal, 1.0% starch, 0.2% ammonia sulfate and 0.3% calcium carbonste, and bacteria str propagated therein by controlling the pH thereof to 7.0 prior to sterilization of the media and culturing the bacteria under aerated agitation for 12- 48hr at 30 deg.C. The respective grown bacteria after the culturing are gathered by a centrifugal separator and the resultant bacteria pastes are freeze-dried in acetone-dry ice and are mixed with each other, whereby a fresh water purifying agent is obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a freshwater purifying agent. Further Details 11 This invention relates to a novel freshwater purifying agent containing one or more non-pathogenic microorganisms.

Water in swimming pools, water tanks, fire prevention tanks, cooling tanks, reservoirs, fountain ponds, irrigation ponds, etc. is susceptible to decay, so the water should be changed at regular intervals and fresh water should be added from time to time. Measures are being taken to add filtrate, weed killer, and other cleaning agents. However, the method of replacing the water and pouring water requires an extremely large amount of labor, and the method of using a purifying agent is a simple method, but the purifying agent itself is harmful. Therefore, there are many restrictions on its use in terms of pollution, availability, and safety.

For this reason, the development of harmless purifying agents is eagerly awaited. In addition, various freshwater fish and other seafood are raised in these ponds and tanks for the purpose of both water purification and aquaculture, but these fish and shellfish are harmless and At present, no purifying agents with high water purification effects are known.

In addition, in aquariums, glass tanks, glass bowls, breeding containers, etc., food fish, ornamental fish, breeding observation fish, breeding experimental fish, and seafood such as shellfish, shrimp, and crabs are kept in large quantities or in small numbers. However, when kept in these artificial environments for long periods of time, a food chain is formed between the fish, shellfish, plants, and microorganisms in the water, and the fish and shellfish have a self-cleaning effect (unlike in the natural environment). excrement, feed residue, etc. accumulate,
Furthermore, as they rot, they pollute the water and produce toxic substances, which can cause seafood to become infected and become infected if left untreated.
Growth stops and even death occurs. Therefore, it is not a fool's errand that it is essential to use the feed amount KJ1B on a daily basis, but it is essential to frequently change the water within a short period of time. However, as mentioned above, even in the case of small-capacity tanks or pots, the task of replacing water is complicated. Furthermore, when the water that the fish and shellfish were accustomed to was replaced, the growth of the fish and shellfish stopped due to slight differences in water temperature, amount of dissolved oxygen, and composition of the water.
, it is undeniable that activity will slow down.

As a result, the growth of fish and shellfish has decreased, fish diseases have occurred due to decreased physical strength, and the yield of farmed fish and shellfish has drastically decreased, leading to mass mortality. purification is hindered. Plus, when you dispose of dirty, used water, you get even more! Secondary pollution is caused, and pollution problems also arise.

The present invention solves these drawbacks all at once, and develops a new water purifying agent that has an extremely excellent water purifying effect, does not have any harmful effects on fish and shellfish, and has long-lasting effects. It was made for the purpose of

As a result of conducting research from various angles to achieve this purpose, we found that using chemical substances would not achieve the desired purpose.
We obtained new knowledge that good results can be obtained by using microorganisms, and based on this knowledge, we continued our research and found that inorganic and organic substances such as ammonia and nitrite ions in the excreta of seafood One type of garden that decomposes nitrogen compounds or! It has been found that administering a combination of two or more species has a significant purifying effect. Therefore, we screened and selected a large number of microorganisms, and further investigated their combinations. By combining a large number of cellulite-degrading bacteria and oil-degrading bacteria to decompose oils and fats, it has an even more amazing purification effect, and is useful not only for purifying fresh water, but also for raising seafood,
It was found that this method is useful for preventing diseases, leading to the present invention.

That is, the advantage of the present invention lies in a freshwater purifying agent characterized by artificially culturing non-pathogenic saprophytic bacteria and keeping it in a dormant state.

These saprophytic bacteria are classified into three groups: inorganic or organic nitrogen compound, cellulose-degrading bacteria, and fat-degrading bacteria, and the 11 that belong to each group are as follows.

Inorganic or organic nitrogen compound decomposition cocoon (1) Domonas denitrificans (Ps
eu113onajdenitrifioans )(
(2) Bacillus subtilis (Baoilw @ubtilig) (Feural Research Institute No. 6114) (3) Bacillus subtilis (Baoilw @ubtilig)
lw *urigi*rwais)
(No. 11s) (4) Arthropacta agilis (Mrthro1wter aaKlisg) (Kyokoken No. 6113) (5) Kurchia zoffii (Kurnobuchipta) (Kyokoken Bacteria No. 6119) Cellulose-degrading bacteria (a) 7 Svergillus terreus (Mu9shoken 1us tarrsw) (Koken Kenkyuyori No. 6112) (7) Bacteroides succinoges (Baota
roidw mooinogenas+ ) (Mooinogenas+) (Mooinogenas+)
(No. 6111) Oil-degrading bacteria (8) Bacteroides lipolyteicum (Baot
(9) Brevibacterium lipolyticum (Br
edlwtsrim 1 voice 1ytium) (Koukou Kenkyoku No. 6118) Shout Tsukasia Kotrina (Tsukasai Oo Parent 1-) (Koukou Kenzonyori No. 6120) Ps, Denitrificans is derived from the soil. Separated cocoons, aerobic and hyperaerobic.

B. subtilis and B. chelingiensis both cocooned in aerobic cocoons on a threshold separated from the soil;
It has spore-forming ability. However, in the latter garden, it does not have the property of causing diseases to insects, whereas N. = Larina has the ability to form chlamydospores in an aerobic garden.

Many hydrocarbons, fats and oils, and phenols can be used as carbon sources. Agilis is a plesiomorphic aerobic cocoon with a wide temperature range. In addition to organic nitrogen compounds, a large number of hydrocarbons can be used as carbon sources. Both Bacteroides succinogenes and Boliticum are cocoons isolated from the rumen of cows and are anaerobic bacteria.

Brew, Libolyticum is a bacterium isolated from whey digestion plants, is an aerobic and facultative aerobe, and has a wide temperature range. And it can quickly break down fat. Furthermore, Zoffii was isolated from the decomposition product of chicken manure, and is aerobic and facultatively aerobic. In other words, Terex is an aerobic filamentous fungus that has spore-forming ability.

The above-mentioned (13-(4) and lI/I'i of trout can be cultured in a stainless steel O large capacity fermentation tank.The medium composition is 3% soybean all, 1.0% starch,
Ammonium sulfate 12%, calcium carbonate O,S%, pH adjusted to 7.0 before cocooning the medium, SO
The surface was grown by aeration and agitation culture at ℃.

The above-mentioned (5) p (7) to (9) can also be cultured in a large capacity fermentation tank made of stainless steel. The medium composition is soybean paste %, starch 10%,
Calcium carbonate concentration is 1%, and the pH of the medium is adjusted to 7.0 before harvesting.Culture is carried out at 30°C, but without aeration or excessive stirring. The above-mentioned (6) II (i.e., Aspergillus 1lI) species, which is carried out under mild conditions, can also be mass-cultured in a stainless steel fermentation tank.

As for the culture medium (υ~(4), the medium using IIK is used, but its OpH is adjusted to 10.
1) to (4) Perform aeration and agitation culture in the same manner as OII to propagate Maro. In addition, regarding the medium of (1) to (to) above, in addition to the medium described above, other ingredients such as minerals, vitamins, yeast extract, fishin meal, plain heart sugar, etc. , NZ-amine,
It is also possible to further add other commonly used ingredients.

In the present invention, although it is effective to purify fresh water even when each of the above three groups is used alone, it is preferable to use one or two of the inorganic or organic nitrogen compound decomposition plants, heptalulose decomposition plant, In combination with oil-decomposed cocoons, and furthermore, the above (1) to (
It is ideal to combine all aspects of (to). When mixing these bacteria, they are usually mixed in equal amounts, but in this case it is not limited to just the ratio. Incubate and then mix. After continuing the culture for 12 to 41 hours, each of the growths was collected by centrifugation, and the image paste thus obtained was freeze-dried in an a+ton dry ice bath, and then transferred to one another. Because it is mixed with.

It is also possible to purely separate only the surrounding body from the bacterial cell paste and dry it, or to leave it wet without drying it.

Ninth, if it is in a naughty state like painting paste, it is not only inconvenient to handle and store, but also causes the cocoon to become overly active. Therefore, in order to solve these problems, it is preferable to mix the one-piece paste with an inert carrier. As inert carriers, any material can be used as long as it is inert to these orchids, for example clays or clays such as diatomaceous earth, montmorillonite, kaolin, burr & light, bentonite. Minerals such as aluna, silica, calcium carbonate, aluminum carbonate, apatite, and attapulgite can be widely used. These inert carriers and microbial cells are mixed at the most suitable ratio depending on the carrier used, the amount of microbial cells, and its moisture content.

In the form of an inert carrier, it acts not only as a desiccant for the surrounding body, but also as a protective agent for one body. Preferably 1:
If mixed at a ratio of 1 to 1:5, the lifespan of the whole will be extended, and germination and proliferation of bacterial cells will also be inhibited. There will no longer be a need for packaging.

There are no special precautions to be taken when storing it, just store it in a cool, dry place.

Although microbial products usually require very delicate storage and preservation precautions, the simple operation of simply mixing the wet cake and the carrier without forming a lyo-7 is sufficient. Can be stored and preserved for a long time. Of course, Lyofile may be used as is.

To use the freshwater purifying agent according to the present invention, it is sufficient to dissolve it in a small amount of water and sprinkle it on the water surface, or it can be simply added in a small amount directly into the water. It is very convenient and does not require complicated usage. It can also be used by mixing it with feed, if necessary. There are no particular restrictions on when to use it, but it is most efficient to use it when the water becomes cloudy.

However, it is of course also possible to use it from the beginning or at any other suitable time. If you are raising seafood, it is best to use it after changing the water.
There is no problem if you use it without changing the water. The amount used may be very small, but as a rough guideline, if you are raising fish and using it in a tank or aquarium, the amount should be approximately 15 to 80%.
F% preferably about 2 when used in a 25-50Ps pond
0 to 80F, preferably 25 to 501 pl, and for maintenance use, 2 to 3 times a year, 3 to 15 pl as necessary.
1. Preferably, it is sufficient to input it at a level of 5 to 10 points, and maintenance and management is extremely simple. When the purpose is to purify water when no fish or shellfish are being raised, the desired purpose can be sufficiently achieved with a smaller amount than the above amount. Of course, the amount used above is a must! It is also possible to adjust the amount depending on the situation.

The purifying agent of the present invention has extremely high safety, and when handling it, there is no irritation to the eyes, and no irritation to the oral cavity or skin before penetration. Moreover, you can also check the usage amount.
At, and the usage itself is not a harmful bacterium, so there is no problem 5 in terms of harm, and no toxicity is observed at all to fish and shellfish. Alternatively, you can, for example, replace the water in a large capacity tank or pot and pour it into tap water that has a faint chlorine odor or neutralized water with hypo, etc. If the purifying agent of the present invention is used, a remarkable effect can be obtained in that the water purification effect is maintained for a long period of time, from sIO days to 10 days, and furthermore, freshwater fish can be reared at high density. Very high economic efficiency can be obtained.

Of course, if necessary, it is also possible to use clear water from which surfaces have been removed using a bacterial filter or the like.

Hereinafter, the present invention will be explained based on examples.

First, the following microorganisms were cultured according to their respective methods.

(1) Pg, denitrificans (2)
B. subtilis (3) B. subtilis (4) Mu.
Agilis (5) K. Zofiii (6) Musip, Tereus (7) Blot, Succinogenes (8)
Baot, Rivoliticum (g) Br.
A medium containing 3% N, Coralina soybean meal, m5IN, 0.2% ammonium sulfate, and 0.3% charcoal was cocooned after adjusting the pH to 7. (1) to (4) and (2)
inoculate with a culture of Each culture was carried out in a 7-armenter made of stainless steel, the temperature was maintained at 30°C, and the culture was continued for 24 hours with aeration and stirring. (s
) t (7) to (9), culture is carried out in a stainless steel 7-armenter in the same manner as above, but the culture is carried out using 6% soybean ζ-le, 10% starch, and charcoal O1s
Each culture was cultured at 30° C. for 24 hours in a medium having a composition of 30% and 30° C., with slow stirring and no aeration. (6) i! ! The culture was carried out in the same manner as in cases (1) to (4), except that the pH of the medium was adjusted to 7.0 and 0. After the cultivation was completed, the culture was centrifuged to obtain a bacterial paste.

This paste was freeze-dried in an acetone-dry ice bath.

The cultures of the 10 types of bacteria thus produced were mixed with equal amounts of j1 (lk weight) of Cannabis earth to obtain mixtures of bacteria and carriers.

Next, the mixtures of these bacteria and carriers were combined to prepare water purifiers having the formulations shown in Examples 1 to g below.In each Example, the number indicates the formulation powder 1? Represents the number of noodles (unit: 100 million).

In addition, I of the above 10 spears! lt-Example 9 combining all
The powder formulation is light pink to brown in color and is a free-flowing powder with a slightly sour odor (this formulation
-600J).

Example 1 Pg, Denitrificans 2.5 Example 2 B, S. subtilis 2.5 Baot, S. subtilis
livolyticum 8.5 Example 4 Ps, denitrificans 2. OK, Zoffii 0.4B, Succinogenes 0.4 N, Coralina 0.4 Br@v, Ribolyticum 04 Based on Example 5 Mu, Agilis 2. OB, T. subtilis α4 B, T. thuringiensis Q, 4 MU, Tereus α4 plexus, :F larina &4 4 p.s. Sout, Lipolyticum O1s Example I Ps+,, denitrificans 1. OB, subtilis 1・OB, thuringiensis +L5 9. Tereus 0.5 Bwt, Succinogenes 15 Br, Libolyticum 0.5 ahead Coralina 0.1 Example 8 Mu, Agilis 0.5, Sophii 04 ps, Denitrificans 'L0 8aot, Succinogenes 1s Byrt, Libolyticum 1s Br Nail, Livoliticum OS N, Coralina O,S Example 9 Mu, Agilis O,S B, Subtilis 1 B, Thuringiensis O,S! , Zoffii 0.1 P (Denitrificans 1 Mu Suppression, Terex O, 5Baot, Succinogenes O, SBB10 Livolicum
0.5 Br・Ma, Lipoliticum O, SN, Coralina 0.1 The effects of each of the formulations of Examples 1 to 9 of the present invention were compared with the untreated formulation in which the formulation was not administered, and the following Test Example 1
~3! :Rated.

Test Example 1 Nishikigoi breeding test 75anX 55anX 3 Two Bam glass tanks each with sterilized river sand at the bottom! Lay it in a thick layer of + ■, and add 1007 - 1007 of tap water that has been pumped day and night, and the average weight is about 75.
20 g-1 were put in each. The same type of internal filter was used in both areas, and aeration was performed using an aerator.

Nosan brand No. 1 food was used for feeding, and care was taken to ensure that there was as little leftover food as possible. The microbial preparation of the present invention, use-see, was added to one of the water tanks by a free-spirited student to a 50-level powder, and the mixture was gently stirred to ensure that it was well distributed throughout the tank, and this was used as a treatment area. Water temperature is 1s℃~24℃
The breeding test period was 60 days. No water changes,
The following results were obtained by replenishing only the naturally evaporated water and not cleaning or replacing the filter (Table 1).

Table 1 1) The filter in the control group was on the verge of clogging, and the water was running at 11 liters.
(I wonder if it smells bad) In the treated area, there was no turbidity, the taste was 1af14, and the odor was very weak.

2) After the test was completed, the ammonia concentration and nitrite ion in both tanks were examined and the following results were obtained (Table 2).

Table 2 3) When the contents of the filter were taken from the treated and control groups into test tubes and left at room temperature, the control group showed further decomposition and fermentation, but the treated group showed no change. In addition to the fact that the amount of contents in the filter in the treated area was overwhelmingly smaller than that in the control area, it was inferred that the contents of the filter in the treated area were converted into -degradable substances due to the application of the symptom-inducing preparation of the present invention.

Test Example 2 Twelve glass aquariums of eel bulk rearing test F5 (711x!!
Numbers 11 to Arashi 11 are assigned to the tanks corresponding to the two comparative examples, and tanks N1 to Pigeon 9! The area was treated with the present invention's symptomatic agent, and Na1O and Pigeon 11 were used as the control area (untreated area) (Table 3).All the tanks were filled with several strips of sand at the depth of 5 countries - day and night. I added 1,001 cups of tap water each. Free-raised eels must be selected to have as small a degree of variation in size as possible, otherwise the feed will be consumed evenly and effectively, resulting in large losses, making it difficult to judge the effectiveness of microbial preparations. Choose a size fish with a weight of 41LOf~47. Of
Twenty fish within this range were placed in each tank. After the release, in the aquariums of the treatment areas from -1 to 9, microbial preparations (1) to (Q) prepared according to the attached formulation example were added to the aquarium -1 using the formulation example (1). As shown in the table below, 5f of each prepared microbial preparation powder was added and stirred (soP level). A filter containing activated carbon of the same type was installed in both the control and treatment areas, and aeration was performed using an air stone, with care being taken to ensure that the dissolved oxygen level did not fall below 7.5p. In addition, in order to reproduce the high temperature conditions when fish production is more likely to occur, a 1 kW heater with an automatic temperature regulator was used to adjust the water temperature to 21°C (±O, S°C). The test period was 180 days, and the amount of water was not replaced, but the amount of natural evaporation was replenished at appropriate times, and efforts were made to maintain a constant amount of water. Only in the treated area, a microbial ψ preparation was additionally applied at a weight ratio of 50 P to the amount of water added during water replenishment. The feed used was Showa Sangyo Eel Marnir B, and initially the same amount was fed to all groups, but after about three weeks, the wax feed in the control group decreased, while the treated group had a strong appetite. The number of feeding groups has increased, but care must be taken to ensure that there is as little leftover food as possible after feeding in all tanks.j1. Ta. The results are shown in Table 4.

In the case of the treatment area shown in Table S, the water is very clear and there is little odor, and when the eel sinks into the sand at the bottom after ingesting food, it drops to -1, but the difference is small. The mixture settled in about 4 to 10 minutes, and the suspension disappeared.

1. In the control plot, about 3 weeks after release, the water became conspicuous, a foul odor was present, the animals began to move slowly, and deaths began to occur. After the eel burrowed into the bottom sand, it became cloudy and 1IifI4 did not subside for more than 30 minutes, and even after that, floating substances remained floating for a long time. The foul odor and suspension continued until the end of the test, and a total of 13 fish died in the tank at 11h10, -1
A total of 16 fish died in one tank.

& Filters in the control plot Both tanks were completely clogged around 1 ton month, and the filters were replaced.After that, the filters were replaced frequently every 20 to 30 days, but there were some differences between the filters in the treatment plot depending on the tank. Although there was a case, the air flow became weaker from around the go day, but it did not completely fail, and I only replaced the cartridge once for each tank from around the -113th day to the 129th day.

4. Observing the contents of the 6I filter, it was found that in the treatment area,
The contents of the control plot were slimy and had a strong odor of putrefaction, and the cleaning of the filter was somewhat difficult. However, the stains were not removed and it took a long time to clean.

The breeding density at the time of pick-up in the treatment area was 2.0712 Kf/1004-31,07 HIKf
/1Q (1t If this is converted to an eel pond with R2m of water, it will be 41.4Kf/M'~411.4Kg/M", and in the case of forced oxidation method, the density will be about 12Kf/M1,
Compared to this, the treatment area L46-B achieves 11 times higher density.

1. The water in the treated area was clearer and the odor was much weaker than in the control area, but the odor in the control area was strong and the water was extremely stale.

λ When we examined the rainbow trout that died in the control area, we found that all of them had adhesion of the secondary valves and dilatation of the capillary capillaries, indicating that they had died due to ammonia poisoning.The three surviving trout were also found not to have been fed. The amount was 172 or less compared to the treated area, and it was presumed that the activity decreased due to blood loss anemia caused by ansonia, and the body weight did not increase as much as in the treated area.

The clogging of the LIll filter was also significantly faster in the control group.
The filter was completely clogged on September 11, 127, and the filter was replaced, but the treated area remained free of blockage until the end of the experiment.

The contents of the filter were placed in test tubes for both the control and treated areas and left for two weeks.The contents of the control area produced small bubbles coming out of the bottom, fermentation, and decomposition progressed, but the contents of the treated area did not. It did not corrode, but instead turned into a substance that is difficult to decompose. Naturally, the content was a little higher in the treated area.

Procedural amendment (spontaneous) October 20, 1975 Kazuo Wakasan, Commissioner of the Patent Office 1, Indication of the case 1978 Patent Application No. 17207
No. 1 & Relationship with the case of the person making the amendment Patent applicant name Tame Co., Ltd. - Varnish Sea Interflatle
Heiichi Ichiban Company 4, Agent 5, Date of amendment order Showa 1920, Month, Day 6
, Number of inventions increased by the amendment 7, Detailed explanation of the r invention in the specification subject to the amendment" Column O - 8, Contents of the amendment Contents of the amendment as attached (Japanese Patent Application No. 172079/1989) (1) Description Page 1, line 12 [and the same Politicam, ``'' is corrected to ``wa''.

(2) Line 13 of the same statement: “It is an anaerobic bacterium.”
Insert the following statement after .

“Bacteroides lipolyticum is a halophilic aerobic bacterium.” (3) Page 8, line 8 “(T) ~ (ω”)
), (-”)〇(4) Insert the following sentence after “Proliferate 0” in the first line of page 9.

``(8) Sono (i.e., Bacteroides lipolyticum) is treated with Nutripros (Difco, USA) K salt for 31s.
OJ (5) same page 14 [line 15 "(a) to 9)” to “of, (9) J” 0 (6) Same page 1S, line 3 “・・・・・・8.OK
Adjusted. Insert the following sentence after j. O “The bacteria in (8) are grown in nutrient broth (Difco, USA) K salt 3
-, inoculated into a medium adjusted to pH 6.5 to 7.0 before sterilization, and incubated at 35°C for aerated shaking or aerated agitation culture.
0”

Claims (1)

[Scope of Claims] 1) A freshwater purification agent characterized by artificially culturing non-pathogenic saprophytes and making them dormant. 2) Fresh water purification characterized in that the saprophyte according to claim 1 contains one or more types of saprophyte left behind from inorganic or organic nitrogen compound decomposing bacteria, hexaleurose decomposition (2), and fat decomposition. agent.