Bacteria Tablets

Abstract

To provide a bacterial tablet that has reduced effect on the bubbling of an aeration device.SOLUTION: A bacterial tablet is in the shape of a tablet comprising the mixture of a bacterial carrier having Bacillus bacteria supported thereon and an excipient primarily composed of sodium sulfate.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a tablet carrying bacteria that purifies water in an aquarium. [Background technology]

[0002] In aquariums where aquarium organisms are kept, bacteria are used to decompose organic matter such as excrement and leftover food, which can cause deterioration of water quality. Methods for supplying bacteria to an aquarium include pouring powdered or liquid bacteria into the water, but this method has various problems, such as the labor required to measure the powder or liquid, the labor required to clean up if the bacteria falls outside the aquarium, and the discomfort caused by the bacteria getting on your hands.

[0003] As a method for solving such problems, a method has been proposed in which bacteria are supported on tablets and the tablets are directly added to water (Non-Patent Document 1). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Product name: "Natto bacteria tablets", manufactured by Ito Sui Co., Ltd., searched on August 7, 2020, Internet<URL:http: / / www.itosui.co.jp / series / 505 / > Summary of the Invention [Problem to be solved by the invention]

[0005] However, when an aeration device that dissolves air into the water in an aquarium containing such tablets is operated, there is a problem in that bubbles form on the water surface.

[0006] Therefore, an object of the present invention is to provide a bacterial tablet that has little effect on foaming caused by an aeration device. [Means for solving the problem]

[0007] The bacterial tablet is formed into a tablet shape by mixing a bacterial support carrying Bacillus bacteria with an excipient mainly composed of sodium sulfate.

[0008] By using sodium sulfate as the main excipient, it is possible to create a bacterial tablet that is less affected by foaming caused by an aeration device. Foaming is a phenomenon that occurs when air is adsorbed to the air-liquid interface of a surface active substance or surfactant. This is because sodium sulfate is an inorganic substance and has less effect on foaming than components that combine organic substances such as polyethylene glycol with hydroxyl groups.

[0009] In the bacterial tablet, the Bacillus bacteria form spores.

[0010] The formation of highly durable spores can improve the shelf life of the bacterial tablets.

[0011] In the bacterial tablet, the bacillus bacterium is Bacillus licheniformis.

[0012] By using Bacillus licheniformis, which has the best ability to decompose organic matter among the Bacillus bacteria, the organic matter decomposition effect of the bacterial tablet can be improved because Bacillus licheniformis has many types of enzymes such as alkaline phosphatase, esterase, esterase lipase, β-galactosidase, and glucosidase.

[0013] The bacterial tablet may also contain magnesium sulfate, and the weight ratio of the magnesium sulfate in the entire bacterial tablet may be 10% or less.

[0014] By adding magnesium sulfate and reducing the ratio of sodium sulfate, the amount of sodium can be reduced, and the increase in the salinity of the water can be suppressed. In addition, by keeping the weight ratio of magnesium sulfate at 10% or less, the increase in water hardness can be suppressed. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 shows the results of an aeration test on the bacterial tablet of the comparative example. [Diagram 2] FIG. 2 shows the results of an aeration test on the bacterial tablet of the embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] <Bacteria Tablet> The bacterial tablet according to the present embodiment is formed into a tablet shape by mixing a bacteria support carrying Bacillus bacteria with an excipient mainly composed of sodium sulfate. This allows the bacterial tablet to be directly put into water, reducing the labor of measuring powder or liquid, the labor of cleaning when it falls outside the water tank, and the discomfort caused when it adheres to the hands. Here, the tablet shape refers to, for example, a cube, a rectangular parallelepiped, a sphere, a cylinder, a prism, a cone, a pyramid, an ellipsoid, a cylinder, or a tablet shape that is a combination of these. In particular, from the viewpoint of moldability and usability of the bacterial tablet, a tablet shape that is approximately triangular is preferable.

[0017] (bacteria carrier) The bacterial support according to this embodiment uses a support substrate and a spore liquid or spore powder of Bacillus bacteria. As the support substrate, an inorganic mineral salt is preferable from the viewpoints of moldability, storage stability, and water solubility. As the inorganic mineral salt, sodium sulfate or magnesium sulfate, which is easily soluble in water and has little effect on the liquid property (pH), is preferable. In particular, sodium sulfate, which has no effect on the total hardness of water, is preferable as the support substrate. For example, a liquid or powder containing Bacillus bacteria (in a non-spore-formed state) may be used as the bacterial support.

[0018] The Bacillus bacteria supported on the bacterial support are preferably Bacillus firmus, Bacillus spumilus, Bacillus licheniformis, Bacillus amyloliquefaciens, or Bacillus licheniformis, which form highly durable spores. By sporulation, the preservation properties of the bacterial tablet (e.g., suppression of bacterial activity during storage) can be improved. In particular, Bacillus licheniformis, which is excellent at decomposing organic matter, is preferred. This is because Bacillus licheniformis has many types of enzymes, such as alkaline phosphatase, esterase, esterase lipase, β-galactosidase, and glucosidase.

[0019] (Excipients) The excipient according to this embodiment is sodium sulfate or a combination of sodium sulfate and magnesium sulfate, which is non-foaming, easily soluble in water, and has little effect on liquidity. In terms of reducing the effect on the total hardness of water, the weight ratio of magnesium sulfate in the entire bacterial tablet is preferably 10% or less. This makes it possible to suppress an increase in the total hardness of water. In addition, by including magnesium sulfate, the amount of sodium can be reduced, and an increase in the salinity of water can be suppressed.

[0020] With the above-mentioned configuration, it is possible to provide a bacterial tablet that has little effect on foaming caused by the aeration device.

[0021] <How to make bacterial tablets> An example of a method for producing a spore liquid, a spore powder, a bacterial carrier, and a bacterial tablet is described below. Note that the methods for producing the spore liquid, the spore powder, the bacterial carrier, and the bacterial tablet are not limited, and bacterial tablets produced by methods and conditions other than those described below are also included in the present invention.

[0022] (Preparation of spore suspension) (1) A Bacillus strain is cultured on an agar medium at 30°C for 18 to 48 hours. (2) The strain cultured in (1) above is transferred to a liquid medium and gradually scaled up at 30°C for 18 to 24 hours. (3) The culture medium scaled up in (2) above is cultured in a jar fermenter at 30°C for 120 to 168 hours to form spores. (4) After confirming that spores have formed in the culture medium in (3) above, the spores are centrifuged and collected from the culture medium using a refrigerated centrifuge. (5) The spores collected in (4) above are washed several times with saline and sterile water. (6) The spores washed in (5) above were suspended in 1 L of sterile water until the number of Bacillus bacteria reached 1.0 × 10 9 By confirming that the concentration is greater than or equal to cfu / mL, a highly concentrated spore solution can be obtained.

[0023] (Preparation of bacterial carriers) The spore liquid or the spore powder obtained by drying the spore liquid obtained above is mixed with a support substrate in an arbitrary ratio, whereby the Bacillus bacteria are supported on the support substrate. Then, when the number of bacteria of the Bacillus bacteria reaches 1.0×10 8 A bacteria carrier can be obtained by confirming that the Bacillus bacteria count is equal to or higher than cfu / g. Note that a bacteria carrier may be prepared by mixing a liquid or powder containing Bacillus bacteria (in a non-sporulated state) with a carrier substrate in any ratio.

[0024] When the bacteria carrier is large (e.g., 1 mm or larger), it is preferable to crush the bacteria carrier with a mixer or crusher from the viewpoint of improving the tableting property described later. In order to incorporate a high concentration of Bacillus bacteria into the bacteria tablet, the spore liquid or spore powder and the carrier substrate are preferably mixed at a weight ratio of 3:7 to 2:8.

[0025] (Mixing of Bacterial Carrier with Excipients) The bacterial support and excipient obtained above are placed in a thick vinyl bag or powder mixer in a weight ratio of 0.1:9.9 to 2:8, and mixed. In order to blend a high concentration of Bacillus bacteria into the bacterial tablet and form a tablet with sufficient hardness, when using a bacterial support with magnesium sulfate as the support base, it is preferable to mix the bacterial support and excipient in a weight ratio of 1 (or less) to 9 (or more). In addition, when using a bacterial support with sodium sulfate as the support base, it is preferable to mix the bacterial support and excipient in a weight ratio of 2:8. In addition, when the bacterial support and excipient are mixed, if the powders of each are solidified, it is preferable to remove the solidified powder with a 1 mm mesh from the viewpoint of improving the tableting property described later.

[0026] (Tablet compression) The mixture of the bacteria carrier and excipient obtained above was compressed into tablets using a tablet press, and the number of Bacillus bacteria was reduced to 1.0 × 10 6 cfu / g or more. This allows bacteria tablets (e.g., 0.5 g per tablet) to be obtained. For example, one tablet is suitable for a 10 L tank of water, and it is preferable to store such bacteria tablets in an aluminum bag to prevent moisture.

[0027] It is preferable to measure the bacterial tablets in groups of 10. If the bacterial tablets are weighed one by one, it is difficult to judge whether the measured value is within the margin of error. In addition, in order to provide a high concentration of Bacillus bacteria in the aquarium by adding one tablet, the number of Bacillus bacteria per 10 tablets is set to 5.0 x 10 6 cfu or more. EXAMPLES

[0028] <Excipient selection test> Bacterial tablets containing Bacillus licheniformis with magnesium sulfate, sodium sulfate or wood chips as the bacterial carrier were prepared by the above process using the excipients shown in Table 1. The weight ratios shown in Table 1 indicate the weight ratios of the entire bacterial tablet. [Table 1]

[0029] Three bacteria tablets of the above selection example were put into a beaker filled with 100 mL of tap water to conduct a selection test for the excipient. In the selection test, the tablet compressibility, water solubility and turbidity of the water, pH, and total hardness of the tablet were evaluated. The tablet compressibility was evaluated by whether the tablet could be compressed without problems using a tablet press, and the water solubility was evaluated by whether the tablet dissolved in water after being put in. In addition, from the viewpoint of suppressing the influence on ornamental fish, the presence or absence of turbidity of the water, whether the pH of the water did not fluctuate by 0.5 or more, and whether the total hardness was 100 mg / L or less were evaluated. Furthermore, the presence or absence of foaming when aeration was applied was evaluated. The evaluation results of the selection test are shown in Table 2 below. For reference, the evaluation results of a comparative example (manufactured by Ito Sui, product name "Natto Bacillus Tablet") are also shown.

[0030] [Table 2]

[0031] From Table 2, it was confirmed that sodium sulfate (Selected Examples 19, 20, 22) or a combination of sodium sulfate and magnesium sulfate (weight ratio 10% or less) (Selected Examples 18, 21) that are good in all respects of tablet compression property, water solubility of the tablet, total water hardness, pH, foaming, and the presence or absence of turbidity are suitable for the bacterial tablet of the present invention. In addition, it was confirmed that when the weight ratio of magnesium sulfate in the entire bacterial tablet exceeds 10% (Selected Examples 6, 7), the total water hardness exceeds 100 mg / L, and is therefore not suitable for the bacterial tablet of the present invention.

[0032] <Aeration test> As an example, Bacillus licheniformis (bacterial count: 1.0 × 10) containing sodium sulfate as the excipient and magnesium sulfate as the bacterial carrier was used. 6 The bacteria tablets containing 100% sucrose (100% cfu / g) were prepared by the above process. Then, an aeration test was carried out for the above comparative examples and examples.

[0033] The bacterial tablet of the comparative example or the example was placed in a beaker filled with 100 mL of tap water, and the bacterial tablet was completely dissolved. Then, an air stone (small round stone made by GEX) attached to an aeration device (e-AIR1000 made by GEX) was placed in the beaker, and the aeration device was operated to check for bubbling of the water. In addition, the bubbling of the water in a blank state where no bacterial tablet was placed was also checked. The experimental results of the comparative example are shown in FIG. 1, and the experimental results of the example are shown in FIG. 2.

[0034] Figure 1(a) shows the state in which the comparative bacterial tablet was added and the aeration device was operated, and Figure 1(b) shows the state in which the aeration device was operated as a blank. As shown in Figures 1(a) and 1(b), the beaker into which the comparative bacterial tablet was added showed foaming (countless small bubbles) caused by the aeration device, unlike the blank beaker. The comparative bacterial tablet dissolved in water in about 30 minutes.

[0035] FIG. 2(a) shows the state in which the bacterial tablet of the embodiment is put in and the aeration device is operated, and FIG. 2(b) shows the state in which the aeration device is operated as a blank. As shown in FIG. 2(a) and FIG. 2(b), the beaker in which the bacterial tablet of the embodiment is put in, like the blank beaker, no foaming caused by the aeration device was observed. This confirmed that the bacterial tablet of the embodiment has little effect on foaming caused by the aeration device. The bacterial tablet of the embodiment dissolved in water in about 60 minutes.

[0036] <Salt concentration test> A salinity test was carried out using the bacterial tablets of the above example.

[0037] 1 kg of soil (Gex Pure Soil Black) and fermented carbon dioxide (Gex Fermented CO2 Starter Set) were placed in an aquarium filled with 3.5 L of tap water, and aquatic plants (Glossostigma, Rotala, Pinnatifida, Myriophyllum) were planted. Then, 20 tablets (approximately 10 g) of the bacterial tablets of the embodiment were added, and a blank was used in which no bacterial tablets were added. The salinity was measured and the condition of the aquatic plants was confirmed after 15 days. The experimental results (salt concentration) are shown in Table 3 below.

[0038] [Table 3]

[0039] As shown in Table 3 above, 15 days after the start of the experiment, the salinity of the water in the tank into which the bacteria tablets of the embodiment were added rose by 0.3%. This confirmed that the salinity of the water, which affects freshwater fish, was below 1.0%, and that the bacteria tablets of the embodiment did not affect freshwater fish. It was also confirmed that there was no change in the condition of the aquatic plants before and after the experiment.

[0040] <Water quality testing> Water quality tests were carried out using the bacteria tablets of the above examples.

[0041] Put 50 ml of sterile water and three carp pellets (Hartz Pond Pellets) into a sterilized Erlenmeyer flask. Then, the Erlenmeyer flask containing one bacterial tablet of the above example and a blank Erlenmeyer flask containing no bacterial tablet were placed on a shaker (speed: 100 min). -1 ) for one month. After that, the water in each Erlenmeyer flask was evaporated in a dryer, the weight of the residue was measured, and the weight loss rate was compared. The experimental results are shown in Table 4 below. [Table 4]

[0042] As shown in Table 4 above, when the bacteria tablets of the embodiment were added, the weight loss rate increased by an additional 7% compared to the blank state. This confirmed that adding the bacteria tablets of the embodiment accelerated the decomposition of carp feed and improved water quality.

Claims

1. A bacterial tablet formed into a tablet shape by mixing a bacterial support in which Bacillus bacteria are supported on an inorganic mineral salt support substrate and an excipient made of an inorganic mineral salt containing sodium sulfate.

2. The bacterial tablet according to claim 1 , wherein the Bacillus bacterium forms spores.

3. The bacterial tablet according to claim 1 or 2, wherein the Bacillus bacterium is Bacillus licheniformis.

4. 4. The bacterial tablet according to claim 1, which contains magnesium sulfate, and the weight ratio of said magnesium sulfate in said bacterial tablet as a whole is 10% or less.

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