JP7592245B2 - Capsule containing thatch-decomposing bacteria and method for preserving lawn land - Google Patents

Description

The present invention relates to capsules containing thatch-decomposing bacteria and a method for preserving lawn land.

On lawns such as golf course greens, athletic fields, and gardens, grass is mowed regularly to keep the grass at an even height. Clippings generated during mowing accumulate in the soil. The layer of accumulated clippings, dead leaves, stems, roots, etc. is called thatch, and when thatch forms, it becomes difficult for water, air fertilizer, and pesticides to penetrate the soil and the roots of the turfgrass. The inhibition of water penetration can lead to poor growth of turfgrass and increased death. Thatch can also act as a breeding ground for putrefactive bacteria.

There are two known methods for removing thatch: physical and biological. As a physical method, vertical work is carried out on lawns. In vertical work, thatch is scraped off using machinery. Aeration is also carried out by drilling holes in the ground to improve the air and water permeability of the soil. Topsoiling is also carried out by covering the lawn with gravel to promote the growth of turfgrass.

As a biological means, microorganisms are used. Patent Document 1 discloses microcapsules containing thatch-decomposing bacteria. The microcapsules are porous and are formed from an environmentally degradable polymer based on poly-ε-caprolactone or alginate-chitosan gel beads.

If thatch-decomposing bacteria alone are sprayed directly onto a lawn without being encapsulated in microcapsules, they are subject to the harsh natural conditions of sunlight, temperature changes, and rainfall, making it difficult for the thatch-decomposing bacteria to settle on the lawn and prone to becoming inactivated within a short period of time. In contrast, in Patent Document 1, by carrying thatch-decomposing bacteria in microcapsules, the thatch-decomposing bacteria can be efficiently established on the lawn, and the thatch-decomposing ability can be obtained more stably for a longer period than thatch-decomposing bacteria alone.

JP 2011-83222 A

Improvements to capsules containing thatch-decomposing bacteria are needed to efficiently preserve lawns. If the preservation performance of thatch-decomposing bacteria could be improved and their activity could be maintained for an even longer period of time, the frequency with which the thatch-decomposing bacteria needs to be sprayed could be reduced, allowing lawns to be efficiently preserved.

The present invention was made in consideration of the above circumstances, and aims to provide a capsule containing thatch-decomposing bacteria that can preserve thatch-decomposing bacteria for a long period of time and maintain their activity, and a method for preserving lawn land.

The capsule containing thatch-decomposing bacteria according to the first aspect of the present invention comprises:
Thatch-decomposing bacteria belonging to Bacillus pumilus;
A capsule having an alginate gel wall material containing the thatch-decomposing bacteria;
A protective agent for suppressing drying of the thatch-decomposing bacteria;
Including,
The protective agent is
Glucose, sucrose or trehalose .

A method for preserving lawn land according to a second aspect of the present invention comprises:
The capsules containing thatch-decomposing bacteria according to the first aspect of the present invention are spread on a lawn.

According to the present invention, thatch-decomposing bacteria can be preserved for long periods of time and their activity can be maintained.

1 shows the morphology of capsules encapsulating thatch-decomposing bacteria, (A) shows the morphology of capsules encapsulating Bacillus subtilis, and (B) shows the morphology of capsules encapsulating Bacillus pumilus. 1 shows the survival rate of bacteria in capsules encapsulating thatch-decomposing bacteria, (A) showing the survival rate of Bacillus subtilis, and (B) showing the survival rate of Bacillus pumilus. 1 shows the morphology of capsules after drying, (A) shows the morphology of capsules encapsulating Bacillus subtilis, and (B) shows the morphology of capsules encapsulating Bacillus pumilus. 1 shows the survival rate of bacteria in a capsule after drying, (A) shows the survival rate of Bacillus subtilis, and (B) shows the survival rate of Bacillus pumilus. Figure 1 shows the cellulolytic activity of capsules after drying, (A), (B) and (C) show the viscosity over time of aqueous sodium carboxymethylcellulose solutions containing capsules on days 0, 7 and 21 of storage, respectively.

The following describes an embodiment of the present invention with reference to the drawings. Note that the present invention is not limited to the following embodiment and drawings.

(Embodiment)
The capsule containing thatch decomposition bacteria according to the present embodiment includes thatch decomposition bacteria, a capsule containing the thatch decomposition bacteria, and a protective agent that suppresses the drying of the thatch decomposition bacteria. The thatch decomposition bacteria can be any bacteria that has the ability to decompose thatch. Examples of the thatch decomposition bacteria include bacteria belonging to Bacillus subtilis and bacteria belonging to Bacillus pumilus. Particularly preferably, the thatch decomposition bacteria is bacteria belonging to Bacillus pumilus.

The thatch-decomposing bacteria are encapsulated in capsules whose wall material is an environmentally degradable polymer. Environmentally decomposable polymers include biodegradable polymers that are decomposed into water and carbon dioxide by microorganisms in soil, and disintegrative polymers that decompose over time in the natural environment due to sunlight and water. Biodegradable polymers are preferred as environmentally decomposable polymers because they serve as food for the thatch-decomposing bacteria.

Specific examples of biodegradable polymers include aliphatic polyesters such as polycaprolactone, polylactic acid, polyglycolic acid, polybutylene succinate, and polyethylene succinate, as well as polyvinyl alcohol, polyphosphoric acid, polyamino acids, chitosan, chitin, cellulose, starch, cellulose acetate, bacterial cellulose, curdlan, pullulan, and biopolyesters, as well as composites of these. Preferably, the environmentally degradable polymer is alginate gel.

The capsule containing thatch-decomposing bacteria according to this embodiment can be dried and used according to the mode of use. In this case, the cell moisture necessary for the survival of the thatch-decomposing bacteria is lost due to drying. In contrast, the capsule containing thatch-decomposing bacteria according to this embodiment contains a protective agent that inhibits the drying of the thatch-decomposing bacteria. The protective agent is not particularly limited as long as it is a substance that inhibits the drying of the thatch-decomposing bacteria, but a substance that does not pollute the environment is preferable. As the protective agent, a substance containing a hydroxyl group is particularly suitable. Specific examples of the protective agent include sugars, polyols, and amino acids. Sugars retain bound water, so that moisture can be sustained on the cell membrane surface. Sugars have a high protective effect on proteins, have almost no impact on the environment, and are extremely low in toxicity to living organisms. The sugars may be monosaccharides, disaccharides, or polysaccharides. Preferably, the sugars are, for example, glucose, trehalose, and sucrose.

The size of the capsules containing thatch-decomposing bacteria is, for example, 0.1 to 10 mm, 0.5 to 8 mm, 1 to 5 mm, or 2 to 4 mm in average particle diameter. The average particle diameter of the capsules containing thatch-decomposing bacteria is preferably 3 to 4 mm.

The form of the capsules containing thatch-decomposing bacteria is not limited, but may be, for example, gel beads. Gel beads have a leather-like surface and a rough sponge-like interior, with thatch-decomposing bacteria contained in the voids of the sponge. As gel bead capsules containing thatch-decomposing bacteria, alginate gel beads or alginate-chitosan gel beads have excellent support and habitat suitability for thatch-decomposing bacteria.

The following is an example of a method for producing capsules containing thatch-decomposing bacteria in the form of gel beads. In this production method, an aqueous solution of an environmentally degradable polymer containing thatch-decomposing bacteria is dropwise mixed as a dispersed phase into a continuous phase containing a gelling agent. This forms capsules containing the thatch-decomposing bacteria and having the environmentally decomposable polymer as a wall material. The formed capsules are then hardened. To harden the capsules, for example, they can be stirred in the above-mentioned newly prepared continuous phase.

The aqueous solution of environmentally degradable polymer containing thatch-degrading bacteria may be prepared by dissolving alginate in a medium containing cultured thatch-degrading bacteria. The concentration of the environmentally degradable polymer in the aqueous solution of environmentally degradable polymer is arbitrary, but when alginate is used as the environmentally degradable polymer, the concentration is, for example, 0.1 to 10%, 0.5 to 5%, or 1 to 3% by weight/volume percentage (w/v). The concentration of the gelling agent in the continuous phase is also arbitrary, but for example, 0.1 to 10%, 0.5 to 5%, or 1 to 3% by weight/volume percentage (w/v). When the environmentally degradable polymer is alginate, the gelling agent preferably contains a polyvalent metal salt. Alginic acid forms a salt with a polyvalent ion. Multiple carboxyl groups in an alginic acid molecule are crosslinked by a polyvalent cation, and the polyvalent metal salt of alginic acid becomes a salt that is insoluble in water. Calcium chloride is preferred as the gelling agent.

Next, the hardened capsules are collected by filtration, and the capsules are immersed in a solution containing a protective agent. The capsules may be dried before being immersed in the solution containing the protective agent. The concentration of the protective agent solution is adjusted as appropriate, but is, for example, 0.1 to 1.0 mol/L. The immersion time is, for example, several hours, preferably 1 to 5 hours or 2 to 4 hours.

The protective agent may be contained in an aqueous solution of environmentally degradable polymer containing thatch-decomposing bacteria (dispersed phase) that is mixed into the continuous phase. This allows the protective agent to be contained in the capsules containing thatch-decomposing bacteria without immersing the hardened capsules in a solution containing the protective agent. Even if the protective agent is contained in the dispersed phase, the hardened capsules may be further immersed in a solution containing the protective agent.

The capsules containing thatch-decomposing bacteria can be recovered by filtration. The recovered capsules containing thatch-decomposing bacteria may be freeze-dried. The capsules containing thatch-decomposing bacteria may also be washed before freeze-drying.

The capsule containing thatch-decomposing bacteria may be in the form of a core shell. In the case of a core shell, the capsule containing thatch-decomposing bacteria has one or more hollow internal cavities, and contains thatch-decomposing bacteria in the internal cavities. In the core shell, the thatch-decomposing bacteria proliferate in the internal cavities without being affected by environmental changes, and the bacteria that have decreased due to release outside the capsule containing thatch-decomposing bacteria are replenished. For this reason, the core shell has the advantage that the thatch-decomposing bacteria can stably exert their thatch-decomposing ability for a longer period of time.

To manufacture a core-shell capsule containing thatch-decomposing bacteria, first, an environmentally degradable aqueous polymer solution containing thatch-decomposing bacteria is mixed into an organic phase O in which an environmentally degradable wall polymer is dissolved in a low-boiling organic solvent, to prepare an S/O emulsion in which droplets of the aqueous polymer solution containing thatch-decomposing bacteria are dispersed as an inner aqueous phase S in the organic phase O. Then, the S/O emulsion is mixed into an aqueous phase and stirred to prepare an S/O/W emulsion in which droplets of the S/O emulsion are dispersed in an outer aqueous phase W. Next, submerged drying is performed by at least one of heating and reducing pressure to remove the organic solvent in the organic phase O of the S/O/W emulsion and gel the wall polymer, thereby producing a capsule in which the inner cavity of the capsule made of the environmentally decomposable polymer is filled with an inner aqueous phase S containing thatch-decomposing bacteria.

Then, like the gel beads, the capsules are collected by filtration and immersed in a solution containing a protective agent. The capsules may be dried before being immersed in the solution containing the protective agent. The protective agent may be added to the aqueous solution of environmentally degradable polymer containing thatch-decomposing bacteria that is mixed into the organic phase O.

As the low-boiling organic solvent for the organic phase O, a non-polar solvent with a boiling point of 85°C or less is preferred for drying in liquid by at least one of heating and reducing pressure, and examples of such solvents include dichloromethane, chloroform, and ethyl acetate. Two or more of these may be used in combination, but it is preferable to use dichloromethane alone or in combination with other solvents as the main component.

Preferably, the wall polymer of the organic phase O is mainly composed of poly-ε-caprolactone. By using poly-ε-caprolactone as the wall polymer, capsules with excellent recovery rate and physical strength can be produced. In addition, poly-ε-caprolactone is particularly compatible with thatch-decomposing bacteria, and the activity of the thatch-decomposing bacteria is maintained at a high level by using poly-ε-caprolactone as food, thereby exhibiting superior thatch-decomposing ability. Preferably, the average molecular weight of poly-ε-caprolactone is 10,000 to 500,000.

It is also advisable to add an emulsion stabilizer to the organic phase O in order to prepare an S/O emulsion by adding the internal aqueous phase S. Examples of emulsion stabilizers include span-based surfactants such as sorbitan monooleate, as well as various surfactants, water-soluble resins, and water-soluble polysaccharides that are generally used in emulsion preparation.

The environmentally degradable polymer used in the internal aqueous phase S may be any water-soluble polymer that does not interfere with the activity and proliferation of thatch-decomposing bacteria, such as alginate, water-soluble polymeric polysaccharides such as κ-carrageenan and chitosan, and polyvinyl alcohol, with alginate being preferred due to its compatibility with thatch-decomposing bacteria. When using sodium alginate as the alginate, the aqueous solution concentration should be about 0.5 to 10% by mass; if it is too high, the dispersion stability of the S/O emulsion decreases and aggregation is likely to occur. In addition, polypeptone, yeast extract, magnesium sulfate, etc. may be appropriately blended into the aqueous polymer solution as a nutrient source for the thatch-decomposing bacteria.

When preparing an S/O emulsion by adding and mixing the inner aqueous phase S into the organic phase O, it is preferable to stir under ice cooling in order to protect the thatch-decomposing bacteria. In addition, it is preferable that the aqueous phase used for the outer aqueous phase W of the S/O/W emulsion contains a dispersion stabilizer. Any dispersion stabilizer generally used in emulsion preparation can be used, but gelatin is particularly suitable because excellent emulsion effects can be obtained by using a high molecular weight emulsion stabilizer. When gelatin is used, the concentration in the outer aqueous phase is preferably about 0.1 to 10 mass%. After drying in liquid, a proteolytic enzyme such as papain may be added to decompose the high molecular weight gelatin into low molecular weight gelatin and improve the filtration characteristics of the prepared capsule and the outer aqueous phase.

In the above-mentioned submerged drying, heating and/or decompression are performed under stirring in order to volatilize and remove the low-boiling organic solvent from the organic phase, but from the viewpoint of processing efficiency, it is preferable to perform heating and decompression simultaneously. In heating, the liquid temperature of the emulsion can be raised stepwise or continuously over several hours, but the maximum temperature reached can be lower than the boiling point of the low-boiling organic solvent. In decompression, the pressure of the atmosphere in contact with the liquid surface of the emulsion can be similarly reduced stepwise or continuously over several hours, but the maximum decompression can be about one-fifth of atmospheric pressure.

The capsules containing thatch-decomposing bacteria according to this embodiment are preferably used in a lawn conservation method. In this lawn conservation method, the capsules containing thatch-decomposing bacteria are spread on the lawn. In spreading, a liquid containing the capsules containing thatch-decomposing bacteria may be spread, or a powder obtained by drying the capsules containing thatch-decomposing bacteria may be spread. The capsules containing thatch-decomposing bacteria may be mixed with a filler and spread. Known fillers are used, and examples of the filler include sand, xerolite, and ammonium sulfate.

As described above in detail, the capsules containing thatch-decomposing bacteria according to this embodiment contain a protective agent, so that the thatch-decomposing bacteria can survive even under dry conditions, and can be stably preserved and maintain their activity for a long period of time, as shown in the following examples. Therefore, the capsules containing thatch-decomposing bacteria can reduce the frequency of spraying the capsules containing thatch-decomposing bacteria, and the lawn can be efficiently preserved. In lawns where the capsules containing thatch-decomposing bacteria are sprayed, the thatch does not accumulate thickly, making it difficult for air and water to penetrate into the soil and the roots of the lawn grass, and the sprayed fertilizers and pesticides do not become absorbed by the thatch and become less effective. In addition, the proliferation of pathogens or pests and the generation of toxic gases due to decay are also prevented, so the lawn grass can be maintained in a healthy state for a long period of time.

The environmentally degradable polymer used as the wall material of capsules containing thatch-decomposing bacteria not only serves as food for the thatch-decomposing bacteria, helping to prevent a decline in their activity, but also decomposes and disappears over time in the natural environment, so there is no environmental burden on the turf where it is sprayed or the surrounding area.

In another embodiment, a turf conservation material is provided that contains the capsules containing the thatch-decomposing bacteria. The turf conservation material may further contain a filler.

The present invention will be explained in more detail with reference to the following examples, but the present invention is not limited to these examples.

(Encapsulation of thatch-decomposing bacteria)
702 medium (140 mL of ultrapure water, 1.4 g of Polypeptone, 0.28 g of Bacto Yeast Extract, and 0.14 g of magnesium sulfate heptahydrate) was placed in a culture flask, plugged with a silicone stopper, and sterilized in an autoclave (ATS-40, manufactured by ALP) at 121°C for 15 minutes. Frozen and stored Bacillus bacteria were thawed in a clean bench, and the cells were administered into a culture flask containing 702 medium. The flask to which the cells were administered was transferred to a bioshaker (TB-12T, manufactured by Takasaki Scientific Instruments Co., Ltd.), and the cells were cultured at 30°C and 150 rpm. The culture times of Bacillus pumilus NBRC12092 (hereinafter simply referred to as "Bp") and Bacillus subtilis NBRC13714 (hereinafter simply referred to as "Bs") were 36 hours and 18 hours, respectively. Bp and Bs were cultured to obtain approximately 10 ⁸ cells/mL of bacterial cells. Bp and Bs were obtained from the National Institute of Technology and Evaluation.

After the culture, sodium alginate was added to the 702 medium to make a 2% (w/v) solution and dissolved to prepare a sodium alginate aqueous solution. This was dropped into the gelling bath of 1.1% (w/v) calcium chloride aqueous solution with a syringe to prepare capsules. The amount of sodium alginate dropped was set so that the weight ratio to the calcium chloride aqueous solution was 1:10. After the capsules were prepared, they were collected by suction filtration and added again to a newly prepared 1.1% (w/v) calcium chloride aqueous solution. The capsules were allowed to harden while being gently stirred with a magnetic stirrer for 30 minutes. After hardening, the capsules were collected by filtering again. The collected capsules were placed in a centrifuge tube and freeze-dried for 12 hours using a freeze dryer (FDU-1200, manufactured by EYELA).

The capsules were hardened, filtered, and collected, then placed on a plastic petri dish and photographed under a stereomicroscope (C-DSD115, Nikon) (magnification: 16.4x).

The number of viable bacteria in the capsule was measured by the dilution plate method. First, the capsule was added to a sodium citrate aqueous solution adjusted to pH 7 and dissolved to prepare a bacterial suspension. The agar medium (1 L of ultrapure water, 10 g of Polypeptone, 2 g of Bacto Yeast Extract, 1 g of magnesium sulfate heptahydrate, and 15 g of agar powder) was transferred to a clean bench, and 10 mL was poured into a petri dish and spread thinly to solidify. 0.9 mL of 0.9 wt% saline was measured and poured into multiple tubes using a pipette. 0.1 mL of the bacterial suspension was measured and poured into a tube containing saline, and suspended. 0.1 mL of the suspension was measured and transferred to the next test tube. This operation was repeated to perform serial dilutions. 0.1 mL of all diluted samples were measured using a pipette, dropped onto the agar medium, and thinly coated. The petri dish was inverted and cultured statically in an incubator (SLI-450N, EYELA) at 30°C for 2 days.

After the culture, the petri dishes were removed from the incubator, and the number of colonies formed on the surface of the agar medium was counted. A colony is a bacterial colony formed by the proliferation of a single bacterial body, and the number of live bacteria on the agar medium can be measured by counting the colonies. If there are too many colonies, the colonies overlap and accurate counting becomes impossible, and conversely, if there are too few colonies, the error increases when the colonies are not formed correctly. The number of colonies can be controlled by the dilution ratio during serial dilution, and samples with 30 to 300 colonies were used to evaluate the number of live bacteria. The number of live bacteria contained in 1 g of capsule was calculated using the following formula.
(Number of colonies x amount of sodium citrate aqueous solution used [cells]) / amount of capsules dissolved in sodium citrate aqueous solution [g])

(result)
Figure 1 (A) and Figure 1 (B) show the morphology of capsules encapsulating Bs and Bp, respectively, before drying. Spherical capsules of 3 to 4 mm were obtained. The number of viable bacteria immobilized in the capsules was approximately 10 ⁷ cells/g-wet.

(Evaluation of survival of thatch-decomposing bacteria in capsules with and without the addition of a protective agent)
Sodium alginate was dissolved in the 702 medium in which the thatch-decomposing bacteria had been cultured to prepare a sodium alginate aqueous solution. This was dropped into a 1.1% (w/v) calcium chloride aqueous solution, which was a gelling bath, using a syringe to prepare capsules. After preparing the capsules and hardening them for 30 minutes, the collected capsules were added to a sample bottle containing a sugar aqueous solution (glucose aqueous solution, sucrose aqueous solution, or trehalose aqueous solution) containing sugar at a specified concentration, and immersed at room temperature for 3 hours. The amount of capsules added at this time was approximately 4 g per 40 mL of sugar aqueous solution. The capsules containing sugar were collected by filtration and then freeze-dried for 12 hours.

To calculate the survival rate of bacteria in the capsules, the viable cell count was measured by the dilution plate method as described above. The survival rate was calculated by calculating the viable cell count (N _i [cells/g]) in the capsules before freeze-drying and the viable cell count (N [cells/g]) after freeze-drying.

(result)
Figures 2(A) and 2(B) show the survival rate of the bacteria in the capsules encapsulating Bs and Bp, respectively. In the case of Bs and Bp, glucose (Glc), sucrose (Suc) and trehalose (Tre) showed a higher survival rate than the case without sugar. Glc, Suc and Tre acted as protective agents and improved the preservation performance of the thatch-decomposing bacteria in the capsules.

(Changes over time in the survival rate of thatch-decomposing bacteria in capsules)
The capsules were immersed in a 0.2 mol/L sucrose solution for 3 hours, collected by filtration, and freeze-dried for 12 hours. After 12 hours of freeze-drying, the day the capsules were prepared was set as the storage day 0, and the capsules stored in a refrigerator at 4 ° C for 0, 7, and 21 days were evaluated. The capsules were added to a sodium citrate solution adjusted to pH 7 and dissolved to prepare a bacterial suspension. The agar medium was transferred to a clean bench, and 10 mL was poured into a petri dish and thinly spread and solidified. The bacterial suspension was serially diluted in the same manner as above. All diluted samples were measured out in 0.1 mL with a pipette, dropped onto the agar medium, and thinly applied. The petri dish was turned upside down and subjected to static culture at 30 ° C for 2 days in an incubator. After culture, the petri dish was removed from the incubator, the number of colonies formed on the surface of the agar medium was counted, and the number of live bacteria contained in 1 g of capsule was calculated. Since the number of encapsulated bacteria decreases over the course of storage, the transition in the number of live bacteria from the start of storage was evaluated as the survival rate. For comparison, a capsule that was not immersed in a sucrose solution (without a protective agent) was also evaluated in the same way.

(result)
3(A) and 3(B) show the morphology of capsules encapsulating Bs and Bp, respectively. The capsules encapsulating Bs or Bp showed no change in appearance even after storage for several days.

Figures 4(A) and 4(B) show the time course of the survival rate of the bacteria in capsules encapsulating Bs and Bp, respectively. By including a protective agent, the capsules encapsulating Bs or Bp had a higher survival rate of the bacteria compared to capsules without a protective agent, and the survival rate of the bacteria was maintained at least until the 21st day.

(Cellulose decomposition test using capsules containing thatch-decomposing bacteria)
The capsules containing Bp immersed in a 0.2 mol/L sucrose solution for 3 hours were collected by filtration and then freeze-dried for 12 hours. The day the capsules were prepared was set as the storage day 0, and the capsules stored in a refrigerator at 4°C for 0, 7, and 21 days were evaluated. 0.1 g of capsules were added to an Erlenmeyer flask containing a HEPES solution in which sodium carboxymethylcellulose (CMC-Na) had been dissolved, and the mixture was stirred (150 rpm) for 3 days under conditions of 30°C. During the 3 days of stirring, the HEPES solution to which the capsules had been added was sampled every other day and the viscosity was measured using a rotational viscometer (LVDV-E, manufactured by BROOKFIELD). The main component of the thatch layer is cellulose, and CMC-Na, a derivative of cellulose, increases the viscosity of the aqueous solution. Therefore, the decomposition ability of cellulose was evaluated by measuring the amount of decrease in viscosity. For comparison, capsules not immersed in a sucrose aqueous solution (without a protective agent) were also evaluated in the same manner.

(result)
Figures 5(A), 5(B) and 5(C) show the cellulolytic activity of capsules encapsulating Bp on days 0, 7 and 21 of storage, respectively. A decrease in viscosity was observed on days 0, 7 and 21. Capsules containing a protective agent maintained cellulolytic activity even after 21 days of storage.

The present invention is suitable for the conservation or management of turfgrass, and in particular for a method for conserving lawn land and a lawn conservation material.

Claims (2)

Thatch-decomposing bacteria belonging to Bacillus pumilus;
A capsule having an alginate gel wall material containing the thatch-decomposing bacteria;
A protective agent for suppressing drying of the thatch-decomposing bacteria;
Including ,
The protective agent is
glucose, sucrose or trehalose;
Capsule containing thatch-decomposing bacteria. The capsule containing thatch-decomposing bacteria according to claim 1 is spread on a lawn.
How to conserve lawns.