JPH074231B2 - Lactic acid bacteria viable agent for silage preparation - Google Patents

Description

TECHNICAL FIELD The present invention relates to a lactic acid bacterium viable agent for silage preparation.

More specifically, the present invention relates to a viable lactic acid bacterium preparation for silage having a high survival rate of lactic acid bacterium.

In general, the silage-preparing lactic acid bacteria viable agent has a low survival rate of lactic acid bacteria, and the effect was not clear, but when the silage-preparing lactic acid bacteria viable agent of the present invention is used, many surviving lactic acid bacteria proliferate well. Since it is possible to prepare good quality silage, it will greatly contribute to the dairy industry.

(Problems to be Solved by Prior Art and Invention) In general, in order to prepare good quality silage, it is important to sufficiently carry out lactic acid fermentation in the initial stage to rapidly lower the pH. However, in reality, depending on the type of plant to be silage, its water content, temperature, etc., sufficient fermentation does not always occur. For this reason, it is recommended that a stable lactic acid fermentation be performed by artificially adding a high concentration of a lactic acid bacterium viable agent suitable for silage fermentation, without relying only on lactic acid bacteria that adhere to plants.
For this reason, various lactic acid bacteria viable agents have been studied. However, even when there is a sufficient amount of live lactic acid bacteria during the production of the viable lactic acid bacteria agent for silage preparation, the lactic acid bacteria will die out during the distribution process to be removed, and when it is actually used for silage preparation, a sufficient amount will occur. The fact is that there is no living lactic acid bacterium. Even if a sufficient amount of live lactic acid bacteria is present, addition is not carried out uniformly unless the shape is such that it can be easily added. As a result, the fact is that the addition of a viable lactic acid bacterium agent at the time of silage preparation does not always lead to an effect.

(Means for Solving Problems) As a result of intensive studies by the present inventors, it may be related to the base that also enhances the survival of lactic acid bacteria. We have found that the survival of lactic acid bacteria can be significantly enhanced by using a product containing 50% or more of Corn grits with a particle size of 10 to 80 mesh.

The present invention relates to a lactic acid bacterium viable agent for silage preparation, which comprises a base having a water content of 4% or less and a particle size of 10 to 80 mesh corn grits of 50% or more.

In general, corn grits means corn rough meal, but those commercially available as corn grits are widely used in the present invention.

In the present invention, since commercially available corn grits have a high water content, it is necessary to dry the water content to 4% or less using a dryer or the like.

The water content of corn grits has a great influence on the survival properties of lactic acid bacteria, and the corn bacteria with a water content of more than 4% has an extremely low survival rate of lactic acid bacteria and cannot be put to practical use.

The particle size of corn grits is an important factor for dispersibility and survival of lactic acid bacteria. If the particle size of the corn grits exceeds 10 mesh, the particle size is too large and the lactic acid bacterium viable powder and the corn grits are easily separated, and even if the lactic acid bacterium viable agent is evenly dispersed, the lactic acid bacteria contained therein are not always uniform. Not added to. Further, generally, the work of adding the lactic acid bacteria viable agent is carried out in the field, so if it is less than 80 mesh, it becomes powdery and easily scattered by the wind, and it becomes difficult to uniformly mix it with the silage-prepared plant.

In the present invention, a base material having a water content of 4% or less and 50% or more of corn grits having a particle size of 10 to 80 mesh is used, but all corn grits may be used, and up to 50% calcium carbonate, zeolite, corn cob, If it is a sufficiently dried substance such as dextrin, it can be appropriately mixed.

In the present invention, lactic acid bacteria are separately prepared. The lactic acid bacterium may be any bacterium used for dairy farming or a bacterium that can be used. Next, a method for culturing lactic acid bacteria used in the present invention will be described.

As the medium, a medium that contains a carbon source, a nitrogen source, an inorganic substance, vitamins, amino acids, and the like and is commonly used for culturing microorganisms can be widely used. The carbon source may be any assimilable carbon compound, for example glucose, sucrose,
Maltose is used. The nitrogen source may be any available nitrogen compound, such as peptone, meat extract,
Casein acid hydrolyzate and the like are used. In addition, salts of phosphates, magnesium, sodium, potassium, calcium, iron, manganese, etc., vitamins, amino acids, antifoaming agents, and surfactants are used as necessary.

As the medium, both liquid medium and solid medium can be used. The initial pH of the medium is pH 5 to 8, preferably pH 6 to 8, the culture temperature is 20 to 45 ° C, preferably 25 to 40 ° C, and the culture time is 12
The culture can be suitably carried out for a period of time to 3 days, preferably 15 to 24 hours.

The culture thus obtained can be used as it is or as washed bacterial cells. In addition, the culture or the microbial cell can be used as it is or after adding an additive, drying, or after being formulated, and then used.

The lactic acid bacterium powder has a water content of 4% or less and is uniformly mixed with a base material containing 50% or more of corn grits having a particle size of 10 to 80 mesh.

Any method may be used as long as they are uniformly mixed.
Mix with a V-type mixer.

Next, production examples and examples of the present invention will be shown.

Production Example 1 200 g of casein peptone, 30 g of yeast extract, 20 g of potassium phosphate 2 g, and 50 g of glucose were dissolved in 10 L of demineralized water, and a medium whose pH was adjusted to 7.5 with 1N sodium hydroxide solution was placed in a jar fermenter. After sterilizing for 15 minutes, the mixture was cooled, inoculated with a culture solution of Lactobacillus casei ATCC 4940 precultured beforehand, and statically cultured at 35 ° C. for 20 hours under aerobic conditions.

The culture broth thus obtained was centrifuged to collect the bacterial cells, which was lyophilized using a 10% skim milk 1% sodium glutamate solution sterilized in advance as a dispersion medium, and 450 g of live bacterial powder of Lactobacillus casei ATCC 4940. Got The viable cell count of lactic acid bacteria contained in this viable cell powder was 1 × 10 ¹⁰ cells / g.

Production Example 2 Streptococcus lactis C-6513.FERM P-9810 was cultured and dried in the same manner as in Production Example 1. However, the culture conditions were 30 ° C. and 18 hours. The amount of Streptococcus lactis C-6513 viable powder obtained was 410 g.
The viable cell count of lactic acid bacteria contained in this viable cell powder was 1 × 10 ¹⁰ cells / g.

Production Example 3 Pediococcus pentosaceus ATCC25744 was cultured and dried in the same manner as in Production Example 1. However,
The culture conditions were 30 ° C. and 23 hours. The amount of live bacterium powder of Pediococcus pentosaceus ATCC25744 obtained was 320 g. The number of viable lactic acid bacteria contained in this viable bacterial powder is 1 × 10 ^10.
It was / g.

Production Example 4 When 20 kg of corn grits (particle size 32 to 60 mesh) was thinly spread on a tray and dried at 100 ° C. for 4 hours with a ventilation type shelf dryer, the water content was 3.0% (w / w).

Production Example 5 When 20 kg of corn grits (particle size: 35 to 80 mesh) was thinly spread on a tray and dried at 100 ° C. for 5 hours with a ventilation type shelf dryer, the water content was 2.7% (w / w).

Production Example 6 When 20 kg of corn grits (particle size: 10 to 32 mesh) was spread thinly on a tray and dried at 100 ° C. for 3 hours with a ventilation type shelf dryer, the water content was 4.0% (w / w).

Production Example 7 (Control Example) 9 parts of commercially available Corn grits was added to 1 part of the dried cells obtained in Production Example 1 and mixed well to obtain a viable cell agent. Immediately after preparation, the viable count of lactic acid bacteria contained in this probiotic agent is 8 × 10 ^8.
It was a piece / g.

Production Example 7-2 (Control Example) 9 parts of bran was added to 1 part of the dried bacterial cells obtained in Production Example 1 and mixed well to obtain a viable agent. Immediately after preparation, the viable cell count of the lactic acid bacteria contained in this probiotic agent was 8 × 10 ⁸ cells / g.

Production Example 8 To 1 part of the dried bacterial cells obtained in Production Example 1 was added 9 parts of Corn grits obtained in Production Example 4 and mixed well to obtain a viable cell agent. Immediately after preparation, the viable cell count of the lactic acid bacteria contained in this probiotic agent was 8 × 10 ⁸ cells / g.

Production Example 9 (Control Example) 5 parts of commercially available Corn grits, 2 parts of calcium carbonate and 2 parts of zeolite were added to 1 part of the dried bacterial cells obtained in Production Example 2 and mixed well to obtain a viable cell agent. . Immediately after preparation, the viable cell count of the lactic acid bacteria contained in this probiotic agent was 8 × 10 ⁸ cells / g.

Production Example 10 5 parts of Corn grits obtained in Production Example 5, 2 parts of calcium carbonate and 2 parts of zeolite were added to 1 part of the dried bacterial cell obtained in Production Example 2 and mixed well to obtain a viable cell agent. It was Immediately after preparation, the number of viable lactic acid bacteria contained in this probiotic agent is 8 × 10 ⁸ /
It was g.

Production Example 11 (Control Example) To 1 part of the dried bacterial cells obtained in Production Example 3, 7 parts of commercially available Corn grits and 2 parts of corn cob were added and mixed well to obtain a viable cell agent. Immediately after preparation, the viable cell count of the lactic acid bacteria contained in this probiotic agent was 8 × 10 ⁸ cells / g.

Production Example 12 7 parts of Corn grits obtained in Production Example 6 and 2 parts of dried corn cob were added to 1 part of the dried bacterial cell obtained in Production Example 3 and mixed well to obtain a viable cell agent. Immediately after preparation, the viable cell count of the lactic acid bacteria contained in this probiotic agent was 8 × 10 ⁸ cells / g.

Example 1 In order to examine the preservability of lactic acid bacteria in the probiotic agent obtained in Production Example, a survival rate test was conducted by leaving it at 45 ° C. for 10 days.

As is clear from the above results, Production Examples 8, 10 and 12 using Cornglitz based on the present invention as a base material were overwhelmingly compared with the respective Control Production Examples 7, 9 and 11 using the commercially available products. Survival was improved.

Example 2 Using the probiotic agent obtained in Production Example 7-2 and Production Example 8, silage of 10 ton alfalfa scale was prepared. The amount of probiotic agent added was 5 kg each, and a can with a hole was hung where the grass was shredded and transported to the silo, and the probiotic agent was placed in the can and continuously dropped by an appropriate amount. did. After foaming for one month, samples were taken from the upper part, middle part and lower part of the silo to measure pH.

As is clear from the above results, in Production Example 8 in which the corn grits used in the present invention was used as a base, the pH was lower than that without addition, regardless of the location of the silo, and the effect of adding the probiotic agent was observed. However, in Production Example 7-2, the effect of addition varies depending on the location of the silo. This is because the viable bacterial agent obtained in Production Example 8 had good fluidity and was added almost uniformly, whereas the viable bacterial agent obtained in Production Example 7-2 had poor fluidity and was added uniformly. It seems that it was not done.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication (C12N 1/20 C12R 1:46) (C12N 1/20 C12R 1:01) (72) Inventor Murata Kazuya 2-4-1 Ebisu-Minami, Shibuya-ku, Tokyo Calpis Food Industry Co., Ltd. Research and Development Center (72) Inventor Hiroshi Miyazaki 2-4-1 Ebisu-Minami, Shibuya-ku, Tokyo Calpis Food Industry Co., Ltd. R & D Center

Claims (1)

[Claims] 1. A viable lactic acid bacterium agent for silage preparation, which comprises a base material having a water content of 4% or less and a particle size of 10 to 80 mesh corn grits of 50% or more.