JP2528055B2 - Intestinal Salmonella bacteria reducing agent for addition to avian feed and its use - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agent for reducing Salmonella intestinal bacterium and a method for using the same. More specifically, it is added to the feed of birds such as chickens for the purpose of reducing harmful bacteria in the intestine. The present invention relates to an agent for reducing intestinal Salmonella bacteria and a method of using the same.

[0002]

2. Description of the Related Art In feeds for livestock such as chickens, weight gain effect,
Antibiotics have been added to feed for the purpose of improving breeding efficiency and preventing bacterial diarrhea.
In order to avoid residues in eggs, the law on the safety of feed (the law on ensuring the safety of feed and improving its quality) has restricted the use of antibiotics and the like.

Therefore, instead of antibiotics and the like, substances that achieve the above-mentioned objects are required, and probiotic agents are drawing attention as such substances. Such probiotic agents include lactobacilli [genus Lactobacillus ], lactic acid cocci [eg Streptococcus faecal
is)], bifidobacteria (Bifidobacterium) or butyric acid bacteria (e.g. cross-tri Jiu scan butyricum (Clostridiu
[ M butyricum )] as an active ingredient is commercially available [Kimura Makoto; Livestock Research Vol. 33, No. 1, 31-36]
(1979)]. In addition, as a viable agent for Bacillus, B. natto BN strain [B. natto is taxonomically B. subtil
It is classified as is; Bajisu Manual of Systematic Bacteriology Biology Vol. II (19
86)] spore preparation [trade name "Growen", manufactured by Eisai Co., Ltd.] [Makoto Kimura; supra, Teruo Ohta; Miso Science and Technology No. 293 (July 1978) Print, 1-9
Other], and Bacillus toyoi [(B. toyoi
Is a taxonomically classified B. cereus ) spore preparation (trade name “Toyoselin”), manufactured by Asahi Kasei Kogyo Co., Ltd.].

On the other hand, Bacillus subtilis C-3102
It is known that administration of a probiotic agent containing microbial cells of (Ministry of Industrial Science and Technology No. 1096) to mammals, birds, fish and other animals produces effects such as weight gain and improvement of feed efficiency. (JP-A-63-209580), and a mixed feed for piglets containing Bacillus subtilis C-3102 as an active ingredient has been proposed by the present applicant (JP-A-62-232343).

[0005]

DISCLOSURE OF THE INVENTION However, when a formulation containing viable cells is ingested after being mixed in feed, the intestinal Salmonella bacteria of birds such as chickens are decreased with time. It is not known to have an effect.

The object of the present invention is to maintain the health of birds by decreasing the intestinal Salmonella bacteria in birds such as chicken over time.
Consequently, it is intended to provide an enteric Salmonella genus-suppressing agent for bird feed addition and a method of using the same to avoid food poisoning caused by this bacterium.

[0007]

According to the present invention, Bacillus subtilis C-3102 (Microtechnical Research Institute No. 1096)
No.) as an active ingredient, an agent for reducing enteric Salmonella bacteria for addition to avian feeds is provided.

According to the present invention, the intestinal Salmonella is
A use of bacteria-reducing agent, intestinal support should be reduced
Containing, as an active ingredient, live cells of Bacillus subtilis C-3102 ( Microtechnical Research Institute No. 1096), which has at least 1/20 the number of bacteria per gram of feed of the bacterium of the genus Lumonella per gram of feed. The intestinal monkey characterized in that the intestinal salmonella genus reducing agent is continuously contained in the feed for at least 7 days before the slaughter of the bird to be ingested by the bird.
Methods of using Monella bacteria reducing agents are provided.

Hereinafter, the present invention will be described in more detail.

In the present invention, the probiotic agent to be contained as an essential active ingredient is Bacillus subtilis C-3102 (known as Microbiology Research Institute No. 1).
No. 096), and its mycological properties have been already disclosed in JP-A-63-209580 and JP-A-62.
No. 232343.

The Bacillus subtilis C-3102
For culturing (Mikori Kenjiro No. 1096), for example, a liquid medium or a solid medium containing a carbon source, a nitrogen source, an inorganic substance or the like, which is usually used for culturing microorganisms, can be used as the medium . The carbon source may be any assimilable carbon source, for example glucose, sucrose, starch, molasses, etc., and the nitrogen source, for example, peptone, meat extract, hydrolyzed caseinate, ammonium sulfate and the like. You can Further, if necessary, phosphates, salts of magnesium, sodium, potassium, calcium, iron, manganese, etc., vitamins, amino acids, antifoaming agents, surfactants and the like can be added. The culture conditions are preferably aerobic conditions, and the initial pH of the medium is 5 to 9, particularly 6 to 8,
The culturing temperature is preferably 20 to 50 ° C., particularly 35 to 40 ° C., and the culturing time is preferably 12 hours to 7 days.

[0012] The culture thus obtained may be used as such or its concentrate, or bacterial cells separated therefrom,
The agent can be used as it is or by adding additives such as an excipient as the intestinal Salmonella genus- reducing agent for bird feed of the present invention. In addition, the above-mentioned culture, concentrate, or microbial cell can be used as it is or after adding additives such as excipients, or can be formulated into a post-proliferative agent.
Excipients are not particularly limited, calcium carbonate, defatted rice sugar,
Corn grits, corn flour, bran, skim milk powder and the like can be used. The probiotic agent of the present invention is C-3102.
It is preferable to contain 10 ⁶ to 10 ¹¹ cells / g of viable cells of the strain (Mikori Kenjo No. 1096), that is, spores and / or vegetative cells.

The bird to be ingested with the intestinal Salmonella genus- reducing agent of the present invention is not particularly limited, but for example, chicken,
It can be added to feeds for livestock such as eagle birds, ducks, turkeys and quails; pet birds; birds raised for appreciation at facilities such as zoos. The dosage form is not particularly limited, but it can be suitably used by adding it to a feed known as powder, particularly fine powder, and mixing it. Addition amount bird species of intestinal Salmonella bacteria reducing agent of the present invention when admixed to a known feed, varies depending day Ordinance, Bacillus subtilis C
The number of viable bacteria of -3102 (Ministry of Industrial Science and Technology No. 1096) is 1
It is preferable to add and mix so as to obtain 0 ³ to 10 ⁹ pieces / g, and particularly 10 ⁴ to 10 ⁶ pieces / g.

The effective ingestion amount of the intestinal salmonella bacterium reducing agent of the present invention depends on the number of harmful intestinal bacteria per gram of feces per gram of Bacillus subtilis C-3102.
No. 096), it is desirable to ingest a large amount of viable cells, and preferably at least 1/20 of the number of intestinal Salmonella bacteria to be reduced per gram of feces.
It is desirable to ingest the viable cells of Bacillus subtilis C-3102 (Microtechnical Laboratory Article No. 1096), and more preferably the same number or more. In addition, the administration period varies depending on the type of bird, the age, etc., but in the case of livestock, the administration should be continued at least 7 days before slaughter. Furthermore, different because the general antimicrobial agents intestinal salmonicida
It is most preferred to administer while increasing the number of B. genus bacteria to prevent an increase in the number of bacteria.

[0015]

INDUSTRIAL APPLICABILITY The reducing agent of the present invention comprises Bacillus subtilis C-3102 (Mikori Kenzo No. 1096) as an active ingredient.
No.) viable cells, the number of Salmonella enterobacteria in birds can be markedly reduced, which can maintain the health of the birds and cause them when the birds are eaten. It can greatly reduce the food poisoning that can occur. Also,
Bacillus subtilis C-3102
Since the viable cells of No. 96) are known bacteria known as feed additives, they are also excellent in safety.

[0016]

EXAMPLES Examples of the present invention will be described below.

Example 1 (Production of live cells) Soybean peptone 200 g and phosphoric acid 2 in 10 liters of tap water
A medium in which 10 g of potassium and 200 g of molasses were dissolved and the pH of which was adjusted to 7.5 with a 1N sodium hydroxide solution was placed in a jar fermenter, sterilized at 121 ° C for 15 minutes, and pre-cultured with Bacillus subtilis. C-3
Inoculated with the culture solution of No. 102 (Microtechnology Research Institute No. 1096),
The culture was performed at 37 ° C. for 40 hours with aeration and stirring.

The thus obtained culture broth is centrifuged to collect the bacterial cells, which are then dried and mixed with calcium carbonate to reduce the intestinal Salmonella genus bacterial reducing agent (hereinafter referred to as reducing agent 1) 20.
0 g was obtained. The viable cell count contained in the reducing agent 1 was 1 × 10 ¹⁰ cells / g.

Example 2 (Salmonella bacterium infection test) A commercially available compounded feed to which the reducing agent 1 obtained in Example 1 was added at a viable cell count of 3 × 10 ⁵ cells / g or not was added. Four-day-old male broilers (cobbs) that were raised from the time of chick insertion were used to produce Salmonella typhimirium ( Salmonel).
la typhimirium SU-27 (Gifu University) 1.6 × 10
Apply ⁷ cells / ml of sterile saline suspension to 1 ml / wing,
It was administered intrathecally using a sonde. As commercially available compounded feed, in the 1 to 7-day age, the 8 to 21-day age, and the 22-28 day age, the trade name "CIA HiBloace HS-1", the trade name "CIA HiBloace HS-2" and the trade name ""CIBROACEG2" (both manufactured by ITOCHU Feed Co., Ltd.) was used. Five birds in each group were killed daily from the time of infection, the contents of the cecum were collected, and the number of Salmonella was measured. The results are shown in FIG.

Example 3 (Production of live cells) Product name "Trypticase Soy broth (Trypticase S
oy Broth) "(manufactured by BBL) 20 g medium of 30 g / l
In a test tube and after sterilization, Bacillus subtilis C-3
1 of 102 (Microtechnical Lab. No. 1096) culture colonies
White ears were inoculated and cultured with shaking at 37 ° C. for 20 hours.

The culture broth thus obtained (hereinafter referred to as reducing agent 2) was used as trypticase soy broth (BBL).
Medium solution, River digest (manufactured by OXIOD) 2.5 g / l, yeast extract (manufactured by OXIOD) 5 g
/ L, medium composition of salt (Pharmacopoeia) 5 g / l (pH about 7.
In the solution of 4), the reducing agent 2 containing the viable cell counts shown in Tables 1 to 4 was prepared.

Example 4 (Test for reducing concentration of Salmonella spp.
Test) Salmonella bacterium isolated from 53-day-old broiler feces is trade name "Trypticase soy broth" (BBL)
After statically culturing at 37 ° C for 20 hours at room temperature, dilute it with the product name "Trypticase Soy Broth" (BBL), and then Table 1 to Table
A Salmonella solution containing 4 bacteria was prepared.

The reducing agent 2 of Example 2 was added to each Salmonella solution, and the mixture was subjected to static mixed culture at 37 ° C. for 20 hours and 44 hours.
The results are shown in Tables 1 to 4 .

Tables 1 and 2 show the results after culturing for 20 hours, and Tables 3 and 4 show the results after culturing for 44 hours. The symbols in the table are as follows.

−: Sterilizing concentration ±: Decreasing concentration (development inhibitory concentration) +: Developmental concentration

[0026]

[Table 1]

[0027]

[Table 2]

[0028]

[Table 3]

[0029]

[Table 4]

In particular, as is clear from Table 4 , it is understood that the growth inhibitory effect is exhibited even after the number of bacteria of the reducing agent 2 to the number of Salmonella bacteria is 3/50 after 44 hours.

[Brief description of drawings]

FIG. 1 is a graph showing the results of Example 2 showing the effect of the intestinal toxic bacterium-reducing agent of the present invention on Salmonella typhimurium SU-27 (Gifu University).

Claims (2)

(57) [Claims] 1. A Bacillus subtilis C-3102 (FERM BP No. 1096 No.) birds feed additive for intestinal Salmonella containing viable cells as an active ingredient (Salmonella) genus fine <br/> bacteria reducing agent . 2. The intestinal Salmonella genus bacterial reduction according to claim 1.
Intestinal Salmonella to be used for reducing drug usage
At least the feed group relative to the number of bacteria of the genus bacteria per gram of feces.
Bacillus subtilis C-3 with 1/20 the number of bacteria per lamb
Active ingredient of 102 (Microtechnical Laboratory Article No. 1096)
The intestinal Salmonella spp.
Feeding small doses continuously from at least 7 days before bird slaughter
Intestinal support characterized by being included in the product and ingested by birds
Use of Lumonella bacteria reducing agent .