JP2506314B2 - Prophylactic / therapeutic agent and feed for sepsis in poultry and pigs - Google Patents

Description

Detailed Description of the Invention

[0001]

The present invention relates to relates to a preventive or therapeutic agent and feed of sepsis of house fowl-pig.

[0002]

Follow <br/> come BACKGROUND OF as well as the invention is to provide a home avian-pigs, stress caused by multi-head feather breeding environment, environmental pollution, in particular, it is dry manure, whirled up in the air as dust, Breathing contaminated air is highly likely to cause the infection of pathogenic bacteria through the trachea, resulting in pneumonia and the systemic disease sepsis and death. Among other things, there is an extremely high mortality rate of the young life of the house fowl, pigs are immature immune function. As a result, the economic loss is large, such as lowering the productivity of the house fowl and pigs, is caused the management on critical issues. Various antibacterial substances have been used to deal with this, but due to the increase in drug-resistant bacteria, the problem of residues in the products of chickens and pigs, and the administration by injection, it is a major problem. in scale production, it has the disadvantage not practical not easily implemented from such a point of effort to be applied to home avian-pigs. The view of the conventional problems, the development of an agent for the prophylaxis or treatment of sepsis practical Suitable home avian-pigs can be administered at low cost production can Thus also easily replaces the antibiotic is desired.

[0003]

The present invention SUMMARY OF THE INVENTION, the above conventional problem persists, intended to provide an agent for the prophylaxis or treatment of sepsis house fowl, pork you meet the above requirements, Rakutobachi poultry derived
Ruth Avialius (Lactobacillus a
viarus) Z-1 strain, FERM P-13855
Cultivated sterilized cells or cell walls of poultry cultures for poultry sepsis
Prevention and cure of septicemia in poultry characterized by being an active ingredient
Medical agent and Bifidobacterium thermophyte derived from pig
Lamb (Bifidobacterium thermo)
cultivation of ZH strain, FERM P-13856
A sterilized product or cell wall of a nutrient is an active ingredient against sepsis in pigs.
A preventive / therapeutic agent for porcine sepsis characterized by
I do.

[0004]

Sterilization body or its cell wall of the working Strains of the respective Z-1 strain · ZH strain, especially home fowl, pork, when administered orally to Yoyowaiki house fowl, pork <br/>, respectively the result in infection protective effect against pathogens of sepsis, decreases significantly mortality sepsis, to be effective in the prevention and treatment of sepsis, the
It was also observed among the various strains belonging to the strains and Bifidobacterium belonging to other La Kutobachirasu genus is effective, especially. Sterilization, or the cell wall of the respective strains, some have this alone is added to the feed of the house fowl, pork,
By salary as feed, can be done easily and large-scale infection protective effect of sepsis of house fowl and pigs, is a practical, is enhanced breeding efficiency and productivity of the house fowl-pig.

[0005]

Next, embodiments of the present invention will be described in detail. In general ,
Lactobacillus avariaus belonging to the genus Lactobacillus,
L. casei, L .; fermentum, L .;
acidophilus, L .; strains belonging to the La Kutobachirasu genus such saviarius are Gram-positive, facultative or obligate anaerobic asporogenic bacilli, usually, catalase-negative, gelatin-decomposed, produce lactic acid by fermentation of glucose, aerobic Develops better under anaerobic conditions than under conditions
It has the following properties . The present inventor examined many strains isolated from fresh adult feces for the presence or absence of the protective effect against infection pathogens of sepsis. As a result, Z-1
Lactobacillus avialius (La)
C. bacillus avariaus) is a gram-positive, aspore-free obligate anaerobic coccobacillus, has the properties of the above-mentioned strains of the genus Lactobacillus, and has a particularly excellent immunopotentiating effect against pathogens of sepsis. However, it was found to have a protective effect against sepsis, and this was deposited as FERM P-13855.
Further, the biochemical properties of the Z-1 strain will be explained.
Non-development at 5 ° C, non-motile, catalase-negative, lactic acid production, and glucose decomposing ability, glucose, mannose, fructose, galactose, sucrose, maltose,
Cellobiose, lactose, melibiose, raffinose, esculin, salicin and amygdalin were positive, and arabinose and xylose were negative.

In general , Bifidobacterium (B
Bifid belonging to the genus ifidobacterium
Obacterium thermophilum,
B. infantis, B. breve, B.I. lo
ngum, B. strain belonging to the genus Bifidobacterium such as bifidum are Gram-positive, with obligate anaerobic asporogenic bacilli, club-shaped, curved, Y-shaped, but exhibit polymorphism, such as V-shaped, long It does not form a chain. Catalase negative, indole negative, no nitrate reduction
It has the following properties . The present inventor has examined many strains isolated from the intestinal contents of pigs for the presence or absence of an infection protective effect against the pathogens of sepsis. As a result, among others, Bifidobacterium thermophilum isolated as ZH strain
is a gram-positive, obligately anaerobic, aspore-free bacillus, is a thermostable bacterium that can grow at 46 ° C, and has the characteristics of the above-mentioned strains belonging to the genus Bifidobacterium. It was found that it has a particularly excellent immunopotentiating effect against the pathogen of Escherichia coli and has an infection protective effect against sepsis, and this was deposited as FERM P-13856. Furthermore, the biochemical properties of the ZH strain will be explained. Catalase-negative, indole-negative, nitrate non-reducing, non-degradation of arabinose, xylose, ribose, etc.
Decomposes mannose, maltose, raffinose, glycogen, etc.

[0007] For culturing each of the above strains, Briggs river broth
Any of various media conventionally used for culturing lactic acid bacteria and bifidobacteria such as h) can be used. The medium is prepared and cultivated according to a conventional method, for example, Moubunsha, 19
Anaerobically according to the method described in "World of Enterobacteria" published in 1980. After the culture is completed, the culture solution is cooled and then the cells are collected by centrifugation. Next, after centrifuging and washing the obtained microbial cells, sterilization treatment such as heating to sterilize, or the separated microbial cells before or after sterilization treatment such as heating, enzymatic decomposition, interface The cell wall is formed by stirring in the activator solution, or mechanical grinding such as grinding or grinding before or after drying. For drying the cells or cell wall, in addition to heating, a freeze drying method, a spray drying method, or the like is used.

According to the present invention, thus obtained
The Z-1 strain, sterilization, or cell wall, or a mixture of these cultures of the ZH strains, as sepsis preventive and therapeutic agent, alone or added to the feed, respectively orally administered at home avian-pigs, such as chickens To do. In particular, when administered orally to immature of infancy house avian-pig immune function, favorably grown without infecting pathogens of sepsis decreased mortality from sepsis, proportional to a conventional home <br/> It is possible to easily and economically improve the breeding productivity of poultry / pigs and improve the conventional disadvantages.

The present invention will be described with reference to more specific examples. Example 1 (Preparation of Prototype) After thoroughly passing carbon dioxide gas, 500 ml of sterilized Briggs River Broth was inoculated with Lactobacillus avialius Z-1 strain and Bifidobacterium thermophilum ZH strain, respectively. Each of the bacterial solutions pre-incubated at 37 ° C for 24 hours was added with 10 liters of Briggs
In addition to river broth, static culture was carried out anaerobically at 37 ° C. for 24 hours. Each culture was ice-cooled, collected by centrifugation, and washed with cold physiological saline. Thus, the white wet bacterium 2 of the Z-1 strain
8 grams and 24 grams of white wet cells of ZH strain were obtained. Z
After treating 16 g of the -1 strain at 100 ° C. for 30 minutes, it was freeze-dried to obtain 4 g of a heat-sterilized body (prototype 1). The remaining 12 g of wet cells were suspended in 200 ml of a 1% Tween 20 aqueous solution, and then stirred and treated at 80 ° C. for 1 hour. After cooling this with ice, it was washed by centrifugation, the obtained cell wall fraction was freeze-dried,
2.6 grams of cell wall (Prototype 2) was obtained. On the other hand, ZH
12 grams of the 24 grams of wet bacterial cells of the strain was
Then, the same treatment was performed to obtain 2.7 g of a heat-disinfected body (Prototype 3). In addition, 12 g of the remaining wet bacterial cells of the ZH strain was treated in the same manner as in Prototype 2 to obtain 2.2 g of cell wall (Prototype 4).

Example 2 (Septic infection protection test according to prototype 1) A prototype consisting of 10-group 5-week-old mice (ICR, male) consisting of the sterilized body of the Z-1 strain prepared in Example 1 Each mouse was orally administered once at 5, 50, 500 and 5000 micrograms (μg). Twenty-four hours after that, 8 × 10 ⁸ (CFU) strains of O-78 known as septic pathogenic Escherichia coli were intravenously inoculated per animal, and mortality due to sepsis was observed. The result is shown in FIG.
As is clear from the figure, the survival rate on the 14th day after inoculation was 10% in the non-administered group, whereas it was improved to 50-70% in the prototype 1-administered group. The protective effect against sepsis by administration was confirmed.

Example 3 (Septic Infection Prevention Test by Prototype 2) Prototype 2 comprising the cell wall of the Z-1 strain prepared in Example 1
Was tested under the same experimental conditions as in Example 2. The result is shown in FIG. As is clear from the figure, the survival rate on day 14 after intravenous inoculation of pathogenic Escherichia coli was 0% in the non-administration group, whereas it was 10% in the prototype 2 administration group.
Above, especially 500 micrograms (μg) per mouse
In the administration group, it increased to 60%, and it was confirmed that the oral administration of the prototype 2 had the effect of preventing sepsis infection.

Example 4 (Septic Infection Prevention Test by Prototype 4) Prototype 4 comprising the cell wall of the ZH strain prepared in Example 1 was tested under the same conditions as in Example 2. The result is shown in FIG. As is clear from the figure, the survival rate on day 7 after intravenous inoculation of pathogenic E. coli was 0 in the non-administered group.
%, The trial 4 administration group improved to 40 to 80%, confirming the effect of prototyping 4 to prevent sepsis infection.

[0013] The prototype 3 consisting of a bactericidal body of the ZH strain did not show the effect of preventing sepsis infection under the same conditions as in Example 2, but it was 5000 µg / mouse, 500 µg.
According to oral administration of 1 / mouse, survival rate on day 14 after infection is 20% and 50%, respectively, which is approximately the same as or slightly inferior to prototype 4, but it was confirmed to be effective in preventing sepsis infection. Was given.

Next, an example in which the agent for preventing and / or treating sepsis of the present invention is applied to domestic chickens will be shown. Example 5 A group of 6 2-week-old broilers (Chunky, female)
After the prototype 2 obtained in Example 1 was suspended in physiological saline, 50
Microgram (μg) / feather was orally administered once. Twenty-four hours later, the O-2 strain known as septic pathogenic Escherichia coli was intravenously inoculated with 1 × 10 ⁷ cells (CFU) per bird, and the survival rate on the 7th day after the inoculation was examined. The results are shown in Table 1 below.

[0015]

[Table 1] As is clear from Table 1 above, on the 7th day after inoculation, the survival rate was 0% in the non-administered group, whereas the survival rate was improved in the prototype 2 administration group by 50%, and the survival rate was confirmed for chickens. The preventive effect of sepsis infection was confirmed.

Example 6 Fifty piglets (LWD) were orally administered with 5 mg (mg) / day of the prototype 4 per animal for 5 days at the time of initiation,
Furthermore, at the age of 3 weeks, the mice were orally administered for 3 days, and the animals were raised normally. For comparison, 50 piglets (LWD) were raised normally without oral administration of the prototype, and some of them died of pneumonia due to sepsis. All of the administration groups grew satisfactorily, and infection preventive effects were observed.

Example 7 1 of the trial product 1 was added to the compounded feed for raising piglets (LWD).
The feed prepared by adding and mixing 0 ppm was administered to 50 piglets (LWD) for 3 weeks from the beginning, and thereafter, they were fed with normal feed. As a result, all grew well without suffering from sepsis.

Example 8 To a chicken feed, 1 ppm of the trial product 4 was added to the feed and mixed, and the mixture was administered to 100 chicks every day from the time of birth to 4 weeks of age. It was fed with feed. As a result, all grew well without suffering from sepsis.

When adding the prototypes 1 to 4 to the feed, the addition amount is preferably in the range of about 0.1 ppm to 100 ppm.

[0020]

As described above, according to the present invention ,
Lactobacillus avialius Z-1 strain, bifidobak
Sterilization thereof or a cell wall of the bacteria were cultured Agrobacterium-thermophilum ZH strain, home to fowl or swine, when mixed alone or feed for oral administration, sterilizing bodies and cell walls house
It has an excellent immune defense effect against the pathogenic bacteria of sepsis in poultry and pigs , that is, it has an infection protection effect, can be raised smoothly and satisfactorily, is practical, and reduces the mortality rate due to sepsis compared to the past. , economically bring about the effect of such results in improved productivity of the house fowl-pig.

[Brief description of drawings]

FIG. 1 is a graph showing the protective effect against sepsis in mice by a prototype consisting of a bactericidal strain of Lactobacillus avialius Z-1 strain of the example of the present invention.

FIG. 2 is a graph showing the protective effect against infection of sepsis by a prototype consisting of the cell wall of Lactobacillus avialius Z-1 strain.

FIG. 3 is a graph showing the protective effect against sepsis by a prototype consisting of the cell wall of Bifidobacterium thermophilum ZH strain.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication (C12N 1/20 (C12N 1/20 C12R 1: 225) C12R 1: 225) (C12N 1/20 (C12N 1/20 C12R 1:01) C12R 1:01) (72) Inventor Shizuo Sato 6-42-2 Ojidai, Sakura City, Chiba Prefecture Sanshiti Ichihara B-102 (72) Inventor Takashi Sasaki Sakura City, Chiba Prefecture 2-15-5 Miyamae (56) Reference JP 61-257930 (JP, A) JP 63-238095 (JP, A) JP 1-242532 (JP, A) JP 3-120222 (JP, A) International Publication No. 93-1823 (WO, A)

Claims (3)

(57) [Claims] 1. A poultry-derived Lactobacillus aviarius.
Z-1 strain, sterilized body of culture of FERM P-13855
Or use the cell wall as an active ingredient against poultry sepsis
A preventive and / or therapeutic agent for poultry sepsis characterized by: 2. A Bifidobacterium thermophilum ( porcine-derived Bifidobacterium thermophilum)
mofilum) ZH strain, FERM P-13856
Of sterilized cell culture or cell wall of pigs against sepsis in pigs
A prophylactic / therapeutic agent for porcine sepsis, which is characterized as an ingredient . 3. A feed for preventing / treating sepsis of poultry / porcine , which comprises the bactericidal body of the strain according to claims 1 or 2 or a cell wall thereof added to the feed.