JPH06192126A - Preventing and treating agent for salmonella infectious disease of animal - Google Patents

Abstract

PURPOSE:To obtain a preventing and treating agent having high preventing and treating effects on Salmonella infectious disease and free from problem of resistance to the medicine and environmental pollution, low in toxicity to animal and hardly remaining in animal bodies and having high safety. CONSTITUTION:This preventing and treating agent for Salmonella infectious disease of animal contains at least one inactivated bacterial strain of Salmonella belonging to O4 group and at least one inactivated bacterial strain of Salmonella belonging to O9 group on classification of Salmonella by O antigen. As Salmonella belonging to O4 group, Salmonella typhimurium, Salmonella paratyphi-B, etc., and as Salmonella belonging to O9 group, Salmonella enteritidis, Salmonella dublin, Salmonella typhi are exemplified and, as the combination thereof, S. typhimurium/S. dublin and S. typhimurium/S. enteritides are preferably used and these Salmonella bacteria have especially large effects on prevention and treatment of cow Salmonella infectious diseases. When this preventing and treating agent is orally administered continuously for >=5 days, further good result is obtained.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a prophylactic / therapeutic agent for Salmonella infectious diseases in animals and a method for preventing / treating Salmonella infectious diseases in animals.

[0002]

2. Description of the Related Art Animal diseases caused by Salmonella include various diseases such as typhoid fever, paratyphoid fever, Salmonella enteritis and other diarrhea, sepsis and the like. Since these diseases caused by Salmonella are infectious diseases, once they occur, they are transmitted one after another, and are likely to cause great damage particularly in the fields of livestock industry and poultry industry, which handle livestock and poultry.

In the prevention and treatment of such Salmonella infectious diseases, conventionally, disinfection of the breeding environment such as a barn, administration of antibiotics, administration of synthetic drugs, simultaneous administration of inactivated Salmonella and specific antibiotics, etc. have been carried out. ing. However, the disinfection method tends to cause disinfection leakage due to a decrease in disinfection power due to the mixture of organic substances. In addition, even when an animal is fortunately recovered from a Salmonella infection, the infected animal often becomes a carrier animal and continues to excrete, which makes it difficult to completely cut off the infection route. In addition, administration of drugs such as antibiotics and synthetic medicines limits the types of effective drugs that have effective susceptibility for the prevention and treatment of diseases.
Tolerance is likely to occur, and as a result of those drugs remaining in the body of animals, there is concern about the effect on the human body that ingests animal meat and the like.

In order to solve these problems, various immunological methods using a vaccine have been studied. As vaccines, there are live vaccines with live bacteria and killed vaccines, and it is said that the immunity given by live vaccines is mainly cell-mediated immunity, and immunity with killed vaccines is mainly involved in humoral immunity, It has been previously reported that cell-mediated immunity is important for protection against Salmonella infection (Us
hiba D. keio J. Medicine 1965; 14: 45-61).

[0005] As a live vaccine, a mutant strain that requires a certain nutrition to grow, which is produced by subjecting a highly virulent strain of Salmonella causing some damage to some treatment or subculturing for a long period of time. Pathogenic plasmid-deficient strains and the like have long been studied as attenuated strains with extremely low or reduced virulence. It has been reported that the live vaccine has a high protective effect and that protection against different bacterial groups, that is, cross immunity is established (Carlos.
E. et al .; Microbial Pathogenesis 1991; 10: 149-15
8) It is said that it is a necessary condition that the administered live bacteria grow in the body and stimulate the living body. If the live bacteria are not sufficiently attenuated, fever, diarrhea, decreased appetite, etc. It is often a problem because it causes side effects. In addition, it is feared that the pathogenicity may be restored by taking in the mutant strain and the virulent plasmid to make them virulent during long-term use of the attenuated bacteria. In addition, since live bacteria proliferate inside the body, they are also discharged outside the body, which causes a problem of environmental pollution.

On the other hand, the killed vaccine is often less effective than the live vaccine, and it is generally said that cross immunity is not established, but it has an advantage that problems such as environmental pollution do not occur. However, even if killed bacteria, when administered parenterally, a polysaccharide lipid called endotoxin, which is contained in large amounts in Salmonella cells, exerts toxicity and causes side effects such as shock death. There is a fear. Attempts to administer killed bacteria orally instead of parenterally, such as by injection, have been made with Shigella, cholera, Salmonella, etc. Although it almost disappeared, there was a problem that the immunogenicity was significantly reduced and the effect was reduced.

Further, it has been reported that three types of dead bacteria of Salmonella, O group, H group and Vi group, are orally administered simultaneously (DuPont HL et al. Bull. Wld. Hlth. Org. 197).
1; 44: 667-672), which is for measuring antibody titer in blood and promoting growth, and no protective effect against infection has been reported.

[0008]

Therefore, the object of the present invention is to have a high prophylactic / therapeutic effect against Salmonella infectious diseases, to be free from drug resistance and environmental pollution, and to have low toxicity to animals. Moreover, it does not easily remain in the body,
A highly safe prophylactic / therapeutic agent for Salmonella infection in animals,
Further, it is to provide a method for preventing and treating Salmonella infection.

[0009]

As a result of continuous research by the present inventors, among the Salmonella, bacteria belonging to two specific groups, that is, the O 4 group and the O 9 group are inactivated and their When the bacteria are combined and administered to animals, a synergistic effect of these two bacteria is produced, and the prevention and treatment of Salmonella infection can be carried out extremely effectively, and the above-mentioned object can be achieved. The present invention has been completed by finding that even better results can be obtained when continuously performed for 5 days or more.

That is, according to the present invention, at least one inactivating bacterium of O. salmonella belonging to O 4 group (O 4 group) and O in terms of classification by O antigen (O antigen) of Salmonella and O.
An agent for preventing / treating Salmonella infectious disease in animals, which comprises at least one inactivating bacterium of Salmonella belonging to Group 9 (Group 9).

Furthermore, the present invention combines at least one inactivating bacterium of Salmonella belonging to the O 4 group and at least one inactivating bacterium of Salmonella belonging to the O 9 group in terms of classification by Salmonella O antigen, A method for preventing or treating Salmonella infection in an animal, which comprises continuously orally administering to the animal for 5 days or more.

[0012] Here, the classification of Salmonella by O antigen means classification by serotype against O antigen which is Salmonella bacterium antigen, and normally Salmonella is classified into O 2 group to O 67 group by this classification method by O antigen. It is classified into 46 groups, and the details are described in "Intestinal Bacteria (II) Specifics 1. Sal.
genus monella "112-182 pages Modern Publishing Co., Ltd. (1985
11 ・ 15).

The prophylactic / therapeutic agent for Salmonella infection according to the present invention comprises, among the above-mentioned Salmonella, at least one inactivating bacterium of Salmonella belonging to the O 4 group and at least one of Salmonella belonging to the O 9 group. It is necessary to contain activated bacteria.

Examples of Salmonella belonging to the O 4 group include Salmonella typhimurium.
murium ), Salmonella paratyphibe ( Salmonella)
paratyphi-B ) etc. can be mentioned.

Further, examples of Salmonella belonging to the O 9 group include, for example, Salmonella enteritidis.
la enteritidis), Salmonella Dublin (Salmonella
dublin ), Salmonella Nesteved ( Salmonella)
naestved ), Salmonella typh
i ) and so on.

The prophylactic / therapeutic agent of the present invention contains inactivated one of the Salmonellas belonging to the O 4 group and inactivated one of Salmonella belonging to the O 9 group; Including two or more inactivated Salmonellas belonging to the group O 9 and one inactivated one of the Salmonellas belonging to the group O 9; one inactivated one of Salmonellas belonging to the group O 4 and an O Inactivated two or more Salmonellas belonging to Group 9; or O
It may include inactivated two or more Salmonellas belonging to the fourth group and inactivated two or more Salmonellas belonging to the O 9 group.

Various kinds of combinations of the O 4 group bacteria and the O 9 group bacteria can be selected, and preferred examples thereof include Salmonella typhimurium / Salmonella Dublin and Salmonella typhimurium. / Salmonella enteritidis, etc., and these combinations are particularly effective for the prevention and treatment of Salmonella infection in cattle, and prevent cattle from dying or aborting due to Salmonella infection. be able to.

Examples of the method for inactivating Salmonella include heat treatment, formalin treatment, γ-ray irradiation and the like, but inactivation by heat treatment or formalin treatment is simple and high drug efficacy is maintained. Is preferred and is preferable. 65 to 130 ° C when inactivating by heating
It is preferable to heat to the above temperature, and when it is inactivated by the formalin treatment, it is preferable to immerse Salmonella in an about 0.1 to 0.5% formalin aqueous solution for the treatment. At that time,
Even if the O 4 group Salmonella and the O 9 group Salmonella are inactivated separately and then used together as the preventive / therapeutic agent of the present invention, the O 4 group Salmonella and the O 9 group Salmonella are combined. Those which are combined and then inactivated may be used as the prophylactic / therapeutic agent of the present invention.

When the prophylactic / therapeutic agent of the present invention is orally administered, its toxicity is suppressed to a very low level even if it is continuously administered, and Salmonella infectious disease can be more effectively prevented and treated. The dosage form in that case is a liquid form;
It can be a semi-solid state such as a paste; a solid state such as powder, granules, tablets, coated tablets, capsules and pills. The preventive / therapeutic agent of the present invention is
If necessary, various components commonly used in this type of drug, such as an excipient, a filler, a binder, a disintegrant, etc., may be contained as long as they do not interfere with the efficacy of the drug.

The preventive / therapeutic agents of the present invention are cow, pig, horse,
Livestock such as sheep and goats; poultry such as chickens, ducks, ducks, quails and turkeys; extremely useful for the prevention and treatment of Salmonella infections in various animals such as dogs, cats, monkeys, pigeons, turtles, lizards and deer It is valid. Particularly when administered to cattle, it is possible to prevent and treat diarrhea and pneumonia due to the Salmonella infection, and serious diseases such as death and abortion due to sepsis associated therewith.

The method and dose for oral administration of the prophylactic / therapeutic agent of the present invention to animals include the type of animal, body weight, age,
It can be changed in various ways depending on the health condition and symptoms.
In general, it is recommended to administer the amount of wet bacteria (water content of about 90%) at a rate of about 1 to 50 mg per kg of animal body weight per day. Especially for calves, weigh 1 per day
Sufficient effects are achieved with a small amount of 1 to 5 mg per kg.
In that case, if it is mixed with milk substitute, oral rehydration solution, drinking water, feed and the like for administration, the administration can be performed easily and it is desirable from the viewpoint of labor saving. In particular, when administered to calves, the prophylactic / therapeutic agent of the present invention can be administered to the rumen when mixed with a carrier that directly enters the abomasum without passing through the rumen, such as liquid milk replacer. Since it is not directly changed, it is highly effective. However, the present invention is not limited thereto, and the prophylactic / therapeutic agent of the present invention may be directly orally administered directly to an animal. The preventive / therapeutic agent of the present invention has no toxicity,
Also, no side effects occur. When the prophylactic / therapeutic agent of the present invention is orally administered, it is desirable that the prophylactic / therapeutic agent is continuously administered for 5 days or more, particularly 5 to 20 days, whereby a high prophylactic / therapeutic effect against Salmonella infection can be achieved. it can. Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited thereto.

[0022]

【Example】

<< Example 1 >> (1) Salmonella typhimurium and Salmonella dubulin separated from diarrheal stool of a calf were cultured in a brain heart infusion liquid medium at 37 ° C. for 24 hours, and then centrifugally washed with sterile physiological saline. did. Then, the cells were resuspended in sterile physiological saline and then heated at 121 ° C. for 15 minutes to inactivate each bacterium. Each inactivated bacterium was mixed in an amount equal to the weight (wet bacterium) in a state of being suspended in sterile physiological saline to prepare a preventive / therapeutic agent for Salmonella infection according to the present invention.

(2) First test: Sixteen Holstein male calves (body weight: about 40 to 50 kg / head) of about 7 days old were prepared for purchase at the livestock market, and 8 cows were randomly divided into 2 wards. It was For each of the calves in the 1st ward, the preventive / therapeutic agent for Salmonella infectious disease produced in (1) above was mixed with a milk replacer to be fed in the morning and evening at a rate of 100 mg per wet dose. It was administered twice a day for 10 consecutive days. on the other hand,
The calves in the second ward were similarly fed with milk substitutes containing no prophylactic / therapeutic agent twice a day for 10 days. After that, a normal milk substitute was fed at a rate of 500 g / 4 liter / head per day, and artificial milk, hay and water were fed as a constant feed. The number of deaths 42 days after the introduction is as shown in Table 1 below. Among the dead calves in the second ward, Salmonella typhimurium from two real organs and the other dead calf. Salmonella Dublin was detected in the parenchymal organs of Salmonella, but Salmonella was not detected in the dead calves in the 1st ward, and the cause of death was protracted diarrhea caused by causes other than Salmonella.

(3) Second test: 16 Holstein male calves (body weight: about 40 to 50 kg / head) before and after 7 days of age were separately prepared for purchase at the livestock market, and 8 of them were divided into 2 wards. Preventing in exactly the same way as the first test in (2) above.
When the therapeutic agent was administered and reared, the number of dead cows in each ward 42 days after the introduction was as shown in Table 1 below, and one of the dead calves in the second ward had Salmonella Typhimurium and Salmonella Dublin were detected in the parenchymal organs of another dead calf.

[0025]

[Table 1] 1st ward 2nd ward Administered prophylactic / therapeutic agent Non-administered prophylactic / therapeutic agent 1st study : Number of trials 8 8 Deaths 1 3 Second test : Number of test animals 8 8 Deaths 0 2 Total results : Number of test animals 16 16 Deaths 1 5 Mortality rate (%) 6 31

From the results shown in Table 1 above, it is understood that the agent for preventing / treating Salmonella infection of the present invention has a high effect on the prevention of Salmonella infection in calves.

Example 2 (1) In the same manner as in (1) of Example 1, an inactivating bacterium of Salmonella typhimurium and an inactivating bacterium of Salmonella Dublin were obtained. (2) First test: 16 Holstein male calves (body weight: about 40 to 50 kg / head) before and after 7 days of age were prepared for purchase at the livestock market and randomly divided into 4 groups of 4 cows each. For each of the calves in the first ward, an equal weight (wet bacterium) mixture of the inactivating bacterium of Salmonella typhimurium and the inactivating bacterium of Salmonella dublin produced in (1) above was used as the wet bacterium amount. It was mixed with a milk replacer, which was fed in the morning and evening, at a rate of 100 mg per dose, and was administered twice a day for 7 consecutive days.

For each of the calves in the 2nd ward, only the inactivating bacterium of Salmonella typhimurium produced in the above (1) was fed as a milk substitute at a rate of 100 mg each time in the morning and evening. Was administered twice a day for 7 consecutive days. For each of the calves in the 3rd ward, only the inactivating bacterium of Salmonella Dublin produced in (1) above was mixed with a milk replacer to be fed in the morning and evening at a rate of 100 mg each time as a wet bacterium amount. It was administered twice daily for 7 consecutive days. To the calves in the fourth ward, the milk replacer was similarly fed twice a day and evening for 7 days without administration of neither the inactivating bacteria of Salmonella typhimurium nor the inactivating bacteria of Salmonella dublin.

Thereafter, all the calves in the 1st to 4th wards were given only normal milk substitutes for 5 days, and then Salmonella Dublin was orally administered at a rate of 10 ⁶ / head. After attacking, and then feeding normal milk substitutes, the number of deaths in each ward at the time of one month was examined, and the results were as shown in Table 2 below.

(3) Second test: Fifteen Holstein male calves (body weight: about 40 to 50 kg / head) before and after 7 days of age were prepared separately for purchase at the livestock market. Each of the 4th ward is divided into 3 animals, and administration of the inactivating bacterium of Salmonella typhimurium and / or inactivating bacterium of Salmonella Dublin in the same manner as the first test of (2) above, breeding and salmonella・ When an attack test was performed using Typhyllium, the number of deaths in each ward at the age of one month was as shown in Table 2 below.

[0031]

[Table 2] 1st Ward 2nd Ward 3rd Ward 3rd Ward 4th Ward Salmonella typhimurium inactivating bacteria 50 100 0 0 (mg / dose) Salmonella Dublin inactivating bacteria 50 0 100 0 (mg / dose) First test : Number of test 4 4 4 4 Deaths 0 2 0 4 Second test : Number of test animals 4 4 4 3 Deaths 0 0 1 3 Overall results : Number of trials 8 8 8 7 Deaths 0 2 1 7 Mortality rate (%) 0 25 13 100

From the results in Table 2 above, in the case of the first ward in which the inactivating bacterium of Salmonella of the O 4 group and the inactivating bacterium of Salmonella of the O 9 group are used in combination, the mortality rate of calves is It is 0%, which shows that it is highly effective in preventing Salmonella infection.

Example 3 (1) In the same manner as in (1) of Example 1, each of an inactivated bacterium of Salmonella typhimurium and an inactivated bacterium of Salmonella Dublin was obtained. (2) First test: 16 Holstein male calves (body weight: about 40 to 50 kg / head) before and after 7 days of age were prepared for purchase at the livestock market and randomly divided into 4 groups of 4 cows each. For each of the calves in the first ward, an equal weight (wet bacterium) mixture of the inactivating bacterium of Salmonella typhimurium and the inactivating bacterium of Salmonella dublin produced in (1) above was used as the wet bacterium amount. Each of the doses was mixed at a rate of 25 mg each in the morning and evening, and the mixture was administered twice a day for 7 consecutive days.

For each of the calves in the second ward, only the inactivating bacterium of Salmonella typhimurium produced in the above (1) was fed as a substitute milk in the amount of 50 mg per wet morning and evening. Was administered twice a day for 7 consecutive days. For each of the calves in the 3rd ward, only the inactivating bacterium of Salmonella Dublin produced in the above (1) was mixed with the milk replacer to be fed in the morning and evening at a rate of 50 mg per time as a wet bacterium amount. It was administered twice daily for 7 consecutive days. To the calves in the fourth ward, the milk replacer was similarly fed twice a day and evening for 7 days without administration of neither the inactivating bacteria of Salmonella typhimurium nor the inactivating bacteria of Salmonella dublin.

Thereafter, to all the calves in the 1st to 4th wards, only normal milk substitutes were fed for 5 days, and then Salmonella Dublin was orally administered at a rate of 10 ⁶ / head. After attacking, and then feeding normal milk substitutes, the number of deaths in each ward at the time of one month was examined, and the results were as shown in Table 2 below.

(3) Second test: 16 Holstein male calves (body weight: about 40 to 50 kg / head) of about 7 days old were prepared separately for purchase at the livestock market, and 4 of them were randomly distributed to 4 wards. Separately, administration, breeding and challenge test with Salmonella typhimurium of Salmonella typhimurium inactivating bacteria and / or Salmonella Dublin inactivating bacteria were performed in exactly the same manner as the first test of (2) above. As a result, the number of deaths in each ward at the time of one month's age was as shown in Table 3 below.

[0037]

[Table 3] 1st Ward 2nd Ward 3rd Ward 3rd Ward 4th Ward Salmonella typhimurium inactivating bacterium 25 50 0 0 (mg / dose) Salmonella Dublin inactivating bacterium 25 0 50 0 (mg / dose) First test : Number of test 4 4 4 4 Deaths 0 4 2 4 Second test : Number of test animals 4 4 4 4 Deaths 0 2 3 4 Overall results : Number of test animals 8 8 8 8 Deaths 0 6 5 8 Mortality rate (%) 0 75 63 100

From the results in Table 3 above, in the case of the 1st ward in which the inactivating bacterium of Salmonella of the O 4 group and the inactivating bacterium of Salmonella of the O 9 group are used in combination, the dose is small. However, the calf mortality rate is 0%, which shows that it is highly effective in preventing Salmonella infection.

Example 4 (1) Salmonella typhimurium [American Type
Culture Collection (hereinafter abbreviated as ATCC) 14028] and Salmonella enteritidis (RB-1 strain) were cultured in a heart infusion agar medium at 37 ° C. for 24 hours, and then scraped to remove wet bacteria. 50 mg
Suspended in sterile saline at a ratio of 1 / ml and 1
Each bacterium was inactivated by heating at 21 ° C. for 20 minutes. Each inactivated bacterium was mixed in an amount equal to the weight (wet bacterium) in a state of being suspended in sterile physiological saline to prepare a preventive / therapeutic agent for Salmonella infection according to the present invention.

(2) Holstein male calves around 7 days old
16 animals (body weight: about 40 to 50 kg / head) were prepared for purchase at the livestock market and were randomly divided into 4 groups of 4 animals each. First
None of the salmonella infectious disease preventive / therapeutic agents of the present invention produced in (1) above was administered to each of the calves in the plot. In the calf in the second ward, the salmonella infectious disease preventive / therapeutic agent of the present invention produced in the above (1) was mixed with a milk replacer to be fed in the morning and evening in a wet bacterial amount of 100 mg per dose. Twice a day for 2 consecutive days.

For calves in the 3rd ward, the agent for preventing and treating Salmonella infectious disease of the present invention produced in the above (1) was mixed with a milk replacer to be fed in the morning and evening at a rate of 100 mg per wet dose. Then, the drug was continuously administered twice a day for 4 days. Then, the calf in the 4th ward is mixed with the salmonella infectious disease preventive / therapeutic agent of the present invention produced in the above (1) in a milk replacer to be fed in the morning and evening at a rate of 100 mg per time. Twice a day for 6 consecutive days.

After that, all the calves in the 1st to 4th wards were given only the normal milk substitute, and then 10 ⁵ Salmonella typhimurium / head at the time of the 20th day of age. Oral administration was carried out in order to attack, and then normal milk substitutes were fed to examine the number of deaths in each ward at the age of 1 month. The results are shown in Table 4 below.

[0043]

[Table 4] Number of days of administration of prophylactic / therapeutic agents for Salmonella infection (days) Number of test animals Death number Death rate (%) 1 ward 0 4 4 100 2 ward 2 4 2 50 3 ward 4 4 1 25 4 ward 6 4 0 0

From the results shown in Table 4 above, it can be seen that continuous administration of the prophylactic / therapeutic agent for Salmonella infection of the present invention for 5 days or longer, particularly 6 days or longer, is effective for the prevention of Salmonella infection.

Example 5 (1) Salmonella typhimurium (ATCC 14028) and Salmonella Dublin (field isolate) were cultured in a brain heart infusion liquid medium at 37 ° C. for 24 hours, and then formalin was added thereto. Each bacterium was inactivated by adding 0.3%. Then, after centrifugal washing with sterile physiological saline, it was resuspended in sterile physiological saline.

(2) 4 weeks of 5 week old C3H / HeN mice
0 animals were prepared, and 8 animals were randomly divided into 5 groups.
The mice in the first ward received normal drinking water for 3 weeks without administering either the inactivating bacterium of Salmonella typhimurium or the inactivating bacterium of Salmonella Dublin produced in (1) above. Feeded. To the mice in the second ward, drinking water containing only the inactivating bacterium of Salmonella typhimurium produced in the above (1) at a rate of 0.5 mg (wet bacterium) / ml was fed with a normal diet for 2 weeks. After that, normal water and food were fed for one week thereafter.

In the third group of mice, only the inactivated bacterium of Salmonella typhimurium produced in the above (1) was treated with 0.
The drinking water added at a rate of 75 mg (wet bacteria) / ml was fed with the normal feed for 2 weeks, and then the drinking water and the feed were fed for 1 week. For the mouse in the 4th ward, 0.5 mg (wet bacterium) / mg of the inactivating bacterium of Salmonella typhimurium and 0.1 mg (wet bacterium) of the inactivating bacterium of Salmonella Dublin were produced in the above-mentioned (1). The drinking water added at a ratio of 1 ml / ml was fed with the normal feed for 2 weeks, and then the drinking water and the feed were fed for 1 week. The mouse in the 5th ward contained Salmonella produced in (1) above.
0.5 mg (wet bacterium) / m of inactivating bacteria of Typhyllium
g and 0.5 mg of Salmonella Dublin inactivating bacteria
Drinking water added at a rate of (wet bacteria) / ml was fed together with the normal feed for 2 weeks, and then for 1 week, the normal drinking water and the feed were fed.

(3) After the lapse of the above 3 weeks, live salmonella typhimurium was administered intraperitoneally to the mice in each group at a rate of 2 × 10 ⁴ cells / mouse. The survival rate of the mice 3 weeks after the challenge was as shown in Table 5 below.

[0049]

[Table 5] Concentration in drinking water (mg / ml) Salmonella / Thai Salmonella / Number of test animals Number of surviving animals Survival rate (%) Fymulium Dublin 1 ward 0 0 8 0 0 2 ward 0.5 0 8 6 75 3 ward 0.75 0 8 7 88 4 wards 0.5 0.1 8 8 100 5 wards 0.5 0.5 8 8 100

From the results in Table 5 above, when the inactivating bacterium of Salmonella typhimurium and the inactivating bacterium of Salmonella dublin were mixed and administered, the inactivating bacterium of Salmonella typhimurium was administered alone. It can be seen that the survival rate of the mouse is extremely higher than that of.

Example 6 (1) Salmonella typhimurium (ATCC 14028) and Salmonella Dublin (field isolate) were cultured in a brain heart infusion liquid medium at 37 ° C. for 24 hours, and then formalin was added. Each bacterium was inactivated by adding 0.3%. Then, after centrifugal washing with sterile physiological saline, it was resuspended in sterile physiological saline.

(2) 3 weeks of 5 week old C3H / HeN mice
Two animals were prepared, and 8 animals were randomly divided into 4 groups.
The mice in the first ward received normal drinking water for 3 weeks without administering either the inactivating bacterium of Salmonella typhimurium or the inactivating bacterium of Salmonella Dublin produced in (1) above. Feeded. For the mice in the second ward, the drinking water containing only the inactivating bacterium of Salmonella Dublin produced in (1) above at a rate of 0.1 mg (wet bacterium) / ml was added.
Feed with regular food for a week and then 1
He was given regular drinking water and food for a week.

In the third group of mice, 0.1 m of the inactivating bacterium of Salmonella typhimurium produced in (1) above was used.
Drinking water containing g (wet bacterium) / ml and Salmonella Dublin inactivating bacteria at a rate of 0.1 mg (wet bacterium) / ml was fed with normal food for 2 weeks, and then for 1 week with normal drinking water. Feeded. For the mice in the 4th ward, 0.5 mg (wet bacterium) / ml of the inactivating bacterium of Salmonella typhimurium and 0.1 mg (wet bacterium) of the inactivating bacterium of Salmonella Dublin produced in (1) above were used. / Ml
The drinking water added at the ratio of 2 was fed with the normal feed for 2 weeks, and then the drinking water and the feed were fed for 1 week.

(3) After the lapse of the above 3 weeks, live salmonella Dublin was administered intraperitoneally to the mice in each group at a rate of 2 × 10 ⁴ cells / mouse. The survival rate of the mice 3 weeks after the challenge was as shown in Table 6 below.

[0055]

[Table 6] Concentration in drinking water (mg / ml) Salmonella / Thai Salmonella / Number of test animals Number of surviving animals Survival rate (%) Fymulium Dublin 1 ward 0 0 8 0 0 2 ward 0 0.1 8 4 50 3 ward 0.1 0.1 8 6 75 4 wards 0.5 0.1 8 8 100

From the results shown in Table 6 above, in the case where the inactivating bacterium of Salmonella typhimurium and the inactivating bacterium of Salmonella dublin were mixed and administered, the inactivating bacterium of Salmonella dublin was administered alone. It can be seen that the survival rate of the mouse is extremely higher than that of.

Example 7 (1) Salmonella typhimurium (ATCC 14028) and Salmonella Dublin (field isolate) were each cultivated in a brain heart infusion liquid medium at 37 ° C. for 24 hours, and then formalin was added thereto. Each bacterium was inactivated by adding 0.3%. Then, after centrifugal washing with sterile physiological saline, it was resuspended in sterile physiological saline.

(2) First test: 5-week old BALB
40 / c mice were prepared, and 10 mice each were divided into 4 groups. In each of the mice in the first section, 0.2 of the inactivating bacterium of Salmonella typhimurium produced in the above (1)
Saline was added at a rate of 5 mg (wet bacterium) / ml and Salmonella Dublin inactivating bacterium 0.25 mg (wet bacterium) / ml at a rate of 1 ml / mouse each day [0.25 mg of the above inactivating bacterium (0.25 mg Wet bacterium / animal; total 0.5 mg (wet bacterium) /
Oral administration was carried out for 10 days at a rate of [rats] [total dose for 10 days: 5 mg (wet bacteria) / animal] normal drinking water and food were fed for 10 days.

In each of the mice in the second group, the inactivating bacterium 1 of Salmonella typhimurium produced in the above (1)
0.2 ml of physiological saline added at a ratio of 2.5 mg (wet bacterium) / ml and 12.5 mg (wet bacterium) inactivating bacterium of Salmonella Dublin [amount of each inactivating bacterium above 2.
5 mg (wet bacterium) / animal; total 5.0 mg (wet bacterium) / animal] was intraperitoneally injected, and then drinking water and food were fed as usual for 10 days. For each mouse in the 3rd ward,
Oral administration of physiological saline at a rate of 1 ml per day for 10 days was followed by normal drinking water and food for 10 days.
Mice in the fourth section were intraperitoneally injected with physiological saline at a rate of 0.2 ml / mouse, and then drinking water and food were fed as usual for 10 days.

Immediately after the 10-day oral administration was completed in the first and third sections, and the day after the intraperitoneal injection in the second and fourth sections, the activity of the mice was decreased, diarrhea and reverse hair coating were observed. The presence or absence of standing and weight loss and the number of deaths were examined. Ten days later, the surviving mice were treated with Salmonella.
Table 7 below shows the number of mouse deaths examined by intraperitoneal challenge with live Dublin cells at a rate of 2 × 10 ⁴ cells / animal, and after 3 weeks.

(3) Second test: 5-week-old BALB
40 / c mice were prepared, and 10 mice each were divided into 4 groups. The day after the same administration and feeding as in the first test was performed on the mice in each of the above-mentioned groups, the activity decrease, diarrhea, hair inversion and weight loss, and the number of deaths were examined. It was Ten days after that, the surviving mice were treated with 2 live strains of Salmonella typhimurium.
It was shown in Table 7 below when the number of mouse deaths was examined after 3 weeks from the challenge by intraperitoneal administration at a rate of × 10 ⁴ / animal.

[0062]

[Table 7]

From the results shown in Table 7 above, the mortality rate of mice was 0% in the case of the first group which was orally administered with the inactivating bacteria of Salmonella of the O 4 group and the inactivating bacteria of Salmonella of the O 9 group. , Is highly effective in the prevention of Salmonella infection, whereas in the second ward, which is administered intraperitoneally, side effects after intraperitoneal administration are stronger and preventive effects than when administered orally. It can be seen that the mortality rate of the mouse is 12%, which is inferior.

[0064]

When the prophylactic / therapeutic agent for Salmonella infectious disease of the present invention is used, adverse effects on the environment due to the spraying of a large amount of conventionally used antiseptics, the use of drugs such as antibiotics and synthetic medicines The problem of the emergence of resistant bacteria due to the drug, the concern that such drugs may remain in animals, the environmental pollution associated with the expulsion of the live vaccine grown in the body, and the strong effects of parenteral administration of dead bacteria Side effects and the like can be prevented, and the Salmonella infection in animals can be prevented and treated extremely safely and effectively. And, a preventive / therapeutic agent for Salmonella infection of the present invention, a milk replacer for feeding animals,
When orally administered in addition to drinking water, food, etc., the prophylactic / therapeutic agent can be easily administered to an animal without any trouble. In particular, prevention of Salmonella infection of the present invention
The therapeutic agent is suitable as a preventive and therapeutic agent for Salmonella infection in calves, and the amount is 1 per 1 kg of calf weight.
Oral administration of small doses of ~ 5 mg / day can prevent and effectively treat calves from Salmonella infections.

Claims (2)

[Claims] 1. In terms of classification by Salmonella O antigen, O
A prophylactic / therapeutic agent for Salmonella infections in animals, which comprises at least one inactivating bacterium of Salmonella belonging to Group 4 and at least one inactivating bacterium of Salmonella belonging to O9 group. 2. According to the classification by Salmonella O antigen, O
A combination of at least one inactivating bacterium of Salmonella belonging to Group 4 and at least one inactivating bacterium of Salmonella belonging to O 9 group, and continuously orally administering to the animal for 5 days or more. A method for preventing or treating Salmonella infection.