JPH1112184A - Formulation for treating bovine salmonellosis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject formulation for removing salmonella bacteria from bovine intestinal tracts by including a lactobacillus, a butyric acid bacterium and a saccharogenic bacterium as active ingredients. SOLUTION: This formulation contains the mixture of three kinds of live bacteria comprising a lactobacillus, a butyric acid bacterium and a saccharogenic bacterium. The lactobacillus includes Streptococcus faecalis T-110 (referred to as SF), is a gram-positive facultative anaerobic micrococcus, and has characters comprising a high lactic acid-producing ability and a large proliferation rate. The butyric acid bacterium includes Clostridium butyricum TO-A (referred to as CB), is a gram-positive strictly aerobic bacillus bacterium, has a spore-forming ability, and produces a short chain fatty acid such as butyric acid or acetic acid. The saccharogenic bacterium includes Bacillus mesentericus TO-A (referred to BM), is a gram-positive aerobic bacillus bacterium, and produces an amylolytic enzyme. The three kinds of bacteria are preferably added in a SF:CB:BM ratio of 10:1:1. The formulation is administered at each dose of 20-50 g for a calf or at each dose of 50-200 g for an adult cattle. The various syndromes of the cattle due to the salmonellosis are improved.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for salmonella infection in cattle.

[0002]

2. Description of the Related Art Since the late 1970s, domestic cattle-producing farmers have been infected with Salmonella, which has become a serious problem in dairy management. In 1994, the number reported to the Ministry of Agriculture, Forestry and Fisheries alone reached 2,000, with 571 deaths.
I'm on my head. Estimated total damage in FY1993 was about 26
In the case of dairy farming, especially in the case of private management, dairy farming has to be abandoned.

Until the 1980s, Salmonella infectious disease was mainly in calves, but since then, adult cattle have become more contaminated, and the contamination of Salmonella continues to increase as long as the current population breeding style is maintained. I think that the.

[0004] On the other hand, in the state of food poisoning in the whole country in fiscal 1994, Salmonella bacteria accounted for half of the contaminating bacteria, reaching 14,410 people. Conventionally, the causal food was mainly chicken eggs and their processed products, but in recent years, it has been reported that beef or milk has been damaged. Considering that the number of victims of enterohemorrhagic Escherichia coli O157 that occurred in Japan in 1996 was about 9,000, it can be understood how much the number of food poisoning damages caused by Salmonella.

[0005] The serotypes of Salmonella bacteria that have been a problem in the livestock field include Salmonella typhimurium (S
T), Salmonella Dublin (SD) and Salmonella Enteritidis (SE) are representative. In particular, ST is the most isolated as a contaminant of cattle. Although the number of cows occurring in SD is smaller than that in ST, the probability of systemic infection from the intestinal tract via the portal vein is high, and there are cases where sepsis and invasion of other organs occur, resulting in death. Unless SD is completely eliminated from feces, many cases will not be approved for shipment and will be delayed before shipment or will die before confirmation of bacterial evacuation. is there. SE is a bacterial species that is more problematic in broilers and laying hens than in cattle, so it is a bacterial species that requires more attention as a source of environmental pollution than cattle infectious diseases.

[0006]

[0005] Administration of antibiotics has been actively performed as a countermeasure against Salmonella infection. Antibiotics are widely used as feed additives other than veterinary drugs because they have been confirmed to have a body weight gain effect. However, due to the problem of persistence in dairy products and the possibility of emergence of resistant bacteria, regulations on the amount of antibiotics used and the fields of application are being tightened. In addition, the development of vaccines for Salmonella has been studied for a long time, but has not yet been commercialized.

[0007] Under such circumstances, in the field of poultry, an attempt was made by Nurumi et al. To formulate intestinal bacteria in the cecal contents of parent chickens and to competitively eliminate pathogenic microorganisms such as Salmonella by administering to chickens. Done and has recently been commercialized.

However, in the field of cattle, there has been almost no attempt at such a competitive exclusion method, and there have been many products mainly comprising a rumen function improving agent in the bovine rumen. Also, attempts have been made to use live bacterial preparations utilizing lactic acid bacteria, bifidobacteria, and the like, but no attempt has been made to actively eliminate Salmonella.

An object of the present invention is to provide a remedy for Salmonella infectious disease for eliminating Salmonella bacteria from bovine intestinal tract.

[0010]

Means for Solving the Problems The present inventors have succeeded in creating an infection model using calves instead of rats and mice that have been conventionally used as a Salmonella infection experimental model. The inventors have found that the mixture shows an effect of eliminating ST and SD in vitro, and have completed the present invention.

That is, the present invention provides (1) a therapeutic agent for salmonella infection in cattle, which comprises a lactic acid bacterium, a butyric bacterium and a saccharifying bacterium as active ingredients, and (2) the cattle are fattening cows or milking cows. (1) The therapeutic agent for salmonella infection in cattle according to the above (1).

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The therapeutic agent for bovine Salmonella infection according to the present invention is a viable bacterial agent characterized by mixing three types of viable bacteria of lactic acid bacteria, butyric acid bacteria and saccharifying bacteria.

The lactic acid bacterium used in the present invention is not particularly limited. For example, Streptococcus faecalis T-110
(Hereinafter abbreviated as “SF”) is a Gram-positive facultative anaerobic bacterium isolated from a healthy adult, having a high lactic acid-producing ability and a high growth rate.

The butyric acid bacterium is not particularly limited. For example, Clostridium butyricum TO-A
Tyricum TO-A (hereinafter abbreviated as "CB") is a Gram-positive obligate anaerobic bacillus isolated from healthy adults and has sporulation ability, and produces short-chain fatty acids such as butyric acid and acetic acid.

[0015] The saccharifying bacteria are not particularly limited. For example, Bacillus mecentericus TO-A (Bacillus mesen)
tericus TO-A (hereinafter abbreviated as “BM”) is a Gram-positive aerobic bacterium isolated from soil and produces amylolytic enzymes.

By administering the mixture of the three bacteria of the present invention to cattle infected with Salmonella, Salmonella can be removed, and the mixture of the three bacteria of the present invention is effective as an agent for treating Salmonella infection in cattle. In addition, the administration of the mixture of three bacteria of the present invention to normal cattle can prevent the onset of Salmonella infection, and the mixture of three bacteria of the present invention is also effective as an agent for preventing Salmonella infection in cattle.

It is unknown why the mixture of the three bacteria of the present invention is effective for Salmonella infectious disease in cattle, but some of the factors are due to the fact that these three bacteria in the present invention are symbiotic. I think that the. That is, when SF and CB are mixed and cultured, CB is about 10 times less than that of the single culture.
The number of viable bacteria increases twice. When SF and BM are cultured in a starch medium, the viable cell count of SF increases about 10 times. In addition, the inhibitory effect of SF or CB on pathogenic Escherichia coli was confirmed by anaerobic continuous flow culture, and the effect of inhibiting the pathogenic Escherichia coli of SF or CB alone was stronger than that of SF or CB under the condition where the symbiotic relationship between SF and CB was maintained. A significant inhibitory effect was observed. On the other hand, when the BM and the bifidobacterium were mixedly cultured, an increase in the growth efficiency of about 10 times compared to the bifidobacterium single culture was observed.

Each of the three species of the present invention can be cultured in an appropriate medium. That is, it can be cultured in a medium having a composition comprising a carbon source such as glucose, sucrose, starch, etc., nitrogen and minerals such as peptone, meat extract, yeast extract and the like. After culturing in each medium, centrifugation is performed to obtain bacterial cells. Thereafter, the obtained cells are dispersed in a skim milk solution or the like and freeze-dried. The obtained dried product is pulverized, and potato starch, corn starch, lactose, acid clay,
It may be mixed with excipients such as natural aluminum silicate to obtain three kinds of bacterial powder.

SF, CB, BM of the mixture of three bacteria of the present invention
Is ideally 10: 1: 1 as a ratio of the number of bacteria, but is not limited to this ratio.

The dose of the mixture of three bacteria of the present invention may be usually administered orally or 20 to 50 g at a time for calves and 50 to 200 g at a time for adults. At the time of feeding, ordinary feed additives such as vitamins and minerals may be added to the feed.

The mixture of the three bacteria according to the present invention is effective as a therapeutic agent for bovine Salmonella infection, but also exerts an excellent effect when administered to cattle to prevent bovine Salmonella infection.

The cows to which the present invention is applied are not particularly limited, but include, for example, fattening cows such as Holstein, Wagyu, F1 (a cross between Holstein and Wagyu), Jersey breeds, and milking cows.

[0023]

【Example】

Example 1 A mixed preparation of three viable bacteria having the composition shown in Table 1 below was prepared. The lactic acid bacteria were Streptococcus faecalis T-110 (FERM P-8936), and the butyric acid bacteria were Clostridium butyricum TO-A (Clostridium butyri).
cum TO-A) (Seiko Kogyo FERM P-8935)
Bacillus bacterium is Bacillus mesentericus TO-A (Bacill
us mesentericus TO-A)
934) was used.

[0024]

[Table 1]

Four 10-day-old Holstein breeds were
Two groups were treated with the inoculum-mixed preparation and two control groups.

In each of the test plots, a milk substitute for lactating calves (trade name: Maxicare) without antibiotics was added twice a morning and evening, 2 L each.
I was paid by In the group receiving the mixed preparation of three viable bacteria, the preparation was mixed with 50 g of milk substitute per animal per day, and injected daily from introduction to necropsy.

7 days after introduction, 10% NaHCO per animal
₃ Salmonella Dublin (Salmo
nella Dublin) (SD) 5230P + strain (number of bacteria: 10 ⁴⁾
/ Ml) was administered intragastrically to all animals in a stomach tube.

As a test method, 1 g of feces was diluted with physiological saline, inoculated into a Harner tetrathionate medium and an SGB sulfa basal medium, and cultured at 37 ° C. for 24 hours. The culture was cultured on a novobiocin-added DHL agar medium, and Salmonella colonies were searched. The type of the isolated Salmonella was confirmed by slide agglutination using anti-Salmonella O9 group serum. Further, for Salmonella-negative ones, the enrichment medium was left at room temperature for 7 days to perform delayed secondary enrichment.

Table 2 shows the fecal findings, and Table 3 shows the state of SD separation from feces.

[0030]

[Table 2]

[0031]

[Table 3]

As is clear from the results shown in Table 2, 3
In the group treated with the inoculum mixed preparation, a tendency to significantly improve the convenience was observed.

As is apparent from the results shown in Table 3, 3
Salmonella bacteria were no longer detected within 5 days after SD challenge in the inoculum-mixed preparation-administered group, whereas 9% in the control group.
The detection of Salmonella continued until the day.

Example 2 A mixed preparation of three viable bacteria shown in Table 4 below was prepared. Lactic acid bacteria, butyric acid bacteria, and saccharifying bacteria used in Example 1 were used.

[0035]

[Table 4]

Eight Holstein breeds 30 days old were given 3
The group receiving the inoculum mixed preparation was 4 animals, and the control group was 4 animals.

In both test plots, lactating milk for calves (pro milk) was fed twice a morning and evening, 2 L each. 3
In the administration group of the inoculum-mixed preparation, the preparation was mixed with milk substitute at a rate of 50 g per animal per day and administered daily from introduction to necropsy. In addition, hay and commercial artificial milk (trade name: Super Hoiku Mullet) were fed constantly.

7 days after introduction, 10% NaHCO / head
₃ Salmonella Typhimurium (ST) suspended in 100 ml (number of bacteria: 10 ⁴
/ Ml) was administered intragastrically to all animals in a stomach tube.

As a test method, 1 g of feces was diluted with physiological saline, inoculated into a Harner tetrathionate medium and an SGB sulfa basal medium, and cultured at 37 ° C. for 24 hours. The culture was cultured on a novobiocin-added DHL agar medium, and Salmonella colonies were searched. The type of the isolated Salmonella was confirmed by slide agglutination using anti-Salmonella O9 group serum. Further, for Salmonella-negative ones, the enrichment medium was left at room temperature for 7 days to perform delayed secondary enrichment.

Table 5 shows hematological findings, and Table 6 shows the state of ST separation from feces.

[0041]

[Table 5]

[0042]

[Table 6]

As is evident from the results shown in Table 5, no significant difference was observed in the control group and the group administered with the mixed preparation of the viable bacteria.

As is clear from the results shown in Table 6, 3
In the group treated with the inoculum mixed preparation, a significant difference was observed in the total score (Total score) after ST challenge as compared to the control group on the 12th day. In addition, all three species administered were frequently detected in feces.

[0045]

As described above, the mixed preparation of three viable bacteria of the present invention containing lactic acid bacteria, butyric acid bacteria, and saccharifying bacteria as active ingredients is effective as a therapeutic agent for bovine Salmonella infectious disease. By administering the mixed preparation of live bacteria to cattle, it is expected to improve various symptoms of cattle due to Salmonella infection, prevent the spread of Salmonella, and purify the dairy environment.

Claims (2)

[Claims] 1. A therapeutic agent for salmonella infectious disease in cattle, comprising lactic acid bacteria, butyric acid bacteria and saccharifying bacteria as active ingredients. 2. The cow according to claim 1, wherein the cow is a fattening cow or a milking cow.
The therapeutic agent for salmonella infectious disease of cattle as described in [1].