JPS62294623A - Preventive for salmonella infectious disease - Google Patents

Abstract

PURPOSE:The titled preventive consisting of a specific antibiotic and an inert Salmonella mold. CONSTITUTION:A preventive for Salmonella infectious diseases containing at least one selected from penicillin antibiotic, polypeptide antibiotic (e.g. colistin, etc.), macrolide (e.g. erythromycin, etc.), cephem antibiotic (e.g. cefalexin, etc.), chloramphenicol antibiotic, tetracycline and phosphorus-containing antibiotic (e.g. Quebemycin, etc.) as an effect enhancer and an inert Salmonella mold. Referring to a dose, the amount of the inert mold is at least 1mg/day/kg, preferably 2-100mg/day/kg and the amount of the antibiotic is at least 1mg/day/kg, preferably 2-100mg/day/kg. The preventive is orally administered. Solution, powder, granule, tablet, coated tablet, capsule, pill, etc., may be used as the dosage form. The preventive is continuously and orally administered every day and a period of the administration is preferably more than a week.

Description

[Detailed description of the invention] 3. Detailed description of the invention [Industrial application field] The present invention relates to an agent for preventing Salmonella infection.

[Conventional technology]

Infectious diseases caused by Salmonella include, for example, typhoid fever or paratyphoid fever or Salmonella enteritis, and have long affected the health of humans and non-human animals. BACKGROUND ART Conventionally, as a preventive method against Salmonella infection, it has been considered to administer live attenuated bacteria orally or parenterally. In addition, inactivated bacteria can be used instead of live bacteria to infect humans [H,L, Dupont].
Bull, Wld, Hlth, Org, J Sat. 4.667-672 (1971)) or animals (Special Publications Showa 4
8-19931) or by parenterally administering typhoid/paratyphoid fever vaccines to humans to prevent Salmonella infections.

In addition, it has been reported in mice that intermittent oral administration of inactivated bacteria and large doses of streptomycin improves the preventive effect [R.H. Waldman et al.
, ImmunityJ 6.58-61 (1972
)].

[Problem that the invention seeks to solve]

These conventional methods have the following drawbacks. Prophylactic methods using attenuated live bacteria are relatively effective, but they are not practical due to safety issues such as reversion of pathogenicity due to changes in the strain and environmental contamination. On the other hand, prophylactic methods using only inactive bacteria are highly safe due to their inactivation, but cell-mediated immunity is deeply involved in the defense of Summonella infection, and inactive bacteria are not sufficient. It was not possible to impart sufficient cell-mediated immunity, and the effect was insufficient.

In the method of using streptomycin together with the inactive fraction, the dosage of the inactive fraction and streptomycin is 10% for the former.
9-1(2), the latter requires a relatively large amount of 2.5M 97 times, and the inactivation takes a long time, such as 3 hours at 63°C. Therefore, it is economically inferior, and its effects on animals other than mice and antibiotics other than streptomycin are unknown.

The present inventors investigated a safe, highly effective, and economical preventive agent for Salmonella infection that does not have the drawbacks of these conventional methods, and discovered the present invention.

polypeptide antibiotics, macrolide antibiotics,
This invention relates to a preventive agent for Salmonella infection comprising at least one selected from the group consisting of cephem antibiotics, chloram-72dicol antibiotics, tetracycline antibiotics, and phosphorus-containing polysaccharide antibiotics, and inactivated Salmonella bacteria.

The Salmonella bacteria used in the present invention include, for example, Salmonella typhi and Salmonella typhimurium.
phimurium) or Salmonella paratyphi A
(8, paratyphi A) and Salmonella enteritidis (8, enteritidis). In the present invention, these bacteria are inactivated by heating, formalin treatment, or the like.

The effect-enhancing substances used in the present invention are more effective than honeycillin antibiotics, polypeptide antibiotics, macrolide antibiotics, cephem antibiotics, chloramphenicol antibiotics, tetracycline antibiotics, and phosphorus-containing polysaccharide antibiotics. At least one selected from the group consisting of:

Examples of R-nigilin antibiotics include penicillin, benzylpenicillin, and phenoxymethyl include nigiline, ampicillin, amoxicillin, bacampicillin, etc.
Examples of polypeptide antibiotics include colistin and bacitracin; examples of macrolide antibiotics include erythromycin, spiramycin, kitasamycin, and rifanviciliy; examples of cephalic antibiotics include cephalexin and cefazolin; Examples of chloramphenicol antibiotics include chloramphenicol and thiamphenicol; examples of tetracycline antibiotics include oxytetracycline and doxycycline;
Examples of antibiotics include kebemycin and macarobomycin. Compounds of these pharmaceutically acceptable salts or esters may also be used.

Among these, penicillin antibiotics are preferred, with penicillin and ampicillin being particularly preferred.

The amount of inactive bacteria used in the present invention is at least 1 V day/bw, preferably 2 to 100 q/day/bw, as a wet bacterial load, while the amount of antibiotics is at least 11 days/bw. body weight, preferably 2 to 100 V' day^ body weight.

The prophylactic agent of the present invention is orally administered by mixing an inactive drug and an antibiotic. As the dosage form for oral administration, conventional dosage forms for oral administration are used. Examples include liquid and solid dosage forms such as liquids, powders, granules, tablets, coated tablets, capsules, and gun powder. Examples of liquid preparations include those in which an inactive dye and an antibiotic are suspended in water. In the production of solid preparations, the inactive suspension is, for example, freeze-dried to form a solid, an antibiotic is added to this, and additives conventionally used in the production of solid preparations, such as excipients and fillers, are added. , a binder, a disintegrant, etc. are added to form a solid agent. In addition, inactive particles and antibiotics can be added to feed or food.

When administering the prophylactic agent of the present invention, favorable results can be obtained by continuous daily oral administration, and the administration period is preferably one week or more.

The prophylactic agent of the present invention exhibits almost no toxicity, and no side effects other than the effects conventionally possessed by the antibiotics used at the same time.

The prophylactic agent of the present invention can be used with humans or non-human animals, such as cows, to achieve excellent effects.

〔Example〕

Next, examples will be shown.

Example 1 (4) Salmonella tephimurium (S, typh
imurium) (American Type Culture Collection 1402B) in a liquid medium at 37C for 24 hours.
After incubation for an hour, the cells were centrifuged, washed, suspended in physiological saline, and heated at 100° C. for 5 minutes. This was made into an inactive image.

(B) The inactive fraction obtained in (4) is 0 as the amount of wet bacteria.
．． 5 to 5 η/- suspended in drinking water, and either ampicillin sodium 0.02 to 0.5 y/- or penicillin G potassium 20 to 500 units/id was added, and nonvital buck teeth and antibiotics were used as controls. Mice [Ba1b/C, female, 5 weeks old] were given ad libitum access to either alone or water alone for 6 weeks.

After the end of the administration, only water was administered for one week, and Salmonella typhimurium was infected at 105/mouse [orally (po) using a sonde].
, or 101-2/mouse [intraperitoneal (ip)]. Mortality after oral (po) challenge was 0.15 to 115 in the combined antibiotics and inactivated bacteria group, v5 in the inactivated bacteria alone group, and 515 in each antibiotic rate/individual group and water alone group. Ta. In addition, the number of deaths after intraperitoneal (IP) challenge was 2710 in the combination antibiotics and inactivation group and 9710 in the antibiotics alone group.
The combination group was effective for both.

Histopathologically, no significant changes were observed in the combination group, but in the antibiotic alone group, findings characteristic of salmonellosis, including necrosis, were observed in the liver tiles.

Example 2 Six Holstein male cows (7 days old) were divided into two groups,
Group A (3 cows) was fed twice in the morning and evening with the inactive extract obtained in the prisoners of Example 1 added at a wet bacterial dose of 50 IIgZ and ampicillin sodium 100m5F/time. It was administered for 10 days. Group B (3 animals) received 1001 g of ampicillin sodium in the milk replacer fed twice in the morning and evening.
The same administration was repeated for 10 days. After completion of administration,
They were fed only regular milk replacer for 3 days and then challenged by orally administering Salmone 2 Chinimurium to ax105/head. The results obtained are shown in Table 1.

Shishi 1 A 2 4 0 B 0
4 3 Example 3 Forty Holstein male cows (7 days old) were divided into two groups, and one group (20 cows) was treated with the inactivated M5om obtained from the prisoners of Example 1.
y/time and 10011 g/time of ampicillin sodium was added to the milk replacer given in the morning and evening, and was orally administered continuously for 7 days. -10,000 and ■ groups (20 animals) were administered in the same manner, with only 1001f/dose of ampicillin sodium added to the milk replacer.

Table 2 shows the body weights and outcomes of cows in each group at the start of administration and after 2 weeks.

(Table 2) 46 49 Survival 44 44
living) sl 54 48 46 (C
) 49 52 45 44 (b) 45
50 1 51 51 (F) 44 50
1 49 49 (G) 52 58
55 52 1 (b) 51 52 #
47 47 1(I) 42 41
46 50 (J) 45 49 45
47 (a) 47 48 43 42 (
g) 49 53 42 -
Death H505, 24447 Survival (goods) 47 48 46 46 (0) 4
7 49. 18 50 I) 45 4
4 53 53 (Q) 49 50
44 45 (published 43 44
46 Death (S) 51 52
49 53 Survival (I')
46 46 47 44 Average 47.5±
2949.6±3.8 470±3.317.5±
6.4 Example 4 Section 0.3 Inactivated bacteria obtained by inactivation with formalin,
Ampicillin (AB-PC) was added to nigiline (Pcχ colistin (CL-containing ampicillin (Rlf)) in the water supply of mice, and AB-PC was added as a control.
Mice (Balb, female, 5 weeks old) were allowed to freely ingest each of c SPc SCL and Rlf alone and water alone for 2 weeks. Next, the animals were given only water for one week, and then challenged by administering Salmonella tefimuricum (105/mouse) using an oral probe.

The results 20 days after the challenge are shown in Table 3.

Table 3 AB-Pc O,02("lrm/) 0.5(
49/m/) 1 to 0.1 #
0150.5 # 0
15Pc 20 (JIIQ/d)
0151 100#
015# 500 015
CL 400 (unit 1*> 05 (+rd)
115a1t 111L05 (1*)
0151 (L5
More than 1 015-PcQ
, 1# 0 515Pc
100 (unit/d 51
5CL 400#
515R1f Q, 05 (%'d) --# 515 Example 5 The inactivated bacteria obtained in Example 1 (A) were freeze-dried.
9 was added and mixed uniformly. 50q thereof was filled into hard gelatin capsules.

Example 6 8a1 monellae heat-inactivated at 100°C for 5 minutes
A mixture of P. teritidis (S, e, ) with ampicillin sodium (AB-Pc) added as an effect enhancer, and AB-PC alone as a control were given to mice (Balb/c) ad libitum via a water bottle for 5 weeks. B, e were given only water for one week after being given body medication.
.

102 live bacteria/mouse were collected intraperitoneally and challenged. After attack 2
Table 4 shows the results on day 0.

Table 4 0.5 0.1
37100 0.1
10/10 Example 7 Benzyl was added to the inactivated bacteria obtained in Example 1 (A), including nigiline potassium (PCG), ampicillin sodium (ABPC), cefazolin sodium (CEZ), and tetracycline hydrochloride ( TCχ Chloramphenicol sodium succinate (CPχ Erythromycin ethylsuccinate (m), Colistin sodium metasulfonate (CL), Rifampicin (RFP)
, were mixed and suspended in the water bottle of a mouse (Balb/C), and as a control, PCG,
ABPC5CEZ STC, CP, EM.

Mice were given CL, RFP, and water ad libitum for 3 weeks, then given only water for 1 week, and at the time of systemic administration, 1057 S, t, and m mice were administered with an oral probe to challenge them. The results 20 days after the attack are shown to the public.

PCG 100 units/-α (C 01
5PC() 500 units/d
015ABPCO, 119/d
015ABPCO, 5 recommended y/ml
015CEZ O,5897at
015TCα1 Tomi 9/ld
015C
P 0.1811/ld O1
5EM O,1s/+d 01
5CL 400 units/j
115RFP 0.051g/ml
115RFP O, 5Q/rd
O15PCo 100 units/rd
0m9/sd 515ABPCO, 111
F/Wai l
515CEZ O, 511g/ml
515TC0, 1111F/d
5150P O,I Q7ml
515KMO, 11g/mg
515CL 400 units/d
515RFP O,05I
f/d #515 Example 8 Salmonellat heat inactivated at 100°C for 5 minutes
yphimurium (8, t, ) with ampicillin sodium (AB-Pc) ad libitum to mice in a water bottle for 3 weeks, while the same mixture was administered by oral probe 1 to 2 times/week. Mice were gavaged for 3 weeks. For one week after the end of the administration, the mice were given only water, and when they nested, they were challenged with 105 S, T, and live bacteria per animal using an oral probe. The results 20 days after the challenge are shown in Table 6.

Table 6 Ba1vc mouse α to /++t/ o, ossv/
- Unlimited 3 weeks 015I 5 capes/times 0.
5η once/week I 2151 5 I
O35I 2 I I 115α5#
0.51 1 1 1 415I
α5 z 0.5-#-2, # #
1151 0*/ml 0.05η/- constant
15150 starch/time 0.5 da/time 1 time/week I
515# 01 0.512 1
1 5150 I OI I
515 As is clear from Table 6 above, ad libitum administration is more effective than forced administration, and despite the fact that the total dose, especially the amount of AB-Pc as an effect enhancer, is smaller in ad libitum administration, the effect is superior. was.

〔effect〕

As can be seen from the examples, the prophylactic agent of the present invention can achieve excellent effects with extremely small amounts of inactive particles and antibiotics compared to conventional methods.

The prophylactic agent of the present invention can be particularly effective when continuously administered for 7 days or more.

Claims (1)

[Claims] At least one selected from the group consisting of penicillin antibiotics, polypeptide antibiotics, macrolide antibiotics, cephem antibiotics, chloramphenicol antibiotics, tetracycline antibiotics, and phosphorous polysaccharide antibiotics. A preventive agent for Salmonella infection consisting of 1 species and inactivated Salmonella bacteria.