JPH0330574B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a poultry growth promoter. Conventionally, antibacterial substances have been generally used as growth promoters for poultry, but these have not been satisfactory due to problems such as bacterial resistance and side effects. Recently, live bacteria preparations such as lactic acid bacteria preparations have been used instead of antibacterial substances, but their effects are insufficient. As a result of intensive research into methods for promoting the growth of poultry, the present inventors found that Clostridium innocuum and Clostridium symbiosum
The present invention was completed by confirming that administration of bacterial cells obtained by culturing M. symbiosum was effective. That is, the present invention is a poultry growth promoter containing as an active ingredient bacterial cells obtained by culturing Clostridium innoculum or Clostridium symbiosum, or a mixture thereof. The Clostridium inocium used in the present invention is “Bergey’s Manual of Determinative”.
Bacteriology, 8th edition (1974). In other words, the morphology of the bacteria is a Gram-positive spore-forming bacillus, which exists singly in a straight line shape with circular ends, and the spores are oval and endemic, and some have swollen ends. It is nonmotile, catalase negative, indole negative, and ferments glucose to produce butyrate, acetic acid, and isoamyl alcohol. In addition, Clostridium symbiosum was recently reported by Kanauchi et al., “International Journal of Systematic
It was confirmed to be a spore-forming bacterium by the ``Bergey's Manual,'' 8th edition (1974), and was transferred from the genus Fusobacterium to the genus Clostridium, and the bacterial name was also Fusobacterium. This strain was renamed from Fusobacterium symbiosum. This bacterium is a Gram-negative spore-forming bacillus, with pointed ends, singly or double-chained, and spores are spherical and ubiquitous, and although the bacterial body is swollen, it is difficult to form spores. Non-motile, catalase negative, indole negative, PYG broth (peptone yeast extract)
produces butyrate and acetate in glucose broth). Since the two bacterial species mentioned above are obligate anaerobic bacteria, their cultivation must be carried out under anaerobic conditions. For example, by adding an appropriate amount of a mixture of L-cysteine hydrochloride and ascorbic acid to a culture medium immediately after heat sterilization to remove dissolved oxygen in the culture medium, anaerobic culture can be easily achieved even in static culture. Inspection” Volume 18, No. 7
See pages 16 to 16 (1974)]. Culture temperature is approximately 30-42℃,
The optimal temperature is around 37°C and the pH is preferably around neutral. After culturing, keep the culture solution at about 20℃.
After cooling back and forth, the cells are separated and collected by continuous centrifugation at about 2±2°C. The recovered bacterial cells are dispersed in a phosphate buffer or water containing amino acids such as monosodium glutamate and lysine, and then freeze-dried. The form of the bacteria to be administered is not particularly limited, such as dry preparations or liquid preparations, and it may be mixed with feed and given. In addition, examples of poultry to which the growth promoter of the present invention is applied include chickens, quail, turkeys, guinea fowl, ducks, geese, and others. The fungal agent of the present invention is preferably used by oral administration. A significant weight gain effect is shown at doses of 10 ⁶ or more per day. These effects of the present invention will be explained below with specific examples. For one test, 0-day-old broilers (Hubbard breed) were divided into 7 groups of 20 birds per group, 1 group was used as a control (physiological saline administration group), and the remaining 6 groups were treated as fungal agent administration groups. Freeze-dried bacterial cells of each bacterial species shown in Table 1 were diluted with physiological saline to a predetermined concentration and used. In other words, the number was adjusted to 10 ⁵ pieces per 0.5 ml of saline in the first test, 10 ⁶ pieces per 0.5 ml of saline in the second test, and 10 ⁸ pieces per 0.5 ml of saline in the third test. It was orally administered once a day, and the weight gain and feed conversion rate (the amount of feed (Kg) required to increase body weight by 1Kg) after 3 weeks were determined. As shown in Table 1, specific effects were observed on two types of Clostridium bacteria, Clostridium inocium and Clostridium symbiosum.

[Table] Furthermore, it was unexpectedly found that the poultry growth promoter of the present invention has preventive and therapeutic effects on poultry salmonellosis. The effect in this regard will be demonstrated by the following test. For one test, 0-day-old broilers (Hubbard breed) were divided into 9 groups of 10 birds each.
The group was classified as a control (physiological saline administration group). The remaining 8 groups were treated as bacterial agent administration groups, and lyophilized bacterial cells of each bacterial species shown in Tables 1 to 3 were diluted with physiological saline to a predetermined concentration and used. In the first test (Table 2), there were ¹⁰⁶ pieces per 0.5ml of physiological saline, in the second test (Table 3) there were ¹⁰⁷ pieces per 0.5ml, and in the third test (Table 4) there were ¹⁰⁸ pieces per 0.5ml of saline. Adjust each freeze-dried fungicide so that
Administered orally. The next day, 0.5 ml of physiological saline containing 10 ⁸ pieces of Salmonella typhimurium was orally administered to infect the mice with Salmonella bacteria. When these chicks were sacrificed one week after infection and the number of Salmonella bacteria in the cecal contents was examined, the results shown in Tables 2 to 4 were obtained. From these tables, it is recognized that among Clostridium genus bacteria, two species, Clostridium inocium and Clostridium symbiosum, are particularly effective.

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[Table] In addition, 0-day-old broilers (Hubbard breed) were divided into 5 groups of 10 birds per group, and ^{10 5} freeze-dried cells of each bacterial species shown in Table 5 were prepared per 0.5 ml of physiological saline. Each freeze-dried fungal preparation was prepared and orally administered to chicks immediately after hatching for 3 consecutive days. The next day 10 saline containing ⁸ Salmonella teifimillium 0.5
ml was orally administered to infect with Salmonella. Thereafter, the same treatment as above was performed, and the results shown in Table 5 were obtained. In other words, even when the amount of bacteria administered at one time is as low as 10 ⁵ bacteria, it is particularly effective against two species, Clostridium innoculum and Clostridium symbiosum.

[Table] Furthermore, we tested for Salmonella in the feces (including cecal stool) of 297 1-week-old broiler chickens (Hubbard breed) and found Salmonella in 39 of the 297 chickens. One group was used as a control (19 birds), and the remaining 20 birds were used as a test group. A freeze-dried fungicide mixture of Clostridium innoculum and Clostridium symbiosum was applied to each bird for 10 days from 11 days of age. ⁸
was administered orally. As a result of examining the detection status of Salmonella bacteria in feces at the age of 3 weeks, the results shown in Table 6 were obtained. As is clear from this table, a decrease in the Salmonella detection rate was observed in the lyophilized bacterial agent administration area.

[Table] As is clear from Tables 2 to 6 above, the fungal agent of the present invention is used once for 10 ⁷ or more, or at least 10 ⁵
It exhibits remarkable preventive and therapeutic effects against poultry salmonellosis at a dose of 1 x 3 days. As is clear from the above description, the poultry growth promoter of the present invention has the advantage of not only merely promoting growth but also having preventive or therapeutic effects on poultry salmonellosis. Therefore, in this specification, the expression "poultry growth promoter" refers to an agent effective for (1) promoting normal poultry growth and (2) preventing or treating poultry salmonellosis, or both. It is something. Next, the method for producing the fungal agent of the present invention will be explained with reference to Examples. Example 1 Clostridium inocium (this strain is
1 platinum loop (deposited with ATCC in 1962 under registration number ATCC 14501) was added to BTI broth (Brain Heart Infusion Broth, manufactured by Difco).
After inoculating and culturing at 37℃ for 20 hours, culture in 100 times the volume of the same medium at 37℃ for 20 hours, and then once again.
Culture in 100 times the volume of the same medium at 37°C for 20 hours. After culturing, the culture solution was rapidly cooled to 20°C, and then
Bacterial cells were separated from the culture solution using a continuous centrifuge at ±2°C. The isolated bacterial cells were suspended in physiological saline in an amount 10 times the weight of the bacteria, and then centrifuged again to wash the bacterial cells. This washing operation was repeated again to obtain 40 m of wet bacterial cells from the 10 culture solutions. The obtained bacterial cells were mixed with 3% sodium glutamate in an amount 10 times the amount of bacterial cells and 5
% soluble starch in a phosphoric acid buffer solution (containing 0.4% potassium dihydrogen phosphate and 1.0% sodium hydrogen phosphate), and then freeze-dried. The obtained freeze-dried cells were ground into powder to obtain the anti-Salmonella agent of the present invention. One gram of this bacterial agent contained 10 ^{to 11} viable Clostridium inocium bacteria. Example 2 An anti-Salmonella agent of the present invention was obtained in the same manner as in Example 1, except that Clostridium symbiosum (this strain has been deposited with the ATCC under registration number ATCC 14940 in 1976) was used instead of Clostridium inocium. One gram of this bacterial agent contained 10 ^{to 11} live Clostridium symbiosum bacteria. Example 3 Clostridium inocium and Clostridium inocium obtained by the methods of Examples 1 and 2
The anti-Salmonella agent of the present invention was obtained by mixing the lyophilized cell powders of Symbiosum at a ratio of 1:1.

Claims (1)

[Claims] 1 Clostridium innocuum or Clostridium symbiosum
1. A growth promoter for poultry, characterized in that the active ingredient is bacterial cells obtained by culturing or a mixture of both bacterial cells.