Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Genetically modified micro-organism – cell – or virus
Patent
1988-08-09
1990-01-09
Schain, Howard E.
Drug, bio-affecting and body treating compositions
Whole live micro-organism, cell, or virus containing
Genetically modified micro-organism, cell, or virus
4352527, 435842, A61K 3908
Patent
active
048927317
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a novel biological intestinal antiseptic. More particularly, this invention relates to a biological intestinal antiseptic which contains, as the active component, cells or endospores of Clostridium butyricum MII588-Sens 1 strain having a sensitivity to a butyric-acid bacteriophage KMl, or cells or endospore of another Clostridium butyricum MII588-Res 1 strain which is a spontaneous mutant derived from the Sens 1 strain and which has gained resistance to the phage KM1. The antiseptic of this invention is useful for the prevention and therapeutic treatment of bacterial food-poisonings.
BACKGROUND ART
Among bacteriophages which infect Clostridium bacteria as obligatory anaerobes, the bacteriophage which was separated from Clostridium tetani by Cowels in 1934 was first known. Since then, bacteriophages capable of lysing various Clostridium bacteria have been reported. There are however rather few reports on such bacteriophages which lyse butyric-acid bacteria. It has been reported by Tahara et al. and Hongo et al. that one species of butyric-acid bacteria is lysed by the enzymes induced by phages HM3 and HM7 which infect Clostridium saccharoperbutylacetonicum. On the other hand, Schlechte described in 1980 such phage 5 which lyses oncolytic Clostridium oncolyticum M55 and Clostridium butyricum H8 {Schlechte, H. et al. "Archiv fur Geschuwulstforschung", 50, 53-57 (1980)}.
The present inventors discovered the new bacteriophage KM1 which is totally different from these reported phages. Starting from this discovery, the present inventors further have found out a special butyric-acid bacterium, namely Clostridium butyricum MII588-Sens 1 strain having a sensitivity to the bacteriophage KM1 from amongst Clostridium butyricum MII588 strains, as well as a mutant, Clostridium butyricum MII588-Res 1 strain which is derived from the Clostridium butyricum MII588-Sens 1 strain and having gained a phage resistance. It has also been found by the present inventors that these strains, i.e., the Sens 1 and Res 1 strains exhibit, both in vitro and in vivo, a strong and stable antagonism against the causative bacteria of acute gastritis and enterocolitis, such as infectious bacteria of food-poisonings and enteric infections, enteropathogenic Escherichia coli and multiple-drug-resistant Shigella strains. Based on these findings, we have found that cells or endospores of the MII588-Sens 1 strain or the MII588-Res 1 strain are effective as an intestinal antiseptic or as a bacteriostatic agent against the causative bacteria of food-poisonings.
Bacteriophage KM1 was isolated from soil. Isolation, purification and biological characteristics of the bacteriophage KM1 have recently been reported in detail by Maeds, Ishii, Tanaka, Mikami and Arai {"J. General Microbiology", 132, 2271-2275 (1986)}.
The characteristics of this phage are as follows.
Shape and host range:
A regular plaque (lysed lesion) of bacteriophage KM1 is transparent and reaches 1-2 mm in diameter 24 hours after incubation. Under electron microscope using the negative staining method, bacteriophage KM1 is in the form of a tadpole and has a hexagonal head whose diameter is 60 nm. A tail having a contractile sheath has a length of 90 nm. Phage 5 of Schlechte et al., which was used for the sake of comparison, has an elliptical head whose diameter is 50.times.80 nm. Its tail is as long as 125 nm, has no sheath and is flexible. The G+C contents were 30.5 mol % for phage KM1 and 35.4 mol % for phage 5, which are both greater than the G+C contents (27-28 mol %) of standard C. butyricum. These two bacteriophages are clearly different from each other, because it is shown that when their DNAs are treated with restriction enzymes Bgl II, Eco RI and Hind III and the resultant DNA fragments are compared by an agarose gel electrophoresis, revealing that their electrophoresis patterns are entirely different. The DNA of phage 5 cannot be digested by Bgl II.
Bacteriophage KM1 cannot lyse entirely any of the butyric-acid bacteria which have been
REFERENCES:
Maeda et al., Journal of General Microbiology (1986), 132, 2271-2275.
Arai Tadashi
Maeda Akio
Tanaka Mamoru
Koh Choon P.
Schain Howard E.
Tadashi Arai
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