I. TYPING METHODS FOR CLOSTRIDIUM
17
Dolman and Chang (1972) described a large number of different phage
morphologies which they ti ied to associate with different toxigenic types of
C. ' botulinum. Perhaps a significant look at bacteriophages in relationship
to the taxonomy of C. botulinum is presented by Sugiyama and King (1972).
Rather than looking at lysogenic cultures as a source of phages, these
workers isolated from bottom sediments phages which were virulent for
wild-type C. botulinum. The host ranges indicated seven different phages
which fell into three morphological groups. The hosts were restricted
to non-proteolytic types B, E and D and to a few non-toxigenic
strains resembling type E; none of the phages was active on proteolytic
types A, B or F or on cultures producing type C or D toxin. This, in
addition to serological evidence (Solomon et al., 1971 ; Batty and Walker,
1965; Takumi and Kawata, 1976), DNA homologies (Lee and Rieman,
1970a, b) and metabolic properties of types C and D, proteolytic A, D and
F and non-proteolytic B, E and F, indicates that serological toxin typing
bears little or no relationship to the natural relationship of these organisms.
In 1973, Mitsui et al. reported an inducible lysin produced after UV
light or Mitomycin C treatment of C. botulinum A190. This lysin was
active against freeze-thawed cells and cells treated in other manners, but
not against viable cells. Kiritani et al. (1973) made some interesting observations concerning numerical taxonomy of C. botulinum and C. sporogenes
strains and their susceptibilities to induced lysins and to Mitomycin C.
Based upon numerical taxonomy and lysin spectra, toxin activity was the
only criterion to differentiate proteolytic C. botulinum types A, B and F
from C. sporogenes. These organisms, all belonging to phenon I, were
susceptible to the C. botulinum A190 lysin. Phenon I1 contained nonproteolytic types B, C, D, E and F. Although a number of botulinum
strains were more sensitive than C. sporogenes to Mitomycin C, the species
cannot be reliably differentiated on this basis. It appears that phage host
ranges of potential typing schemes of C. botulinum have not yet been
described. With the soft agar overlay method of Eklund et al. (1972), it
should be possible to examine strains with a view of typing.
2. Bacteriophages of C. tetani
Little has been reported on the phages of C. tetani. Cowles (1934)
isolated phage from crude sewage by an enrichment technique using five
bacterial strains. Two of these were sensitive to phage and lysogenisation of
the sensitive bacteria was demonstrated. Cowles examined toxin production in C. tetani without demonstrating that bacteriophages were responsible for toxin production.
Prescott and Altenbern (1967a) described lysis of C. tetani following
treatment with Mitomycin C or UV irradiation. A subsequent paper
17
Dolman and Chang (1972) described a large number of different phage
morphologies which they ti ied to associate with different toxigenic types of
C. ' botulinum. Perhaps a significant look at bacteriophages in relationship
to the taxonomy of C. botulinum is presented by Sugiyama and King (1972).
Rather than looking at lysogenic cultures as a source of phages, these
workers isolated from bottom sediments phages which were virulent for
wild-type C. botulinum. The host ranges indicated seven different phages
which fell into three morphological groups. The hosts were restricted
to non-proteolytic types B, E and D and to a few non-toxigenic
strains resembling type E; none of the phages was active on proteolytic
types A, B or F or on cultures producing type C or D toxin. This, in
addition to serological evidence (Solomon et al., 1971 ; Batty and Walker,
1965; Takumi and Kawata, 1976), DNA homologies (Lee and Rieman,
1970a, b) and metabolic properties of types C and D, proteolytic A, D and
F and non-proteolytic B, E and F, indicates that serological toxin typing
bears little or no relationship to the natural relationship of these organisms.
In 1973, Mitsui et al. reported an inducible lysin produced after UV
light or Mitomycin C treatment of C. botulinum A190. This lysin was
active against freeze-thawed cells and cells treated in other manners, but
not against viable cells. Kiritani et al. (1973) made some interesting observations concerning numerical taxonomy of C. botulinum and C. sporogenes
strains and their susceptibilities to induced lysins and to Mitomycin C.
Based upon numerical taxonomy and lysin spectra, toxin activity was the
only criterion to differentiate proteolytic C. botulinum types A, B and F
from C. sporogenes. These organisms, all belonging to phenon I, were
susceptible to the C. botulinum A190 lysin. Phenon I1 contained nonproteolytic types B, C, D, E and F. Although a number of botulinum
strains were more sensitive than C. sporogenes to Mitomycin C, the species
cannot be reliably differentiated on this basis. It appears that phage host
ranges of potential typing schemes of C. botulinum have not yet been
described. With the soft agar overlay method of Eklund et al. (1972), it
should be possible to examine strains with a view of typing.
2. Bacteriophages of C. tetani
Little has been reported on the phages of C. tetani. Cowles (1934)
isolated phage from crude sewage by an enrichment technique using five
bacterial strains. Two of these were sensitive to phage and lysogenisation of
the sensitive bacteria was demonstrated. Cowles examined toxin production in C. tetani without demonstrating that bacteriophages were responsible for toxin production.
Prescott and Altenbern (1967a) described lysis of C. tetani following
treatment with Mitomycin C or UV irradiation. A subsequent paper