Jpn. J. Med. Sci. Biol., 44, 141-146, 1991.

Short Communication

PERITRICHOUS FLAGELLATION IN PLESIOMONAS SHIGELLOIDES

STRAINS

Kohaku INOUE, Yoshimasa KOSAKO1, Kenji SUZUKI2

and Toshio SHIMADA3*

Social Welfare Corporation Colony Ranzangoh Clinical Laboratory, Ranzan-

machi, Hiki-gun, Saitama 355-02, 1Japan Collection of Microorganisms, RIKEN,
Wako-shi, Saitama 351-01, 2Laboratory of Technology and 3Department of
Bacteriology, National Institute of Health, Kamiosaki, Shinagawa-ku, Tokyo141

(Received April 8, 1991. Accepted July 25, 1991)

SUMMARY: In total, 131 strains of Plesiomonas shigelloides isolated from

various sources were tested for peritrichous flagella by a flagella staining
method. When incubated on a solid medium for 18 hr at 25 C, peritrichous
flagella were demonstrated in 89(68%) of them. With an electron microscope, the
peritrichous flagella were clearly distinguished from the lophotrichous ones by
their wavelength.

Plesiomonas shigelloides is a gram-negative and facultatively anaerobic

rod that has commonly been associated with sporadic episodes and rare outbreaks
of diarrhea in humans (1). In recent years much interest has been stimulated in
regard to their enteropathogenicity (1,2).

( 1848)
( 2-1)
( )
( )
*Address reprint requests to: T . Shimada, Department of Bacteriology,
National Institute of Health, Kamiosaki, Shinagawa-ku, Tokyo141, Japan.

141
 The shape and arrangement of flagella on the bacterial cell have been used

as a significant criterion in the taxonomy of bacteria. Plesiomonas shigelloides is

known to produce lophotrichous flagella (1,3). Ewing et al. (4), however, reported

mixed lophotrichous and peritrichous flagella in young cultures of certain strains.

In the present study, the incidence of peritrichous flagella in strains of P.

shigelloides isolated from various sources was investigated.

In total, 131 strains of P. shigelloides were studied. Of these strains, 80

were isolated from the feces of patients with gastroenteritis or their contacts and

the remaining 51 strains from animals, river water, river fish, birds or mud.

Two media were used to investigate the development of flagella; nutrient

agar containing 1% Lab-Lemco Powder, 1% Bacto-Peptone, 0.5% NaCl, 0.1%

glucose and 1.3% agar (Difco) in distilled water at pH 7.0 and Nutrient Broth No.

2 (Oxoid).

Motility was determined by direct microscopic observation of a wet-mount

preparation.

For staining, all cultures were incubated at 25 C for 18 hr. Organisms

were stained for flagella with gFlagella Staining Solution-Shionogih (5). The

bacterial suspension and smear were prepared as described by Leifson (6). Cells

of P. shigelloides showing peritrichous flagella by the method were also shadowed

with palladium and examined by electron microscopy.

Peritrichous flagella were demonstrated in 89(68%) of the 131 strains of P.

shigelloides when incubated on a solid medium for 18 hr at 25 C (Fig. 1a). The

peritrichous flagella were easily removed mechanically from the cells during the

course of harvesting and suspending the cells. In broth, all 131 strains showed

lophotrichous flagella (Fig. 1c). The peritrichous flagella in the 89 strains of P.

shigelloides were observed also by electron microscopy. Electron micrographs of

the strains showed the presence of unsheathed lophotrichous and peritrichous

flagella. The peritrichous flagella were clearly distinguishable from the

lophotrichous flagella in that the peritrichous flagella had a definitely shorter

wavelength than the lophotrichous undulant flagella (Figs.lb, id, 2a and 2b). No

significant correlation was observed between the ability to produce peritrichous

flagella and the serovars (7) of the strains or their sources.

In some strains of mesophilic Aeromonas spp., peritrichous flagella are not

clearly distinguishable from monotrichous flagella by their morphological

structure, because their wavelengths are similar (8). There was, however, clear

142
 Fig. 1a

Fig. 1b

Fig. 1a. Peritrichous flagella of P. shigelloides strain P1-656 (O31:H3)

grown on a solid medium at 25 C for 18 hr. Flagella stain with

g Flagella Staining Solution-Shionogih.~ 1,000.

Fig. lb. An electron micrograph of P . shigelloides strain P1-656

(O31:H3) grown on a solid medium at 25 C for 18 hr. Palladium

shadowed. ~ 15,000.

difference in the morphological characteristics among the strains of P .

shigelloides examined.
Ewing et al. (2) described that young cultures of P . shigelloides have
peritrichous flagella. From the results obtained herein, it is evident that many

143
 Fig. 1c

Fig. 1d

Fig. 1c. Lophotrichous flagella of P. shigelloides strain P1-656 (O31:H3)

grown in broth at 25 C for 18 hr. Flagella stain. ~1,000.

Fig. 1d. An electron micrograph of P. shigelloides strain P1-656

(O31:H3) grown in broth at 25 C for 18 hr. ~15,000.

stains of P. shigelloides defined as lophotrichous flagellated organisms (1,3) can

produce peritrichous flagella when grown on solid media. The antigenic
properties of the lophotrichous and peritrichous flagella were not examined.
Further study is needed to clarify whether they are antigenically identical or not.

144
 Fig. 2a

Fig. 2b

Fig. 2a. An electron micrograph of P. shigelloides strain 11173

(O38:H5) grown on a solid medium at 25 C for 18 hr. ~ 15,000.

Fig. 2b. An electron micrograph of P. shigelloides strain 11173

(O38:H5) grown in broth at 25 C for 18 hr. ~ 15,000.

145
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