The occurrence and characterization of toxigenic vibrios in surface water and sediment samples of the fresh water environment of the Ohta River were studied. The membrane filter, pad preenrichment technique, followed by the placement of membranes onto thiosulfate citrate-bile salt-sucrose agar, was used for the enumeration of total vibrios. Qualitative examination of pathogenic vibrios was also attempted. In addition, a survey was conducted to determine the incidence of Clostridium botulinum in sediment samples of the Ohta River and the Hiroshima coast. In the identification of 361 strains, 12 species of Vibrio and two species of Listonella were observed. Non-Ol Vibrio chokrae was prevalent among the members of the genus Vibrio. Vibrio parahaemolyticus (serotype 04:K34), isolated in fresh water, is significant and suggests that some still unknown conditions promote the survival of these organisms in fresh water. An estimated 132 strains were hemolytic by a simple agar method, and further characterization revealed that 82% of the hemolytic vibrios (107 strains) produced various toxins. About 71% (93 strains) elaborated cytotoxin, 55% (72 strains) produced hemolysin, and 44% (58 strains) responded for both cytotoxin and hemolysin in the crude toxin extracts. All the non-Ol V. cholerae showed cytotoxic activity, and the virulent strains of Vibrio fluvialis and Vibrio spp. showed cytotonic responses in RK-13 cells. Of 36 sediment samples tested, 10 harbored C. botulinum spores (28%) and were isolated invariably in all the regions of the Hiroshima coast and in the Ohta River, except the upper region of the Ohta River.
Hypermastus tokunagai is a eulimid gastropod infecting the sand dollar Scaphechinus mirabilis. The eulimid is easily detached from the host and thus regarded as a temporary parasite. In this study, the growth and reproductive cycle of H. tokunagai were investigated using specimens collected from S. mirabilis in the western Seto Inland Sea, Japan. Newly recruited individuals of H. tokunagai appeared in early autumn and grew rapidly, while growth was retarded from mid-autumn to winter months. A second rapid growth occurred the following summer. Based on the growth pattern, the life span of H. tokunagai was estimated to be more than 20 months. This longer life span compared to other eulimids could be due to the accessibility and population stability of its host S. mirabilis. Maturation of H. tokunagai occurred within a short period in the summer, and was synchronized with the second growth phase. Hypermastus tokunagai showed size dimorphism between sexes and an investigation on change in shell shape with growth revealed that this characteristic can be attributed to the female's acquisition of a larger shell within a short time during its maturation period.
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