Plasma-dependent chemotactic activity of hemocytes derived from a juvenile estuarine gastropod mollusc, Clithon retropictus, to Vibrio parahaemolyticus and Escherichia coli strains.

Hemocytes of adult Clithon retropictus were attracted chemotactically to live Vibrio parahaemolyticus and Escherichia coli strains. The chemotaxis was stimulated by the plasma of adult C. retropictus. Hemocytes of the juvenile specimen were attracted chemotactically to V. parahaemolyticus and E. coli strains in the presence of the plasma of the juvenile and only to E. coli strain in the absence of the plasma. These evidences suggest that hemocytes of juvenile C. retropictus might be defective to recognize V. parahaemolyticus strains and that the hemocytes would display full activities in the presence of the plasma factor(s).     

Migratory responses of hemocytes to Vibrio parahaemolyticus in the alimentary tract of an estuarine neritid gastropod, Clithon retropictus.

Migratory responses of hemocytes to Vibrio parahaemolyticus strain D3 in the alimentary tracts of an estuarine neritid gastropod, Clithon retropictus, and a related marine neritid, Nerita albicilla, were examined under the scanning electron microscope. After ingesting the strain, active responses were seen at the esophagus, stomach and anterior intestine of adult C. retropictus and at the middle and posterior intestines of adult N. albicilla. When the alimentary tracts were isolated from the gastropod and incubated in vitro with strain D3, active response was induced at the most parts of the tract of the adult gastropods and at the stomach and the anterior intestine of juvenile C. retropictus. The responding hemocytes were confirmed to be granulocytes in the semi-thin sections of the tract of adult C. retropictus. The poor hemocyte responses at the middle and posterior intestines of juvenile C. retropictus might support the colonization of the organism there.     

Migratory deficiency of Clithon retropictus hemocytes to Vibrio parahaemolyticus and Escherichia coli

Hemocytes of a marine gastropod, Nerita albicilla, but not those of an estuarine gastropod, Clithon retropictus, were observed to migrate to live and heat-killed cells of Vibrio parahaemolyticus and Escherichia coli through Nucleopore membrane in Blind well chamber. The defective migration of C. retropictus hemocytes might reflect, at least in part, the survival of V. parahaemolyticus in the estuarine gastropod.     

An estuarine neritid gastropod, Clithon corona, a potential reservoir of thermostable direct hemolysin-producing Vibrio parahaemolyticus

An estuarine neritid gastropod, Clithon corona, maintained in UV-irradiated recirculating artificial seawater with a salinity of 15 per mil (%o) was found to retain thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus in the gut at significantly higher levels than TDH-non-producing one for at least 14 days. Another estuarine neritid gastropod, C. sowerbianus, was not able to support the preferential survival of TDH-producing organisms. This evidence suggests that, if TDH-producing vibrios are brought to estuaries inhabited by C. corona, repeated ingestion of V. parahaemolyticus by this gastropod could lead to accumulation of TDH-producing vibrios in the estuaries.     

Geographical features of estuaries for neritid gastropods including Clithon retropictus to preserve thermostable direct hemolysin-producing Vibrio parahaemolyticus.

Thermostable direct hemolysin-producing strain of Vibrio parahaemolyticus was not detected from the alimentary tract of 7 neritid gastropods including Clithon retropictus at 9 estuaries of Southwest Islands in Japan in the present study. The strain has been detected from C. retropictus at 2 estuaries facing The Sea of Japan but not at 2 estuaries facing The Seto Inland Sea and The Pacific Ocean in Western Japan in our previous studies. In comparison with geographical features of the estuaries where the strain was detected and not, thick accumulation of muddy sediments at the riverbed and stagnation of brackish water at low tide seem to be essential for the strain to survive in neritid gastropods including C. retropictus.     

Survivals of Vibrio parahaemolyticus and Escherichia coli in a gastropod mollusc, Heminerita japonica.

Vibrio parahaemolyticus strains D-3 and R-13 were found to be cleared within 7 days from a marine neritid gastropod mollusc, Heminerita japonica, maintained in artificial seawater with salinities of 15, 25 and 35 permil (%) at 25 degrees C. Escherichia coli strain YS-2 survived at a level of 10(2) colony forming units per gram in the mollusc maintained in 15% water for up to 14 days and fell to non-detectable level within 7 days in a 35% salinity group. The ability of H. japonica to clear these organisms seems to be less active than that of a marine species. Nerita albicilla, and more active than that of an estuarine species. Clithon retropictus.     

In vitro attachment of Vibrio parahaemolyticus to hemocytes of two gastropod molluscs.

Vibrio parahaemolyticus D-3 was observed to attach to hemocytes of a marine gastropod mollusc, Nerita albicilla, regardless of the presence of N. albicilla serum. The organism attached to hemocytes of an estuarine gastropod, Clithon retropictus, in the presence of C. retropictus serum while the attachment to the hemocytes was decreased significantly in the absence of the serum. These evidences suggest that N. albicilla hemocytes would facilitate the clearance of V. parahaemolyticus from the alimentary tract of the mollusc and that C. retropictus hemocytes would protect C. retropictus against the invasion of V. parahaemolyticus to hemocoel of the mollusc.     

Chemotactic activity of hemocytes derived from two marine neritid gastropod molluscs, Nerita albicilla and Heminerita japonica, to Vibrio parahaemolyticus and Escherichia coli strains.

Hemocytes of two marine neritid gastropods, Nerita albicilla and Heminerita japonica, were attracted chemotactically to live Vibrio parahaemolyticus and Escherichia coli strains. Chemotactic attraction of N. albicilla hemocytes was enhanced in the presence of N. albicilla plasma, while that of H. japonica hemocytes was not enhanced in the presence of H. japonica plasma. Chemotactic activity of the hemocytes seems to participate in the rapid elimination of V. parahaemolyticus from these gastropods.