Meeting Abstract
P1.40 Tuesday, Jan. 4 Immune Properties of the Hemolymph of the Giant Keyhole Limpet, Megathura crenulata MARTIN, Gary G.; MARTIN, Alanna M.; TSAI, Whitney L.E.*; WATANABE, Kristie; Occidental College, Los Angeles; Occidental College, Los Angeles; Occidental College, Los Angeles; Occidental College, Los Angeles gmartin@oxy.edu
The giant keyhole limpet, Megathura crenulata is being cultured for extraction of its respiratory pigment, keyhole limpet hemocyanin (KLH), which serves as an adjuvant in immunological research on the treatment of cancer and allergy. We are interested in features of the innate immune system that keep the blood and body of limpets pathogen free. Two gram negative bacteria, E. coli and Vibro fisheri, were injected into limpets and their clearance from the blood was followed. Prior to each experiment 100 µm of blood was obtained from each animal and spread on agar plates to insure that test animals were not infected. Hemolymph volumes of limpets were determined as approximately 50% of the body weight and bacteria were injected to obtain an initial concentration of 1×109 bacteria per ml. After 1, 2, 6, and 24 hr, samples (<1 ml) of blood were collected and used to determine the total hemocyte counts and spread on plates to determine colony-forming units. Results for both bacterial species were similar; bacterial levels were high in the blood at 1 and 2 hr after injection, dropped significantly by 6 hr, and were gone by 24 hr. These results provide the timetable for ongoing tests for changes in lectins, lysins and lysozyme in the blood of the giant keyhole limpets following bacterial injections. These proteins have been shown to serve important roles in the recognition and clearance of bacteria from the blood of numerous mollusks and our data will be compared with these previous studies.
