Storage of osmotically treated entomopathogenic nematode Steinernema carpocapsae

摘要

The infective juveniles (IJs) of Steinernema carpocapsae 'All' were osmotically stressed by a mixture of ionic (fortified artificial seawater) and non-ionic (3.2 mol/L glycerol) solutions to establish a method for osmotic storage of entomopathogenic nematodes. Seven combinations (termed solution A to G) with different proportions of these two solutions were tested, with sterile extra pure water (sepH_2O, termed solution H) as a control. The mortality of the IJs at a concentration of 5 x 10~5 IJ/mL in the solutions A to G, and H were 13.2%, 16.2%, 16.7%, 13.5%, 25.2%, 31.6%, 44.6%, and 1.0%, respectively, after 21 days storage at 25℃. Most of the IJs shrunk and stopped motility after 6—9 hours incubation at 25℃ in solutions A to D. Based on the results, solutions A to D and H were chosen to further test the osmotic survival of the IJs at different IJ concentrations (5 x 10~5, 2.5 x 10~5, 2 000 IJ/mL) and incubation temperature (30℃, 25℃, 10℃). The resulting IJs were exposed to a high temperature assay (45℃ for 4 h, HTA). Osmotically stressed IJs showed improved heat tolerance. The mortality of the IJs increased with the increasing concentrations of the test IJs and the storage temperatures after exposing to the HTA. More than 88.4%, 62.3% or 2.4% of the treated IJs died at the above three IJ concentrations, respectively. At the three IJ concentrations (2000 IJs/mL, 2.5 x 10~5 IJs/mL or 5 x 10~5 IJs/mL), the highest mortality was recorded in solution D (11.6%, 85.9% or 98.0%, respectively), and the lowest mortality in solution B (2. 4%, 62.3% or 86.6%, respectively). No untreated IJs survived after the heat treatment. During 42 days storage at 10℃, the IJs mortality in the solution A to D and H were 7.19%, 5.97%, 4.41%, 4.34%, and 4.34% respectively, and showed no significant differences. In conclusion, osmotic treatment of the IJs of S. carpocapsae ' All' in a mixture of ionic and non-ionic solutions enhances the heat tolerance. The mortality of the IJs after HTA increased with the increasing concentrations of the test IJs and the storage temperatures after exposure to the HTA. The result is promising for the osmotic storage of the entomopathogenic nematodes.