In vitro development of third- and fourth-stage larvae of Dirofilaria immitis: comparison of basal culture media, serum levels and possible serum substitutes.

In vitro development and survival of third-stage larvae of Dirofilaria immitis were compared in four different culture media and in the presence of varying concentrations of four different medium supplements. Motility and the incidence of third- to fourth-stage molting were used as criteria for evaluating different culture conditions. No significant differences in either motility or molting response were detected between larvae cultured in NCTC-135, F12(K), CMRL 1066 or Dulbecco's Modified Eagle's Medium. Fetal calf serum enhanced development and survival of the cultured larvae in dose-dependent fashion. Its effects were maximal at a concentration of 20 percent of the total medium volume. Addition of a commercial medium supplement, NuSerum, also gave a dose-related increase in larval development and viability. The activity of NuSerum in this respect was comparable to that of fetal calf serum. The tripeptide glycylhistidyllysine and bovine serum albumin, fraction V both failed to stimulate development of third-stage D. immitis larvae in vitro.