In vitro rearing of Edovum puttleri, an egg parasitoid of the Colorado potato beetle, from egg to pupal stage in artificial diets devoid of insect sources: Effects of dietary amino acid and carbohydrate levels

Abstract

Previous studies have demonstrated that Edovum puttleri, an egg parasitoid of the Colorado potato beetle, can be reared through the pupal stage in an artificial diet containing either hemolymph from Manduca sexta or CPB embryonic cell line-conditioned medium. In order to improve the diet and reduce production costs, i.e., eliminate the insect-derived components, the effects of 13 amino acids and 20 carbohydrates on the growth and development of the parasitoid were determined. In the presence of any one of five of the amino acids (arginine, glutamine, lysine, threonine and valine) at a concentration of 1%, or eight of the carbohydrates (cellobiose, fructose, gentiobiose, glucose, lactose, melinbiose, sorbitol and threhalose), also at a concentration of 1%, more than 75% of the larvae molted to the second instar. At higher concentrations (2 and 3%), all five of the amino acids and most of the eight sugars promoted third instar formation. Prepupal formation was also observed. In the presence of threonine (at 2 and 3%) or glutamine (at 3%), between 22 and 29% of the parasitoids formed prepupae. Of the six sugars (at 3%) that promoted prepupal formation, lactose and sorbitol which promoted 18 and 20% prepupation, respectively, were the most effective. In the presence of all 13 amino acids (each at 3%) or all 20 carbohydrates (each at 3%), 36 and 39%, respectively, of the parasitoids formed prepupae. At lower concentrations, reduced percentages of prepupal formation were observed. Since the amino acids, glutamine and threonine, and the sugars, fructose, gentiobiose, glucose, lactose, sorbitol and trehalose, when added individually to the diet (at a concentration of 3%) were most effective in promoting prepupal formation, the effect of one of these two amino acids in combination with each of the six sugars was tested. Glutamine in combination with lactose or sorbitol was able to promote pupation (≤6.3%), and threonine in combination with any one of five of the sugars (lactose and sorbitol were most effective) supported pupal formation (≤6.7%). Our study demonstrated that E. puttleri can be successfully reared from the egg through the pupal stage in an artificial diet devoid of insect materials.