Effect of inoculum age and physical parameters on in vitro culture of the entomopathogenic nematode Steinernema feltiae

Author(s)

Leite, L. G. ^[1] ; Shapiro-Ilan, D. I. ^[2] ; Hazir, S. ^[3] ; Jackson, M. A. ^[4]

Affiliation

^[1] APTA, Inst Biol, CP 70, BR-13001970 Campinas, SP - Brazil

^[2] USDA ARS, SEFTNRL, Byron, GA 31008 - USA

^[3] Adnan Menderes Univ, Fac Arts & Sci, Dept Biol, Aydin - Turkey

^[4] USDA ARS, NCAUR, Peoria, IL 61604 - USA

• Document type: Journal article
• Source: JOURNAL OF HELMINTHOLOGY; v. 91, n. 6, p. 686-695, NOV 2017.
• Web of Science Citations: 2

Abstract

Entomopathogenic nematodes (EPNs) of the families Steinernematidae and Heterorhabditidae have a symbiotic association with bacteria which makes them virulent against insects. EPNs have been mass produced using in vivo and in vitro methods, including both solid and liquid fermentation. This study assessed the effect of nematode inoculum age on the production of Steinernema feltiae in liquid, solid and biphasic processes. Several physical parameters were also assessed: the effect of medium viscosity, flask size and aeration speed on the recovery and yield of infective juveniles (IJs). Inoculum age treatments included inoculum liquid cultures that were 7, 14, 21 and 28 days old. Nematodes from the same inoculum were added to one liquid medium (liquid culture), one solid medium with bacteria previously grown in sponge (solid culture) and a variation of the solid medium (a biphasic culture), in which the bacteria were first grown in liquid and, then, soaked into the sponges, with the purpose of providing a more homogeneous bacterial culture before nematode inoculation. Experiments were conducted in Erlenmeyer flasks. Eight treatments were established involving combinations of three variables: two media (with and without 0.2% agar), two flask sizes (250 and 150 ml) and two agitation speeds (180 and 280 rpm). The study showed increases in nematode yield for liquid cultures, but not for solid or biphasic cultures, with the advance of the inoculum age up to 28 days of growth. Furthermore, the addition of 0.2% agar to the liquid medium and increasing the aeration rate by using larger flasks with higher agitation speed may increase nematode recovery and final yield. The experiments were conducted using shake flasks but the results may also be applicable for bioreactors. (AU)

• FAPESP's process: 14/00651-4 - In vitro mass production of entomopathogenic nematodes: selection of media, biphasic production, formulation and use of the industrial waste
• Grantee: Luís Garrigós Leite
• Support Opportunities: Scholarships abroad - Research