MASS PRODUCTION OF THE BENEFICIAL NEMATODE Heterorhabditis bacteriophora ON SOLID MEDIA USING SOLID STATE FERMENTATION TECHNOLOGY

MARY TESS JOHNSON¹, DEVANG UPADHYAY²*, SIVANADANE MANDJINY³, REBECCA BULLARD DILLARD⁴, FREDERICK JEFF⁵, HOLMES LEONARD^6
1Sartorius Stedim Biotechnology Laboratory, Biotechnology Research and Training Center, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
²Sartorius Stedim Biotechnology Laboratory, Biotechnology Research and Training Center, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
³Department of Chemistry and Physics, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
⁴School of Graduate Studies and Research, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
⁵College of Arts & Sciences, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
⁶Sartorius Stedim Biotechnology Laboratory, Biotechnology Research and Training Center, The University of North Carolina at Pembroke, Pembroke, NC 28372 USA
* Corresponding Author : danny.uncp@gmail.com

Received : 25-10-2016     Accepted : 03-11-2016     Published : 12-11-2016
Volume : 8     Issue : 55       Pages : 3030 - 3032
Int J Agr Sci 8.55 (2016):3030-3032

Keywords : Heterorhabditis bacteriophora, Photorhabdus luminescens, Entomopathogenic nematodes, Solid state fermentation
Academic Editor : Dr Satyanarayana C
Conflict of Interest : None declared
Acknowledgements/Funding : A warm thank you to Farm Bureau of Robeson, North Carolina; the University of North Carolina at Pembroke Chemistry and Physics Department and the UNCP Biotechnology Center for financial assistance.
Author Contribution : Conceptualization and experimental work: Mary Tess Johnson, Devang Upadhyay and Leonard Holmes; Manuscript preparation: Mary Tess Johnson, Devang Upadhyay, Sivanadane Mandjiny, Rebecca Bullard-Dillard, Jeff Frederick and Leonard Holmes

The focus of this study was to mass produce the entomopathogenic nematode (EPN), Heterorhabditis bacteriophora as a bio-control agent (bio-pesticide) using its symbiont bacteria Photorhabdus luminescens on a solid media surface. The process of growing these nematodes was to upscale the surface area of a solid agar media, thus increasing the yield of the beneficial nematodes. The solid agar media was adjusted to conditions, which provided an ideal growing environment for these nematodes to maintain vitality for an entire life cycle. The bacterial symbiont was then inoculated by an in-vitro culture 24 hours prior to nematode inoculation and furthermore leading to the inoculation of Heterorhabditis bacteriophora. The inoculated entomopathogenic nematodes have a 7-8 day life cycle .Once the nematodes develop into infective juveniles, the hermaphrodites can begin to self-fertilize its eggs and produce new offspring. Numbers of offspring then maximize after approximately seven days post-nematode inoculation. After harvesting, the nematodes are sanitized and stored for further use. The yield of H. bacteriophora was 5.5x105 IJs/gram with variation of 10%. The scale-up technology used in this study can be further improved by altering solid media and to optimize the growth conditions using larger surface area.