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In vitro liquid culture of the mollusc-parasitic nematode Phasmarhabditis (Rhabditida: Rhabditidae)

Published online by Cambridge University Press:  15 November 2022

A. Pieterse Open the ORCID record for A. Pieterse [Opens in a new window] ,
S. Haukeland Open the ORCID record for S. Haukeland [Opens in a new window] ,
V. Půža ,
J. L. Ross Open the ORCID record for J. L. Ross [Opens in a new window]  and
A. P. Malan Open the ORCID record for A. P. Malan [Opens in a new window]
A. Pieterse
Affiliation:
Department of Conservation Ecology and Entomology, Faculty of AgriSciences, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
S. Haukeland
Affiliation:
International Centre of Insect Physiology and Ecology (icipe), PO Box 30772-00100, Nairobi, Kenya Norwegian Institute of Bioeconomy Research (NIBIO), PO Box 115, NO-1431 Ås, Norway
V. Půža
Affiliation:
Biology Centre CAS, Institute of Entomology ASCR, Laboratory of Entomopathogenic Nematodes, České Budějovice, Czech Republic
J. L. Ross*
Affiliation:
Department of Conservation Ecology and Entomology, Faculty of AgriSciences, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa School of Biological Sciences, University of Aberdeen AB24 3UU, United Kingdom
A. P. Malan
Affiliation:
Department of Conservation Ecology and Entomology, Faculty of AgriSciences, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa
*
Author for correspondence: J. L. Ross, E-mail: [email protected]
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Abstract

The success of the mollusc-parasitic nematode, Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae), as a biological control agent in Europe has led to worldwide interest in phasmarhabditids as biocontrol agents. In this study, the mass culture potential of three phasmarhabditids, namely Phasmarhabditis papillosa, Phasmarhabditis kenyaensis and Phasmarhabditis bohemica, was assessed. In addition, ten bacterial candidates, consisting of seven associated with slugs and three associated with entomopathogenic nematodes, were investigated. The bacteria were tested for their ability to cause mortality to Deroceras invadens, as well as to support nematode growth. Initial mortality studies demonstrated that Kluyvera, Aeromonas and Pseudomonas spp. (AP3) caused 100% mortality when they were injected into the haemocoel of D. invadens. However, in growth studies, Pseudomonas sp. (AP4) was found to be the most successful bacterium, leading to recovery and reproduction in almost all nematode species, except for P. kenyaensis. In flask studies, P. bohemica, which showed exceptional growth with Pseudomonas sp. (AP1), was chosen for further investigation. The effect of inoculating flasks with different concentrations of Pseudomonas sp. (AP1), as well as with different concentrations of P. bohemica, was evaluated by assessing the nematode populations for 14 days. The results indicated that the lowest, 1% (v/v), bacteria inoculation led to higher total nematode and to infective juvenile (IJ) yield, with flasks with the highest IJ inoculum (3000 IJs/ml) having a positive effect on the total number of nematodes and IJs in cultures of P. bohemica. This study presents improvements for the mass-culturing of nematodes associated with molluscs.

Keywords

Phasmarhabditismass productionbacteriainfective juvenilespathogenicity
Type
Research Paper
Information
Journal of Helminthology , Volume 96 , 2022 , e84
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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Footnotes

*

Current address: Crop Health and Protection (CHAP), National Agri-Food Innovation Campus, Sand Hutton, York. YO411LZ, United Kingdom.

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