Plant parasitic nematode survival and detection to inform biosecurity risk assessment

Plant parasitic nematodes (PPN) are known to survive periods of desiccation, an ability that increases the risk of them surviving unintentional transport between countries. To investigate nematode survival in soil subject to prolonged storage, soil collected from a native forest and an organic orchard was stored separately in cupboards at ambient temperature for 36 months. Subsamples were taken at 0, 3, 6, 12, 13, 24 and 36 months to determine the presence of plant parasitic and total nematodes using a standard misting technique. Pratylenchus was used as a model to determine if PPNs that had been under prolonged storage were able to infect plant hosts at 13, 24 and 36 months. Overall, the total number of nematodes recovered from stored soil declined over time, with differences in species diversity determined by molecular methods, related to soil origin. No PPN were recovered in soil stored beyond 13 months using the three-day misting technique. By comparison, Pratylenchus nematodes, using a baiting method, were found to successfully invade host plant roots (ryegrass and white clover) even after 36 months storage and were observed to produce offspring at 13 months. Baiting was not effective for Pratylenchus found in soil originally collected from the forest but was for orchard soil, a result attributed to the lack of suitable host plants for the Pratylenchus species found in forest soil. This study demonstrated, that in protected environments, nematodes could survive for at least 36 months and were observed to produce offspring at 13 months. Baiting with a host plant was more sensitive in detecting nematodes than using the misting extraction technique, although this approach only works where the host plant is known. Without a priori knowledge of the nematode-plant host association, plant baiting may also produce false negatives. In the context of plant biosecurity and providing an accurate risk assessment in soil contaminants, the development of a generic test for PPN that induces nematodes in a resting stage to emerge and respond to a cue would enhance the probability of detection. However, as assessments at the border are often time limited, a molecular based bioassay that can be used to indicate the presence of multiple species of live PPN species may be a more feasible option for risk assessments